Comments
I would like to add one more fact about Dr. Matapurkar's patent from US Patent office in 2003. He got patent on stem cells of Endoderm to grow Urinary Bladder and Bile Duct. One of the article has been published in Ind. J of Etl Biology in 2010 regarding Urinary bladder regeneration using endoderm cells from colon. This is based on Matapurkar's hypothesis on the principles of embryology.
asha.[protected]@gmail.com
asha.[protected]@gmail.com
Hi Guys, Plz Guys don't waste your golden time in these kind of institutes. They will call you like, you have an interview. But, When you reach their office they will take F2F interview and further they will convince you to join for the course. It will take by some lady, you won't see that lady during your progression of course. After that, there will be one manager round like discussing about money for course and she will tell job assurance by Nucot is 200%. But everything is Fake / Acting.
Once you registered for the course, they will give the course date and timings will be as per their convince to do some fraud's. After 1 or 2 days, you can see that interviewer lady, then she won't come and one more person will be added to that group and his name is Fraud Sandeep. He will make you to pay remaining amount by speaking like amazing, but he is one of the bustard in the world.
After completion of course, they won't conduct any test. But they'll conduct in paper's like school. Note: They don't have single computer for student's.
Don't believe their words, if they said anything ask them to give some document's related that. It is not registered in any where.
It's one of the worst institute in the world. Don't believe Manager XXXXX word (Lady) and fraud Sandeep. Since she is lady, I don't want to mention her name.
************Don't waste your golden times guys*****Never join this kind of institute*******Don't believe their words*************
Once you registered for the course, they will give the course date and timings will be as per their convince to do some fraud's. After 1 or 2 days, you can see that interviewer lady, then she won't come and one more person will be added to that group and his name is Fraud Sandeep. He will make you to pay remaining amount by speaking like amazing, but he is one of the bustard in the world.
After completion of course, they won't conduct any test. But they'll conduct in paper's like school. Note: They don't have single computer for student's.
Don't believe their words, if they said anything ask them to give some document's related that. It is not registered in any where.
It's one of the worst institute in the world. Don't believe Manager XXXXX word (Lady) and fraud Sandeep. Since she is lady, I don't want to mention her name.
************Don't waste your golden times guys*****Never join this kind of institute*******Don't believe their words*************
****FRAUD SANDEEP******FRAUD RASHMI******FRAUD NUCOT***FAKE EMPLOYEES***EVERYTHING IS FAKE & FRAUD***
Hi friends .. I had got call from nucot. one of the hr contacted me n told that ur resume has been shortlisted for company com down for interview. ven i went there they told me vil provide u vit ITIL foundation training n provide 100% job opportunites. they charged 11, 030 for the training. those dogs brain wash the mind of students in a such a way as if they hav kept offer letter n our hands.In the beginning they told only 2 rounds of interview vil b there after completeing course ie technical n hr. package would be 2.8 to 3.2 lac per annum. n vil nt provide vit any BPO jobs. But after completeing they dint provide even single IT jobs.
JUST FOR SAKE they vil send mails that to all BPO jobs. ven v all batchmates went n asked abt this that sandeep told v hav not told like this . He is just cheating the students n making money for himself. ALL THE NUCOT PEOPLE ARE CHEATERS. they r just playing vit students life. Many students hav got cheated by them including me n my friends. V have suffered alot EVERYONE WILL BE EAGER TO GET JOB AND THIS NUCOT PEOPLE R MISUSING THIS N CHEATING STUDENTS N THE NAME OF JOB.
SO PLZ GUYS DONT EVER JOIN THIS FAKE INSTITUTE.
******Never Join this fraud institute***** Never believe their words********Don't waste your Golden Time**********************
Hi friends .. I had got call from nucot. one of the hr contacted me n told that ur resume has been shortlisted for company com down for interview. ven i went there they told me vil provide u vit ITIL foundation training n provide 100% job opportunites. they charged 11, 030 for the training. those dogs brain wash the mind of students in a such a way as if they hav kept offer letter n our hands.In the beginning they told only 2 rounds of interview vil b there after completeing course ie technical n hr. package would be 2.8 to 3.2 lac per annum. n vil nt provide vit any BPO jobs. But after completeing they dint provide even single IT jobs.
JUST FOR SAKE they vil send mails that to all BPO jobs. ven v all batchmates went n asked abt this that sandeep told v hav not told like this . He is just cheating the students n making money for himself. ALL THE NUCOT PEOPLE ARE CHEATERS. they r just playing vit students life. Many students hav got cheated by them including me n my friends. V have suffered alot EVERYONE WILL BE EAGER TO GET JOB AND THIS NUCOT PEOPLE R MISUSING THIS N CHEATING STUDENTS N THE NAME OF JOB.
SO PLZ GUYS DONT EVER JOIN THIS FAKE INSTITUTE.
******Never Join this fraud institute***** Never believe their words********Don't waste your Golden Time**********************
This article is about the cell type. For the medical therapy, see Stem cell therapy.
Stem cell
MSC high magnification.jpg
Transmission electron micrograph of an adult stem cell displaying typical ultrastructural characteristics.
Details
Latin Cellula praecursoria
Identifiers
Code TH H2.00.01.0.00001
TH H1.00.01.0.00028, H2.00.01.0.00001
FMA 63368
Anatomical terminology
Stem cells are undifferentiated biological cells that can differentiate into specialized cells and can divide (through mitosis) to produce more stem cells. They are found in multicellular organisms. In mammals, there are two broad types of stem cells: embryonic stem cells, which are isolated from the inner cell mass of blastocysts, and adult stem cells, which are found in various tissues. In adult organisms, stem cells and progenitor cells act as a repair system for the body, replenishing adult tissues. In a developing embryo, stem cells can differentiate into all the specialized cells—ectoderm, endoderm and mesoderm (see induced pluripotent stem cells)—but also maintain the normal turnover of regenerative organs, such as blood, skin, or intestinal tissues.
There are three known accessible sources of autologous adult stem cells in humans:
Bone marrow, which requires extraction by harvesting, that is, drilling into bone (typically the femur or iliac crest).
Adipose tissue (lipid cells), which requires extraction by liposuction.
Blood, which requires extraction through apheresis, wherein blood is drawn from the donor (similar to a blood donation), and passed through a machine that extracts the stem cells and returns other portions of the blood to the donor.
Stem cells can also be taken from umbilical cord blood just after birth. Of all stem cell types, autologous harvesting involves the least risk. By definition, autologous cells are obtained from one's own body, just as one may bank his or her own blood for elective surgical procedures.
Adult stem cells are frequently used in medical therapies, for example in bone marrow transplantation. Stem cells can now be artificially grown and transformed (differentiated) into specialized cell types with characteristics consistent with cells of various tissues such as muscles or nerves. Embryonic cell lines and autologous embryonic stem cells generated through Somatic-cell nuclear transfer or dedifferentiation have also been proposed as promising candidates for future therapies.[1] Research into stem cells grew out of findings by Ernest A. McCulloch and James E. Till at the University of Toronto in the 1960s.[2][3]
Contents [hide]
1 Properties
1.1 Self-renewal
1.2 Potency definition
1.3 Identification
2 Embryonic
3 Fetal
4 Adult
5 Amniotic
6 Induced pluripotent
7 Lineage
8 Treatments
8.1 Disadvantages
9 Research patents
10 See also
11 References
12 External links
Properties
The classical definition of a stem cell requires that it possess two properties:
Self-renewal: the ability to go through numerous cycles of cell division while maintaining the undifferentiated state.
Potency: the capacity to differentiate into specialized cell types. In the strictest sense, this requires stem cells to be either totipotent or pluripotent—to be able to give rise to any mature cell type, although multipotent or unipotent progenitor cells are sometimes referred to as stem cells. Apart from this it is said that stem cell function is regulated in a feed back mechanism.
Self-renewal
Two mechanisms exist to ensure that a stem cell population is maintained:
Obligatory asymmetric replication: a stem cell divides into one mother cell that is identical to the original stem cell, and another daughter cell that is differentiated.
Stochastic differentiation: when one stem cell develops into two differentiated daughter cells, another stem cell undergoes mitosis and produces two stem cells identical to the original.
Potency definition
Main article: Cell potency
Pluripotent, embryonic stem cells originate as inner cell mass (ICM) cells within a blastocyst. These stem cells can become any tissue in the body, excluding a placenta. Only cells from an earlier stage of the embryo, known as the morula, are totipotent, able to become all tissues in the body and the extraembryonic placenta.
Human embryonic stem cells
A: Stem cell colonies that are not yet differentiated.
B: Nerve cells, an example of a cell type after differentiation.
Potency specifies the differentiation potential (the potential to differentiate into different cell types) of the stem cell.[4]
Totipotent (a.k.a. omnipotent) stem cells can differentiate into embryonic and extraembryonic cell types. Such cells can construct a complete, viable organism.[4] These cells are produced from the fusion of an egg and sperm cell. Cells produced by the first few divisions of the fertilized egg are also totipotent.[5]
Pluripotent stem cells are the descendants of totipotent cells and can differentiate into nearly all cells, [4] i.e. cells derived from any of the three germ layers.[6]
Multipotent stem cells can differentiate into a number of cell types, but only those of a closely related family of cells.[4]
Oligopotent stem cells can differentiate into only a few cell types, such as lymphoid or myeloid stem cells.[4]
Unipotent cells can produce only one cell type, their own, [4] but have the property of self-renewal, which distinguishes them from non-stem cells (e.g. progenitor cells, muscle stem cells).
Identification
In practice, stem cells are identified by whether they can regenerate tissue. For example, the defining test for bone marrow or hematopoietic stem cells (HSCs) is the ability to transplant the cells and save an individual without HSCs. This demonstrates that the cells can produce new blood cells over a long term. It should also be possible to isolate stem cells from the transplanted individual, which can themselves be transplanted into another individual without HSCs, demonstrating that the stem cell was able to self-renew.
Properties of stem cells can be illustrated in vitro, using methods such as clonogenic assays, in which single cells are assessed for their ability to differentiate and self-renew.[7][8] Stem cells can also be isolated by their possession of a distinctive set of cell surface markers. However, in vitro culture conditions can alter the behavior of cells, making it unclear whether the cells will behave in a similar manner in vivo. There is considerable debate as to whether some proposed adult cell populations are truly stem cells.
Embryonic
Main article: Embryonic stem cell
Embryonic stem (ES) cells are stem cells derived from the inner cell mass of a blastocyst, an early-stage embryo.[9] Human embryos reach the blastocyst stage 4–5 days post fertilization, at which time they consist of 50–150 cells. ES cells are pluripotent and give rise during development to all derivatives of the three primary germ layers: ectoderm, endoderm and mesoderm. In other words, they can develop into each of the more than 200 cell types of the adult body when given sufficient and necessary stimulation for a specific cell type. They do not contribute to the extra-embryonic membranes or the placenta.
Nearly all research to date has made use of mouse embryonic stem cells (mES) or human embryonic stem cells (hES). Both have the essential stem cell characteristics, yet they require very different environments in order to maintain an undifferentiated state. Mouse ES cells are grown on a layer of gelatin as an extracellular matrix (for support) and require the presence of leukemia inhibitory factor (LIF). Human ES cells are grown on a feeder layer of mouse embryonic fibroblasts (MEFs) and require the presence of basic fibroblast growth factor (bFGF or FGF-2).[10] Without optimal culture conditions or genetic manipulation, [11] embryonic stem cells will rapidly differentiate.
A human embryonic stem cell is also defined by the expression of several transcription factors and cell surface proteins. The transcription factors Oct-4, Nanog, and Sox2 form the core regulatory network that ensures the suppression of genes that lead to differentiation and the maintenance of pluripotency.[12] The cell surface antigens most commonly used to identify hES cells are the glycolipids stage specific embryonic antigen 3 and 4 and the keratan sulfate antigens Tra-1-60 and Tra-1-81. The molecular definition of a stem cell includes many more proteins and continues to be a topic of research.[13]
There are currently no approved treatments using embryonic stem cells. The first human trial was approved by the US Food and Drug Administration in January 2009.[14] However, the human trial was not initiated until October 13, 2010 in Atlanta for spinal injury victims. On November 14, 2011 the company conducting the trial announced that it will discontinue further development of its stem cell programs.[15] ES cells, being pluripotent cells, require specific signals for correct differentiation—if injected directly into another body, ES cells will differentiate into many different types of cells, causing a teratoma. Differentiating ES cells into usable cells while avoiding transplant rejection are just a few of the hurdles that embryonic stem cell researchers still face.[16] Many nations currently have moratoria on either ES cell research or the production of new ES cell lines. Because of their combined abilities o[censored]nlimited expansion and pluripotency, embryonic stem cells remain a theoretically potential source for regenerative medicine and tissue replacement after injury or disease.
Mouse embryonic stem cells with fluorescent marker
Human embryonic stem cell colony on mouse embryonic fibroblast feeder layer
Fetal
The primitive stem cells located in the organs of fetuses are referred to as fetal stem cells.[17] There are two types of fetal stem cells:
Fetal proper stem cells come from the tissue of the fetus proper, and are generally obtained after an abortion. These stem cells are not immortal but have a high level of division and are multipotent.
Extraembryonic fetal stem cells come from extraembryonic membranes, and are generally not distinguished from adult stem cells. These stem cells are acquired after birth, they are not immortal but have a high level of cell division, and are pluripotent.[18]
Adult
Main article: Adult stem cell
Stem cell division and differentiation. A: stem cell; B: progenitor cell; C: differentiated cell; 1: symmetric stem cell division; 2: asymmetric stem cell division; 3: progenitor division; 4: terminal differentiation
Adult stem cells, also called somatic (from Greek σωματικóς, "of the body") stem cells, are stem cells which maintain and repair the tissue in which they are found.[19] They can be found in children, as well as adults.[20]
Pluripotent adult stem cells are rare and generally small in number, but they can be found in umbilical cord blood and other tissues.[21] Bone marrow is a rich source of adult stem cells, [22] which have been used in treating several conditions including spinal cord injury, [23] liver cirrhosis, [24] chronic limb ischemia [25] and endstage heart failure.[26] The quantity of bone marrow stem cells declines with age and is greater in males than females during reproductive years.[27] Much adult stem cell research to date has aimed to characterize their potency and self-renewal capabilities.[28] DNA damage accumulates with age in both stem cells and the cells that comprise the stem cell environment. This accumulation is considered to be responsible, at least in part, for increasing stem cell dysfunction with aging (see DNA damage theory of aging).[29]
Most adult stem cells are lineage-restricted (multipotent) and are generally referred to by their tissue origin (mesenchymal stem cell, adipose-derived stem cell, endothelial stem cell, dental pulp stem cell, etc.).[30][31]
Adult stem cell treatments have been successfully used for many years to treat leukemia and related bone/blood cancers through bone marrow transplants.[32] Adult stem cells are also used in veterinary medicine to treat tendon and ligament injuries in horses.[33]
The use of adult stem cells in research and therapy is not as controversial as the use of embryonic stem cells, because the production of adult stem cells does not require the destruction of an embryo. Additionally, in instances where adult stem cells are obtained from the intended recipient (an autograft), the risk of rejection is essentially non-existent. Consequently, more US government funding is being provided for adult stem cell research.[34]
Amniotic
Multipotent stem cells are also found in amniotic fluid. These stem cells are very active, expand extensively without feeders and are not tumorigenic. Amniotic stem cells are multipotent and can differentiate in cells of adipogenic, osteogenic, myogenic, endothelial, hepatic and also neuronal lines.[35] Amniotic stem cells are a topic of active research.
Use of stem cells from amniotic fluid overcomes the ethical objections to using human embryos as a source of cells. Roman Catholic teaching forbids the use of embryonic stem cells in experimentation; accordingly, the Vatican newspaper "Osservatore Romano" called amniotic stem cells "the future of medicine".[36]
It is possible to collect amniotic stem cells for donors or for autologuous use: the first US amniotic stem cells bank [37][38] was opened in 2009 in Medford, MA, by Biocell Center Corporation[39][40][41] and collaborates with various hospitals and universities all over the world.[42]
Induced pluripotent
Main article: Induced pluripotent stem cell
These are not adult stem cells, but rather adult cells (e.g. epithelial cells) reprogrammed to give rise to pluripotent capabilities. Using genetic reprogramming with protein transcription factors, pluripotent stem cells equivalent to embryonic stem cells have been derived from human adult skin tissue.[43][44][45] Shinya Yamanaka and his colleagues at Kyoto University used the transcription factors Oct3/4, Sox2, c-Myc, and Klf4[43] in their experiments on cells from human faces. Junying Yu, James Thomson, and their colleagues at the University of Wisconsin–Madison used a different set of factors, Oct4, Sox2, Nanog and Lin28, [43] and carried out their experiments using cells from human foreskin.
As a result of the success of these experiments, Ian Wilmut, who helped create the first cloned animal Dolly the Sheep, has announced that he will abandon somatic cell nuclear transfer as an avenue of research.[46]
Frozen blood samples can be used as a source of induced pluripotent stem cells, opening a new avenue for obtaining the valued cells.[47]
Lineage
Main article: Stem cell line
To ensure self-renewal, stem cells undergo two types of cell division (see Stem cell division and differentiation diagram). Symmetric division gives rise to two identical daughter cells both endowed with stem cell properties. Asymmetric division, on the other hand, produces only one stem cell and a progenitor cell with limited self-renewal potential. Progenitors can go through several rounds of cell division before terminally differentiating into a mature cell. It is possible that the molecular distinction between symmetric and asymmetric divisions lies in differential segregation of cell membrane proteins (such as receptors) between the daughter cells.[48]
An alternative theory is that stem cells remain undifferentiated due to environmental cues in their particular niche. Stem cells differentiate when they leave that niche or no longer receive those signals. Studies in Drosophila germarium have identified the signals decapentaplegic and adherens junctions that prevent germarium stem cells from differentiating.[49][50]
Treatments
Main article: Stem cell therapy
Diseases and conditions where stem cell treatment is being investigated.
Diseases and conditions where stem cell treatment is being investigated include:
Diabetes[51]
Rheumatoid arthritis[51]
Parkinson's disease[51]
Alzheimer's disease[51]
Osteoarthritis[51]
Stroke and traumatic brain injury repair[52]
Learning disability due to congenital disorder [53]
Spinal cord injury repair [54]
Heart infarction [55]
Anti-cancer treatments [56]
Baldness reversal[57]
Replace missing teeth [58]
Repair hearing [59]
Restore vision [60]
Amyotrophic lateral sclerosis [61]
Crohn's disease [62]
Wound healing [63]
Stem cell therapy is the use of stem cells to treat or prevent a disease or condition. Bone marrow transplant is a crude form of stem cell therapy that has been used clinically for many years without controversy. No stem cell therapies other than bone marrow transplant are widely used.[64][65]
Research is underway to develop various sources for stem cells, and to apply stem cell treatments for neurodegenerative diseases and conditions, diabetes, heart disease, and other conditions.[66]
In more recent years, with the ability of scientists to isolate and culture embryonic stem cells, and with scientists' growing ability to create stem cells using somatic cell nuclear transfer and techniques to created induced pluripotent stem cells, controversy has crept in, both related to abortion politics and to human cloning.
Disadvantages
Stem cell treatments may require immunosuppression because of a requirement for radiation before the transplant to remove the patient's previous cells, or because the patient's immune system may target the stem cells. One approach to avoid the second possibility is to use stem cells from the same patient who is being treated.
Pluripotency in certain stem cells could also make it difficult to obtain a specific cell type. It is also difficult to obtain the exact cell type needed, because not all cells in a population differentiate uniformly. Undifferentiated cells can create tissues other than desired types.[67]
Some stem cells form tumors after transplantation[68]; pluripotency is linked to tumor formation especially in embryonic stem cells, fetal proper stem cells, induced pluripotent stem cells. Fetal proper stem cells form tumors despite multipotency.[citation needed]
Hepatotoxicity and drug-induced liver injury account for a substantial number of failures of new drugs in development and market withdrawal, highlighting the need for screening assays such as stem cell-derived hepatocyte-like cells, that are capable of detecting toxicity early in the drug development process.[69]
Research patents
Further information: Consumer Watchdog vs. Wisconsin Alumni Research Foundation
Some of the fundamental patents covering human embryonic stem cells are owned by the Wisconsin Alumni Research Foundation (WARF) - they are patents 5, 843, 780, 6, 200, 806, and 7, 029, 913 invented by James A. Thomson. WARF does not enforce these patents against academic scientists, but does enforce them against companies.[70]
In 2006, a request for the US Patent and Trademark Office (USPTO) to re-examine the three patents was filed by the Public Patent Foundation on behalf of its client, the non-profit patent-watchdog group Consumer Watchdog (formerly the Foundation for Taxpayer and Consumer Rights).[70] In the re-examination process, which involves several rounds of discussion between the USTPO and the parties, the USPTO initially agreed with Consumer Watchdog and rejected all the claims in all three patents, [71] however in response, WARF amended the claims of all three patents to make them more narrow, and in 2008 the USPTO found the amended claims in all three patents to be patentable. The decision on one of the patents (7, 029, 913) was appealable, while the decisions on the other two were not.[72][73] Consumer Watchdog appealed the granting of the '913 patent to the USTPO's Board of Patent Appeals and Interferences (BPAI) which granted the appeal, and in 2010 the BPAI decided that the amended claims of the '913 patent were not patentable.[74] However, WARF was able to re-open prosecution of the case and did so, amending the claims of the '913 patent again to make them more narrow, and in January 2013 the amended claims were allowed.[75]
In July 2013, Consumer Watchdog announced that it would appeal the decision to allow the claims of the '913 patent to the US Court of Appeals for the Federal Circuit (CAFC), the federal appeals court that hears patent cases.[76] At a hearing in December 2013, the CAFC raised the question of whether Consumer Watchdog had legal standing to appeal; the case could not proceed until that issue was resolved.[77]
See also
Cell bank
Human genome
Meristem
Partial cloning
Plant stem cell
Stem cell controversy
Stem cell marker
Shinya Yamanaka
References
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Stem cell
MSC high magnification.jpg
Transmission electron micrograph of an adult stem cell displaying typical ultrastructural characteristics.
Details
Latin Cellula praecursoria
Identifiers
Code TH H2.00.01.0.00001
TH H1.00.01.0.00028, H2.00.01.0.00001
FMA 63368
Anatomical terminology
Stem cells are undifferentiated biological cells that can differentiate into specialized cells and can divide (through mitosis) to produce more stem cells. They are found in multicellular organisms. In mammals, there are two broad types of stem cells: embryonic stem cells, which are isolated from the inner cell mass of blastocysts, and adult stem cells, which are found in various tissues. In adult organisms, stem cells and progenitor cells act as a repair system for the body, replenishing adult tissues. In a developing embryo, stem cells can differentiate into all the specialized cells—ectoderm, endoderm and mesoderm (see induced pluripotent stem cells)—but also maintain the normal turnover of regenerative organs, such as blood, skin, or intestinal tissues.
There are three known accessible sources of autologous adult stem cells in humans:
Bone marrow, which requires extraction by harvesting, that is, drilling into bone (typically the femur or iliac crest).
Adipose tissue (lipid cells), which requires extraction by liposuction.
Blood, which requires extraction through apheresis, wherein blood is drawn from the donor (similar to a blood donation), and passed through a machine that extracts the stem cells and returns other portions of the blood to the donor.
Stem cells can also be taken from umbilical cord blood just after birth. Of all stem cell types, autologous harvesting involves the least risk. By definition, autologous cells are obtained from one's own body, just as one may bank his or her own blood for elective surgical procedures.
Adult stem cells are frequently used in medical therapies, for example in bone marrow transplantation. Stem cells can now be artificially grown and transformed (differentiated) into specialized cell types with characteristics consistent with cells of various tissues such as muscles or nerves. Embryonic cell lines and autologous embryonic stem cells generated through Somatic-cell nuclear transfer or dedifferentiation have also been proposed as promising candidates for future therapies.[1] Research into stem cells grew out of findings by Ernest A. McCulloch and James E. Till at the University of Toronto in the 1960s.[2][3]
Contents [hide]
1 Properties
1.1 Self-renewal
1.2 Potency definition
1.3 Identification
2 Embryonic
3 Fetal
4 Adult
5 Amniotic
6 Induced pluripotent
7 Lineage
8 Treatments
8.1 Disadvantages
9 Research patents
10 See also
11 References
12 External links
Properties
The classical definition of a stem cell requires that it possess two properties:
Self-renewal: the ability to go through numerous cycles of cell division while maintaining the undifferentiated state.
Potency: the capacity to differentiate into specialized cell types. In the strictest sense, this requires stem cells to be either totipotent or pluripotent—to be able to give rise to any mature cell type, although multipotent or unipotent progenitor cells are sometimes referred to as stem cells. Apart from this it is said that stem cell function is regulated in a feed back mechanism.
Self-renewal
Two mechanisms exist to ensure that a stem cell population is maintained:
Obligatory asymmetric replication: a stem cell divides into one mother cell that is identical to the original stem cell, and another daughter cell that is differentiated.
Stochastic differentiation: when one stem cell develops into two differentiated daughter cells, another stem cell undergoes mitosis and produces two stem cells identical to the original.
Potency definition
Main article: Cell potency
Pluripotent, embryonic stem cells originate as inner cell mass (ICM) cells within a blastocyst. These stem cells can become any tissue in the body, excluding a placenta. Only cells from an earlier stage of the embryo, known as the morula, are totipotent, able to become all tissues in the body and the extraembryonic placenta.
Human embryonic stem cells
A: Stem cell colonies that are not yet differentiated.
B: Nerve cells, an example of a cell type after differentiation.
Potency specifies the differentiation potential (the potential to differentiate into different cell types) of the stem cell.[4]
Totipotent (a.k.a. omnipotent) stem cells can differentiate into embryonic and extraembryonic cell types. Such cells can construct a complete, viable organism.[4] These cells are produced from the fusion of an egg and sperm cell. Cells produced by the first few divisions of the fertilized egg are also totipotent.[5]
Pluripotent stem cells are the descendants of totipotent cells and can differentiate into nearly all cells, [4] i.e. cells derived from any of the three germ layers.[6]
Multipotent stem cells can differentiate into a number of cell types, but only those of a closely related family of cells.[4]
Oligopotent stem cells can differentiate into only a few cell types, such as lymphoid or myeloid stem cells.[4]
Unipotent cells can produce only one cell type, their own, [4] but have the property of self-renewal, which distinguishes them from non-stem cells (e.g. progenitor cells, muscle stem cells).
Identification
In practice, stem cells are identified by whether they can regenerate tissue. For example, the defining test for bone marrow or hematopoietic stem cells (HSCs) is the ability to transplant the cells and save an individual without HSCs. This demonstrates that the cells can produce new blood cells over a long term. It should also be possible to isolate stem cells from the transplanted individual, which can themselves be transplanted into another individual without HSCs, demonstrating that the stem cell was able to self-renew.
Properties of stem cells can be illustrated in vitro, using methods such as clonogenic assays, in which single cells are assessed for their ability to differentiate and self-renew.[7][8] Stem cells can also be isolated by their possession of a distinctive set of cell surface markers. However, in vitro culture conditions can alter the behavior of cells, making it unclear whether the cells will behave in a similar manner in vivo. There is considerable debate as to whether some proposed adult cell populations are truly stem cells.
Embryonic
Main article: Embryonic stem cell
Embryonic stem (ES) cells are stem cells derived from the inner cell mass of a blastocyst, an early-stage embryo.[9] Human embryos reach the blastocyst stage 4–5 days post fertilization, at which time they consist of 50–150 cells. ES cells are pluripotent and give rise during development to all derivatives of the three primary germ layers: ectoderm, endoderm and mesoderm. In other words, they can develop into each of the more than 200 cell types of the adult body when given sufficient and necessary stimulation for a specific cell type. They do not contribute to the extra-embryonic membranes or the placenta.
Nearly all research to date has made use of mouse embryonic stem cells (mES) or human embryonic stem cells (hES). Both have the essential stem cell characteristics, yet they require very different environments in order to maintain an undifferentiated state. Mouse ES cells are grown on a layer of gelatin as an extracellular matrix (for support) and require the presence of leukemia inhibitory factor (LIF). Human ES cells are grown on a feeder layer of mouse embryonic fibroblasts (MEFs) and require the presence of basic fibroblast growth factor (bFGF or FGF-2).[10] Without optimal culture conditions or genetic manipulation, [11] embryonic stem cells will rapidly differentiate.
A human embryonic stem cell is also defined by the expression of several transcription factors and cell surface proteins. The transcription factors Oct-4, Nanog, and Sox2 form the core regulatory network that ensures the suppression of genes that lead to differentiation and the maintenance of pluripotency.[12] The cell surface antigens most commonly used to identify hES cells are the glycolipids stage specific embryonic antigen 3 and 4 and the keratan sulfate antigens Tra-1-60 and Tra-1-81. The molecular definition of a stem cell includes many more proteins and continues to be a topic of research.[13]
There are currently no approved treatments using embryonic stem cells. The first human trial was approved by the US Food and Drug Administration in January 2009.[14] However, the human trial was not initiated until October 13, 2010 in Atlanta for spinal injury victims. On November 14, 2011 the company conducting the trial announced that it will discontinue further development of its stem cell programs.[15] ES cells, being pluripotent cells, require specific signals for correct differentiation—if injected directly into another body, ES cells will differentiate into many different types of cells, causing a teratoma. Differentiating ES cells into usable cells while avoiding transplant rejection are just a few of the hurdles that embryonic stem cell researchers still face.[16] Many nations currently have moratoria on either ES cell research or the production of new ES cell lines. Because of their combined abilities o[censored]nlimited expansion and pluripotency, embryonic stem cells remain a theoretically potential source for regenerative medicine and tissue replacement after injury or disease.
Mouse embryonic stem cells with fluorescent marker
Human embryonic stem cell colony on mouse embryonic fibroblast feeder layer
Fetal
The primitive stem cells located in the organs of fetuses are referred to as fetal stem cells.[17] There are two types of fetal stem cells:
Fetal proper stem cells come from the tissue of the fetus proper, and are generally obtained after an abortion. These stem cells are not immortal but have a high level of division and are multipotent.
Extraembryonic fetal stem cells come from extraembryonic membranes, and are generally not distinguished from adult stem cells. These stem cells are acquired after birth, they are not immortal but have a high level of cell division, and are pluripotent.[18]
Adult
Main article: Adult stem cell
Stem cell division and differentiation. A: stem cell; B: progenitor cell; C: differentiated cell; 1: symmetric stem cell division; 2: asymmetric stem cell division; 3: progenitor division; 4: terminal differentiation
Adult stem cells, also called somatic (from Greek σωματικóς, "of the body") stem cells, are stem cells which maintain and repair the tissue in which they are found.[19] They can be found in children, as well as adults.[20]
Pluripotent adult stem cells are rare and generally small in number, but they can be found in umbilical cord blood and other tissues.[21] Bone marrow is a rich source of adult stem cells, [22] which have been used in treating several conditions including spinal cord injury, [23] liver cirrhosis, [24] chronic limb ischemia [25] and endstage heart failure.[26] The quantity of bone marrow stem cells declines with age and is greater in males than females during reproductive years.[27] Much adult stem cell research to date has aimed to characterize their potency and self-renewal capabilities.[28] DNA damage accumulates with age in both stem cells and the cells that comprise the stem cell environment. This accumulation is considered to be responsible, at least in part, for increasing stem cell dysfunction with aging (see DNA damage theory of aging).[29]
Most adult stem cells are lineage-restricted (multipotent) and are generally referred to by their tissue origin (mesenchymal stem cell, adipose-derived stem cell, endothelial stem cell, dental pulp stem cell, etc.).[30][31]
Adult stem cell treatments have been successfully used for many years to treat leukemia and related bone/blood cancers through bone marrow transplants.[32] Adult stem cells are also used in veterinary medicine to treat tendon and ligament injuries in horses.[33]
The use of adult stem cells in research and therapy is not as controversial as the use of embryonic stem cells, because the production of adult stem cells does not require the destruction of an embryo. Additionally, in instances where adult stem cells are obtained from the intended recipient (an autograft), the risk of rejection is essentially non-existent. Consequently, more US government funding is being provided for adult stem cell research.[34]
Amniotic
Multipotent stem cells are also found in amniotic fluid. These stem cells are very active, expand extensively without feeders and are not tumorigenic. Amniotic stem cells are multipotent and can differentiate in cells of adipogenic, osteogenic, myogenic, endothelial, hepatic and also neuronal lines.[35] Amniotic stem cells are a topic of active research.
Use of stem cells from amniotic fluid overcomes the ethical objections to using human embryos as a source of cells. Roman Catholic teaching forbids the use of embryonic stem cells in experimentation; accordingly, the Vatican newspaper "Osservatore Romano" called amniotic stem cells "the future of medicine".[36]
It is possible to collect amniotic stem cells for donors or for autologuous use: the first US amniotic stem cells bank [37][38] was opened in 2009 in Medford, MA, by Biocell Center Corporation[39][40][41] and collaborates with various hospitals and universities all over the world.[42]
Induced pluripotent
Main article: Induced pluripotent stem cell
These are not adult stem cells, but rather adult cells (e.g. epithelial cells) reprogrammed to give rise to pluripotent capabilities. Using genetic reprogramming with protein transcription factors, pluripotent stem cells equivalent to embryonic stem cells have been derived from human adult skin tissue.[43][44][45] Shinya Yamanaka and his colleagues at Kyoto University used the transcription factors Oct3/4, Sox2, c-Myc, and Klf4[43] in their experiments on cells from human faces. Junying Yu, James Thomson, and their colleagues at the University of Wisconsin–Madison used a different set of factors, Oct4, Sox2, Nanog and Lin28, [43] and carried out their experiments using cells from human foreskin.
As a result of the success of these experiments, Ian Wilmut, who helped create the first cloned animal Dolly the Sheep, has announced that he will abandon somatic cell nuclear transfer as an avenue of research.[46]
Frozen blood samples can be used as a source of induced pluripotent stem cells, opening a new avenue for obtaining the valued cells.[47]
Lineage
Main article: Stem cell line
To ensure self-renewal, stem cells undergo two types of cell division (see Stem cell division and differentiation diagram). Symmetric division gives rise to two identical daughter cells both endowed with stem cell properties. Asymmetric division, on the other hand, produces only one stem cell and a progenitor cell with limited self-renewal potential. Progenitors can go through several rounds of cell division before terminally differentiating into a mature cell. It is possible that the molecular distinction between symmetric and asymmetric divisions lies in differential segregation of cell membrane proteins (such as receptors) between the daughter cells.[48]
An alternative theory is that stem cells remain undifferentiated due to environmental cues in their particular niche. Stem cells differentiate when they leave that niche or no longer receive those signals. Studies in Drosophila germarium have identified the signals decapentaplegic and adherens junctions that prevent germarium stem cells from differentiating.[49][50]
Treatments
Main article: Stem cell therapy
Diseases and conditions where stem cell treatment is being investigated.
Diseases and conditions where stem cell treatment is being investigated include:
Diabetes[51]
Rheumatoid arthritis[51]
Parkinson's disease[51]
Alzheimer's disease[51]
Osteoarthritis[51]
Stroke and traumatic brain injury repair[52]
Learning disability due to congenital disorder [53]
Spinal cord injury repair [54]
Heart infarction [55]
Anti-cancer treatments [56]
Baldness reversal[57]
Replace missing teeth [58]
Repair hearing [59]
Restore vision [60]
Amyotrophic lateral sclerosis [61]
Crohn's disease [62]
Wound healing [63]
Stem cell therapy is the use of stem cells to treat or prevent a disease or condition. Bone marrow transplant is a crude form of stem cell therapy that has been used clinically for many years without controversy. No stem cell therapies other than bone marrow transplant are widely used.[64][65]
Research is underway to develop various sources for stem cells, and to apply stem cell treatments for neurodegenerative diseases and conditions, diabetes, heart disease, and other conditions.[66]
In more recent years, with the ability of scientists to isolate and culture embryonic stem cells, and with scientists' growing ability to create stem cells using somatic cell nuclear transfer and techniques to created induced pluripotent stem cells, controversy has crept in, both related to abortion politics and to human cloning.
Disadvantages
Stem cell treatments may require immunosuppression because of a requirement for radiation before the transplant to remove the patient's previous cells, or because the patient's immune system may target the stem cells. One approach to avoid the second possibility is to use stem cells from the same patient who is being treated.
Pluripotency in certain stem cells could also make it difficult to obtain a specific cell type. It is also difficult to obtain the exact cell type needed, because not all cells in a population differentiate uniformly. Undifferentiated cells can create tissues other than desired types.[67]
Some stem cells form tumors after transplantation[68]; pluripotency is linked to tumor formation especially in embryonic stem cells, fetal proper stem cells, induced pluripotent stem cells. Fetal proper stem cells form tumors despite multipotency.[citation needed]
Hepatotoxicity and drug-induced liver injury account for a substantial number of failures of new drugs in development and market withdrawal, highlighting the need for screening assays such as stem cell-derived hepatocyte-like cells, that are capable of detecting toxicity early in the drug development process.[69]
Research patents
Further information: Consumer Watchdog vs. Wisconsin Alumni Research Foundation
Some of the fundamental patents covering human embryonic stem cells are owned by the Wisconsin Alumni Research Foundation (WARF) - they are patents 5, 843, 780, 6, 200, 806, and 7, 029, 913 invented by James A. Thomson. WARF does not enforce these patents against academic scientists, but does enforce them against companies.[70]
In 2006, a request for the US Patent and Trademark Office (USPTO) to re-examine the three patents was filed by the Public Patent Foundation on behalf of its client, the non-profit patent-watchdog group Consumer Watchdog (formerly the Foundation for Taxpayer and Consumer Rights).[70] In the re-examination process, which involves several rounds of discussion between the USTPO and the parties, the USPTO initially agreed with Consumer Watchdog and rejected all the claims in all three patents, [71] however in response, WARF amended the claims of all three patents to make them more narrow, and in 2008 the USPTO found the amended claims in all three patents to be patentable. The decision on one of the patents (7, 029, 913) was appealable, while the decisions on the other two were not.[72][73] Consumer Watchdog appealed the granting of the '913 patent to the USTPO's Board of Patent Appeals and Interferences (BPAI) which granted the appeal, and in 2010 the BPAI decided that the amended claims of the '913 patent were not patentable.[74] However, WARF was able to re-open prosecution of the case and did so, amending the claims of the '913 patent again to make them more narrow, and in January 2013 the amended claims were allowed.[75]
In July 2013, Consumer Watchdog announced that it would appeal the decision to allow the claims of the '913 patent to the US Court of Appeals for the Federal Circuit (CAFC), the federal appeals court that hears patent cases.[76] At a hearing in December 2013, the CAFC raised the question of whether Consumer Watchdog had legal standing to appeal; the case could not proceed until that issue was resolved.[77]
See also
Cell bank
Human genome
Meristem
Partial cloning
Plant stem cell
Stem cell controversy
Stem cell marker
Shinya Yamanaka
References
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Jump up ^ Chambers I, Colby D, Robertson M, Nichols J, Lee S, Tweedie S, Smith A (2003). "Functional expression cloning of Nanog, a pluripotency sustaining factor in embryonic stem cells". Cell 113 (5): 643–55. doi:10.1016/S[protected]. PMID 12787505.
Jump up ^ Boyer LA, Lee TI, Cole MF, Johnstone SE, Levine SS, Zucker JP, Guenther MG, Kumar RM, Murray HL, Jenner RG, Gifford DK, Melton DA, Jaenisch R, Young RA (2005). "Core transcriptional regulatory circuitry in human embryonic stem cells". Cell 122 (6): 947–56. doi:10.1016/j.cell.2005.08.020. PMC 3006442. PMID 16153702.
Jump up ^ Adewumi O, Aflatoonian B, Ahrlund-Richter L, Amit M, Andrews PW, Beighton G, Bello PA, Benvenisty N, Berry LS, Bevan S, Blum B, Brooking J, Chen KG, Choo AB, Churchill GA, Corbel M, Damjanov I, Draper JS, Dvorak P, Emanuelsson K, Fleck RA, Ford A, Gertow K, Gertsenstein M, Gokhale PJ, Hamilton RS, Hampl A, Healy LE, Hovatta O, Hyllner J, Imreh MP, Itskovitz-Eldor J, Jackson J, Johnson JL, Jones M, Kee K, King BL, Knowles BB, Lako M, Lebrin F, Mallon BS, Manning D, Mayshar Y, McKay RD, Michalska AE, Mikkola M, Mileikovsky M, Minger SL, Moore HD, Mummery CL, Nagy A, Nakatsuji N, O'Brien CM, Oh SK, Olsson C, Otonkoski T, Park KY, Passier R, Patel H, Patel M, Pedersen R, Pera MF, Piekarczyk MS, Pera RA, Reubinoff BE, Robins AJ, Rossant J, Rugg-Gunn P, Schulz TC, Semb H, Sherrer ES, Siemen H, Stacey GN, Stojkovic M, Suemori H, Szatkiewicz J, Turetsky T, Tuuri T, van den Brink S, Vintersten K, Vuoristo S, Ward D, Weaver TA, Young LA, Zhang W (2007). "Characterization of human embryonic stem cell lines by the International Stem Cell Initiative". Nat. Biotechnol 25 (7): 803–16. doi:10.1038/nbt1318. PMID 17572666.
Jump up ^ Ron Winslow (2009). "First Embryonic Stem-Cell Trial Gets Approval from the FDA". The Wall Street Journal. 23. January 2009.
Jump up ^ "Embryonic Stem Cell Therapy At Risk? Geron Ends Clinical Trial". ScienceDebate.com. Retrieved[protected].
Jump up ^ Wu DC, Boyd AS, Wood KJ (2007). "Embryonic stem cell transplantation: potential applicability in cell replacement therapy and regenerative medicine". Front Biosci 12 (8–12): 4525–35. doi:10.2741/2407. PMID 17485394.
Jump up ^ Ariff Bongso; Eng Hin Lee, ed. (2005). "Stem cells: their definition, classification and sources". Stem Cells: From Benchtop to Bedside. World Scientific. p. 5. ISBN[protected]. OCLC[protected].
Jump up ^ Moore, K.L., T.V.N. Persaud, and A.G. Torchia. Before We Are Born: Essentials of Embryology and Birth Defects. Philadelphia, PA: Saunders, Elsevier. 2013. Print
Jump up ^ "Stem Cells" Mayo Clinic. Mayo foundation for medical education and research n.d Web. March 23, 2013
Jump up ^ Jiang Y, Jahagirdar BN, Reinhardt RL, Schwartz RE, Keene CD, Ortiz-Gonzalez XR, Reyes M, Lenvik T, Lund T, Blackstad M, Du J, Aldrich S, Lisberg A, Low WC, Largaespada DA, Verfaillie CM (2002). "Pluripotency of mesenchymal stem cells derived from adult marrow". Nature[protected]: 41–9. doi:10.1038/nature00870. PMID 12077603.
Jump up ^ Ratajczak MZ, Machalinski B, Wojakowski W, Ratajczak J, Kucia M (2007). "A hypothesis for an embryonic origin of pluripotent Oct-4(+) stem cells in adult bone marrow and other tissues". Leukemia 21 (5): 860–7. doi:10.1038/sj.leu.2404630. PMID 17344915.
Jump up ^ Narasipura SD, Wojciechowski JC, Charles N, Liesveld JL, King MR (2008). "P-Selectin coated microtube for enrichment of CD34+ hematopoietic stem and progenitor cells from human bone marrow". Clin Chem 54 (1): 77–85. doi:10.1373/clinchem.2007.089896. PMID 18024531.
Jump up ^ William JB, Prabakaran R, Ayyappan S, Puskhinraj H, Rao D, Manjunath SR, Thamaraikannan P, Dedeepiya VD, Kuroda S, Yoshioka H, Mori Y, Preethy SK, Abraham SJK (2011). "Functional Recovery of Spinal Cord Injury Following Application of Intralesional Bone Marrow Mononuclear Cells Embedded in Polymer Scaffold – Two Year Follow-up in a Canine". Journal of Stem Cell Research & Therapy 1 (3). doi:10.4172/[protected].1000110.
Jump up ^ Terai S, Ishikawa T, Omori K, Aoyama K, Marumoto Y, Urata Y, Yokoyama Y, Uchida K, Yamasaki T, Fujii Y, Okita K, Sakaida I (2006). "Improved liver function in patients with liver cirrhosis after autologous bone marrow cell infusion therapy". Stem Cells 24 (10): 2292–8. doi:10.1634/stemcells.[protected]. PMID 16778155.
Jump up ^ Subrammaniyan R, Amalorpavanathan J, Shankar R et al. (September 2011). "Application of autologous bone marrow mononuclear cells in six patients with advanced chronic critical limb ischemia as a result of diabetes: our experience". Cytotherapy 13 (8): 993–9. doi:10.3109/14653249.2011.579961. PMID 21671823.
Jump up ^ Madhusankar N. "Use of Bone Marrow derived Stem Cells in Patients with Cardiovascular Disorders". Journal of Stem Cells and Regenerative Medicine.
Jump up ^ Dedeepiya VD, Rao YY, Jayakrishnan GA, Parthiban JK, Baskar S, Manjunath SR, Senthilkumar R, Abraham SJ (2012). "Index of CD34+ Cells and Mononuclear Cells in the Bone Marrow of Spinal Cord Injury Patients of Different Age Groups: A Comparative Analysis". Bone Marrow Res 2012: 787414. doi:10.1155/2012/787414. PMC 3398573. PMID 22830032.
Jump up ^ Gardner RL (2002). "Stem cells: potency, plasticity and public perception". Journal of Anatomy 200 (3): 277–82. doi:10.1046/j.[protected].2002.00029.x. PMC 1570679. PMID 12033732.
Jump up ^ Behrens A, van Deursen JM, Rudolph KL, Schumacher B (2014). "Impact of genomic damage and ageing on stem cell function". Nat. Cell Biol. 16 (3): 201–7. doi:10.1038/ncb2928. PMC 4214082. PMID 24576896.
Jump up ^ Barrilleaux B, Phinney DG, Prockop DJ, O'Connor KC (2006). "Review: ex vivo engineering of living tissues with adult stem cells". Tissue Eng 12 (11): 3007–19. doi:10.1089/ten.2006.12.3007. PMID 17518617.
Jump up ^ Gimble JM, Katz AJ, Bunnell BA (2007). "Adipose-derived stem cells for regenerative medicine". Circ Res 100 (9): 1249–60. doi:10.1161/01.RES.[protected].83288.09. PMID 17495232.
Jump up ^ "Bone Marrow Transplant".
Jump up ^ Kane, Ed[protected]. "Stem-cell therapy shows promise for horse soft-tissue injury, disease". DVM Newsmagazine. Retrieved[protected].
Jump up ^ "Stem Cell FAQ". US Department of Health and Human Services.[protected]. Archived from the original on[protected].
Jump up ^ De Coppi P, Bartsch G, Siddiqui MM, Xu T, Santos CC, Perin L, Mostoslavsky G, Serre AC, Snyder EY, Yoo JJ, Furth ME, Soker S, Atala A (2007). "Isolation of amniotic stem cell lines with potential for therapy". Nature Biotechnology 25 (5): 100–106. doi:10.1038/nbt1274. PMID 17206138.
Jump up ^ "Vatican newspaper calls new stem cell source 'future of medicine' :: Catholic News Agency (CNA)". Catholic News Agency.[protected]. Retrieved[protected].
Jump up ^ "European Biotech Company Biocell Center Opens First U.S. Facility for Preservation of Amniotic Stem Cells in Medford, Massachusetts". Reuters.[protected]. Retrieved[protected].
Jump up ^ "Europe's Biocell Center opens Medford office – Daily Business Update". The Boston Globe.[protected]. Retrieved[protected].
Jump up ^ "The Ticker". BostonHerald.com.[protected]. Retrieved[protected].
Jump up ^ "Biocell Center opens amniotic stem cell bank in Medford". Mass High Tech Business News.[protected]. Retrieved[protected].
Jump up ^ "News » World’s First Amniotic Stem Cell Bank Opens In Medford". wbur.org. Retrieved[protected].
Jump up ^ "Biocell Center Corporation Partners with New England's Largest Community-Based Hospital Network to Offer a Unique... – MEDFORD, Mass., March 8 /PRNewswire/". Massachusetts: Prnewswire.com. Retrieved[protected].
^ Jump up to: a b c "Making human embryonic stem cells". The Economist.[protected].
Jump up ^ Brand, Madeleine; Palca, Joe and Cohen, Alex[protected]. "Skin Cells Can Become Embryonic Stem Cells". National Public Radio.
Jump up ^ "Breakthrough Set to Radically Change Stem Cell Debate". News Hour with Jim Lehrer.[protected].
Jump up ^ "His inspiration comes from the research by Prof Shinya Yamanaka at Kyoto University, which suggests a way to create human embryo stem cells without the need for human eggs, which are in extremely short supply, and without the need to create and destroy human cloned embryos, which is bitterly opposed by the pro life movement."Highfield, Roger[protected]. "Dolly creator Prof Ian Wilmut shuns cloning". London: The Telegraph.
Jump up ^ Frozen blood a source of stem cells, study finds. newsdaily.com[protected]
Jump up ^ Beckmann J, Scheitza S, Wernet P, Fischer JC, Giebel B (2007). "Asymmetric cell division within the human hematopoietic stem and progenitor cell compartment: identification of asymmetrically segregating proteins". Blood 109 (12): 5494–501. doi:10.1182/blood[protected]. PMID 17332245.
Jump up ^ Xie T, Spradling AC (1998). "decapentaplegic is essential for the maintenance and division of germline stem cells in the Drosophila ovary". Cell 94 (2): 251–60. doi:10.1016/S[protected]. PMID 9695953.
Jump up ^ Song X, Zhu CH, Doan C, Xie T (2002). "Germline stem cells anchored by adherens junctions in the Drosophila ovary niches". Science[protected]: 1855–7. Bibcode:2002Sci...296.1855S. doi:10.1126/science.1069871. PMID 12052957.
^ Jump up to: a b c d e Stem Cell Basics: What are the potential uses of human stem cells and the obstacles that must be overcome before these potential uses will be realized?. In Stem Cell Information World Wide Web site. Bethesda, MD: National Institutes of Health, U.S. Department of Health and Human Services, 2009. cited Sunday, April 26, 2009
Jump up ^ Steinberg, Douglas (November 2000) Stem Cells Tapped to Replenish Organs thescientist.com
Jump up ^ ISRAEL21c: Israeli scientists reverse brain birth defects using stem cells December 25, 2008. (Researchers from the Hebrew University of Jerusalem-Hadassah Medical led by Prof. Joseph Yanai)
Jump up ^ Kang KS, Kim SW, Oh YH, Yu JW, Kim KY, Park HK, Song CH, Han H (2005). "A 37-year-old spinal cord-injured female patient, transplanted of multipotent stem cells from human UC blood, with improved sensory perception and mobility, both functionally and morphologically: a case study". Cytotherapy 7 (4): 368–73. doi:10.1080/[protected]. PMID 16162459.
Jump up ^ Strauer BE, Schannwell CM, Brehm M (2009). "Therapeutic potentials of stem cells in cardiac diseases". Minerva Cardioangiol 57 (2): 249–67. PMID 19274033.
Jump up ^ Stem Cells Tapped to Replenish Organs thescientist.com, Nov 2000. By Douglas Steinberg
Jump up ^ Hair Cloning Nears Reality as Baldness Cure WebMD November 2004
Jump up ^ Yen AH, Sharpe PT (2008). "Stem cells and tooth tissue engineering". Cell Tissue Res. 331 (1): 359–72. doi:10.1007/s[protected]. PMID 17938970.
Jump up ^ "Gene therapy is first deafness 'cure'". New Scientist. February 14, 2005.
Jump up ^ "BBC NEWS - UK - England - Southern Counties - Stem cells used to restore vision".
Jump up ^ Vastag B (2001). "Stem Cells Step Closer to the Clinic: Paralysis Partially Reversed in Rats with ALS-like Disease". JAMA: the Journal of the American Medical Association 285 (13): 1691–1693. doi:10.1001/jama.285.13.1691. PMID 11277806.
Jump up ^ Anderson, Querida[protected]. "Osiris Trumpets Its Adult Stem Cell Product". Genetic Engineering & Biotechnology News (Mary Ann Liebert, Inc.). p. 13. Retrieved[protected]. (subtitle) Procymal is being developed in many indications, GvHD being the most advanced
Jump up ^ Gurtner GC, Callaghan MJ, Longaker MT (2007). "Progress and potential for regenerative medicine". Annu. Rev. Med. 58: 299–312. doi:10.1146/annurev.med.58.082405.095329. PMID 17076602. Bone marrow transplantation is, as of 2009, the only established use of stem cells.
Jump up ^ Ian Murnaghan for Explore Stem Cells. Updated: 16 December 2013 Why Perform a Stem Cell Transplant?
Jump up ^ Bone Marrow Transplantation and Peripheral Blood Stem Cell Transplantation In National Cancer Institute Fact Sheet web site. Bethesda, MD: National Institutes of Health, U.S. Department of Health and Human Services, 2010. Cited August 24, 2010
Jump up ^ Bubela T, Li MD, Hafez M, Bieber M, Atkins H (2012). "Is belief larger than fact: Expectations, optimism and reality for translational stem cell research". BMC Med 10: 133. doi:10.1186/[protected]. PMC 3520764. PMID 23131007.
Jump up ^ Moore, Keith L., T.V.N. Persaud, and Mark G. Torchia. Before We Are Born: Essentials of Embryology and Birth Defects. Philadelphia, PA: Saunders, Elsevier. 2013 Print.
Jump up ^ Bernadine Healy, M.D.. "Why Embryonic Stem Cells are obsolete" US News and world report. Retrieved on Aug 17, 2015.
Jump up ^ Greenhough S, Hay DC. (2012). "Stem Cell-Based Toxicity Screening: Recent Advances in Hepatocyte Generation". Pharm Med 26 (2): 85–89. doi:10.1007/BF03256896.
^ Jump up to: a b Regalado, Antonio, David P. Hamilton (July 2006). "How a University's Patents May Limit Stem-Cell Researcher." The Wall Street Journal. Retrieved on July 24, 2006.
Jump up ^ Stephen Jenei for Patent Baristas, April 3, 2007 WARF Stem Cell Patents Knocked Down in Round One
Jump up ^ Stephen Jenei for Patent Baristas, March 3, 2008 Ding! WARF Wins Round 2 As Stem Cell Patent Upheld
Jump up ^ Constance Holden for Science Now. March 12, 2008 WARF Goes 3 for 3 on Patents
Jump up ^ Stephen G. Kunin for Patents Post Grant. May 10, 2010 BPAI Rejects WARF Stem Cell Patent Claims in Inter Partes Reexamination Appeal
Jump up ^ United States Patent And Trademark Office. Board Of Patent Appeals and Interferences. The Foundation For Taxpayer & Consumer Rights, Requester And Appellant V. Patent Of Wisconsin Alumni Research Foundation, Patent Owner And Respondent. Appeal[protected], Reexamination Control 95/000, 154. Patent 7, 029, 913 Decision on Appeal
Jump up ^ GenomeWeb staff, July 03, 2013 Consumer Watchdog, PPF Seek Invalidation of WARF's Stem Cell Patent
Jump up ^ Antoinette Konski for Personalized Medicine Bulletin. February 3, 2014 U.S. Government and USPTO Urges Federal Circuit to Dismiss Stem Cell Appeal
features of individual sequences. However, it is possible to create a mixture of smaller probes that are specific to a particular region (locus) of DNA; these mixtures are used to detect deletion mutations. When combined with a specific color, a locus-specific probe mixture is used to detect very specific translocations. Special locus-specific probe mixtures are often used to count chromosomes, by binding to the centromeric regions of chromosomes, which are distinctive enough to identify each chromosome (with the exception of Chromosome 13, 14, 21, 22.)
A variety of other techniques use mixtures of differently colored probes. A range of colors in mixtures of fluorescent dyes can be detected, so each human chromosome can be identified by a characteristic color using whole-chromosome probe mixtures and a variety of ratios of colors. Although there are more chromosomes than easily distinguishable fluorescent dye colors, ratios of probe mixtures can be used to create secondary colors. Similar to comparative genomic hybridization, the probe mixture for the secondary colors is created by mixing the correct ratio of two sets of differently colored probes for the same chromosome. This technique is sometimes called M-FISH.
The same physics that make a variety of colors possible for M-FISH can be used for the detection of translocations. That is, colors that are adjacent appear to overlap; a secondary color is observed. Some assays are designed so that the secondary color will be present or absent in cases of interest. An example is the detection of BCR/ABL translocations, where the secondary color indicates disease. This variation is often called double-fusion FISH or D-FISH. In the opposite situation—where the absence of the secondary color is pathological—is illustrated by an assay used to investigate translocations where only one of the breakpoints is known or constant. Locus-specific probes are made for one side of the breakpoint and the other intact chromosome. In normal cells, the secondary color is observed, but only the primary colors are observed when the translocation occurs. This technique is sometimes called "break-apart FISH".
Stellaris(R) RNA FISH probes[edit]
Stellaris RNA FISH, formerly known as Single Molecule RNA FISH, is a method of detecting and quantifying mRNA and other long RNA molecules in a thin layer of tissue sample. Targets can be reliably imaged through the application of multiple short singly labeled oligonucleotide probes.[12] The binding o[censored]p to 48 fluorescent labeled oligos to a single molecule of mRNA provides sufficient fluorescence to accurately detect and localize each target mRNA in a wide-field fluorescent microscopy image. Probes not binding to the intended sequence do not achieve sufficient localized fluorescence to be distinguished from background.[13]
Single-molecule RNA FISH assays can be performed in simplex or multiplex, and can be used as a follow-up experiment to quantitative PCR, or imaged simultaneously with a fluorescent antibody assay. The technology has potential applications in cancer diagnosis, [14] neuroscience, gene expression analysis, [15] and companion diagnostics.
Fiber FISH[edit]
In an alternative technique to interphase or metaphase preparations, fiber FISH, interphase chromosomes are attached to a slide in such a way that they are stretched out in a straight line, rather than being tightly coiled, as in conventional FISH, or adopting a chromosome territory conformation, as in interphase FISH. This is accomplished by applying mechanical shear along the length of the slide, either to cells that have been fixed to the slide and then lysed, or to a solution of purified DNA. A technique known as chromosome combing is increasingly used for this purpose. The extended conformation of the chromosomes allows dramatically higher resolution – even down to a few kilobases. The preparation of fiber FISH samples, although conceptually simple, is a rather skilled art, and only specialized laboratories use the technique routinely.
Q-FISH[edit]
Q-FISH combines FISH with PNAs and computer software to quantify fluorescence intensity. This technique is used routinely in telomere length research.
Flow-FISH[edit]
Flow-FISH uses flow cytometry to perform FISH automatically using per-cell fluorescence measurements.
Medical applications[edit]
Often parents of children with a developmental disability want to know more about their child's conditions before choosing to have another child. These concerns can be addressed by analysis of the parents' and child's DNA. In cases where the child's developmental disability is not understood, the cause of it can potentially be determined using FISH and cytogenetic techniques. Examples of diseases that are diagnosed using FISH include Prader-Willi syndrome, Angelman syndrome, 22q13 deletion syndrome, chronic myelogenous leukemia, acute lymphoblastic leukemia, Cri-du-chat, Velocardiofacial syndrome, and Down syndrome. FISH on sperm cells is indicated for men with an abnormal somatic or meiotic karyotype as well as those with oligozoospermia, since approximately 50% of oligozoospermic men have an increased rate of sperm chromosome abnormalities.[16] The analysis of chromosomes 21, X, and Y is enough to identify oligozoospermic individuals at risk.[16]
In medicine, FISH can be used to form a diagnosis, to evaluate prognosis, or to evaluate remission of a disease, such as cancer. Treatment can then be specifically tailored. A traditional exam involving metaphase chromosome analysis is often unable to identify features that distinguish one disease from another, due to subtle chromosomal features; FISH can elucidate these differences. FISH can also be used to detect diseased cells more easily than standard Cytogenetic methods, which require dividing cells and requires labor and time-intensive manual preparation and analysis of the slides by a technologist. FISH, on the other hand, does not require living cells and can be quantified automatically, a computer counts the fluorescent dots present. However, a trained technologist is required to distinguish subtle differences in banding patterns on bent and twisted metaphase chromosomes. FISH can be incorporated into Lab-on-a-chip microfluidic device. This technology is still in a developmental stage but, like other lab on a chip methods, it may lead to more portable diagnostic techniques.[17][18]
Species identification[edit]
FISH is often used in clinical studies. If a patient is infected with a suspected pathogen, bacteria, from the patient's tissues or fluids, are typically grown on agar to determine the identity of the pathogen. Many bacteria, however, even well-known species, do not grow well under laboratory conditions. FISH can be used to detect directly the presence of the suspect on small samples of patient's tissue.
FISH can also be used to compare the genomes of two biological species, to deduce evolutionary relationships. A similar hybridization technique is called a zoo blot. Bacterial FISH probes are often primers for the 16s rRNA region.
FISH is widely used in the field of microbial ecology, to identify microorganisms. Biofilms, for example, are composed of complex (often) multi-species bacterial organizations. Preparing DNA probes for one species and performing FISH with this probe allows one to visualize the distribution of this specific species within the biofilm. Preparing probes (in two different colors) for two species allows to visualize/study co-localization of these two species in the biofilm, and can be useful in determining the fine architecture of the biofilm.
Comparative genomic hybridization[edit]
Comparative genomic hybridization can be described as a method that uses FISH in a parallel manner with the comparison of the hybridization strength to recall any major disruptions in the duplication process of the DNA sequences in the genome of the nucleus.[19]
Virtual karyotype[edit]
Virtual karyotyping is another cost-effective, clinically available alternative to FISH panels using thousands to millions of probes on a single array to detect copy number changes, genome-wide, at unprecedented resolution. Currently, this type of analysis will only detect gains and losses of chromosomal material and will not detect balanced rearrangements, such as translocations and inversions which are hallmark aberrations seen in many types of leukemia and lymphoma.
Spectral karyotype[edit]
Spectral karyotyping is an image of colored chromosomes. Spectral karyotyping involves FISH using multiple forms of many types of probes with the result to see each chromosome labeled through its metaphase stage. This type of karyotyping is used specifically when seeking out chromosome arrangements.
features of individual sequences. However, it is possible to create a mixture of smaller probes that are specific to a particular region (locus) of DNA; these mixtures are used to detect deletion mutations. When combined with a specific color, a locus-specific probe mixture is used to detect very specific translocations. Special locus-specific probe mixtures are often used to count chromosomes, by binding to the centromeric regions of chromosomes, which are distinctive enough to identify each chromosome (with the exception of Chromosome 13, 14, 21, 22.)
A variety of other techniques use mixtures of differently colored probes. A range of colors in mixtures of fluorescent dyes can be detected, so each human chromosome can be identified by a characteristic color using whole-chromosome probe mixtures and a variety of ratios of colors. Although there are more chromosomes than easily distinguishable fluorescent dye colors, ratios of probe mixtures can be used to create secondary colors. Similar to comparative genomic hybridization, the probe mixture for the secondary colors is created by mixing the correct ratio of two sets of differently colored probes for the same chromosome. This technique is sometimes called M-FISH.
The same physics that make a variety of colors possible for M-FISH can be used for the detection of translocations. That is, colors that are adjacent appear to overlap; a secondary color is observed. Some assays are designed so that the secondary color will be present or absent in cases of interest. An example is the detection of BCR/ABL translocations, where the secondary color indicates disease. This variation is often called double-fusion FISH or D-FISH. In the opposite situation—where the absence of the secondary color is pathological—is illustrated by an assay used to investigate translocations where only one of the breakpoints is known or constant. Locus-specific probes are made for one side of the breakpoint and the other intact chromosome. In normal cells, the secondary color is observed, but only the primary colors are observed when the translocation occurs. This technique is sometimes called "break-apart FISH".
Stellaris(R) RNA FISH probes[edit]
Stellaris RNA FISH, formerly known as Single Molecule RNA FISH, is a method of detecting and quantifying mRNA and other long RNA molecules in a thin layer of tissue sample. Targets can be reliably imaged through the application of multiple short singly labeled oligonucleotide probes.[12] The binding o[censored]p to 48 fluorescent labeled oligos to a single molecule of mRNA provides sufficient fluorescence to accurately detect and localize each target mRNA in a wide-field fluorescent microscopy image. Probes not binding to the intended sequence do not achieve sufficient localized fluorescence to be distinguished from background.[13]
Single-molecule RNA FISH assays can be performed in simplex or multiplex, and can be used as a follow-up experiment to quantitative PCR, or imaged simultaneously with a fluorescent antibody assay. The technology has potential applications in cancer diagnosis, [14] neuroscience, gene expression analysis, [15] and companion diagnostics.
Fiber FISH[edit]
In an alternative technique to interphase or metaphase preparations, fiber FISH, interphase chromosomes are attached to a slide in such a way that they are stretched out in a straight line, rather than being tightly coiled, as in conventional FISH, or adopting a chromosome territory conformation, as in interphase FISH. This is accomplished by applying mechanical shear along the length of the slide, either to cells that have been fixed to the slide and then lysed, or to a solution of purified DNA. A technique known as chromosome combing is increasingly used for this purpose. The extended conformation of the chromosomes allows dramatically higher resolution – even down to a few kilobases. The preparation of fiber FISH samples, although conceptually simple, is a rather skilled art, and only specialized laboratories use the technique routinely.
Q-FISH[edit]
Q-FISH combines FISH with PNAs and computer software to quantify fluorescence intensity. This technique is used routinely in telomere length research.
Flow-FISH[edit]
Flow-FISH uses flow cytometry to perform FISH automatically using per-cell fluorescence measurements.
Medical applications[edit]
Often parents of children with a developmental disability want to know more about their child's conditions before choosing to have another child. These concerns can be addressed by analysis of the parents' and child's DNA. In cases where the child's developmental disability is not understood, the cause of it can potentially be determined using FISH and cytogenetic techniques. Examples of diseases that are diagnosed using FISH include Prader-Willi syndrome, Angelman syndrome, 22q13 deletion syndrome, chronic myelogenous leukemia, acute lymphoblastic leukemia, Cri-du-chat, Velocardiofacial syndrome, and Down syndrome. FISH on sperm cells is indicated for men with an abnormal somatic or meiotic karyotype as well as those with oligozoospermia, since approximately 50% of oligozoospermic men have an increased rate of sperm chromosome abnormalities.[16] The analysis of chromosomes 21, X, and Y is enough to identify oligozoospermic individuals at risk.[16]
In medicine, FISH can be used to form a diagnosis, to evaluate prognosis, or to evaluate remission of a disease, such as cancer. Treatment can then be specifically tailored. A traditional exam involving metaphase chromosome analysis is often unable to identify features that distinguish one disease from another, due to subtle chromosomal features; FISH can elucidate these differences. FISH can also be used to detect diseased cells more easily than standard Cytogenetic methods, which require dividing cells and requires labor and time-intensive manual preparation and analysis of the slides by a technologist. FISH, on the other hand, does not require living cells and can be quantified automatically, a computer counts the fluorescent dots present. However, a trained technologist is required to distinguish subtle differences in banding patterns on bent and twisted metaphase chromosomes. FISH can be incorporated into Lab-on-a-chip microfluidic device. This technology is still in a developmental stage but, like other lab on a chip methods, it may lead to more portable diagnostic techniques.[17][18]
Species identification[edit]
FISH is often used in clinical studies. If a patient is infected with a suspected pathogen, bacteria, from the patient's tissues or fluids, are typically grown on agar to determine the identity of the pathogen. Many bacteria, however, even well-known species, do not grow well under laboratory conditions. FISH can be used to detect directly the presence of the suspect on small samples of patient's tissue.
FISH can also be used to compare the genomes of two biological species, to deduce evolutionary relationships. A similar hybridization technique is called a zoo blot. Bacterial FISH probes are often primers for the 16s rRNA region.
FISH is widely used in the field of microbial ecology, to identify microorganisms. Biofilms, for example, are composed of complex (often) multi-species bacterial organizations. Preparing DNA probes for one species and performing FISH with this probe allows one to visualize the distribution of this specific species within the biofilm. Preparing probes (in two different colors) for two species allows to visualize/study co-localization of these two species in the biofilm, and can be useful in determining the fine architecture of the biofilm.
Comparative genomic hybridization[edit]
Comparative genomic hybridization can be described as a method that uses FISH in a parallel manner with the comparison of the hybridization strength to recall any major disruptions in the duplication process of the DNA sequences in the genome of the nucleus.[19]
Virtual karyotype[edit]
Virtual karyotyping is another cost-effective, clinically available alternative to FISH panels using thousands to millions of probes on a single array to detect copy number changes, genome-wide, at unprecedented resolution. Currently, this type of analysis will only detect gains and losses of chromosomal material and will not detect balanced rearrangements, such as translocations and inversions which are hallmark aberrations seen in many types of leukemia and lymphoma.
Spectral karyotype[edit]
Spectral karyotyping is an image of colored chromosomes. Spectral karyotyping involves FISH using multiple forms of many types of probes with the result to see each chromosome labeled through its metaphase stage. This type of karyotyping is used specifically when seeking out chromosome arrangements.
features of individual sequences. However, it is possible to create a mixture of smaller probes that are specific to a particular region (locus) of DNA; these mixtures are used to detect deletion mutations. When combined with a specific color, a locus-specific probe mixture is used to detect very specific translocations. Special locus-specific probe mixtures are often used to count chromosomes, by binding to the centromeric regions of chromosomes, which are distinctive enough to identify each chromosome (with the exception of Chromosome 13, 14, 21, 22.)
A variety of other techniques use mixtures of differently colored probes. A range of colors in mixtures of fluorescent dyes can be detected, so each human chromosome can be identified by a characteristic color using whole-chromosome probe mixtures and a variety of ratios of colors. Although there are more chromosomes than easily distinguishable fluorescent dye colors, ratios of probe mixtures can be used to create secondary colors. Similar to comparative genomic hybridization, the probe mixture for the secondary colors is created by mixing the correct ratio of two sets of differently colored probes for the same chromosome. This technique is sometimes called M-FISH.
The same physics that make a variety of colors possible for M-FISH can be used for the detection of translocations. That is, colors that are adjacent appear to overlap; a secondary color is observed. Some assays are designed so that the secondary color will be present or absent in cases of interest. An example is the detection of BCR/ABL translocations, where the secondary color indicates disease. This variation is often called double-fusion FISH or D-FISH. In the opposite situation—where the absence of the secondary color is pathological—is illustrated by an assay used to investigate translocations where only one of the breakpoints is known or constant. Locus-specific probes are made for one side of the breakpoint and the other intact chromosome. In normal cells, the secondary color is observed, but only the primary colors are observed when the translocation occurs. This technique is sometimes called "break-apart FISH".
Stellaris(R) RNA FISH probes[edit]
Stellaris RNA FISH, formerly known as Single Molecule RNA FISH, is a method of detecting and quantifying mRNA and other long RNA molecules in a thin layer of tissue sample. Targets can be reliably imaged through the application of multiple short singly labeled oligonucleotide probes.[12] The binding o[censored]p to 48 fluorescent labeled oligos to a single molecule of mRNA provides sufficient fluorescence to accurately detect and localize each target mRNA in a wide-field fluorescent microscopy image. Probes not binding to the intended sequence do not achieve sufficient localized fluorescence to be distinguished from background.[13]
Single-molecule RNA FISH assays can be performed in simplex or multiplex, and can be used as a follow-up experiment to quantitative PCR, or imaged simultaneously with a fluorescent antibody assay. The technology has potential applications in cancer diagnosis, [14] neuroscience, gene expression analysis, [15] and companion diagnostics.
Fiber FISH[edit]
In an alternative technique to interphase or metaphase preparations, fiber FISH, interphase chromosomes are attached to a slide in such a way that they are stretched out in a straight line, rather than being tightly coiled, as in conventional FISH, or adopting a chromosome territory conformation, as in interphase FISH. This is accomplished by applying mechanical shear along the length of the slide, either to cells that have been fixed to the slide and then lysed, or to a solution of purified DNA. A technique known as chromosome combing is increasingly used for this purpose. The extended conformation of the chromosomes allows dramatically higher resolution – even down to a few kilobases. The preparation of fiber FISH samples, although conceptually simple, is a rather skilled art, and only specialized laboratories use the technique routinely.
Q-FISH[edit]
Q-FISH combines FISH with PNAs and computer software to quantify fluorescence intensity. This technique is used routinely in telomere length research.
Flow-FISH[edit]
Flow-FISH uses flow cytometry to perform FISH automatically using per-cell fluorescence measurements.
Medical applications[edit]
Often parents of children with a developmental disability want to know more about their child's conditions before choosing to have another child. These concerns can be addressed by analysis of the parents' and child's DNA. In cases where the child's developmental disability is not understood, the cause of it can potentially be determined using FISH and cytogenetic techniques. Examples of diseases that are diagnosed using FISH include Prader-Willi syndrome, Angelman syndrome, 22q13 deletion syndrome, chronic myelogenous leukemia, acute lymphoblastic leukemia, Cri-du-chat, Velocardiofacial syndrome, and Down syndrome. FISH on sperm cells is indicated for men with an abnormal somatic or meiotic karyotype as well as those with oligozoospermia, since approximately 50% of oligozoospermic men have an increased rate of sperm chromosome abnormalities.[16] The analysis of chromosomes 21, X, and Y is enough to identify oligozoospermic individuals at risk.[16]
In medicine, FISH can be used to form a diagnosis, to evaluate prognosis, or to evaluate remission of a disease, such as cancer. Treatment can then be specifically tailored. A traditional exam involving metaphase chromosome analysis is often unable to identify features that distinguish one disease from another, due to subtle chromosomal features; FISH can elucidate these differences. FISH can also be used to detect diseased cells more easily than standard Cytogenetic methods, which require dividing cells and requires labor and time-intensive manual preparation and analysis of the slides by a technologist. FISH, on the other hand, does not require living cells and can be quantified automatically, a computer counts the fluorescent dots present. However, a trained technologist is required to distinguish subtle differences in banding patterns on bent and twisted metaphase chromosomes. FISH can be incorporated into Lab-on-a-chip microfluidic device. This technology is still in a developmental stage but, like other lab on a chip methods, it may lead to more portable diagnostic techniques.[17][18]
Species identification[edit]
FISH is often used in clinical studies. If a patient is infected with a suspected pathogen, bacteria, from the patient's tissues or fluids, are typically grown on agar to determine the identity of the pathogen. Many bacteria, however, even well-known species, do not grow well under laboratory conditions. FISH can be used to detect directly the presence of the suspect on small samples of patient's tissue.
FISH can also be used to compare the genomes of two biological species, to deduce evolutionary relationships. A similar hybridization technique is called a zoo blot. Bacterial FISH probes are often primers for the 16s rRNA region.
FISH is widely used in the field of microbial ecology, to identify microorganisms. Biofilms, for example, are composed of complex (often) multi-species bacterial organizations. Preparing DNA probes for one species and performing FISH with this probe allows one to visualize the distribution of this specific species within the biofilm. Preparing probes (in two different colors) for two species allows to visualize/study co-localization of these two species in the biofilm, and can be useful in determining the fine architecture of the biofilm.
Comparative genomic hybridization[edit]
Comparative genomic hybridization can be described as a method that uses FISH in a parallel manner with the comparison of the hybridization strength to recall any major disruptions in the duplication process of the DNA sequences in the genome of the nucleus.[19]
Virtual karyotype[edit]
Virtual karyotyping is another cost-effective, clinically available alternative to FISH panels using thousands to millions of probes on a single array to detect copy number changes, genome-wide, at unprecedented resolution. Currently, this type of analysis will only detect gains and losses of chromosomal material and will not detect balanced rearrangements, such as translocations and inversions which are hallmark aberrations seen in many types of leukemia and lymphoma.
Spectral karyotype[edit]
Spectral karyotyping is an image of colored chromosomes. Spectral karyotyping involves FISH using multiple forms of many types of probes with the result to see each chromosome labeled through its metaphase stage. This type of karyotyping is used specifically when seeking out chromosome arrangements.
features of individual sequences. However, it is possible to create a mixture of smaller probes that are specific to a particular region (locus) of DNA; these mixtures are used to detect deletion mutations. When combined with a specific color, a locus-specific probe mixture is used to detect very specific translocations. Special locus-specific probe mixtures are often used to count chromosomes, by binding to the centromeric regions of chromosomes, which are distinctive enough to identify each chromosome (with the exception of Chromosome 13, 14, 21, 22.)
A variety of other techniques use mixtures of differently colored probes. A range of colors in mixtures of fluorescent dyes can be detected, so each human chromosome can be identified by a characteristic color using whole-chromosome probe mixtures and a variety of ratios of colors. Although there are more chromosomes than easily distinguishable fluorescent dye colors, ratios of probe mixtures can be used to create secondary colors. Similar to comparative genomic hybridization, the probe mixture for the secondary colors is created by mixing the correct ratio of two sets of differently colored probes for the same chromosome. This technique is sometimes called M-FISH.
The same physics that make a variety of colors possible for M-FISH can be used for the detection of translocations. That is, colors that are adjacent appear to overlap; a secondary color is observed. Some assays are designed so that the secondary color will be present or absent in cases of interest. An example is the detection of BCR/ABL translocations, where the secondary color indicates disease. This variation is often called double-fusion FISH or D-FISH. In the opposite situation—where the absence of the secondary color is pathological—is illustrated by an assay used to investigate translocations where only one of the breakpoints is known or constant. Locus-specific probes are made for one side of the breakpoint and the other intact chromosome. In normal cells, the secondary color is observed, but only the primary colors are observed when the translocation occurs. This technique is sometimes called "break-apart FISH".
Stellaris(R) RNA FISH probes[edit]
Stellaris RNA FISH, formerly known as Single Molecule RNA FISH, is a method of detecting and quantifying mRNA and other long RNA molecules in a thin layer of tissue sample. Targets can be reliably imaged through the application of multiple short singly labeled oligonucleotide probes.[12] The binding o[censored]p to 48 fluorescent labeled oligos to a single molecule of mRNA provides sufficient fluorescence to accurately detect and localize each target mRNA in a wide-field fluorescent microscopy image. Probes not binding to the intended sequence do not achieve sufficient localized fluorescence to be distinguished from background.[13]
Single-molecule RNA FISH assays can be performed in simplex or multiplex, and can be used as a follow-up experiment to quantitative PCR, or imaged simultaneously with a fluorescent antibody assay. The technology has potential applications in cancer diagnosis, [14] neuroscience, gene expression analysis, [15] and companion diagnostics.
Fiber FISH[edit]
In an alternative technique to interphase or metaphase preparations, fiber FISH, interphase chromosomes are attached to a slide in such a way that they are stretched out in a straight line, rather than being tightly coiled, as in conventional FISH, or adopting a chromosome territory conformation, as in interphase FISH. This is accomplished by applying mechanical shear along the length of the slide, either to cells that have been fixed to the slide and then lysed, or to a solution of purified DNA. A technique known as chromosome combing is increasingly used for this purpose. The extended conformation of the chromosomes allows dramatically higher resolution – even down to a few kilobases. The preparation of fiber FISH samples, although conceptually simple, is a rather skilled art, and only specialized laboratories use the technique routinely.
Q-FISH[edit]
Q-FISH combines FISH with PNAs and computer software to quantify fluorescence intensity. This technique is used routinely in telomere length research.
Flow-FISH[edit]
Flow-FISH uses flow cytometry to perform FISH automatically using per-cell fluorescence measurements.
Medical applications[edit]
Often parents of children with a developmental disability want to know more about their child's conditions before choosing to have another child. These concerns can be addressed by analysis of the parents' and child's DNA. In cases where the child's developmental disability is not understood, the cause of it can potentially be determined using FISH and cytogenetic techniques. Examples of diseases that are diagnosed using FISH include Prader-Willi syndrome, Angelman syndrome, 22q13 deletion syndrome, chronic myelogenous leukemia, acute lymphoblastic leukemia, Cri-du-chat, Velocardiofacial syndrome, and Down syndrome. FISH on sperm cells is indicated for men with an abnormal somatic or meiotic karyotype as well as those with oligozoospermia, since approximately 50% of oligozoospermic men have an increased rate of sperm chromosome abnormalities.[16] The analysis of chromosomes 21, X, and Y is enough to identify oligozoospermic individuals at risk.[16]
In medicine, FISH can be used to form a diagnosis, to evaluate prognosis, or to evaluate remission of a disease, such as cancer. Treatment can then be specifically tailored. A traditional exam involving metaphase chromosome analysis is often unable to identify features that distinguish one disease from another, due to subtle chromosomal features; FISH can elucidate these differences. FISH can also be used to detect diseased cells more easily than standard Cytogenetic methods, which require dividing cells and requires labor and time-intensive manual preparation and analysis of the slides by a technologist. FISH, on the other hand, does not require living cells and can be quantified automatically, a computer counts the fluorescent dots present. However, a trained technologist is required to distinguish subtle differences in banding patterns on bent and twisted metaphase chromosomes. FISH can be incorporated into Lab-on-a-chip microfluidic device. This technology is still in a developmental stage but, like other lab on a chip methods, it may lead to more portable diagnostic techniques.[17][18]
Species identification[edit]
FISH is often used in clinical studies. If a patient is infected with a suspected pathogen, bacteria, from the patient's tissues or fluids, are typically grown on agar to determine the identity of the pathogen. Many bacteria, however, even well-known species, do not grow well under laboratory conditions. FISH can be used to detect directly the presence of the suspect on small samples of patient's tissue.
FISH can also be used to compare the genomes of two biological species, to deduce evolutionary relationships. A similar hybridization technique is called a zoo blot. Bacterial FISH probes are often primers for the 16s rRNA region.
FISH is widely used in the field of microbial ecology, to identify microorganisms. Biofilms, for example, are composed of complex (often) multi-species bacterial organizations. Preparing DNA probes for one species and performing FISH with this probe allows one to visualize the distribution of this specific species within the biofilm. Preparing probes (in two different colors) for two species allows to visualize/study co-localization of these two species in the biofilm, and can be useful in determining the fine architecture of the biofilm.
Comparative genomic hybridization[edit]
Comparative genomic hybridization can be described as a method that uses FISH in a parallel manner with the comparison of the hybridization strength to recall any major disruptions in the duplication process of the DNA sequences in the genome of the nucleus.[19]
Virtual karyotype[edit]
Virtual karyotyping is another cost-effective, clinically available alternative to FISH panels using thousands to millions of probes on a single array to detect copy number changes, genome-wide, at unprecedented resolution. Currently, this type of analysis will only detect gains and losses of chromosomal material and will not detect balanced rearrangements, such as translocations and inversions which are hallmark aberrations seen in many types of leukemia and lymphoma.
Spectral karyotype[edit]
Spectral karyotyping is an image of colored chromosomes. Spectral karyotyping involves FISH using multiple forms of many types of probes with the result to see each chromosome labeled through its metaphase stage. This type of karyotyping is used specifically when seeking out chromosome arrangements.
features of individual sequences. However, it is possible to create a mixture of smaller probes that are specific to a particular region (locus) of DNA; these mixtures are used to detect deletion mutations. When combined with a specific color, a locus-specific probe mixture is used to detect very specific translocations. Special locus-specific probe mixtures are often used to count chromosomes, by binding to the centromeric regions of chromosomes, which are distinctive enough to identify each chromosome (with the exception of Chromosome 13, 14, 21, 22.)
A variety of other techniques use mixtures of differently colored probes. A range of colors in mixtures of fluorescent dyes can be detected, so each human chromosome can be identified by a characteristic color using whole-chromosome probe mixtures and a variety of ratios of colors. Although there are more chromosomes than easily distinguishable fluorescent dye colors, ratios of probe mixtures can be used to create secondary colors. Similar to comparative genomic hybridization, the probe mixture for the secondary colors is created by mixing the correct ratio of two sets of differently colored probes for the same chromosome. This technique is sometimes called M-FISH.
The same physics that make a variety of colors possible for M-FISH can be used for the detection of translocations. That is, colors that are adjacent appear to overlap; a secondary color is observed. Some assays are designed so that the secondary color will be present or absent in cases of interest. An example is the detection of BCR/ABL translocations, where the secondary color indicates disease. This variation is often called double-fusion FISH or D-FISH. In the opposite situation—where the absence of the secondary color is pathological—is illustrated by an assay used to investigate translocations where only one of the breakpoints is known or constant. Locus-specific probes are made for one side of the breakpoint and the other intact chromosome. In normal cells, the secondary color is observed, but only the primary colors are observed when the translocation occurs. This technique is sometimes called "break-apart FISH".
Stellaris(R) RNA FISH probes[edit]
Stellaris RNA FISH, formerly known as Single Molecule RNA FISH, is a method of detecting and quantifying mRNA and other long RNA molecules in a thin layer of tissue sample. Targets can be reliably imaged through the application of multiple short singly labeled oligonucleotide probes.[12] The binding o[censored]p to 48 fluorescent labeled oligos to a single molecule of mRNA provides sufficient fluorescence to accurately detect and localize each target mRNA in a wide-field fluorescent microscopy image. Probes not binding to the intended sequence do not achieve sufficient localized fluorescence to be distinguished from background.[13]
Single-molecule RNA FISH assays can be performed in simplex or multiplex, and can be used as a follow-up experiment to quantitative PCR, or imaged simultaneously with a fluorescent antibody assay. The technology has potential applications in cancer diagnosis, [14] neuroscience, gene expression analysis, [15] and companion diagnostics.
Fiber FISH[edit]
In an alternative technique to interphase or metaphase preparations, fiber FISH, interphase chromosomes are attached to a slide in such a way that they are stretched out in a straight line, rather than being tightly coiled, as in conventional FISH, or adopting a chromosome territory conformation, as in interphase FISH. This is accomplished by applying mechanical shear along the length of the slide, either to cells that have been fixed to the slide and then lysed, or to a solution of purified DNA. A technique known as chromosome combing is increasingly used for this purpose. The extended conformation of the chromosomes allows dramatically higher resolution – even down to a few kilobases. The preparation of fiber FISH samples, although conceptually simple, is a rather skilled art, and only specialized laboratories use the technique routinely.
Q-FISH[edit]
Q-FISH combines FISH with PNAs and computer software to quantify fluorescence intensity. This technique is used routinely in telomere length research.
Flow-FISH[edit]
Flow-FISH uses flow cytometry to perform FISH automatically using per-cell fluorescence measurements.
Medical applications[edit]
Often parents of children with a developmental disability want to know more about their child's conditions before choosing to have another child. These concerns can be addressed by analysis of the parents' and child's DNA. In cases where the child's developmental disability is not understood, the cause of it can potentially be determined using FISH and cytogenetic techniques. Examples of diseases that are diagnosed using FISH include Prader-Willi syndrome, Angelman syndrome, 22q13 deletion syndrome, chronic myelogenous leukemia, acute lymphoblastic leukemia, Cri-du-chat, Velocardiofacial syndrome, and Down syndrome. FISH on sperm cells is indicated for men with an abnormal somatic or meiotic karyotype as well as those with oligozoospermia, since approximately 50% of oligozoospermic men have an increased rate of sperm chromosome abnormalities.[16] The analysis of chromosomes 21, X, and Y is enough to identify oligozoospermic individuals at risk.[16]
In medicine, FISH can be used to form a diagnosis, to evaluate prognosis, or to evaluate remission of a disease, such as cancer. Treatment can then be specifically tailored. A traditional exam involving metaphase chromosome analysis is often unable to identify features that distinguish one disease from another, due to subtle chromosomal features; FISH can elucidate these differences. FISH can also be used to detect diseased cells more easily than standard Cytogenetic methods, which require dividing cells and requires labor and time-intensive manual preparation and analysis of the slides by a technologist. FISH, on the other hand, does not require living cells and can be quantified automatically, a computer counts the fluorescent dots present. However, a trained technologist is required to distinguish subtle differences in banding patterns on bent and twisted metaphase chromosomes. FISH can be incorporated into Lab-on-a-chip microfluidic device. This technology is still in a developmental stage but, like other lab on a chip methods, it may lead to more portable diagnostic techniques.[17][18]
Species identification[edit]
FISH is often used in clinical studies. If a patient is infected with a suspected pathogen, bacteria, from the patient's tissues or fluids, are typically grown on agar to determine the identity of the pathogen. Many bacteria, however, even well-known species, do not grow well under laboratory conditions. FISH can be used to detect directly the presence of the suspect on small samples of patient's tissue.
FISH can also be used to compare the genomes of two biological species, to deduce evolutionary relationships. A similar hybridization technique is called a zoo blot. Bacterial FISH probes are often primers for the 16s rRNA region.
FISH is widely used in the field of microbial ecology, to identify microorganisms. Biofilms, for example, are composed of complex (often) multi-species bacterial organizations. Preparing DNA probes for one species and performing FISH with this probe allows one to visualize the distribution of this specific species within the biofilm. Preparing probes (in two different colors) for two species allows to visualize/study co-localization of these two species in the biofilm, and can be useful in determining the fine architecture of the biofilm.
Comparative genomic hybridization[edit]
Comparative genomic hybridization can be described as a method that uses FISH in a parallel manner with the comparison of the hybridization strength to recall any major disruptions in the duplication process of the DNA sequences in the genome of the nucleus.[19]
Virtual karyotype[edit]
Virtual karyotyping is another cost-effective, clinically available alternative to FISH panels using thousands to millions of probes on a single array to detect copy number changes, genome-wide, at unprecedented resolution. Currently, this type of analysis will only detect gains and losses of chromosomal material and will not detect balanced rearrangements, such as translocations and inversions which are hallmark aberrations seen in many types of leukemia and lymphoma.
Spectral karyotype[edit]
Spectral karyotyping is an image of colored chromosomes. Spectral karyotyping involves FISH using multiple forms of many types of probes with the result to see each chromosome labeled through its metaphase stage. This type of karyotyping is used specifically when seeking out chromosome arrangements.
A variety of other techniques use mixtures of differently colored probes. A range of colors in mixtures of fluorescent dyes can be detected, so each human chromosome can be identified by a characteristic color using whole-chromosome probe mixtures and a variety of ratios of colors. Although there are more chromosomes than easily distinguishable fluorescent dye colors, ratios of probe mixtures can be used to create secondary colors. Similar to comparative genomic hybridization, the probe mixture for the secondary colors is created by mixing the correct ratio of two sets of differently colored probes for the same chromosome. This technique is sometimes called M-FISH.
The same physics that make a variety of colors possible for M-FISH can be used for the detection of translocations. That is, colors that are adjacent appear to overlap; a secondary color is observed. Some assays are designed so that the secondary color will be present or absent in cases of interest. An example is the detection of BCR/ABL translocations, where the secondary color indicates disease. This variation is often called double-fusion FISH or D-FISH. In the opposite situation—where the absence of the secondary color is pathological—is illustrated by an assay used to investigate translocations where only one of the breakpoints is known or constant. Locus-specific probes are made for one side of the breakpoint and the other intact chromosome. In normal cells, the secondary color is observed, but only the primary colors are observed when the translocation occurs. This technique is sometimes called "break-apart FISH".
Stellaris(R) RNA FISH probes[edit]
Stellaris RNA FISH, formerly known as Single Molecule RNA FISH, is a method of detecting and quantifying mRNA and other long RNA molecules in a thin layer of tissue sample. Targets can be reliably imaged through the application of multiple short singly labeled oligonucleotide probes.[12] The binding o[censored]p to 48 fluorescent labeled oligos to a single molecule of mRNA provides sufficient fluorescence to accurately detect and localize each target mRNA in a wide-field fluorescent microscopy image. Probes not binding to the intended sequence do not achieve sufficient localized fluorescence to be distinguished from background.[13]
Single-molecule RNA FISH assays can be performed in simplex or multiplex, and can be used as a follow-up experiment to quantitative PCR, or imaged simultaneously with a fluorescent antibody assay. The technology has potential applications in cancer diagnosis, [14] neuroscience, gene expression analysis, [15] and companion diagnostics.
Fiber FISH[edit]
In an alternative technique to interphase or metaphase preparations, fiber FISH, interphase chromosomes are attached to a slide in such a way that they are stretched out in a straight line, rather than being tightly coiled, as in conventional FISH, or adopting a chromosome territory conformation, as in interphase FISH. This is accomplished by applying mechanical shear along the length of the slide, either to cells that have been fixed to the slide and then lysed, or to a solution of purified DNA. A technique known as chromosome combing is increasingly used for this purpose. The extended conformation of the chromosomes allows dramatically higher resolution – even down to a few kilobases. The preparation of fiber FISH samples, although conceptually simple, is a rather skilled art, and only specialized laboratories use the technique routinely.
Q-FISH[edit]
Q-FISH combines FISH with PNAs and computer software to quantify fluorescence intensity. This technique is used routinely in telomere length research.
Flow-FISH[edit]
Flow-FISH uses flow cytometry to perform FISH automatically using per-cell fluorescence measurements.
Medical applications[edit]
Often parents of children with a developmental disability want to know more about their child's conditions before choosing to have another child. These concerns can be addressed by analysis of the parents' and child's DNA. In cases where the child's developmental disability is not understood, the cause of it can potentially be determined using FISH and cytogenetic techniques. Examples of diseases that are diagnosed using FISH include Prader-Willi syndrome, Angelman syndrome, 22q13 deletion syndrome, chronic myelogenous leukemia, acute lymphoblastic leukemia, Cri-du-chat, Velocardiofacial syndrome, and Down syndrome. FISH on sperm cells is indicated for men with an abnormal somatic or meiotic karyotype as well as those with oligozoospermia, since approximately 50% of oligozoospermic men have an increased rate of sperm chromosome abnormalities.[16] The analysis of chromosomes 21, X, and Y is enough to identify oligozoospermic individuals at risk.[16]
In medicine, FISH can be used to form a diagnosis, to evaluate prognosis, or to evaluate remission of a disease, such as cancer. Treatment can then be specifically tailored. A traditional exam involving metaphase chromosome analysis is often unable to identify features that distinguish one disease from another, due to subtle chromosomal features; FISH can elucidate these differences. FISH can also be used to detect diseased cells more easily than standard Cytogenetic methods, which require dividing cells and requires labor and time-intensive manual preparation and analysis of the slides by a technologist. FISH, on the other hand, does not require living cells and can be quantified automatically, a computer counts the fluorescent dots present. However, a trained technologist is required to distinguish subtle differences in banding patterns on bent and twisted metaphase chromosomes. FISH can be incorporated into Lab-on-a-chip microfluidic device. This technology is still in a developmental stage but, like other lab on a chip methods, it may lead to more portable diagnostic techniques.[17][18]
Species identification[edit]
FISH is often used in clinical studies. If a patient is infected with a suspected pathogen, bacteria, from the patient's tissues or fluids, are typically grown on agar to determine the identity of the pathogen. Many bacteria, however, even well-known species, do not grow well under laboratory conditions. FISH can be used to detect directly the presence of the suspect on small samples of patient's tissue.
FISH can also be used to compare the genomes of two biological species, to deduce evolutionary relationships. A similar hybridization technique is called a zoo blot. Bacterial FISH probes are often primers for the 16s rRNA region.
FISH is widely used in the field of microbial ecology, to identify microorganisms. Biofilms, for example, are composed of complex (often) multi-species bacterial organizations. Preparing DNA probes for one species and performing FISH with this probe allows one to visualize the distribution of this specific species within the biofilm. Preparing probes (in two different colors) for two species allows to visualize/study co-localization of these two species in the biofilm, and can be useful in determining the fine architecture of the biofilm.
Comparative genomic hybridization[edit]
Comparative genomic hybridization can be described as a method that uses FISH in a parallel manner with the comparison of the hybridization strength to recall any major disruptions in the duplication process of the DNA sequences in the genome of the nucleus.[19]
Virtual karyotype[edit]
Virtual karyotyping is another cost-effective, clinically available alternative to FISH panels using thousands to millions of probes on a single array to detect copy number changes, genome-wide, at unprecedented resolution. Currently, this type of analysis will only detect gains and losses of chromosomal material and will not detect balanced rearrangements, such as translocations and inversions which are hallmark aberrations seen in many types of leukemia and lymphoma.
Spectral karyotype[edit]
Spectral karyotyping is an image of colored chromosomes. Spectral karyotyping involves FISH using multiple forms of many types of probes with the result to see each chromosome labeled through its metaphase stage. This type of karyotyping is used specifically when seeking out chromosome arrangements.
features of individual sequences. However, it is possible to create a mixture of smaller probes that are specific to a particular region (locus) of DNA; these mixtures are used to detect deletion mutations. When combined with a specific color, a locus-specific probe mixture is used to detect very specific translocations. Special locus-specific probe mixtures are often used to count chromosomes, by binding to the centromeric regions of chromosomes, which are distinctive enough to identify each chromosome (with the exception of Chromosome 13, 14, 21, 22.)
A variety of other techniques use mixtures of differently colored probes. A range of colors in mixtures of fluorescent dyes can be detected, so each human chromosome can be identified by a characteristic color using whole-chromosome probe mixtures and a variety of ratios of colors. Although there are more chromosomes than easily distinguishable fluorescent dye colors, ratios of probe mixtures can be used to create secondary colors. Similar to comparative genomic hybridization, the probe mixture for the secondary colors is created by mixing the correct ratio of two sets of differently colored probes for the same chromosome. This technique is sometimes called M-FISH.
The same physics that make a variety of colors possible for M-FISH can be used for the detection of translocations. That is, colors that are adjacent appear to overlap; a secondary color is observed. Some assays are designed so that the secondary color will be present or absent in cases of interest. An example is the detection of BCR/ABL translocations, where the secondary color indicates disease. This variation is often called double-fusion FISH or D-FISH. In the opposite situation—where the absence of the secondary color is pathological—is illustrated by an assay used to investigate translocations where only one of the breakpoints is known or constant. Locus-specific probes are made for one side of the breakpoint and the other intact chromosome. In normal cells, the secondary color is observed, but only the primary colors are observed when the translocation occurs. This technique is sometimes called "break-apart FISH".
Stellaris(R) RNA FISH probes[edit]
Stellaris RNA FISH, formerly known as Single Molecule RNA FISH, is a method of detecting and quantifying mRNA and other long RNA molecules in a thin layer of tissue sample. Targets can be reliably imaged through the application of multiple short singly labeled oligonucleotide probes.[12] The binding o[censored]p to 48 fluorescent labeled oligos to a single molecule of mRNA provides sufficient fluorescence to accurately detect and localize each target mRNA in a wide-field fluorescent microscopy image. Probes not binding to the intended sequence do not achieve sufficient localized fluorescence to be distinguished from background.[13]
Single-molecule RNA FISH assays can be performed in simplex or multiplex, and can be used as a follow-up experiment to quantitative PCR, or imaged simultaneously with a fluorescent antibody assay. The technology has potential applications in cancer diagnosis, [14] neuroscience, gene expression analysis, [15] and companion diagnostics.
Fiber FISH[edit]
In an alternative technique to interphase or metaphase preparations, fiber FISH, interphase chromosomes are attached to a slide in such a way that they are stretched out in a straight line, rather than being tightly coiled, as in conventional FISH, or adopting a chromosome territory conformation, as in interphase FISH. This is accomplished by applying mechanical shear along the length of the slide, either to cells that have been fixed to the slide and then lysed, or to a solution of purified DNA. A technique known as chromosome combing is increasingly used for this purpose. The extended conformation of the chromosomes allows dramatically higher resolution – even down to a few kilobases. The preparation of fiber FISH samples, although conceptually simple, is a rather skilled art, and only specialized laboratories use the technique routinely.
Q-FISH[edit]
Q-FISH combines FISH with PNAs and computer software to quantify fluorescence intensity. This technique is used routinely in telomere length research.
Flow-FISH[edit]
Flow-FISH uses flow cytometry to perform FISH automatically using per-cell fluorescence measurements.
Medical applications[edit]
Often parents of children with a developmental disability want to know more about their child's conditions before choosing to have another child. These concerns can be addressed by analysis of the parents' and child's DNA. In cases where the child's developmental disability is not understood, the cause of it can potentially be determined using FISH and cytogenetic techniques. Examples of diseases that are diagnosed using FISH include Prader-Willi syndrome, Angelman syndrome, 22q13 deletion syndrome, chronic myelogenous leukemia, acute lymphoblastic leukemia, Cri-du-chat, Velocardiofacial syndrome, and Down syndrome. FISH on sperm cells is indicated for men with an abnormal somatic or meiotic karyotype as well as those with oligozoospermia, since approximately 50% of oligozoospermic men have an increased rate of sperm chromosome abnormalities.[16] The analysis of chromosomes 21, X, and Y is enough to identify oligozoospermic individuals at risk.[16]
In medicine, FISH can be used to form a diagnosis, to evaluate prognosis, or to evaluate remission of a disease, such as cancer. Treatment can then be specifically tailored. A traditional exam involving metaphase chromosome analysis is often unable to identify features that distinguish one disease from another, due to subtle chromosomal features; FISH can elucidate these differences. FISH can also be used to detect diseased cells more easily than standard Cytogenetic methods, which require dividing cells and requires labor and time-intensive manual preparation and analysis of the slides by a technologist. FISH, on the other hand, does not require living cells and can be quantified automatically, a computer counts the fluorescent dots present. However, a trained technologist is required to distinguish subtle differences in banding patterns on bent and twisted metaphase chromosomes. FISH can be incorporated into Lab-on-a-chip microfluidic device. This technology is still in a developmental stage but, like other lab on a chip methods, it may lead to more portable diagnostic techniques.[17][18]
Species identification[edit]
FISH is often used in clinical studies. If a patient is infected with a suspected pathogen, bacteria, from the patient's tissues or fluids, are typically grown on agar to determine the identity of the pathogen. Many bacteria, however, even well-known species, do not grow well under laboratory conditions. FISH can be used to detect directly the presence of the suspect on small samples of patient's tissue.
FISH can also be used to compare the genomes of two biological species, to deduce evolutionary relationships. A similar hybridization technique is called a zoo blot. Bacterial FISH probes are often primers for the 16s rRNA region.
FISH is widely used in the field of microbial ecology, to identify microorganisms. Biofilms, for example, are composed of complex (often) multi-species bacterial organizations. Preparing DNA probes for one species and performing FISH with this probe allows one to visualize the distribution of this specific species within the biofilm. Preparing probes (in two different colors) for two species allows to visualize/study co-localization of these two species in the biofilm, and can be useful in determining the fine architecture of the biofilm.
Comparative genomic hybridization[edit]
Comparative genomic hybridization can be described as a method that uses FISH in a parallel manner with the comparison of the hybridization strength to recall any major disruptions in the duplication process of the DNA sequences in the genome of the nucleus.[19]
Virtual karyotype[edit]
Virtual karyotyping is another cost-effective, clinically available alternative to FISH panels using thousands to millions of probes on a single array to detect copy number changes, genome-wide, at unprecedented resolution. Currently, this type of analysis will only detect gains and losses of chromosomal material and will not detect balanced rearrangements, such as translocations and inversions which are hallmark aberrations seen in many types of leukemia and lymphoma.
Spectral karyotype[edit]
Spectral karyotyping is an image of colored chromosomes. Spectral karyotyping involves FISH using multiple forms of many types of probes with the result to see each chromosome labeled through its metaphase stage. This type of karyotyping is used specifically when seeking out chromosome arrangements.
features of individual sequences. However, it is possible to create a mixture of smaller probes that are specific to a particular region (locus) of DNA; these mixtures are used to detect deletion mutations. When combined with a specific color, a locus-specific probe mixture is used to detect very specific translocations. Special locus-specific probe mixtures are often used to count chromosomes, by binding to the centromeric regions of chromosomes, which are distinctive enough to identify each chromosome (with the exception of Chromosome 13, 14, 21, 22.)
A variety of other techniques use mixtures of differently colored probes. A range of colors in mixtures of fluorescent dyes can be detected, so each human chromosome can be identified by a characteristic color using whole-chromosome probe mixtures and a variety of ratios of colors. Although there are more chromosomes than easily distinguishable fluorescent dye colors, ratios of probe mixtures can be used to create secondary colors. Similar to comparative genomic hybridization, the probe mixture for the secondary colors is created by mixing the correct ratio of two sets of differently colored probes for the same chromosome. This technique is sometimes called M-FISH.
The same physics that make a variety of colors possible for M-FISH can be used for the detection of translocations. That is, colors that are adjacent appear to overlap; a secondary color is observed. Some assays are designed so that the secondary color will be present or absent in cases of interest. An example is the detection of BCR/ABL translocations, where the secondary color indicates disease. This variation is often called double-fusion FISH or D-FISH. In the opposite situation—where the absence of the secondary color is pathological—is illustrated by an assay used to investigate translocations where only one of the breakpoints is known or constant. Locus-specific probes are made for one side of the breakpoint and the other intact chromosome. In normal cells, the secondary color is observed, but only the primary colors are observed when the translocation occurs. This technique is sometimes called "break-apart FISH".
Stellaris(R) RNA FISH probes[edit]
Stellaris RNA FISH, formerly known as Single Molecule RNA FISH, is a method of detecting and quantifying mRNA and other long RNA molecules in a thin layer of tissue sample. Targets can be reliably imaged through the application of multiple short singly labeled oligonucleotide probes.[12] The binding o[censored]p to 48 fluorescent labeled oligos to a single molecule of mRNA provides sufficient fluorescence to accurately detect and localize each target mRNA in a wide-field fluorescent microscopy image. Probes not binding to the intended sequence do not achieve sufficient localized fluorescence to be distinguished from background.[13]
Single-molecule RNA FISH assays can be performed in simplex or multiplex, and can be used as a follow-up experiment to quantitative PCR, or imaged simultaneously with a fluorescent antibody assay. The technology has potential applications in cancer diagnosis, [14] neuroscience, gene expression analysis, [15] and companion diagnostics.
Fiber FISH[edit]
In an alternative technique to interphase or metaphase preparations, fiber FISH, interphase chromosomes are attached to a slide in such a way that they are stretched out in a straight line, rather than being tightly coiled, as in conventional FISH, or adopting a chromosome territory conformation, as in interphase FISH. This is accomplished by applying mechanical shear along the length of the slide, either to cells that have been fixed to the slide and then lysed, or to a solution of purified DNA. A technique known as chromosome combing is increasingly used for this purpose. The extended conformation of the chromosomes allows dramatically higher resolution – even down to a few kilobases. The preparation of fiber FISH samples, although conceptually simple, is a rather skilled art, and only specialized laboratories use the technique routinely.
Q-FISH[edit]
Q-FISH combines FISH with PNAs and computer software to quantify fluorescence intensity. This technique is used routinely in telomere length research.
Flow-FISH[edit]
Flow-FISH uses flow cytometry to perform FISH automatically using per-cell fluorescence measurements.
Medical applications[edit]
Often parents of children with a developmental disability want to know more about their child's conditions before choosing to have another child. These concerns can be addressed by analysis of the parents' and child's DNA. In cases where the child's developmental disability is not understood, the cause of it can potentially be determined using FISH and cytogenetic techniques. Examples of diseases that are diagnosed using FISH include Prader-Willi syndrome, Angelman syndrome, 22q13 deletion syndrome, chronic myelogenous leukemia, acute lymphoblastic leukemia, Cri-du-chat, Velocardiofacial syndrome, and Down syndrome. FISH on sperm cells is indicated for men with an abnormal somatic or meiotic karyotype as well as those with oligozoospermia, since approximately 50% of oligozoospermic men have an increased rate of sperm chromosome abnormalities.[16] The analysis of chromosomes 21, X, and Y is enough to identify oligozoospermic individuals at risk.[16]
In medicine, FISH can be used to form a diagnosis, to evaluate prognosis, or to evaluate remission of a disease, such as cancer. Treatment can then be specifically tailored. A traditional exam involving metaphase chromosome analysis is often unable to identify features that distinguish one disease from another, due to subtle chromosomal features; FISH can elucidate these differences. FISH can also be used to detect diseased cells more easily than standard Cytogenetic methods, which require dividing cells and requires labor and time-intensive manual preparation and analysis of the slides by a technologist. FISH, on the other hand, does not require living cells and can be quantified automatically, a computer counts the fluorescent dots present. However, a trained technologist is required to distinguish subtle differences in banding patterns on bent and twisted metaphase chromosomes. FISH can be incorporated into Lab-on-a-chip microfluidic device. This technology is still in a developmental stage but, like other lab on a chip methods, it may lead to more portable diagnostic techniques.[17][18]
Species identification[edit]
FISH is often used in clinical studies. If a patient is infected with a suspected pathogen, bacteria, from the patient's tissues or fluids, are typically grown on agar to determine the identity of the pathogen. Many bacteria, however, even well-known species, do not grow well under laboratory conditions. FISH can be used to detect directly the presence of the suspect on small samples of patient's tissue.
FISH can also be used to compare the genomes of two biological species, to deduce evolutionary relationships. A similar hybridization technique is called a zoo blot. Bacterial FISH probes are often primers for the 16s rRNA region.
FISH is widely used in the field of microbial ecology, to identify microorganisms. Biofilms, for example, are composed of complex (often) multi-species bacterial organizations. Preparing DNA probes for one species and performing FISH with this probe allows one to visualize the distribution of this specific species within the biofilm. Preparing probes (in two different colors) for two species allows to visualize/study co-localization of these two species in the biofilm, and can be useful in determining the fine architecture of the biofilm.
Comparative genomic hybridization[edit]
Comparative genomic hybridization can be described as a method that uses FISH in a parallel manner with the comparison of the hybridization strength to recall any major disruptions in the duplication process of the DNA sequences in the genome of the nucleus.[19]
Virtual karyotype[edit]
Virtual karyotyping is another cost-effective, clinically available alternative to FISH panels using thousands to millions of probes on a single array to detect copy number changes, genome-wide, at unprecedented resolution. Currently, this type of analysis will only detect gains and losses of chromosomal material and will not detect balanced rearrangements, such as translocations and inversions which are hallmark aberrations seen in many types of leukemia and lymphoma.
Spectral karyotype[edit]
Spectral karyotyping is an image of colored chromosomes. Spectral karyotyping involves FISH using multiple forms of many types of probes with the result to see each chromosome labeled through its metaphase stage. This type of karyotyping is used specifically when seeking out chromosome arrangements.
features of individual sequences. However, it is possible to create a mixture of smaller probes that are specific to a particular region (locus) of DNA; these mixtures are used to detect deletion mutations. When combined with a specific color, a locus-specific probe mixture is used to detect very specific translocations. Special locus-specific probe mixtures are often used to count chromosomes, by binding to the centromeric regions of chromosomes, which are distinctive enough to identify each chromosome (with the exception of Chromosome 13, 14, 21, 22.)
A variety of other techniques use mixtures of differently colored probes. A range of colors in mixtures of fluorescent dyes can be detected, so each human chromosome can be identified by a characteristic color using whole-chromosome probe mixtures and a variety of ratios of colors. Although there are more chromosomes than easily distinguishable fluorescent dye colors, ratios of probe mixtures can be used to create secondary colors. Similar to comparative genomic hybridization, the probe mixture for the secondary colors is created by mixing the correct ratio of two sets of differently colored probes for the same chromosome. This technique is sometimes called M-FISH.
The same physics that make a variety of colors possible for M-FISH can be used for the detection of translocations. That is, colors that are adjacent appear to overlap; a secondary color is observed. Some assays are designed so that the secondary color will be present or absent in cases of interest. An example is the detection of BCR/ABL translocations, where the secondary color indicates disease. This variation is often called double-fusion FISH or D-FISH. In the opposite situation—where the absence of the secondary color is pathological—is illustrated by an assay used to investigate translocations where only one of the breakpoints is known or constant. Locus-specific probes are made for one side of the breakpoint and the other intact chromosome. In normal cells, the secondary color is observed, but only the primary colors are observed when the translocation occurs. This technique is sometimes called "break-apart FISH".
Stellaris(R) RNA FISH probes[edit]
Stellaris RNA FISH, formerly known as Single Molecule RNA FISH, is a method of detecting and quantifying mRNA and other long RNA molecules in a thin layer of tissue sample. Targets can be reliably imaged through the application of multiple short singly labeled oligonucleotide probes.[12] The binding o[censored]p to 48 fluorescent labeled oligos to a single molecule of mRNA provides sufficient fluorescence to accurately detect and localize each target mRNA in a wide-field fluorescent microscopy image. Probes not binding to the intended sequence do not achieve sufficient localized fluorescence to be distinguished from background.[13]
Single-molecule RNA FISH assays can be performed in simplex or multiplex, and can be used as a follow-up experiment to quantitative PCR, or imaged simultaneously with a fluorescent antibody assay. The technology has potential applications in cancer diagnosis, [14] neuroscience, gene expression analysis, [15] and companion diagnostics.
Fiber FISH[edit]
In an alternative technique to interphase or metaphase preparations, fiber FISH, interphase chromosomes are attached to a slide in such a way that they are stretched out in a straight line, rather than being tightly coiled, as in conventional FISH, or adopting a chromosome territory conformation, as in interphase FISH. This is accomplished by applying mechanical shear along the length of the slide, either to cells that have been fixed to the slide and then lysed, or to a solution of purified DNA. A technique known as chromosome combing is increasingly used for this purpose. The extended conformation of the chromosomes allows dramatically higher resolution – even down to a few kilobases. The preparation of fiber FISH samples, although conceptually simple, is a rather skilled art, and only specialized laboratories use the technique routinely.
Q-FISH[edit]
Q-FISH combines FISH with PNAs and computer software to quantify fluorescence intensity. This technique is used routinely in telomere length research.
Flow-FISH[edit]
Flow-FISH uses flow cytometry to perform FISH automatically using per-cell fluorescence measurements.
Medical applications[edit]
Often parents of children with a developmental disability want to know more about their child's conditions before choosing to have another child. These concerns can be addressed by analysis of the parents' and child's DNA. In cases where the child's developmental disability is not understood, the cause of it can potentially be determined using FISH and cytogenetic techniques. Examples of diseases that are diagnosed using FISH include Prader-Willi syndrome, Angelman syndrome, 22q13 deletion syndrome, chronic myelogenous leukemia, acute lymphoblastic leukemia, Cri-du-chat, Velocardiofacial syndrome, and Down syndrome. FISH on sperm cells is indicated for men with an abnormal somatic or meiotic karyotype as well as those with oligozoospermia, since approximately 50% of oligozoospermic men have an increased rate of sperm chromosome abnormalities.[16] The analysis of chromosomes 21, X, and Y is enough to identify oligozoospermic individuals at risk.[16]
In medicine, FISH can be used to form a diagnosis, to evaluate prognosis, or to evaluate remission of a disease, such as cancer. Treatment can then be specifically tailored. A traditional exam involving metaphase chromosome analysis is often unable to identify features that distinguish one disease from another, due to subtle chromosomal features; FISH can elucidate these differences. FISH can also be used to detect diseased cells more easily than standard Cytogenetic methods, which require dividing cells and requires labor and time-intensive manual preparation and analysis of the slides by a technologist. FISH, on the other hand, does not require living cells and can be quantified automatically, a computer counts the fluorescent dots present. However, a trained technologist is required to distinguish subtle differences in banding patterns on bent and twisted metaphase chromosomes. FISH can be incorporated into Lab-on-a-chip microfluidic device. This technology is still in a developmental stage but, like other lab on a chip methods, it may lead to more portable diagnostic techniques.[17][18]
Species identification[edit]
FISH is often used in clinical studies. If a patient is infected with a suspected pathogen, bacteria, from the patient's tissues or fluids, are typically grown on agar to determine the identity of the pathogen. Many bacteria, however, even well-known species, do not grow well under laboratory conditions. FISH can be used to detect directly the presence of the suspect on small samples of patient's tissue.
FISH can also be used to compare the genomes of two biological species, to deduce evolutionary relationships. A similar hybridization technique is called a zoo blot. Bacterial FISH probes are often primers for the 16s rRNA region.
FISH is widely used in the field of microbial ecology, to identify microorganisms. Biofilms, for example, are composed of complex (often) multi-species bacterial organizations. Preparing DNA probes for one species and performing FISH with this probe allows one to visualize the distribution of this specific species within the biofilm. Preparing probes (in two different colors) for two species allows to visualize/study co-localization of these two species in the biofilm, and can be useful in determining the fine architecture of the biofilm.
Comparative genomic hybridization[edit]
Comparative genomic hybridization can be described as a method that uses FISH in a parallel manner with the comparison of the hybridization strength to recall any major disruptions in the duplication process of the DNA sequences in the genome of the nucleus.[19]
Virtual karyotype[edit]
Virtual karyotyping is another cost-effective, clinically available alternative to FISH panels using thousands to millions of probes on a single array to detect copy number changes, genome-wide, at unprecedented resolution. Currently, this type of analysis will only detect gains and losses of chromosomal material and will not detect balanced rearrangements, such as translocations and inversions which are hallmark aberrations seen in many types of leukemia and lymphoma.
Spectral karyotype[edit]
Spectral karyotyping is an image of colored chromosomes. Spectral karyotyping involves FISH using multiple forms of many types of probes with the result to see each chromosome labeled through its metaphase stage. This type of karyotyping is used specifically when seeking out chromosome arrangements.
features of individual sequences. However, it is possible to create a mixture of smaller probes that are specific to a particular region (locus) of DNA; these mixtures are used to detect deletion mutations. When combined with a specific color, a locus-specific probe mixture is used to detect very specific translocations. Special locus-specific probe mixtures are often used to count chromosomes, by binding to the centromeric regions of chromosomes, which are distinctive enough to identify each chromosome (with the exception of Chromosome 13, 14, 21, 22.)
A variety of other techniques use mixtures of differently colored probes. A range of colors in mixtures of fluorescent dyes can be detected, so each human chromosome can be identified by a characteristic color using whole-chromosome probe mixtures and a variety of ratios of colors. Although there are more chromosomes than easily distinguishable fluorescent dye colors, ratios of probe mixtures can be used to create secondary colors. Similar to comparative genomic hybridization, the probe mixture for the secondary colors is created by mixing the correct ratio of two sets of differently colored probes for the same chromosome. This technique is sometimes called M-FISH.
The same physics that make a variety of colors possible for M-FISH can be used for the detection of translocations. That is, colors that are adjacent appear to overlap; a secondary color is observed. Some assays are designed so that the secondary color will be present or absent in cases of interest. An example is the detection of BCR/ABL translocations, where the secondary color indicates disease. This variation is often called double-fusion FISH or D-FISH. In the opposite situation—where the absence of the secondary color is pathological—is illustrated by an assay used to investigate translocations where only one of the breakpoints is known or constant. Locus-specific probes are made for one side of the breakpoint and the other intact chromosome. In normal cells, the secondary color is observed, but only the primary colors are observed when the translocation occurs. This technique is sometimes called "break-apart FISH".
Stellaris(R) RNA FISH probes[edit]
Stellaris RNA FISH, formerly known as Single Molecule RNA FISH, is a method of detecting and quantifying mRNA and other long RNA molecules in a thin layer of tissue sample. Targets can be reliably imaged through the application of multiple short singly labeled oligonucleotide probes.[12] The binding o[censored]p to 48 fluorescent labeled oligos to a single molecule of mRNA provides sufficient fluorescence to accurately detect and localize each target mRNA in a wide-field fluorescent microscopy image. Probes not binding to the intended sequence do not achieve sufficient localized fluorescence to be distinguished from background.[13]
Single-molecule RNA FISH assays can be performed in simplex or multiplex, and can be used as a follow-up experiment to quantitative PCR, or imaged simultaneously with a fluorescent antibody assay. The technology has potential applications in cancer diagnosis, [14] neuroscience, gene expression analysis, [15] and companion diagnostics.
Fiber FISH[edit]
In an alternative technique to interphase or metaphase preparations, fiber FISH, interphase chromosomes are attached to a slide in such a way that they are stretched out in a straight line, rather than being tightly coiled, as in conventional FISH, or adopting a chromosome territory conformation, as in interphase FISH. This is accomplished by applying mechanical shear along the length of the slide, either to cells that have been fixed to the slide and then lysed, or to a solution of purified DNA. A technique known as chromosome combing is increasingly used for this purpose. The extended conformation of the chromosomes allows dramatically higher resolution – even down to a few kilobases. The preparation of fiber FISH samples, although conceptually simple, is a rather skilled art, and only specialized laboratories use the technique routinely.
Q-FISH[edit]
Q-FISH combines FISH with PNAs and computer software to quantify fluorescence intensity. This technique is used routinely in telomere length research.
Flow-FISH[edit]
Flow-FISH uses flow cytometry to perform FISH automatically using per-cell fluorescence measurements.
Medical applications[edit]
Often parents of children with a developmental disability want to know more about their child's conditions before choosing to have another child. These concerns can be addressed by analysis of the parents' and child's DNA. In cases where the child's developmental disability is not understood, the cause of it can potentially be determined using FISH and cytogenetic techniques. Examples of diseases that are diagnosed using FISH include Prader-Willi syndrome, Angelman syndrome, 22q13 deletion syndrome, chronic myelogenous leukemia, acute lymphoblastic leukemia, Cri-du-chat, Velocardiofacial syndrome, and Down syndrome. FISH on sperm cells is indicated for men with an abnormal somatic or meiotic karyotype as well as those with oligozoospermia, since approximately 50% of oligozoospermic men have an increased rate of sperm chromosome abnormalities.[16] The analysis of chromosomes 21, X, and Y is enough to identify oligozoospermic individuals at risk.[16]
In medicine, FISH can be used to form a diagnosis, to evaluate prognosis, or to evaluate remission of a disease, such as cancer. Treatment can then be specifically tailored. A traditional exam involving metaphase chromosome analysis is often unable to identify features that distinguish one disease from another, due to subtle chromosomal features; FISH can elucidate these differences. FISH can also be used to detect diseased cells more easily than standard Cytogenetic methods, which require dividing cells and requires labor and time-intensive manual preparation and analysis of the slides by a technologist. FISH, on the other hand, does not require living cells and can be quantified automatically, a computer counts the fluorescent dots present. However, a trained technologist is required to distinguish subtle differences in banding patterns on bent and twisted metaphase chromosomes. FISH can be incorporated into Lab-on-a-chip microfluidic device. This technology is still in a developmental stage but, like other lab on a chip methods, it may lead to more portable diagnostic techniques.[17][18]
Species identification[edit]
FISH is often used in clinical studies. If a patient is infected with a suspected pathogen, bacteria, from the patient's tissues or fluids, are typically grown on agar to determine the identity of the pathogen. Many bacteria, however, even well-known species, do not grow well under laboratory conditions. FISH can be used to detect directly the presence of the suspect on small samples of patient's tissue.
FISH can also be used to compare the genomes of two biological species, to deduce evolutionary relationships. A similar hybridization technique is called a zoo blot. Bacterial FISH probes are often primers for the 16s rRNA region.
FISH is widely used in the field of microbial ecology, to identify microorganisms. Biofilms, for example, are composed of complex (often) multi-species bacterial organizations. Preparing DNA probes for one species and performing FISH with this probe allows one to visualize the distribution of this specific species within the biofilm. Preparing probes (in two different colors) for two species allows to visualize/study co-localization of these two species in the biofilm, and can be useful in determining the fine architecture of the biofilm.
Comparative genomic hybridization[edit]
Comparative genomic hybridization can be described as a method that uses FISH in a parallel manner with the comparison of the hybridization strength to recall any major disruptions in the duplication process of the DNA sequences in the genome of the nucleus.[19]
Virtual karyotype[edit]
Virtual karyotyping is another cost-effective, clinically available alternative to FISH panels using thousands to millions of probes on a single array to detect copy number changes, genome-wide, at unprecedented resolution. Currently, this type of analysis will only detect gains and losses of chromosomal material and will not detect balanced rearrangements, such as translocations and inversions which are hallmark aberrations seen in many types of leukemia and lymphoma.
Spectral karyotype[edit]
Spectral karyotyping is an image of colored chromosomes. Spectral karyotyping involves FISH using multiple forms of many types of probes with the result to see each chromosome labeled through its metaphase stage. This type of karyotyping is used specifically when seeking out chromosome arrangements.
worst it is
Nucot is a fraud company, folks please note that nucot is fraud company and the people who works there are completely fraud, it is family based company,
1. They will call for an interview.
2. They will ask us to join for a course after they will provide us the job.
3. After course no certification will be given.
4. No placement at all. Maximum they may get call center or other bpo jobs.
5. People who manages are completely uneducated.
6. They will keep our contacts and after the course, they will ask us for a money in a friendship basis and cheat us.
7. All cheaters are there in nucot, these people has to be kept behind the bars, very dangerous.
8. Be careful guys.. They are actually dangerous dealers. If you give them money they will not return is secondary, and also they will threaten you, all activities they are carrying.
Please note : the above mentioned is very true about them, they all are of same family, as i have seen them very closely. Sanoj, sandeep big fraudsters and they are a dangerous dealers, do not believe them, if there are some good reviews on them, then they should be on the same family. They spolied my life.
My also friend got cheated from them very badly, as his money with them and they are making fool of him. If money is asked back they are threatening him dangerously. These f... Rs should be beaten black and blue.
1. They will call for an interview.
2. They will ask us to join for a course after they will provide us the job.
3. After course no certification will be given.
4. No placement at all. Maximum they may get call center or other bpo jobs.
5. People who manages are completely uneducated.
6. They will keep our contacts and after the course, they will ask us for a money in a friendship basis and cheat us.
7. All cheaters are there in nucot, these people has to be kept behind the bars, very dangerous.
8. Be careful guys.. They are actually dangerous dealers. If you give them money they will not return is secondary, and also they will threaten you, all activities they are carrying.
Please note : the above mentioned is very true about them, they all are of same family, as i have seen them very closely. Sanoj, sandeep big fraudsters and they are a dangerous dealers, do not believe them, if there are some good reviews on them, then they should be on the same family. They spolied my life.
My also friend got cheated from them very badly, as his money with them and they are making fool of him. If money is asked back they are threatening him dangerously. These f... Rs should be beaten black and blue.
Comment marked as Resolved
Thanks to Sandeep and Sanoj as the problem is resolved ,as i had a discussion with them .Finally we had a good time and they have promised to solve the problem,please do not mind if I was cruel and wrong on my comments. I Sincerely apologize for the act as I was desperate in seeking to solve my problems,Since I had good talks with them where I received a statements of hopes.finally am thankful for them,once again am sorry .Thank you. HI... folks!! such a worst and fraud institute of this nucot.. please guys dont believe their words and activites.. for me frst one lady took up the frst round of interview.. she mesmarizes me with her talks about that fraud insitute. she did'nt ask even a qstn about my technical skills and communication. juss she passed me for further round of interview with manager.he explained me about their process of training(fraudlent training)and he said 14k for the training.. and also he promised me like we ll not sent you out frm my institute without job .. after i realizing myself by reading this reviews of my senoirs(who faced this before me).. its correct. Fraud staff, Fraud manager, Fraud receptionist, Fraud Lady... TOTALLY FRAUD.
Nucot is a training service sales company. Nucot sells training services to fresh engg grads. Nucot is not at all transparent in its business. And according to me they are practicing unethical way of doing business. Trying to waste time & playing with unemployed fresh Engg grads sentiments.
Suggestion to Nucot (you may take it or leave it) – Be proud of what you do. Be proud of your business. Never ever play with sentiments & feeling o[censored]nemployed fresh engineers. Be truthful & give them full details about your service & then call them for an interview. Don’t just call them for a Job interview & then try to sell your training service.
You guys are trying to cut down your sales cost by getting your target audience ( i.e; Fresh engineering grads ) to your doorsteps by giving false information. You should be letting them know in advance that you are a training center & trying to sell training service to them. Make it clear guys & have some courage to tell the truth to everyone in advance. This might help people to asses you & decide whether to discuss with you or not.
Suggestion to Nucot (you may take it or leave it) – Be proud of what you do. Be proud of your business. Never ever play with sentiments & feeling o[censored]nemployed fresh engineers. Be truthful & give them full details about your service & then call them for an interview. Don’t just call them for a Job interview & then try to sell your training service.
You guys are trying to cut down your sales cost by getting your target audience ( i.e; Fresh engineering grads ) to your doorsteps by giving false information. You should be letting them know in advance that you are a training center & trying to sell training service to them. Make it clear guys & have some courage to tell the truth to everyone in advance. This might help people to asses you & decide whether to discuss with you or not.
Early history[edit]
Prior to European settlement, the area was inhabited by several groups of Ohlone Native Americans.[38]
T
(17.3) 55.1
(12.8) 50.0
(10) 60.5
(15.8)
Average low °F (°C) 42.0
(5.6) 44.7
(7.1) 46.6
(8.1) 48.6
(9.2) 52.4
(11.3) 56.0
(13.3) 58.1
(14.5) 58.3
(14.6) 56.8
(13.8) 52.5
(11.4) 46.0
(7.8) 41.9
(5.5) 50.3
(10.2)
Record low °F (°C) 18
(−8) 24
(−4) 25
(−4) 26
(−3) 32
(0) 33
(1) 40
(4) 39
(4) 35
(2) 30
(−1) 21
(−6) 19
(−7) 18
(−8)
Average rainfall inches (mm) 3.07
(78) 3.11
(79) 2.54
(64.5) 1.18
(30) 0.51
(13) 0.10
(2.5) 0.02
(0.5) 0.02
(0.5) 0.18
(4.6) 0.80
(20.3) 1.68
(42.7) 2.61
(66.3) 15.82
(401.9)
Average rainy days (≥ 0.01 in) 10.2 10.3 9.4 5.6 3.2 0.8 0.2 0.3 1.3 3.2 7.2 10.2 61.9
Source: NOAA[68][69]
With the light rainfall, San Jose and its suburbs experience about 300 fully or partly sunny days a year. Rain occurs primarily in the months from November through April or May. During the winter and spring, hillsides and fields turn green with grasses and vegetation, although deciduous trees are few. With the coming of the annual hot summer dry period, the vegetation dies and dries, giving the hills a golden cover which, unfortunately, also provides fuel for frequent grass fires.
Measurable precipitation falls in downtown San Jose on an average of 59 days a year.[citation needed] “Rain year” precipitation has ranged from 4.83 inches (122.7 mm) between July 1876 and June 1877 to 30.30 inches (769.6 mm) between July 1889 and June 1890, although at the current site since 1893 the range is from 5.77 inches (146.6 mm) in “rain year” 1975–76 to 30.25 inches (768.3 mm) in “rain year” 1982–83. The most precipitation in one month was 12.38 inches (314.5 mm) in January 1911. The maximum 24-hour rainfall was 3.60 inches (91.4 mm) on January 30, 1968. Although summer is normally quite dry in San Jose, a very heavy thunderstorm on August 21, 1968, brought 1.92 inches (48.8 mm) of rain, causing some flooding.[70]
The snow level drops as low as 2, 000 ft (610 m) above sea level, or lower, occasionally coating nearby Mount Hamilton and, less frequently, the Santa Cruz Mountains, with snow that normally lasts a few days. This sometimes snarls traffic traveling on State Route 17 towards Santa Cruz. Snow rarely falls in San Jose; the most recent snow to remain on the ground was on February 5, 1976, when many residents around the city saw as much as 3 inches (0.076 m) on car and roof tops. The official observation station measured only 0.5 inches (0.013 m) of snow.[71]
Demographics[edit]
Historical population
Census Pop. %±
1870 9, 089 —
1880 12, 567 38.3%
1890 18, 060 43.7%
1900 21, 500 19.0%
1910 28, 946 34.6%
1920 39, 642 37.0%
1930 57, 651 45.4%
1940 68, 457 18.7%
1950 95, 280 39.2%
1960 204, 196 114.3%
1970 459, 913 125.2%
1980 629, 400 36.9%
1990 782, 248 24.3%
2000 894, 943 14.4%
2010 945, 942 5.7%
Est. 2016 1, 025, 350 [12] 8.4%
U.S. Decennial Census[72]
In 2014, the U.S. Census Bureau released its new population estimates. With a total population of 1, 015, 785, [73] San Jose became the 11th U.S. city to hit the 1 million mark, even though it is currently the 10th most populous city. In 1930, Detroit had reached 1, 568, 662, but by 2000, it had dropped below 1, 000, 000.
[hide]Racial composition 2010[74] 1990[43] 1970[43] 1940[43]
White 42.8% 62.8% 93.6% 98.5%
—Non-Hispanic 28.7% 49.6% 75.7%[75] n/a
Black or African American 3.2% 4.7% 2.5% 0.4%
Hispanic or Latino (of any race) 33.2% 26.6% 19.1%[75] n/a
Asian 32.0% 19.5% 2.7% 1.1%
Other race 15.7% 12.3% 0.8% (X)
Two or more races 5.0% n/a n/a n/a
2010[edit]
Map of racial distribution in San Jose, 2010 U.S. Census. Each dot is 25 people: White, Black, Asian, Hispanic or Other (yellow).
Thematic map showing median household income across central Santa Clara County as of 2014; the darker the color, the more affluent the area.
The 2010 United States Census[76] reported that San Jose had a population of 945, 942. The population density was 5, 256.2 people per square mile (2, 029.4/km²). The racial makeup of San Jose was 404, 437 (42.8%) White, 303, 138 (32.0%) Asian (10.4% Vietnamese, 6.7% Chinese, 5.6% Filipino, 4.6% Indian, 1.2% Korean, 1.2% Japanese, 0.3% Cambodian, 0.2% Thai, 0.2% Pakistani, 0.2% Laotian), 30, 242 (3.2%) African American, 8, 297 (0.9%) Native American, 4, 017 (0.4%) Pacific Islander, 148, 749 (15.7%) from other races, and 47, 062 (5.0%) from two or more races. Hispanic or Latino of any race were 313, 636 persons (33.2%). 28.2% of the city's population were of Mexican descent; the next largest Hispanic groups were those of Salvadoran (0.7%) and Puerto Rican (0.5%) heritage. Non-Hispanic Whites were 28.7% of the population in 2010, [77] down from 75.7% in 1970.[43]
The census reported that 932, 620 people (98.6% of the population) lived in households, 9, 542 (1.0%) lived in non-institutionalized group quarters, and 3, 780 (0.4%) were institutionalized. There were 301, 366 households, out of which 122, 958 (40.8%) had children under the age of 18 living in them, 162, 819 (54.0%) were opposite-sex married couples living together, 37, 988 (12.6%) had a female householder with no husband present, 18, 702 (6.2%) had a male householder with no wife present. There were 16, 900 (5.6%) unmarried opposite-sex partnerships, and 2, 458 (0.8%) same-sex married couples or partnerships. 59, 385 households (19.7%) were made up of individuals and 18, 305 (6.1%) had someone living alone who was 65 years of age or older. The average household size was 3.09. There were 219, 509 families (72.8% of all households); the average family size was 3.54.
The age distribution of the city was as follows: 234, 678 people (24.8%) were under the age of 18, 89, 457 people (9.5%) aged 18 to 24, 294, 399 people (31.1%) aged 25 to 44, 232, 166 people (24.5%) aged 45 to 64, and 95, 242 people (10.1%) who were 65 years of age or older. The median age was 35.2 years. For every 100 females there were 101.1 males. For every 100 females age 18 and over, there were 99.8 males.
There were 314, 038 housing units at an average density of 1, 745.0 per square mile (673.7/km²), of which 176, 216 (58.5%) were owner-occupied, and 125, 150 (41.5%) were occupied by renters. The homeowner vacancy rate was 1.6%; the rental vacancy rate was 4.3%. 553, 436 people (58.5% of the population) lived in owner-occupied housing units and 379, 184 people (40.1%) lived in rental housing units.
2000[edit]
As of the census[78] of 2000, there were 894, 943 people, 276, 598 households, and 203, 576 families residing in the city.
The population density was 5, 117.9 people per square mile (1, 976.1/km²). There were 281, 841 housing units at an average density of 1, 611.8 per square mile (622.3/km²). Of the 276, 598 households, 38.3% had children under the age of 18 living with them, 56.0% were married couples living together, 11.7% had a female householder with no husband present, and 26.4% were non-families. 18.4% of all households were made up of individuals and 4.9% had someone living alone who was 65 years of age or older. The average household size was 3.20 and the average family size was 3.62.
In the city, the population was spread out with 26.4% under the age of 18, 9.9% from 18 to 24, 35.4% from 25 to 44, 20.0% from 45 to 64, and 8.3% who were 65 years of age or older. The median age was 33 years. For every 100 females there were 103.3 males. For every 100 females age 18 and over, there were 102.5 males.
According to a 2007 estimate, the median income for a household in the city was the highest in the U.S. for any city with more than a quarter million residents with $76, 963 annually. The median income for a family was $86, 822.[79] Males had a median income of $49, 347 versus $36, 936 for females. The per capita income for the city was $26, 697. About 6.0% of families and 8.8% of the population were below the poverty line, including 10.3% of those under age 18 and 7.4% of those age 65 or over.
San Jose and the rest of the Bay Area is home to many Christian congregations, including large Roman Catholic and Eastern Orthodox churches, Mormons, and Jehovah's Witnesses, alongside centers of Jewish, Hindu, Islamic, Buddhist and Sikh faiths, among numerous other religious communities.
A high percentage of foreign-born residents (39.0% of the population) live in the city. These include many high-tech workers from East and South Asia, Eastern European immigrants, as well as poorer immigrants from Latin America, many of whom can be found in the large, multi-generational barrio Alum Rock district. San Jose has the largest Vietnamese population of any city in the world outside of Vietnam.[80] The people from these countries have settled in the city and across the Santa Clara Valley primarily during the last three or four decades.
Economy[edit]
The large concentration of high-technology engineering, computer, and microprocessor companies around San Jose has led the area to be known as Silicon Valley. As the largest city in the valley, San Jose has billed itself "the capital of Silicon Valley." Area schools such as the University of California, Berkeley, University of California, Santa Cruz, San Jose State University, San Francisco State University, California State University, East Bay, Santa Clara University, and Stanford University pump thousands of engineering and computer science graduates into the local economy every year.
High economic growth during the tech bubble caused employment, housing prices, and traffic congestion to peak in the late 1990s. As the economy slowed in the early 2000s, employment and traffic congestion diminished somewhat[citation needed]. In the mid-2000s, traffic along major highways again began to worsen as the economy improved. San Jose had 405, 000 jobs within its city limits in 2006, and an unemployment rate of 4.6%. In 2000, San Jose residents had the highest median household income of any city in the United States with a population over 300, 000, and currently has the highest median income of any U.S. city with over 280, 000 people.
The cost of living in San Jose and the surrounding areas is among the highest in California and the nation, according to 2004 data.[81] Housing costs are the primary reason for the high cost of living, although the costs in all areas tracked by the ACCRA Cost of Living Index are above the national average. Households in the city limits have the highest disposable income of any city in the U.S. with over 500, 000 residents.[82][83]
San Jose lists many companies with 1, 000 employees or more, including the headquarters of Adobe, Altera, Brocade Communications Systems, Cadence Design Systems, Cisco Systems, eBay, Lee's Sandwiches, Lumileds, PayPal, Rosendin Electric, Sanmina-SCI, and Xilinx, as well as major facilities for Becton Dickinson, Ericsson, Hewlett-Packard, Hitachi, IBM, Kaiser Permanente, KLA Tencor, Lockheed Martin, Nippon Sheet Glass, Qualcomm, and AF Media Group. The North American headquarters of Samsung Semiconductor are located in San Jose.[84][85] Approximately 2000 employees will work at the new Samsung campus which opened in 2015. Other large companies based in San Jose include Altera, Atmel, CEVA, Cypress Semiconductor, Echelon, Extreme Networks, Harmonic, Integrated Device Technology, Maxim Integrated, Micrel, Move, Netgear, Novellus Systems, Oclaro, OCZ, Online Trading Academy, Quantum, SunPower, Sharks Sports and Entertainment, Supermicro, Tessera Technologies, TiVo, Ultratech, and VeriFone. Sizable government employers include the city government, Santa Clara County, and San Jose State University.[86] Acer's United States division has its offices in San Jose.[87] Prior to its closing, Netcom had its headquarters in San Jose.[88][89]
In January 2014 Forbes Magazine reported that Careerbliss.com had ranked San Jose-Sunnyvale-Santa Clara metro area as the happiest place to work in the USA. The report cited a large concentration of technology jobs that typically offer a high salary and opportunity for growth, in addition to companies providing "fun and innovative work environments" as some of the reasons for the ranking.[90]
Technology[edit]
Main article: Silicon Valley
San Jose residents produce more U.S. patents than any other city.[91] On October 15, 2015, the United States Patent and Trademark Office opened a satellite office in San Jose to serve Silicon Valley and the Western U.S. States.[92][93] Thirty-five percent of all venture capital funds in the U.S. are invested in San Jose and Silicon Valley companies.[91]
On July 31, 2015, Cupertino-based Apple Inc. purchased a 40-acre site in San Jose.[citation needed] The site, which is bare land, will be the site of an office and research campus where it is estimated that up to 16, 000 employees will be located. Apple paid $138.2 million in cash for the site.[citation needed] The seller, Connecticut-based Five Mile Capital Partners, paid $40 million for the site in 2010.[94] Real estate experts expect that other tech companies currently located in Silicon Valley will also follow in Apple's path by purchasing land or property in San Jose.[95]
Top employers[edit]
Left-to-right: eBay Headquarters, Adobe World Headquarters, Cisco's San Jose Main Campus, PayPal's Headquarters
As of June 30, 2015, the top employers in the city are:[96]
# Employer Employees
1 County of Santa Clara 17, 476
2 Cisco Systems 15, 470
3 eBay 6, 130
4 City of San Jose 5, 928
5 San Jose State University 4, 480
6 U.S. Postal Service 3, 900
7 Western Digital 2, 660
8 IBM 2, 360
9 San Jose Unified School District 2, 320
10 Kaiser Permanente 2, 290
11 Good Samaritan Hospital 2, 110
12 Adobe Systems, Inc. 2, 010
13 Target Corporation 2, 000
14 Brocade Communications 1, 780
15 Cadence Design Systems 1, 460
Arts and architecture[edit]
City arts and architecture[edit]
Pictured is the San Jose Center for the Performing Arts at night, a general-purpose venue for several performing arts organizations in San Jose
SJC's new consolidated parking and rental facility, CONRAC, has been fitted with new public art featuring hands of people in Silicon Valley.
Iglesia ni Cristo chapel of San Jose, an icon and gathering place of some immigrants
Because the downtown area is in the flight path to nearby Mineta San Jose International Airport (also evidenced in the above panoramic), there is a height limit for buildings in the downtown area, which is underneath the final approach corridor to the airport. The height limit is dictated by local ordinances, driven by the distance from the runway and a slope defined by Federal Aviation Administration regulations. Core downtown buildings are limited to approximately 300 feet (91 m) but can get taller farther from the airport.[97]
There has been broad criticism over the past few decades of the city's architecture.[98] Citizens have complained that San Jose is lacking in aesthetically pleasing architectural styles. Blame for this lack of architectural "beauty" can be assigned to the re-development of the downtown area from the 1950s onward, in which whole blocks of historic commercial and residential structures were demolished.[99] Exceptions to this
Prior to European settlement, the area was inhabited by several groups of Ohlone Native Americans.[38]
T
(17.3) 55.1
(12.8) 50.0
(10) 60.5
(15.8)
Average low °F (°C) 42.0
(5.6) 44.7
(7.1) 46.6
(8.1) 48.6
(9.2) 52.4
(11.3) 56.0
(13.3) 58.1
(14.5) 58.3
(14.6) 56.8
(13.8) 52.5
(11.4) 46.0
(7.8) 41.9
(5.5) 50.3
(10.2)
Record low °F (°C) 18
(−8) 24
(−4) 25
(−4) 26
(−3) 32
(0) 33
(1) 40
(4) 39
(4) 35
(2) 30
(−1) 21
(−6) 19
(−7) 18
(−8)
Average rainfall inches (mm) 3.07
(78) 3.11
(79) 2.54
(64.5) 1.18
(30) 0.51
(13) 0.10
(2.5) 0.02
(0.5) 0.02
(0.5) 0.18
(4.6) 0.80
(20.3) 1.68
(42.7) 2.61
(66.3) 15.82
(401.9)
Average rainy days (≥ 0.01 in) 10.2 10.3 9.4 5.6 3.2 0.8 0.2 0.3 1.3 3.2 7.2 10.2 61.9
Source: NOAA[68][69]
With the light rainfall, San Jose and its suburbs experience about 300 fully or partly sunny days a year. Rain occurs primarily in the months from November through April or May. During the winter and spring, hillsides and fields turn green with grasses and vegetation, although deciduous trees are few. With the coming of the annual hot summer dry period, the vegetation dies and dries, giving the hills a golden cover which, unfortunately, also provides fuel for frequent grass fires.
Measurable precipitation falls in downtown San Jose on an average of 59 days a year.[citation needed] “Rain year” precipitation has ranged from 4.83 inches (122.7 mm) between July 1876 and June 1877 to 30.30 inches (769.6 mm) between July 1889 and June 1890, although at the current site since 1893 the range is from 5.77 inches (146.6 mm) in “rain year” 1975–76 to 30.25 inches (768.3 mm) in “rain year” 1982–83. The most precipitation in one month was 12.38 inches (314.5 mm) in January 1911. The maximum 24-hour rainfall was 3.60 inches (91.4 mm) on January 30, 1968. Although summer is normally quite dry in San Jose, a very heavy thunderstorm on August 21, 1968, brought 1.92 inches (48.8 mm) of rain, causing some flooding.[70]
The snow level drops as low as 2, 000 ft (610 m) above sea level, or lower, occasionally coating nearby Mount Hamilton and, less frequently, the Santa Cruz Mountains, with snow that normally lasts a few days. This sometimes snarls traffic traveling on State Route 17 towards Santa Cruz. Snow rarely falls in San Jose; the most recent snow to remain on the ground was on February 5, 1976, when many residents around the city saw as much as 3 inches (0.076 m) on car and roof tops. The official observation station measured only 0.5 inches (0.013 m) of snow.[71]
Demographics[edit]
Historical population
Census Pop. %±
1870 9, 089 —
1880 12, 567 38.3%
1890 18, 060 43.7%
1900 21, 500 19.0%
1910 28, 946 34.6%
1920 39, 642 37.0%
1930 57, 651 45.4%
1940 68, 457 18.7%
1950 95, 280 39.2%
1960 204, 196 114.3%
1970 459, 913 125.2%
1980 629, 400 36.9%
1990 782, 248 24.3%
2000 894, 943 14.4%
2010 945, 942 5.7%
Est. 2016 1, 025, 350 [12] 8.4%
U.S. Decennial Census[72]
In 2014, the U.S. Census Bureau released its new population estimates. With a total population of 1, 015, 785, [73] San Jose became the 11th U.S. city to hit the 1 million mark, even though it is currently the 10th most populous city. In 1930, Detroit had reached 1, 568, 662, but by 2000, it had dropped below 1, 000, 000.
[hide]Racial composition 2010[74] 1990[43] 1970[43] 1940[43]
White 42.8% 62.8% 93.6% 98.5%
—Non-Hispanic 28.7% 49.6% 75.7%[75] n/a
Black or African American 3.2% 4.7% 2.5% 0.4%
Hispanic or Latino (of any race) 33.2% 26.6% 19.1%[75] n/a
Asian 32.0% 19.5% 2.7% 1.1%
Other race 15.7% 12.3% 0.8% (X)
Two or more races 5.0% n/a n/a n/a
2010[edit]
Map of racial distribution in San Jose, 2010 U.S. Census. Each dot is 25 people: White, Black, Asian, Hispanic or Other (yellow).
Thematic map showing median household income across central Santa Clara County as of 2014; the darker the color, the more affluent the area.
The 2010 United States Census[76] reported that San Jose had a population of 945, 942. The population density was 5, 256.2 people per square mile (2, 029.4/km²). The racial makeup of San Jose was 404, 437 (42.8%) White, 303, 138 (32.0%) Asian (10.4% Vietnamese, 6.7% Chinese, 5.6% Filipino, 4.6% Indian, 1.2% Korean, 1.2% Japanese, 0.3% Cambodian, 0.2% Thai, 0.2% Pakistani, 0.2% Laotian), 30, 242 (3.2%) African American, 8, 297 (0.9%) Native American, 4, 017 (0.4%) Pacific Islander, 148, 749 (15.7%) from other races, and 47, 062 (5.0%) from two or more races. Hispanic or Latino of any race were 313, 636 persons (33.2%). 28.2% of the city's population were of Mexican descent; the next largest Hispanic groups were those of Salvadoran (0.7%) and Puerto Rican (0.5%) heritage. Non-Hispanic Whites were 28.7% of the population in 2010, [77] down from 75.7% in 1970.[43]
The census reported that 932, 620 people (98.6% of the population) lived in households, 9, 542 (1.0%) lived in non-institutionalized group quarters, and 3, 780 (0.4%) were institutionalized. There were 301, 366 households, out of which 122, 958 (40.8%) had children under the age of 18 living in them, 162, 819 (54.0%) were opposite-sex married couples living together, 37, 988 (12.6%) had a female householder with no husband present, 18, 702 (6.2%) had a male householder with no wife present. There were 16, 900 (5.6%) unmarried opposite-sex partnerships, and 2, 458 (0.8%) same-sex married couples or partnerships. 59, 385 households (19.7%) were made up of individuals and 18, 305 (6.1%) had someone living alone who was 65 years of age or older. The average household size was 3.09. There were 219, 509 families (72.8% of all households); the average family size was 3.54.
The age distribution of the city was as follows: 234, 678 people (24.8%) were under the age of 18, 89, 457 people (9.5%) aged 18 to 24, 294, 399 people (31.1%) aged 25 to 44, 232, 166 people (24.5%) aged 45 to 64, and 95, 242 people (10.1%) who were 65 years of age or older. The median age was 35.2 years. For every 100 females there were 101.1 males. For every 100 females age 18 and over, there were 99.8 males.
There were 314, 038 housing units at an average density of 1, 745.0 per square mile (673.7/km²), of which 176, 216 (58.5%) were owner-occupied, and 125, 150 (41.5%) were occupied by renters. The homeowner vacancy rate was 1.6%; the rental vacancy rate was 4.3%. 553, 436 people (58.5% of the population) lived in owner-occupied housing units and 379, 184 people (40.1%) lived in rental housing units.
2000[edit]
As of the census[78] of 2000, there were 894, 943 people, 276, 598 households, and 203, 576 families residing in the city.
The population density was 5, 117.9 people per square mile (1, 976.1/km²). There were 281, 841 housing units at an average density of 1, 611.8 per square mile (622.3/km²). Of the 276, 598 households, 38.3% had children under the age of 18 living with them, 56.0% were married couples living together, 11.7% had a female householder with no husband present, and 26.4% were non-families. 18.4% of all households were made up of individuals and 4.9% had someone living alone who was 65 years of age or older. The average household size was 3.20 and the average family size was 3.62.
In the city, the population was spread out with 26.4% under the age of 18, 9.9% from 18 to 24, 35.4% from 25 to 44, 20.0% from 45 to 64, and 8.3% who were 65 years of age or older. The median age was 33 years. For every 100 females there were 103.3 males. For every 100 females age 18 and over, there were 102.5 males.
According to a 2007 estimate, the median income for a household in the city was the highest in the U.S. for any city with more than a quarter million residents with $76, 963 annually. The median income for a family was $86, 822.[79] Males had a median income of $49, 347 versus $36, 936 for females. The per capita income for the city was $26, 697. About 6.0% of families and 8.8% of the population were below the poverty line, including 10.3% of those under age 18 and 7.4% of those age 65 or over.
San Jose and the rest of the Bay Area is home to many Christian congregations, including large Roman Catholic and Eastern Orthodox churches, Mormons, and Jehovah's Witnesses, alongside centers of Jewish, Hindu, Islamic, Buddhist and Sikh faiths, among numerous other religious communities.
A high percentage of foreign-born residents (39.0% of the population) live in the city. These include many high-tech workers from East and South Asia, Eastern European immigrants, as well as poorer immigrants from Latin America, many of whom can be found in the large, multi-generational barrio Alum Rock district. San Jose has the largest Vietnamese population of any city in the world outside of Vietnam.[80] The people from these countries have settled in the city and across the Santa Clara Valley primarily during the last three or four decades.
Economy[edit]
The large concentration of high-technology engineering, computer, and microprocessor companies around San Jose has led the area to be known as Silicon Valley. As the largest city in the valley, San Jose has billed itself "the capital of Silicon Valley." Area schools such as the University of California, Berkeley, University of California, Santa Cruz, San Jose State University, San Francisco State University, California State University, East Bay, Santa Clara University, and Stanford University pump thousands of engineering and computer science graduates into the local economy every year.
High economic growth during the tech bubble caused employment, housing prices, and traffic congestion to peak in the late 1990s. As the economy slowed in the early 2000s, employment and traffic congestion diminished somewhat[citation needed]. In the mid-2000s, traffic along major highways again began to worsen as the economy improved. San Jose had 405, 000 jobs within its city limits in 2006, and an unemployment rate of 4.6%. In 2000, San Jose residents had the highest median household income of any city in the United States with a population over 300, 000, and currently has the highest median income of any U.S. city with over 280, 000 people.
The cost of living in San Jose and the surrounding areas is among the highest in California and the nation, according to 2004 data.[81] Housing costs are the primary reason for the high cost of living, although the costs in all areas tracked by the ACCRA Cost of Living Index are above the national average. Households in the city limits have the highest disposable income of any city in the U.S. with over 500, 000 residents.[82][83]
San Jose lists many companies with 1, 000 employees or more, including the headquarters of Adobe, Altera, Brocade Communications Systems, Cadence Design Systems, Cisco Systems, eBay, Lee's Sandwiches, Lumileds, PayPal, Rosendin Electric, Sanmina-SCI, and Xilinx, as well as major facilities for Becton Dickinson, Ericsson, Hewlett-Packard, Hitachi, IBM, Kaiser Permanente, KLA Tencor, Lockheed Martin, Nippon Sheet Glass, Qualcomm, and AF Media Group. The North American headquarters of Samsung Semiconductor are located in San Jose.[84][85] Approximately 2000 employees will work at the new Samsung campus which opened in 2015. Other large companies based in San Jose include Altera, Atmel, CEVA, Cypress Semiconductor, Echelon, Extreme Networks, Harmonic, Integrated Device Technology, Maxim Integrated, Micrel, Move, Netgear, Novellus Systems, Oclaro, OCZ, Online Trading Academy, Quantum, SunPower, Sharks Sports and Entertainment, Supermicro, Tessera Technologies, TiVo, Ultratech, and VeriFone. Sizable government employers include the city government, Santa Clara County, and San Jose State University.[86] Acer's United States division has its offices in San Jose.[87] Prior to its closing, Netcom had its headquarters in San Jose.[88][89]
In January 2014 Forbes Magazine reported that Careerbliss.com had ranked San Jose-Sunnyvale-Santa Clara metro area as the happiest place to work in the USA. The report cited a large concentration of technology jobs that typically offer a high salary and opportunity for growth, in addition to companies providing "fun and innovative work environments" as some of the reasons for the ranking.[90]
Technology[edit]
Main article: Silicon Valley
San Jose residents produce more U.S. patents than any other city.[91] On October 15, 2015, the United States Patent and Trademark Office opened a satellite office in San Jose to serve Silicon Valley and the Western U.S. States.[92][93] Thirty-five percent of all venture capital funds in the U.S. are invested in San Jose and Silicon Valley companies.[91]
On July 31, 2015, Cupertino-based Apple Inc. purchased a 40-acre site in San Jose.[citation needed] The site, which is bare land, will be the site of an office and research campus where it is estimated that up to 16, 000 employees will be located. Apple paid $138.2 million in cash for the site.[citation needed] The seller, Connecticut-based Five Mile Capital Partners, paid $40 million for the site in 2010.[94] Real estate experts expect that other tech companies currently located in Silicon Valley will also follow in Apple's path by purchasing land or property in San Jose.[95]
Top employers[edit]
Left-to-right: eBay Headquarters, Adobe World Headquarters, Cisco's San Jose Main Campus, PayPal's Headquarters
As of June 30, 2015, the top employers in the city are:[96]
# Employer Employees
1 County of Santa Clara 17, 476
2 Cisco Systems 15, 470
3 eBay 6, 130
4 City of San Jose 5, 928
5 San Jose State University 4, 480
6 U.S. Postal Service 3, 900
7 Western Digital 2, 660
8 IBM 2, 360
9 San Jose Unified School District 2, 320
10 Kaiser Permanente 2, 290
11 Good Samaritan Hospital 2, 110
12 Adobe Systems, Inc. 2, 010
13 Target Corporation 2, 000
14 Brocade Communications 1, 780
15 Cadence Design Systems 1, 460
Arts and architecture[edit]
City arts and architecture[edit]
Pictured is the San Jose Center for the Performing Arts at night, a general-purpose venue for several performing arts organizations in San Jose
SJC's new consolidated parking and rental facility, CONRAC, has been fitted with new public art featuring hands of people in Silicon Valley.
Iglesia ni Cristo chapel of San Jose, an icon and gathering place of some immigrants
Because the downtown area is in the flight path to nearby Mineta San Jose International Airport (also evidenced in the above panoramic), there is a height limit for buildings in the downtown area, which is underneath the final approach corridor to the airport. The height limit is dictated by local ordinances, driven by the distance from the runway and a slope defined by Federal Aviation Administration regulations. Core downtown buildings are limited to approximately 300 feet (91 m) but can get taller farther from the airport.[97]
There has been broad criticism over the past few decades of the city's architecture.[98] Citizens have complained that San Jose is lacking in aesthetically pleasing architectural styles. Blame for this lack of architectural "beauty" can be assigned to the re-development of the downtown area from the 1950s onward, in which whole blocks of historic commercial and residential structures were demolished.[99] Exceptions to this
LUCKNOW CENTRAL STORY: Kishan Girhotra's (Farhan) dreams of becoming a singer are shattered when he is falsely accused of murdering an IAS officer in Muradabad. Pronounced guilty, he is jailed for the crime he never committed. Deprived of the freedom he rightfully deserves, the sun shines on him once again even in captivity, as a bunch of prisoners and an NGO worker (Diana Penty) become his voice and the wind beneath his wings. Can he accomplish his dream of forming a band in the prison or does he manage to escape the dungeon?
LUCKNOW CENTRAL REVIEW: Based on true life events, Lucknow Central is a feel-good, human, prison-break drama that succeeds to manipulate you emotionally, despite being predictable and filmy in portions. While the peg is somewhat similar (barring the undertrial element) to the recent Qaidi Band, Lucknow Central has a distinct execution and is way more evolved.
What essentially works for LC is its classic 'winning against all odds' theme. The inmates finding a reason to live and a sense of belonging within the four walls of the Lucknow Central jail instead of their homes is what tugs at your heartstrings. Shunned as misfits and outcasts by the society including their families, the prisoners find solace in each other's company. Ranjit Tiwari handles this aspect of human alienation and friendship, beautifully. His understated sensibility resonates with his talented star cast, starting with the very impressive Ronit Roy. The actor stands out as the curt, conniving and clever jailer. Sadly, his character is reduced to being a toothless tiger eventually.
Farhan ably carries the film on his shoulders but his efforts to nail his desi character are partially visible. For instance, his flawed English pronunciation seems a tad artificial and inconsistent. (Read Corut for court, rohk for Rock and naan existent for non-existent). Nevertheless, he gets the mannerisms and innate innocence of a small town guy right and makes you weep for his misfortune. Deepak Dobriyal is excellent as always and so are Rajesh Sharma, Inaamulhaq and Ravi Kishan. Diana Penty and Gippy Grewal play their parts well.
Considering the film revolves around music, Rangdaari is a beautiful composition that sums up this fascinating tale of dreamers and fighters, who refuse to give up on life or faith.
LUCKNOW CENTRAL REVIEW: Based on true life events, Lucknow Central is a feel-good, human, prison-break drama that succeeds to manipulate you emotionally, despite being predictable and filmy in portions. While the peg is somewhat similar (barring the undertrial element) to the recent Qaidi Band, Lucknow Central has a distinct execution and is way more evolved.
What essentially works for LC is its classic 'winning against all odds' theme. The inmates finding a reason to live and a sense of belonging within the four walls of the Lucknow Central jail instead of their homes is what tugs at your heartstrings. Shunned as misfits and outcasts by the society including their families, the prisoners find solace in each other's company. Ranjit Tiwari handles this aspect of human alienation and friendship, beautifully. His understated sensibility resonates with his talented star cast, starting with the very impressive Ronit Roy. The actor stands out as the curt, conniving and clever jailer. Sadly, his character is reduced to being a toothless tiger eventually.
Farhan ably carries the film on his shoulders but his efforts to nail his desi character are partially visible. For instance, his flawed English pronunciation seems a tad artificial and inconsistent. (Read Corut for court, rohk for Rock and naan existent for non-existent). Nevertheless, he gets the mannerisms and innate innocence of a small town guy right and makes you weep for his misfortune. Deepak Dobriyal is excellent as always and so are Rajesh Sharma, Inaamulhaq and Ravi Kishan. Diana Penty and Gippy Grewal play their parts well.
Considering the film revolves around music, Rangdaari is a beautiful composition that sums up this fascinating tale of dreamers and fighters, who refuse to give up on life or faith.
Mugulu Nage is a 2017 Indian romantic comedy Kannada film directed and co-produced by Yogaraj Bhat and jointly produced by Ganesh and Syed Salaam.[1] It features Ganesh, Apoorva Arora, Nikitha Narayan and Ashika Ranganath in the lead roles.[2] Amulya, and Jaggesh also feature in cameo roles.[3] Whilst the soundtrack and score is by V. Harikrishna, the cinematography is by Sugnan. The first look of the film was released on 14 February 2017 coinciding the Valentines Day.[4]
The project marks the third film in the combination of Yogaraj Bhat and Ganesh after Mungaru Male (2006) and Gaalipata (2008). The filming began on 8 December 2016 in Bengaluru.[5] Further, the shooting took place in Puducherry, [6] Mysuru, Yaana and Sirsi, Karnataka.
The film was released on September 1, 2017 with mixed reviews. Movie is declared as avg at box office [7]
Contents [hide]
1 Cast
2 Production
2.1 Filming
2.2 Casting
3 Soundtrack
4 References
5 External links
Cast[edit]
Ganesh as Pulakeshi
Apoorva Arora as Chaaru
Nikitha Narayan as Siri
Ashika Ranganath as Vaishali Hande
Ananth Nag
Achyuth Kumar as Achyuth
Rangayana Raghu
Jaggesh in a special appearance
Amulya in a guest appearance
Chandan Achar
Niharika
Dharmanna Kadur
Production[edit]
Filming[edit]
In August 2016, it was reported that the successful combination of Yogaraj Bhat and Ganesh are teaming up together again for a new romantic venture.[8] While Bhat was taking care of the script, wrote story and take up the direction, Ganesh was cast as the lead actor apart from co-producing the film and V. Harikrishna was roped in to compose the music. On 30 November 2016, the film was reported to have titled as Mugulu Nage.[9] On 8 December 2016, the filming began with the first schedule officially canned at ISRO Layout in Bengaluru.[10] The second schedule was held at Mysuru followed by the third schedule being shot in Puducherry.[11] It was also reported that team expected the filming would be completed by the end of February 2017. However, the shooting was officially concluded in April 2017.[12]
Casting[edit]
Ganesh and Yogaraj Bhat shooting in Pondicherry for the film Mugulu Nage
Ganesh and Yogaraj Bhat shooting in Pondicherry for the film
After signing in Ganesh for the lead role, actress Amulya was signed in for one of the female leads.[13] It was also reported that the film would feature three more leading female characters, with a total of four different romantic tracks. Actresses Nabha Natesh and Nikitha Narayan were approached to play the other lead roles.[14] However, later Nabha was replaced by model turned actress Ashika Ranganath, playing her first role for a film. Apart from these leading actresses. In February 2017, it was announced that actor Jaggesh would be appearing in a special song sequence penned by Bhat himself.[15] Later in March 2017, Bhat made some last-minute changes by replacing Amulya with actress Apoorva Arora since Amulya got engaged and her marriage dates were clashing with the film schedule.[16] Apoorva joined the team on 18 March and shot her scenes at Barkur. It was also reported that Amulya would still make a guest appearance in the film.
Soundtrack[edit]
Mugulu Nage
Soundtrack album by V. Harikrishna
Released 11 July 2017
Recorded 2017
Genre Feature film soundtrack
Label D Beats
V. Harikrishna scored the film's background and for its soundtrack.[17] The soundtrack album was released starting with the track "Hodi Ombattu" on 11 July 2017 in Hubli and each song in a different city of Karnataka subsequently on every alternate day to "honour each of Harikrishna's songs", who scored for his 100th film.[18][19] The distribution rights procured by D Beats.[20] The album consists of six tracks, the lyrics for which were written by Yogaraj Bhat and Jayanth Kaikini.
Track list
No. Title Lyrics Singer(s) Length
1. "Ninna Snehadinda" Yogaraj Bhat Shreya Ghoshal 4:07
2. "Hodi Ombath" Yogaraj Bhat Vijay Prakash 4:30
3. "Roopasi" Jayanth Kaikini Sonu Nigam 4:17
4. "Kere Yeri" Yogaraj Bhat Sonu Nigam 4:40
5. "Mugulu Nage" Yogaraj Bhat Sonu Nigam 4:15
6. "Kannadi Illada Orinali" Jayanth Kaikini Shreya Ghoshal 4:19
7. "Amara Hale Nenapu" Yogaraj Bhat Vijay Prakash 4:32
Total length: 30:00
References[edit]
Jump up ^
The project marks the third film in the combination of Yogaraj Bhat and Ganesh after Mungaru Male (2006) and Gaalipata (2008). The filming began on 8 December 2016 in Bengaluru.[5] Further, the shooting took place in Puducherry, [6] Mysuru, Yaana and Sirsi, Karnataka.
The film was released on September 1, 2017 with mixed reviews. Movie is declared as avg at box office [7]
Contents [hide]
1 Cast
2 Production
2.1 Filming
2.2 Casting
3 Soundtrack
4 References
5 External links
Cast[edit]
Ganesh as Pulakeshi
Apoorva Arora as Chaaru
Nikitha Narayan as Siri
Ashika Ranganath as Vaishali Hande
Ananth Nag
Achyuth Kumar as Achyuth
Rangayana Raghu
Jaggesh in a special appearance
Amulya in a guest appearance
Chandan Achar
Niharika
Dharmanna Kadur
Production[edit]
Filming[edit]
In August 2016, it was reported that the successful combination of Yogaraj Bhat and Ganesh are teaming up together again for a new romantic venture.[8] While Bhat was taking care of the script, wrote story and take up the direction, Ganesh was cast as the lead actor apart from co-producing the film and V. Harikrishna was roped in to compose the music. On 30 November 2016, the film was reported to have titled as Mugulu Nage.[9] On 8 December 2016, the filming began with the first schedule officially canned at ISRO Layout in Bengaluru.[10] The second schedule was held at Mysuru followed by the third schedule being shot in Puducherry.[11] It was also reported that team expected the filming would be completed by the end of February 2017. However, the shooting was officially concluded in April 2017.[12]
Casting[edit]
Ganesh and Yogaraj Bhat shooting in Pondicherry for the film Mugulu Nage
Ganesh and Yogaraj Bhat shooting in Pondicherry for the film
After signing in Ganesh for the lead role, actress Amulya was signed in for one of the female leads.[13] It was also reported that the film would feature three more leading female characters, with a total of four different romantic tracks. Actresses Nabha Natesh and Nikitha Narayan were approached to play the other lead roles.[14] However, later Nabha was replaced by model turned actress Ashika Ranganath, playing her first role for a film. Apart from these leading actresses. In February 2017, it was announced that actor Jaggesh would be appearing in a special song sequence penned by Bhat himself.[15] Later in March 2017, Bhat made some last-minute changes by replacing Amulya with actress Apoorva Arora since Amulya got engaged and her marriage dates were clashing with the film schedule.[16] Apoorva joined the team on 18 March and shot her scenes at Barkur. It was also reported that Amulya would still make a guest appearance in the film.
Soundtrack[edit]
Mugulu Nage
Soundtrack album by V. Harikrishna
Released 11 July 2017
Recorded 2017
Genre Feature film soundtrack
Label D Beats
V. Harikrishna scored the film's background and for its soundtrack.[17] The soundtrack album was released starting with the track "Hodi Ombattu" on 11 July 2017 in Hubli and each song in a different city of Karnataka subsequently on every alternate day to "honour each of Harikrishna's songs", who scored for his 100th film.[18][19] The distribution rights procured by D Beats.[20] The album consists of six tracks, the lyrics for which were written by Yogaraj Bhat and Jayanth Kaikini.
Track list
No. Title Lyrics Singer(s) Length
1. "Ninna Snehadinda" Yogaraj Bhat Shreya Ghoshal 4:07
2. "Hodi Ombath" Yogaraj Bhat Vijay Prakash 4:30
3. "Roopasi" Jayanth Kaikini Sonu Nigam 4:17
4. "Kere Yeri" Yogaraj Bhat Sonu Nigam 4:40
5. "Mugulu Nage" Yogaraj Bhat Sonu Nigam 4:15
6. "Kannadi Illada Orinali" Jayanth Kaikini Shreya Ghoshal 4:19
7. "Amara Hale Nenapu" Yogaraj Bhat Vijay Prakash 4:32
Total length: 30:00
References[edit]
Jump up ^
Srinivasa Kalyana
IndiaGlitz [Saturday, February 25, 2017 • Kannada] 0 Comments
Kannada Movie Reviews
Mugulu Nage - Mugulu Nage
Eleyaru Naavu Geleyaru - Eleyaru Naavu Geleyaru
Pataki - Pataki
Chakravarthy - Chakravarthy
Rogue - Rogue
Rajakumara - Rajakumara
Jilebi - Jilebi
Hebbuli - Hebbuli
Santhu Straight Forward - Santhu Straight Forward
The talented team at work is visible in contents. The young team with philosophical thought ‘Arishadwargas’ are stitched for a neat and tidy narration. The school age to stage of professional career, different phases are focused and at the end the connectivity to ‘Kama, Kroda, Lobha, Moha, Mada, Matsarya plus Moksha’ is different angle to the ups and downs in love life of protagonist.
The film looks visually good, dialogues are funny, technically new attempts are seen and in 128 minutes of the film ‘Srinivasa Kalyana’ director cum hero of the film MG Srinivas has churned out his best.
The beginning of the film with cartoons and commentary prepare the audience on the developments. When the narration starts flowing from the point of view of elderly actor Dattanna traveling in a car, audience also get a feel of journey towards ‘Moksha’. The ‘Moksha’ in this case is a cool and decent watch from usual shoddy films. From the point of view of protagonist, it is a girl ‘Moksha’ who enters in his life after three ditches in his life.
The teenage tantrums and obsession of LK Balu (Srinivas) is what would have happened in innumerable cases. The mistake of love and later getting transferred in to obsession is the first phase. The ‘Matsara’ also comes in this phase of life as another girl comes friendly with a boy of her (Akshara) liking. Akshara (Kavitha Gowda) becomes possessive and she does not want to see Balu with another girl. After years going out from school she remembers Balu to invite him for her marriage is a shocker. What happened to this huge gap between Balu and Akshara, we don’t find answer.
In the next phase we have Radha (Nikila Rao) coming in the life of Balu. For Radha it is a costly mistake. She has almost fallen in love with Balu who is grown up today. The chance for Balu to explain his situation does not come and that lead to breakup in love. By this time Balu passes all stages of ‘Arishadwargas’ of life for ultimate in life ‘Moksha’. This is not that philosophical ‘Mokha’ – after bumps on the road like how a nice road comes – Balu settles with Moksha (Priyanka Rao) in life.
MG Srinivas in different age groups in this film looks fit and his emotions, dialogue delivery style is convincing. He is a good dancer. The performance of Nikila Rao gathers good attention and plum Kavitha Gowda is naughty from her looks – both have great future ahead. Sujay Shastry is likely to be another Sharan of future. His comedy timing and dialogue delivery is first class. Achyuthkumar and Dattanna are of course flawless actors.
Ashwin Kadambur cinematography is on par with any senior cameramen of the cinema industry. The framing, lighting, angles and new techniques used are brilliant. With natural light he has given splendid visuals to eyes.
The music is another adorable part of this film. Midun Mukundan and Raghu Thane given lilting tunes and some of the lines are well written in the film. Editing is crisp and going back and coming forward in the pattern there is nothing short.
Srinivasa Kalyana is a film for all ages because it takes you back to earlier days and gives a good feel at the end.
Rating: 4 / 5.0
IndiaGlitz [Saturday, February 25, 2017 • Kannada] 0 Comments
Kannada Movie Reviews
Mugulu Nage - Mugulu Nage
Eleyaru Naavu Geleyaru - Eleyaru Naavu Geleyaru
Pataki - Pataki
Chakravarthy - Chakravarthy
Rogue - Rogue
Rajakumara - Rajakumara
Jilebi - Jilebi
Hebbuli - Hebbuli
Santhu Straight Forward - Santhu Straight Forward
The talented team at work is visible in contents. The young team with philosophical thought ‘Arishadwargas’ are stitched for a neat and tidy narration. The school age to stage of professional career, different phases are focused and at the end the connectivity to ‘Kama, Kroda, Lobha, Moha, Mada, Matsarya plus Moksha’ is different angle to the ups and downs in love life of protagonist.
The film looks visually good, dialogues are funny, technically new attempts are seen and in 128 minutes of the film ‘Srinivasa Kalyana’ director cum hero of the film MG Srinivas has churned out his best.
The beginning of the film with cartoons and commentary prepare the audience on the developments. When the narration starts flowing from the point of view of elderly actor Dattanna traveling in a car, audience also get a feel of journey towards ‘Moksha’. The ‘Moksha’ in this case is a cool and decent watch from usual shoddy films. From the point of view of protagonist, it is a girl ‘Moksha’ who enters in his life after three ditches in his life.
The teenage tantrums and obsession of LK Balu (Srinivas) is what would have happened in innumerable cases. The mistake of love and later getting transferred in to obsession is the first phase. The ‘Matsara’ also comes in this phase of life as another girl comes friendly with a boy of her (Akshara) liking. Akshara (Kavitha Gowda) becomes possessive and she does not want to see Balu with another girl. After years going out from school she remembers Balu to invite him for her marriage is a shocker. What happened to this huge gap between Balu and Akshara, we don’t find answer.
In the next phase we have Radha (Nikila Rao) coming in the life of Balu. For Radha it is a costly mistake. She has almost fallen in love with Balu who is grown up today. The chance for Balu to explain his situation does not come and that lead to breakup in love. By this time Balu passes all stages of ‘Arishadwargas’ of life for ultimate in life ‘Moksha’. This is not that philosophical ‘Mokha’ – after bumps on the road like how a nice road comes – Balu settles with Moksha (Priyanka Rao) in life.
MG Srinivas in different age groups in this film looks fit and his emotions, dialogue delivery style is convincing. He is a good dancer. The performance of Nikila Rao gathers good attention and plum Kavitha Gowda is naughty from her looks – both have great future ahead. Sujay Shastry is likely to be another Sharan of future. His comedy timing and dialogue delivery is first class. Achyuthkumar and Dattanna are of course flawless actors.
Ashwin Kadambur cinematography is on par with any senior cameramen of the cinema industry. The framing, lighting, angles and new techniques used are brilliant. With natural light he has given splendid visuals to eyes.
The music is another adorable part of this film. Midun Mukundan and Raghu Thane given lilting tunes and some of the lines are well written in the film. Editing is crisp and going back and coming forward in the pattern there is nothing short.
Srinivasa Kalyana is a film for all ages because it takes you back to earlier days and gives a good feel at the end.
Rating: 4 / 5.0
வெச்சூர் மாடு (மலையாளம்: വെച്ചൂര് പശു ) என்பது கேரளத்தின் கோட்டையம் மாவட்டத்தில் உள்ள வெச்சூர் என்ற ஊரின் பெயரால் அழைக்கப்படுகிறது. இவை சராசரியாக 87 செமீ உயரத்துடனும், 124 செ.மீ. சராசரி நீளமுடனும் இருக்கும். இது உலகின் சிறிய மாடாக கின்னஸ் சாதனை புத்தகத்தில் இடம்பித்துள்ளது.[1] மேலும் இது குறைந்த உணவில் நிறைய பால் கறக்கக்கூடியது. [2] வெச்சூர் மாடுகள் விலங்கு வளர்ப்பு பேராசிரியரான சோசம்மா லைப் மற்றும் அவரது மாணவர்கள் கொண்ட குழு இணைந்து செய்த பணியின் காரணமாக இந்த மாடுகள் மரபியலாக அழிவிலிருந்து காக்கப்பட்டன. [3] இவர்களால் 1989 ஆம் ஆண்டில், ஒரு பாதுகாப்பு அலகு தொடங்கப்பட்டது. 1998 இல் பாதுகாப்பு அறக்கட்டளை அமைப்பு விவசாயிகளின் பங்களிப்புடன் அமைக்கப்பட்டது. [4] வெச்சூர் மாடுகள் 1960 வரை கேரளத்தில் பிரபலமாக இருந்தன, ஆனால் இந்த மாடுகள் கலப்பினத்துக்கு ஆளானதால் அரிய மாடுகளாயின. [5] 2000 ஆம் ஆண்டில், வெச்சூர் மாடு உள்நாட்டு விலங்கு வேறுபாட்டுக்கான FAO வின் உலக கண்காணிப்பு பட்டியலில், உடனடியாக அழியக்கூடிய விலங்கு பட்டியலில் இடம் பெற்றிருந்தது. ஒரு இன விலங்கில் ஆண் பெண் விலங்குகளின் எண்ணிக்கை மிகவும் குறையும்போது இந்த பட்டியலில் சேர்க்கப்பட்டும் . [6] இந்த மாடுகள் சுமார் 200 மாடுகள் இருக்கலாம் என கருதப்படுகிறது, இவற்றில் கிட்டத்தட்ட 100 மாடுகள் கால்நடை மருத்துவக் கல்லூரிகளில் இருக்கும். இந்த மாடுகளின் சராசரி உயரம் 90 செ மீ ஆகவும், சராசரி எடை 130 கோலோவாகவும் இருக்கும். ஒரு நாளைக்கு மூன்று லிட்டர் பால் கறக்கக்கூடியது.
மேற்கோள்கள்[தொகு]
Jump up ↑ "Smallest cow (Height)". Guinness World Records. பார்த்த நாள் January 6, 2012.
Jump up ↑ Krishnakumar, R. (9 April 1999). "A cow and controversy", Frontline (magazine).
Jump up ↑ Animal genetics Resource Bulletin 1997 (FAO)
Jump up ↑ Proceedings of the National Conference on Native Livestock Breeds and their Sustainable Use 2010.
Jump up ↑ Prabu, M. J.[protected]. "Vechur cattle: ideal for household rearing". The Hindu. பார்த்த நாள் January 6, 2012.
Jump up ↑ Sainath, P. (January 5, 2012). "Holy cow! Small is beautiful". The Hindu.
வெளி இணைப்புகள்[தொகு]
Search Wikimedia Commons விக்கிமீடியா பொதுவகத்தில் Vechur Cattle என்னும் தலைப்புடன் தொடர்புடைய பல ஊடகக் கோப்புகள் உள்ளன.
Vechur Conservation Trust
பகுப்புகள்: இந்தியாவில் தோன்றிய மாட்டு இனங்கள்கோட்டயம் மாவட்டம்
குப்தப் பேரரசு (ஆட்சிக் காலம்: கி பி 320 – 550) இந்தியத் துணைக்கண்டத்தின் பெரும் பகுதிகளை ஆண்ட பேரரசுகளில் ஒன்றாக விளங்கியது. குப்தப் பேரரசை நிறுவியவர் ஸ்ரீகுப்தர் ஆவார். கி பி 320 முதல் 550 வரை, குப்தர் எனும் அரச மரபினரால் ஆளப்பட்ட இப்பேரரசு அதன் உச்சக்கட்டத்தில், அக்கால வட இந்தியாவின் பெரும் பகுதியை உள்ளடக்கி இருந்தது. [1] இப்பேரரசின் பகுதிகளாக இன்றைய பாகிஸ்தான், இந்தியா, வங்காளதேசம் ஆகிய நாடுகள் அமைந்திருந்தன.
அறிவியல், கணிதம், வானியல், சமயம், இந்திய தத்துவம் ஆகிய துறைகளில் சிறந்து விளங்கியதால், குப்தப் பேரசின் காலம் இந்தியாவின் பொற்காலம் எனக் குறிப்பிடப்படுவது உண்டு. [2] குப்தர்களின் ஆட்சியில் ஏற்பட்டிருந்த அமைதியும், வளமும் அறிவியல் மற்றும் கலைத் துறைகளில் வளர்ச்சி ஏற்படுவதை ஊக்குவித்தன. பதின்ம எண்முறை, இந்திய எண் முறை மற்றும் பூஜ்ஜியம் குப்தப் பேரரசுக் காலத்துக் கண்டுபிடிப்புக்களே. வரலாற்றாளர்கள், செந்நெறி நாகரிகத்தின் ஒரு மாதிரியாக குப்தப் பேரசை, ஹான் பேரரசு, தாங் பேரரசு மற்றும் ரோமப் பேரரசுடன் ஒன்றாக வைத்து எண்ணுகிறார்கள்.[3][4]
குப்தப் பேரரசர்களில் மிகவும் புகழ் பெற்றவர்கள் முதலாம் சந்திரகுப்தர், சமுத்திரகுப்தர் இரண்டாம் சந்திரகுப்தர் மற்றும் முதலாம் குமாரகுப்தர் மற்றும் ஸ்கந்தகுப்தர் ஆவார்கள்.
மேலும் குப்தர்கள் காலத்தில் அறிவியல், தொழில்நுட்பம், தருக்கம், கணிதம், வானவியல், இந்தியத் தத்துவம், சோதிடம், இந்து தொன்மவியல், இந்து சமயம், பௌத்தம் மற்றும் சமணம் போன்ற சமயங்கள் செழித்ததுடன், இந்துப் பண்பாடு, சமசுகிருத மொழி இலக்கியங்கள் வளர்ந்தது.
நான்காம் நூற்றாண்டின் சமசுகிருத மொழியின் மகாகவி காளிதாசன் தமது இரகுவம்சம் எனும் காவியத்தில், குப்த ஆட்சியாளர்கள், நடு ஆசியாவின் ஆமூ தாரியா ஆறு பாயும் இடங்களில் வாழும் சகர்கள் ஹூணர்கள், காம்போஜர்கள், கிராதர்கள் மற்றும் கிண்ணர நாடுகளையும் சேர்த்து, இந்தியத் துணைக்கண்டத்தின் 21 நாடுகளை வென்றதாக குறிப்பிட்டுள்ளார். [5]
இரண்டாம் சந்திரகுப்தர் ஆட்சிக் காலத்தில் பண்பாட்டு, நாகரீகம், கலைகள், இலக்கியங்கள் நன்கு வளர்ச்சி அடைந்தது. சமஸ்கிருத மொழியில் புராணங்கள், இராமாயணம் மற்றும் மகாபாரதம் போன்ற இதிகாச இலக்கியங்கள் ஓலைச் சுவடிகளில் எழுத்து வடிவம் பெற்றது. [6]
குப்தர்களின் ஆட்சியில் கவிஞர் காளிதாசன், வானிலை மற்றும் கணித அறிஞர்களான ஆரியபட்டர் மற்றும் வராகமிகிரர், பஞ்சதந்திர நூலை எழுதிய விஷ்ணு குப்தர், காம சூத்திரம் நூலை எழுதிய வாத்சாயனர், ஆயுர்வேத மருத்துவரான சுஸ்ருதர் போன்ற பல்கலை அறிஞர்கள் வாழ்ந்தனர். [7]
குப்தர்கள் ஆட்சிக் காலத்தில்அறிவியல் மற்றும் அரசியல் நிர்வாகம் செழித்தோங்கி உச்சகட்டத்தை அடைந்தது. குப்தர்கள் தென்கிழக்கு ஆசியா நாடுகளுடனும் குறிப்பாக இலங்கை மற்றும் பர்மா போன்ற அண்டை நாடுகளுடன் பலமான வணிக உறவுகளைக் கொண்டிருந்தனர். [8]
குப்தப் பேரரசர் விஷ்ணு குப்தர் காலத்தில், குப்தப் பேரரசின் ஆட்சிப் பகுதிகள் சிறிது சிறிதாக, நடு ஆசியாவின் ஹெப்தலைட்டு ஹூணர்களின் தொடர் ஆக்கிரமிப்புகளாலும், அண்டை நாட்டவர்களாலும் ஆக்கிரமிக்கப்பட்டு, இறுதியில் கி பி 550-இல் குப்தப் பேரரசு வீழ்ச்சியுற்றது. [9][10]
குப்தப் பேரரசின் வீழ்ச்சிக்குப் பின்னர் இந்தியத் துணைக்கண்டத்தில் புதிய சிறிய, பெரிய நாடுகள் உருவானது. விஷ்ணு குப்தருக்குப் பின் வந்த பிற்கால குப்த அரசர்கள் மகதத்தின் பகுதிகளை மட்டும் ஆண்டனர்.
மேற்கோள்கள்[தொகு]
Jump up ↑ "Smallest cow (Height)". Guinness World Records. பார்த்த நாள் January 6, 2012.
Jump up ↑ Krishnakumar, R. (9 April 1999). "A cow and controversy", Frontline (magazine).
Jump up ↑ Animal genetics Resource Bulletin 1997 (FAO)
Jump up ↑ Proceedings of the National Conference on Native Livestock Breeds and their Sustainable Use 2010.
Jump up ↑ Prabu, M. J.[protected]. "Vechur cattle: ideal for household rearing". The Hindu. பார்த்த நாள் January 6, 2012.
Jump up ↑ Sainath, P. (January 5, 2012). "Holy cow! Small is beautiful". The Hindu.
வெளி இணைப்புகள்[தொகு]
Search Wikimedia Commons விக்கிமீடியா பொதுவகத்தில் Vechur Cattle என்னும் தலைப்புடன் தொடர்புடைய பல ஊடகக் கோப்புகள் உள்ளன.
Vechur Conservation Trust
பகுப்புகள்: இந்தியாவில் தோன்றிய மாட்டு இனங்கள்கோட்டயம் மாவட்டம்
குப்தப் பேரரசு (ஆட்சிக் காலம்: கி பி 320 – 550) இந்தியத் துணைக்கண்டத்தின் பெரும் பகுதிகளை ஆண்ட பேரரசுகளில் ஒன்றாக விளங்கியது. குப்தப் பேரரசை நிறுவியவர் ஸ்ரீகுப்தர் ஆவார். கி பி 320 முதல் 550 வரை, குப்தர் எனும் அரச மரபினரால் ஆளப்பட்ட இப்பேரரசு அதன் உச்சக்கட்டத்தில், அக்கால வட இந்தியாவின் பெரும் பகுதியை உள்ளடக்கி இருந்தது. [1] இப்பேரரசின் பகுதிகளாக இன்றைய பாகிஸ்தான், இந்தியா, வங்காளதேசம் ஆகிய நாடுகள் அமைந்திருந்தன.
அறிவியல், கணிதம், வானியல், சமயம், இந்திய தத்துவம் ஆகிய துறைகளில் சிறந்து விளங்கியதால், குப்தப் பேரசின் காலம் இந்தியாவின் பொற்காலம் எனக் குறிப்பிடப்படுவது உண்டு. [2] குப்தர்களின் ஆட்சியில் ஏற்பட்டிருந்த அமைதியும், வளமும் அறிவியல் மற்றும் கலைத் துறைகளில் வளர்ச்சி ஏற்படுவதை ஊக்குவித்தன. பதின்ம எண்முறை, இந்திய எண் முறை மற்றும் பூஜ்ஜியம் குப்தப் பேரரசுக் காலத்துக் கண்டுபிடிப்புக்களே. வரலாற்றாளர்கள், செந்நெறி நாகரிகத்தின் ஒரு மாதிரியாக குப்தப் பேரசை, ஹான் பேரரசு, தாங் பேரரசு மற்றும் ரோமப் பேரரசுடன் ஒன்றாக வைத்து எண்ணுகிறார்கள்.[3][4]
குப்தப் பேரரசர்களில் மிகவும் புகழ் பெற்றவர்கள் முதலாம் சந்திரகுப்தர், சமுத்திரகுப்தர் இரண்டாம் சந்திரகுப்தர் மற்றும் முதலாம் குமாரகுப்தர் மற்றும் ஸ்கந்தகுப்தர் ஆவார்கள்.
மேலும் குப்தர்கள் காலத்தில் அறிவியல், தொழில்நுட்பம், தருக்கம், கணிதம், வானவியல், இந்தியத் தத்துவம், சோதிடம், இந்து தொன்மவியல், இந்து சமயம், பௌத்தம் மற்றும் சமணம் போன்ற சமயங்கள் செழித்ததுடன், இந்துப் பண்பாடு, சமசுகிருத மொழி இலக்கியங்கள் வளர்ந்தது.
நான்காம் நூற்றாண்டின் சமசுகிருத மொழியின் மகாகவி காளிதாசன் தமது இரகுவம்சம் எனும் காவியத்தில், குப்த ஆட்சியாளர்கள், நடு ஆசியாவின் ஆமூ தாரியா ஆறு பாயும் இடங்களில் வாழும் சகர்கள் ஹூணர்கள், காம்போஜர்கள், கிராதர்கள் மற்றும் கிண்ணர நாடுகளையும் சேர்த்து, இந்தியத் துணைக்கண்டத்தின் 21 நாடுகளை வென்றதாக குறிப்பிட்டுள்ளார். [5]
இரண்டாம் சந்திரகுப்தர் ஆட்சிக் காலத்தில் பண்பாட்டு, நாகரீகம், கலைகள், இலக்கியங்கள் நன்கு வளர்ச்சி அடைந்தது. சமஸ்கிருத மொழியில் புராணங்கள், இராமாயணம் மற்றும் மகாபாரதம் போன்ற இதிகாச இலக்கியங்கள் ஓலைச் சுவடிகளில் எழுத்து வடிவம் பெற்றது. [6]
குப்தர்களின் ஆட்சியில் கவிஞர் காளிதாசன், வானிலை மற்றும் கணித அறிஞர்களான ஆரியபட்டர் மற்றும் வராகமிகிரர், பஞ்சதந்திர நூலை எழுதிய விஷ்ணு குப்தர், காம சூத்திரம் நூலை எழுதிய வாத்சாயனர், ஆயுர்வேத மருத்துவரான சுஸ்ருதர் போன்ற பல்கலை அறிஞர்கள் வாழ்ந்தனர். [7]
குப்தர்கள் ஆட்சிக் காலத்தில்அறிவியல் மற்றும் அரசியல் நிர்வாகம் செழித்தோங்கி உச்சகட்டத்தை அடைந்தது. குப்தர்கள் தென்கிழக்கு ஆசியா நாடுகளுடனும் குறிப்பாக இலங்கை மற்றும் பர்மா போன்ற அண்டை நாடுகளுடன் பலமான வணிக உறவுகளைக் கொண்டிருந்தனர். [8]
குப்தப் பேரரசர் விஷ்ணு குப்தர் காலத்தில், குப்தப் பேரரசின் ஆட்சிப் பகுதிகள் சிறிது சிறிதாக, நடு ஆசியாவின் ஹெப்தலைட்டு ஹூணர்களின் தொடர் ஆக்கிரமிப்புகளாலும், அண்டை நாட்டவர்களாலும் ஆக்கிரமிக்கப்பட்டு, இறுதியில் கி பி 550-இல் குப்தப் பேரரசு வீழ்ச்சியுற்றது. [9][10]
குப்தப் பேரரசின் வீழ்ச்சிக்குப் பின்னர் இந்தியத் துணைக்கண்டத்தில் புதிய சிறிய, பெரிய நாடுகள் உருவானது. விஷ்ணு குப்தருக்குப் பின் வந்த பிற்கால குப்த அரசர்கள் மகதத்தின் பகுதிகளை மட்டும் ஆண்டனர்.
Hi All - Please don't believe this kind of persons and fake companies who ask you money in demand. NUCOT is a coaching institute after receiving the money they will cheat the students saying that will get placed. Please don't believe.
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Fetal proper stem cells come from the tissue of the fetus proper, and are generally obtained after an abortion. These stem cells are not immortal but have a high level of division and are multipotent.Extraembryonic fetal stem cells come from extraembryonic membranes, and are generally not distinguished from adult stem cells. These stem cells are acquired after birth, they are not immortal but have a high level of cell division, and are pluripotent.[18]AdultMain article: Adult stem cell
Stem cell division and differentiation. A: stem cell; B: progenitor cell; C: differentiated cell; 1: symmetric stem cell division; 2: asymmetric stem cell division; 3: progenitor division; 4: terminal differentiation
Adult stem cells, also called somatic (from Greek Σωματικóς, "of the body") stem cells, are stem cells which maintain and repair the tissue in which they are found.[19]They can be found in children, as well as adults.[20]
Pluripotent adult stem cells are rare and generally small in number, but they can be found in umbilical cord blood and other tissues.[21] Bone marrow is a rich source of adult stem cells, [22] which have been used in treating several conditions including spinal cord injury, [23] liver cirrhosis, [24] chronic limb ischemia [25] and endstage heart failure.[26] The quantity of bone marrow stem cells declines with age and is greater in males than females during reproductive years.[27] Much adult stem cell research to date has aimed to characterize their potency and self-renewal capabilities.[28] In mice, pluripotent stem cells are directly generated from adult fibroblast cultures. However, mice do not live long with stem cell organs.[29]
Most adult stem cells are lineage-restricted (multipotent) and are generally referred to by their tissue origin (mesenchymal stem cell, adipose-derived stem cell, endothelial stem cell, dental pulp stem cell, etc.).[30][31]
Adult stem cell treatments have been successfully used for many years to treat leukemia and related bone/blood cancers through bone marrow transplants.[32] Adult stem cells are also used in veterinary medicine to treat tendon and ligament injuries in horses.[33]
The use of adult stem cells in research and therapy is not as controversial as the use of embryonic stem cells, because the production of adult stem cells does not require the destruction of an embryo. Additionally, in instances where adult stem cells are obtained from the intended recipient (an autograft), the risk of rejection is essentially non-existent. Consequently, more US government funding is being provided for adult stem cell research.[34]
AmnioticMultipotent stem cells are also found in amniotic fluid. These stem cells are very active, expand extensively without feeders and are not tumorigenic. Amniotic stem cells are multipotent and can differentiate in cells of adipogenic, osteogenic, myogenic, endothelial, hepatic and also neuronal lines.[35] Amniotic stem cells are a topic of active research.
Use of stem cells from amniotic fluid overcomes the ethical objections to using human embryos as a source of cells. Roman Catholic teaching forbids the use of embryonic stem cells in experimentation; accordingly, the Vatican newspaper "Osservatore Romano" called amniotic stem cells "the future of medicine".[36]
It is possible to collect amniotic stem cells for donors or for autologuous use: the first US amniotic stem cells bank [37][38] was opened in 2009 in Medford, MA, by Biocell Center Corporation[39][40][41] and collaborates with various hospitals and universities all over the world.[42]
Cord bloodMain article: Cord blood-derived multipotent stem cell
A certain kind of cord blood stem cell (CB-SC) is multipotent and displays embryonic and hematopoietic characteristics. Phenotypic characterization demonstrates that (CB-SCs) display embryonic cell markers (e.g., transcription factors OCT-4 and Nanog, stage-specific embryonic antigen (SSEA)-3, and SSEA-4) and leukocyte common antigen CD45, but that they are negative for blood cell lineage markers (e.g., CD1a, CD3, CD4, CD8, CD11b, CD11c, CD13, CD14, CD19, CD20, CD34, CD41a, CD41b, CD83, CD90, CD105, and CD133).[43][44]
Additionally, CB-SCs display very low immunogenicity as indicated by expression of a very low level of major histocompatibility complex (MHC) antigens and failure to stimulate the proliferation of allogeneic lymphocytes.[43][45] They can give rise to three embryonic layer-derived cells in the presence of different inducers.[43][46]
More specifically, CB-SCs tightly adhere to culture dishes with a large rounded morphology and are resistant to common detaching methods (trypsin/EDTA).[43][45][46] CB-SCs are the active agent in stem cell educator therapy, which has therapeutic potential against autoimmune diseases like type 1 diabetes according to studies by Yong Zhao et al.[44][47][48][49][unreliable medical source?]
Induced pluripotentMain article: Induced pluripotent stem cell
These are not adult stem cells, but rather adult cells (e.g. epithelial cells) reprogrammed to give rise to pluripotent capabilities. Using genetic reprogramming with protein transcription factors, pluripotent stem cells equivalent to embryonic stem cells have been derived from human adult skin tissue.[50][51][52] Shinya Yamanaka and his colleagues at Kyoto University used the transcription factors Oct3/4, Sox2, c-Myc, and Klf4[50] in their experiments on cells from human faces.Junying Yu, James Thomson, and their colleagues at the University of Wisconsin–Madison used a different set of factors, Oct4, Sox2, Nanog and Lin28, [50] and carried out their experiments using cells from human foreskin.
As a result of the success of these experiments, Ian Wilmut, who helped create the first cloned animal Dolly the Sheep, has announced that he will abandon somatic cell nuclear transfer as an avenue of research.[53]
Frozen blood samples can be used as a source of induced pluripotent stem cells, opening a new avenue for obtaining the valued cells.[54]
LineageMain article: Stem cell line
To ensure self-renewal, stem cells undergo two types of cell division (see Stem cell division and differentiation diagram). Symmetric division gives rise to two identical daughter cells both endowed with stem cell properties. Asymmetric division, on the other hand, produces only one stem cell and a progenitor cell with limited self-renewal potential. Progenitors can go through several rounds of cell division before terminally differentiating into a mature cell. It is possible that the molecular distinction between symmetric and asymmetric divisions lies in differential segregation of cell membrane proteins (such asreceptors) between the daughter cells.[55]
An alternative theory is that stem cells remain undifferentiated due to environmental cues in their particular niche. Stem cells differentiate when they leave that niche or no longer receive those signals. Studies in Drosophila germarium have identified the signals decapentaplegic and adherens junctions that prevent germarium stem cells from differentiating.[56][57]
TreatmentsMain article: Stem cell therapy
[[File:Stem cell treatments.svg|thumb|330px|Diseases and conditions where stem cell treatment is promising or emerging. Diabetes, rheumatoid arthritis, Parkinson's, Alzheimer's disease, osteoarthritis:
Stem Cell Basics: What are the potential uses of human stem cells and the obstacles that must be overcome before these potential uses will be realized?. In Stem Cell Information World Wide Web site. Bethesda, MD: National Institutes of Health, U.S. Department of Health and Human Services, 2009. cited Sunday, April 26, 2009Stroke and traumatic brain injury repair:
Steinberg, Douglas (November 2000) Stem Cells Tapped to Replenish Organs thescientist.comLearning defects:
ISRAEL21c: Israeli scientists reverse brain birth defects using stem cells December 25, 2008. (Researchers from the Hebrew University of Jerusalem-Hadassah Medical led by Prof. Joseph Yanai)Spinal cord injury repair:
Kang KS, Kim SW, Oh YH, Yu JW, Kim KY, Park HK, Song CH, Han H (2005). "A 37-year-old spinal cord-injured female patient, transplanted of multipotent stem cells from human UC blood, with improved sensory perception and mobility, both functionally and morphologically: a case study". Cytotherapy 7 (4): 368–73. doi:10.1080/[protected].PMID 16162459.Heart infarction:
Strauer BE, Schannwell CM, Brehm M (2009). "Therapeutic potentials of stem cells in cardiac diseases". Minerva Cardioangiol 57 (2): 249–67. PMID 19274033.Anti-cancer:
Stem Cells Tapped to Replenish Organs thescientist.com, Nov 2000. By Douglas SteinbergBaldness:
Hair Cloning Nears Reality as Baldness Cure WebMD November 2004Replace missing teeth:
Yen AH, Sharpe PT (2008). "Stem cells and tooth tissue engineering". Cell Tissue Res. 331 (1): 359–72.doi:10.1007/s[protected]. PMID 17938970.Repair hearing:
Gene therapy is first deafness 'cure' – health – 14 February 2005 – New ScientistRestore vision:
BBC NEWS | England | Southern Counties | Stem cells used to restore visionAmyotrophic lateral sclerosis:
Vastag, B. (2001). "Stem Cells Step Closer to the Clinic: Paralysis Partially Reversed in Rats with ALS-like Disease".JAMA: the Journal of the American Medical Association 285 (13): 1691–1693. doi:10.1001/jama.285.13.1691.PMID 11277806. editCrohn's disease:
Anderson, Querida[protected]. "Osiris Trumpets Its Adult Stem Cell Product". Genetic Engineering & Biotechnology News (Mary Ann Liebert, Inc.). p. 13. Retrieved[protected]. "(subtitle) Procymal is being developed in many indications, GvHD being the most advanced"Wound healing:
Gurtner GC, Callaghan MJ, Longaker MT (2007). "Progress and potential for regenerative medicine". Annu. Rev. Med.58: 299–312. doi:10.1146/annurev.med.58.082405.095329. PMID 17076602. Bone marrow transplantation is, as of 2009, the only established use of stem cells.Stem cell therapy it is the use of stem cells to treat or prevent a disease or condition. Bone marrow transplant is a crude form of stem cell therapy that has been used clinically for many years without controversy. No stem cell therapies other than bone marrow transplant are widely used.[58][59]
Research is underway to develop various sources for stem cells, and to apply stem cell treatments for neurodegenerative diseases and conditions, diabetes, heart disease, and other conditions.[60]
In more recent years, with the ability of scientists to isolate and culture embryonic stem cells, and with scientists' growing ability to create stem cells using somatic cell nuclear transfer and techniques to created induced pluripotent stem cells, controversy has crept in, both related to abortion politics and to human cloning.
DisadvantagesStem cell treatments may require immunosuppression because of a requirement for radiation before the transplant to remove the patient's previous cells, or because the patient's immune system may target the stem cells. One approach to avoid the second possibility is to use cells from the same patient that is being treated.
Pluripotency in certain stem cells could also make it difficult to obtain a specific cell type. It is also difficult to obtain the exact cell type needed, because not all cells in a population differentiate uniformly. Undifferentiated cells can create tissues other than desired types.[61]
Some stem cells form tumors after transplantation; pluripotency is linked to tumor formation especially in embryonic stem cells, fetal proper stem cells, induced pluripotent stem cells. Fetal proper stem cells form tumors despite multipotency.[citation needed]
Hepatotoxicity and drug-induced liver injury account for a substantial number of failures of new drugs in development and market withdrawal, highlighting the need for screening assays such as stem cell-derived hepatocyte-like cells, that are capable of detecting toxicity early in the drug development process.[62]
Research patentsSome of the fundamental patents covering human embryonic stem cells are owned by the Wisconsin Alumni Research Foundation (WARF) - they are patents 5, 843, 780, 6, 200, 806, and 7, 029, 913 invented by James A. Thomson. WARF does not enforce these patents against academic scientists, but does enforce them against companies.[63]
In 2006, a request for the US Patent and Trademark Office (USPTO) to re-examine the three patents was filed by the Public Patent Foundation on behalf of its client, the non-profit patent-watchdog group Consumer Watchdog(formerly the Foundation for Taxpayer and Consumer Rights).[63] In the re-examination process, which involves several rounds of discussion between the USTPO and the parties, the USPTO initially agreed with Consumer Watchdog and rejected all the claims in all three patents, [64] however in response, WARF amended the claims of all three patents to make them more narrow, and in 2008 the USPTO found the amended claims in all three patents to be patentable. The decision on one of the patents (7, 029, 913) was appealable, while the decisions on the other two were not.[65][66] Consumer Watchdog appealed the granting of the '913 patent to the USTPO's Board of Patent Appeals and Interferences (BPAI) which granted the appeal, and in 2010 the BPAI decided that the amended claims of the '913 patent were not patentable.[67] However, WARF was able to re-open prosecution of the case and did so, amending the claims of the '913 patent again to make them more narrow, and in January 2013 the amended claims were allowed.[68]
In July 2013, Consumer Watchdog announced that it would appeal the decision to allow the claims of the '913 patent to the US Court of Appeals for the Federal Circuit (CAFC), the federal appeals court that hears patent cases.[69] At a hearing in December 2013, the CAFC raised the question of whether Consumer Watchdog had legal standing to appeal; the case could not proceed until that issue was resolved.[70]
Key research events
This section may be too long to read and navigate comfortably. Please considersplitting content into sub-articles or condensing it. (December[protected]: The term "stem cell" was proposed for scientific use by the Russian histologist Alexander Maksimov (1874–1928) at congress of hematologic society in Berlin. It postulated existence of haematopoietic stem cells.1960s: Joseph Altman and Gopal Das present scientific evidence of adult neurogenesis, ongoing stem cell activity in the brain; their reports contradict Cajal's "no new neurons" dogma and are largely ignored.1963: McCulloch and Till illustrate the presence of self-renewing cells in mouse bone marrow.1968: Bone marrow transplant between two siblings successfully treats SCID.1978: Haematopoietic stem cells are discovered in human cord blood.1981: Mouse embryonic stem cells are derived from the inner cell mass by scientists Martin Evans, Matthew Kaufman, and Gail R. Martin. Gail Martin is attributed for coining the term "Embryonic Stem Cell".[71]1992: Neural stem cells are cultured in vitro as neurospheres.1995: Dr. B.G. Matapurkar pioneers in adult stem-cell research with clinical utilization of research in the body and neo-regeneration of tissues and organs in the body. Received International Patent from US Patent Office (USA) in 2001 (effective from 1995). Clinical utilization in human body also demonstrated and patented in 60 patients (World Journal of Surgery-1999[72] and 1991[73]).1997: Dr. B.G. Matapurkar's surgical technique on regeneration of tissues and organs is published.[74] Regeneration of fallopian tube and uterus is published.[75]1997: Leukemia is shown to originate from a haematopoietic stem cell, the first direct evidence for cancer stem cells.1998: James Thomson and coworkers derive the first human embryonic stem cell line at the University of Wisconsin–Madison.[76]1998: John Gearhart (Johns Hopkins University) extracted germ cells from fetal gonadal tissue (primordial germ cells) before developing pluripotent stem cell lines from the original extract.2000s: Several reports of adult stem cell plasticity are published.2001: Scientists at Advanced Cell Technology clone first early (four- to six-cell stage) human embryos for the purpose of generating embryonic stem cells.[77]2003: Dr. Songtao Shi of NIH discovers new source of adult stem cells in children's primary teeth.[78]2004–2005: Korean researcher Hwang Woo-Suk claims to have created several human embryonic stem cell lines from unfertilised human oocytes. The lines were later shown to be fabricated.2005: Researchers at Kingston University in England claim to have discovered a third category of stem cell, dubbed cord-blood-derived embryonic-like stem cells (CBEs), derived from umbilical cord blood. The group claims these cells are able to differentiate into more types of tissue than adult stem cells.2005: Researchers at UC Irvine's Reeve-Irvine Research Center are able to partially restore the ability of rats with paralyzed spines to walk through the injection of human neural stem cells.[79]
Yong Zhao, University of Illinois at Chicago
April 2006 Scientists at the University of Illinois at Chicago identified novel stem cells from the umbilical cord blood with embryonic and hematopoietic characteristics.[43]August 2006: Mouse Induced pluripotent stem cells: the journal Cell publishes Kazutoshi Takahashi and Shinya Yamanaka.[29]November 2006: Yong Zhao et al. revealed the immune regulation of T lymphocytes byCord Blood-Derived Multipotent Stem Cells (CB-SCs).[45]October 2006: Scientists at Newcastle University in England create the first ever artificial liver cells using umbilical cord blood stem cells.[80][81]January 2007: Scientists at Wake Forest University led by Dr. Anthony Atala and Harvard University report discovery of a new type of stem cell in amniotic fluid.[82] This may potentially provide an alternative to embryonic stem cells for use in research and therapy.[83]June 2007: Research reported by three different groups shows that normal skin cells can be reprogrammed to an embryonic state in mice.[84] In the same month, scientist Shoukhrat Mitalipov reports the first successful creation of a primate stem cell line through somatic cell nuclear transfer[85]
Martin Evans, a co-winner of the Nobel Prize in recognition of his gene targeting work.
October 2007: Mario Capecchi, Martin Evans, and Oliver Smithies win the 2007 Nobel Prize for Physiology or Medicine for their work on embryonic stem cells from mice using gene targeting strategies producing genetically engineered mice (known as knockout mice) for gene research.[86]November 2007: Human induced pluripotent stem cells: Two similar papers released by their respective journals prior to formal publication: in Cell by Kazutoshi Takahashi andShinya Yamanaka, "Induction of pluripotent stem cells from adult human fibroblasts by defined factors", [87] and in Science by Junying Yu, et al., from the research group ofJames Thomson, "Induced pluripotent stem cell lines derived from human somatic cells":[88] pluripotent stem cells generated from mature human fibroblasts. It is possible now to produce a stem cell from almost any other human cell instead o[censored]sing embryos as needed previously, albeit the risk of tumorigenesis due to c-myc and retroviral gene transfer remains to be determined.January 2008: Robert Lanza and colleagues at Advanced Cell Technology and UCSF create the first human embryonic stem cells without destruction of the embryo[89]January 2008: Development of human cloned blastocysts following somatic cell nuclear transfer with adult fibroblasts[90]February 2008: Generation of pluripotent stem cells from adult mouse liver and stomach: these iPS cells seem to be more similar to embryonic stem cells than the previously developed iPS cells and not tumorigenic, moreover genes that are required for iPS cells do not need to be inserted into specific sites, which encourages the development of non-viral reprogramming techniques.[91]March 2008-The first published study of successful cartilage regeneration in the human knee using autologous adult mesenchymal stem cells is published by clinicians from Regenerative Sciences[92]October 2008: Sabine Conrad and colleagues at Tübingen, Germany generate pluripotent stem cells from spermatogonial cells of adult human testis by culturing the cells in vitro under leukemia inhibitory factor (LIF) supplementation.[93]30 October 2008: Embryonic-like stem cells from a single human hair.[94]January 2009: Yong Zhao and colleagues confirmed the reversal of autoimmune-caused type 1 diabetes by Cord Blood-Derived Multipotent Stem Cells (CB-SCs) in an animal experiment.[44][47]1 March 2009: Andras Nagy, Keisuke Kaji, et al. discover a way to produce embryonic-like stem cells from normal adult cells by using a novel "wrapping" procedure to deliver specific genes to adult cells to reprogram them into stem cells without the risks o[censored]sing a virus to make the change.[95][96][97] The use of electroporation is said to allow for the temporary insertion of genes into the cell.[98][98][99][100]28 May 2009 Kim et al. announced that they had devised a way to manipulate skin cells to create patient specific "induced pluripotent stem cells" (iPS), claiming it to be the 'ultimate stem cell solution'.[101]11 October 2010 First trial of embryonic stem cells in humans.[102]25 October 2010: Ishikawa et al. write in the Journal of Experimental Medicine that research shows that transplanted cells that contain their new host's nuclear DNA could still be rejected by the invidual's immune system due to foreignmitochondrial DNA. Tissues made from a person's stem cells could therefore be rejected, because mitochondrial genomes tend to accumulate mutations.[103]2011: Israeli scientist Inbar Friedrich Ben-Nun led a team which produced the first stem cells from endangered species, a breakthrough that could save animals in danger of extinction.[104]January 2012: The human clinical trial of treating type 1 diabetes with lymphocyte modification using Cord Blood-Derived Multipotent Stem Cells (CB-SCs) achieved an improvement of C-peptide levels, reduced the median glycated hemoglobin A1C (HbA1c) values, and decreased the median daily dose of insulin in both human patient groups with and without residual beta cell function.[48][49] Yong Zhao's Stem Cell Educator Therapy appears "so simple and so safe"[105]October 2012: Positions of nucleosomes in mouse embryonic stem cells and the changes in their positions during differentiation to neural progenitor cells and embryonic fibroblasts are determined with single-nucleotide resolution.[106]2012: Katsuhiko Hayashi used mouse skin cells to create stem cells and then used these stem cells to create mouse eggs. These eggs were then fertilized and produced healthy baby offspring. These latter mice were able to have their own babies.[107]2013: First time lab grown meat made from muscle stem-cells has been cooked and tasted.[108]2013: First time mice adult cells were reprogrammed into stem cells in vivo.[109]2013: Scientists at Scotland's Heriot-Watt University developed a 3D printer that can produce clusters of living humanembryonic stem cells, potentially allowing complete organs to be printed on demand in the future.[110]2014: Adult mouse cells reprogrammed to pluripotent stem cells using stimulus-triggered acquisition of pluripotency (STAP);[111] a process which involved bathing blood cells in an acid bath (pH 5.7) for 30minutes at 37 °C.[112]See alsoCell bankHuman genomeMeristemPartial cloningPlant stem cellStem cell controversyStem cell markerReferencesJump up^ Tuch BE (2006). "Stem cells—a clinical update". Australian Family Physician 35 (9): 719–21. 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External linksWikimedia Commons has media related to Stem cells.GeneralStem Cell Basics Courtesy of the National Institutes of HealthNature Reports Stem Cells: Introductory material, research advances and debates concerning stem cell research.Understanding Stem Cells: A View of the Science and Issues from the National AcademiesScientific American Magazine (June 2004 Issue) The Stem Cell ChallengeScientific American Magazine (July 2006 Issue) Stem Cells: The Real Culprits in Cancer?Ethics of Stem Cell Research entry by Andrew Siegel in the Stanford Encyclopedia of PhilosophyIsolation of amniotic stem cell lines with potential for therapyBoston Children's Hospital Stem Cell ResearchStem Cell Research and Industry DirectoryCorneal endothelial and epithelial stem cell research and applicationStem Cell Consumer Progress and ResearchStem Cell Research at Johns HopkinsWhat Are Stem Cells?7 Things You Should Know About Cord Blood BankingParent's Guide to Cord Blood Foundation, a non-profit cord blood educational foundationPeer-reviewed journalsCytotherapyCloning and Stem CellsJournal of Stem Cells and Regenerative MedicineStem Cells and DevelopmentRegenerative MedicineStem Cell ResearchStemBook[show]vte
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