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S Whittaker - One of the best experts on this subject based on the ideXlab platform.
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Diagnostic and prognostic importance of T-cell receptor Gene Analysis in patients with Sézary syndrome.
Cancer, 2001Co-Authors: M.r.c.p. Elisabeth A. Fraser-andrews, R Russell-jones, Alison J. Woolford, R.a. Wolstencroft, Alan J. Dean, S WhittakerAbstract:BACKGROUND Sezary syndrome (SS) is characterized by erythroderma, peripheral lymphadenopathy, and circulating Sezary cells and is clinically heteroGeneous. METHODS T-cell receptor (TCR) Gene Analysis was performed using DNA extracted from peripheral blood mononuclear cells from 74 patients, and the results were correlated with a variety of other diagnostic parameters and patient outcomes. RESULTS Two groups were identified: 66 patients with clonal TCR Gene rearrangement (clonal patients) detected with Southern blot Analysis and/or polymerase chain reaction/single-strand conformational polymorphism Analysis and 8 patients with no clonal rearrangement detected (nonclonal patients) using either technique. Clonal patients were compared with nonclonal patients. The following median blood parameters were significantly greater in the clonal group: total white cell count (13.7 109/L vs. 9.6 109/L), lymphocyte count (4.9 109/L vs. 2.2 109/L), absolute Sezary count (3.22 109/L vs. 0.99 109/L), CD4 count (3.17 109/L vs. 1.36 109/L), and CD4:CD8 ratio (15.86 vs. 3.21). An expanded population of T-cells of a specific TCR variable β subset was detected in 7 of 36 clonal patients and in 1 of 4 nonclonal patients. CytoGenetic Analysis of peripheral blood from 1 nonclonal patient and 6 clonal patients was normal. The median survival from the time of diagnosis was 45 months in the clonal group, and 40 of 49 deaths were cutaneous T-cell lymphoma (CTCL)-related, whereas 3 deaths in the nonclonal group were unrelated to CTCL (P < 0.01; log-rank test). Multivariate proportional hazards Analysis showed that the absolute Sezary count and lymph node status were independent prognostic variables (P = 0.016 and P = 0.036, respectively). CONCLUSIONS TCR Gene Analysis defines a distinct clinicopathologic group of patients with SS. Clonal patients have a poor prognosis and are likely to die from leukemia/lymphoma, whereas nonclonal patients may have a reactive, inflammatory T-cell disorder. The authors suggest that the definitive diagnostic criteria for patients with SS should include the presence of a clonal TCR Gene rearrangement. Cancer 2001;92:1745–52. © 2001 American Cancer Society.
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T-cell receptor Gene Analysis in the diagnosis of Sézary syndrome.
Journal of the American Academy of Dermatology, 1999Co-Authors: R Russell-jones, S WhittakerAbstract:Diagnosing Sézary syndrome (SS) on clinicopathological grounds alone is far from straightforward, particularly in the early stages of the disease. Atypical lymphocytes may be seen in the peripheral blood of patients with reactive forms of erythroderma, so additional criteria are needed to establish the diagnosis of a T-cell leukemia/lymphoma. A wide variety of confirmatory tests have been proposed in the literature, but there has been no systematic attempt to compare the specificity and sensitivity of these different methods. Recent data indicate that T-cell receptor (TCR) Gene Analysis is the most useful test currently available and that methods based on polymerase chain reaction are more sensitive than Southern blot Analysis. We propose that the diagnostic criteria for SS should include erythroderma, atypical circulating mononuclear cells, and evidence of a clonal T-cell population in the peripheral blood. Clonality can be established with certainty by cytoGenetic or TCR Gene Analysis, but only the latter is sufficiently sensitive to be of value in routine diagnosis. Immunophenotypic data showing an expanded CD4(+)/CD7(-) population, an elevated CD4/CD8 ratio, or restricted V beta expression are not specific to T-cell malignancy and should not be used as a sole diagnostic criteria in SS. Entry criteria for future clinical studies will need to be more rigorous if meaningful comparisons are to be made between different treatment options.
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T-cell receptor Gene Analysis in the diagnosis of Se ́zary syndrome
Journal of the American Academy of Dermatology, 1999Co-Authors: R Russell-jones, S WhittakerAbstract:Abstract Diagnosing Sezary syndrome (SS) on clinicopathological grounds alone is far from straightforward, particularly in the early stages of the disease. Atypical lymphocytes may be seen in the peripheral blood of patients with reactive forms of erythroderma, so additional criteria are needed to establish the diagnosis of a T-cell leukemia/lymphoma. A wide variety of confirmatory tests have been proposed in the literature, but there has been no systematic attempt to compare the specificity and sensitivity of these different methods. Recent data indicate that T-cell receptor (TCR) Gene Analysis is the most useful test currently available and that methods based on polymerase chain reaction are more sensitive than Southern blot Analysis. We propose that the diagnostic criteria for SS should include erythroderma, atypical circulating mononuclear cells, and evidence of a clonal T-cell population in the peripheral blood. Clonality can be established with certainty by cytoGenetic or TCR Gene Analysis, but only the latter is sufficiently sensitive to be of value in routine diagnosis. Immunophenotypic data showing an expanded CD4 + /CD7 – population, an elevated CD4/CD8 ratio, or restricted V beta expression are not specific to T-cell malignancy and should not be used as a sole diagnostic criteria in SS. Entry criteria for future clinical studies will need to be more rigorous if meaningful comparisons are to be made between different treatment options. (J Am Acad Dermatol 1999;41:254-9.)
Teruo Fujii - One of the best experts on this subject based on the ideXlab platform.
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Microfabricated flow-through device for DNA amplification—towards in situ Gene Analysis
Chemical Engineering Journal, 2004Co-Authors: Tatsuhiro Fukuba, Takatoki Yamamoto, Takeshi Naganuma, Teruo FujiiAbstract:Abstract This study presents a microfabricated device for polymerase chain reaction (PCR) with a flow-through manner for use in environmental microbiology. The device was developed utilizing photolithography and softlithography techniques and evaluated as one component of a totally integrated in situ Gene Analysis system. The developed device was composed of a glass-based “temperature control chip” and polydimethylsiloxane (PDMS)-based “microchannel chip”. For the temperature control chip, six heaters made from indium tin oxide (ITO) are placed on a glass substrate to define three uniform temperature zones for flow-through PCR. On the each heater, a platinum (Pt) line was placed as a temperature sensor. The PDMS microchannel structure was fabricated by using a molding method with a negatively patterned mold master. The width and depth of folded microchannel was 100 μm and total length for 30 cycles of flow-through PCR was approximately 3.0 m. With the flow-through PCR device, 580 and 1450 bp (base pairs) of DNA fragments were successfully amplified from Escherichia coli genomic DNA and directly from untreated cells.
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Microfabricated devices for DNA extraction toward realization of deep-sea in situ Gene Analysis
Oceans '04 MTS IEEE Techno-Ocean '04 (IEEE Cat. No.04CH37600), 1Co-Authors: M. Matsunaga, Tatsuhiro Fukuba, Takatoki Yamamoto, Teruo FujiiAbstract:This study presents microfabricated devices for in situ Gene Analysis in deep-sea environments. Two kinds of microfabricated devices, one for cell lysis and the other for DNA purification, are developed and evaluated using living bacterial cells, Escherichia coli (E. coli), and /spl lambda/DNA, respectively, through laboratory experiments. Both devices are composed of a silicone-elastomer based "microchannel chip" and a glass based "temperature control chip". In the cell lysis device, E. coli cells are lysed by applying a chemical or an enzyme, and can be broken into DNA and impurities. And in the DNA purification device, /spl lambda/DNA is selectively bound to glass beads in the solution with higher ionic concentration. Then, the DNA is eluted by the solution with lower ionic concentration, such as TE buffer. As a result, obtained DNA can be applied to various molecular biological analyses including PCR (polymerase chain reaction). By further integration and automation, the developed devices will contribute to in situ Gene Analysis systems to be mounted on ROVs and AUVs.
R Russell-jones - One of the best experts on this subject based on the ideXlab platform.
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Diagnostic and prognostic importance of T-cell receptor Gene Analysis in patients with Sézary syndrome.
Cancer, 2001Co-Authors: M.r.c.p. Elisabeth A. Fraser-andrews, R Russell-jones, Alison J. Woolford, R.a. Wolstencroft, Alan J. Dean, S WhittakerAbstract:BACKGROUND Sezary syndrome (SS) is characterized by erythroderma, peripheral lymphadenopathy, and circulating Sezary cells and is clinically heteroGeneous. METHODS T-cell receptor (TCR) Gene Analysis was performed using DNA extracted from peripheral blood mononuclear cells from 74 patients, and the results were correlated with a variety of other diagnostic parameters and patient outcomes. RESULTS Two groups were identified: 66 patients with clonal TCR Gene rearrangement (clonal patients) detected with Southern blot Analysis and/or polymerase chain reaction/single-strand conformational polymorphism Analysis and 8 patients with no clonal rearrangement detected (nonclonal patients) using either technique. Clonal patients were compared with nonclonal patients. The following median blood parameters were significantly greater in the clonal group: total white cell count (13.7 109/L vs. 9.6 109/L), lymphocyte count (4.9 109/L vs. 2.2 109/L), absolute Sezary count (3.22 109/L vs. 0.99 109/L), CD4 count (3.17 109/L vs. 1.36 109/L), and CD4:CD8 ratio (15.86 vs. 3.21). An expanded population of T-cells of a specific TCR variable β subset was detected in 7 of 36 clonal patients and in 1 of 4 nonclonal patients. CytoGenetic Analysis of peripheral blood from 1 nonclonal patient and 6 clonal patients was normal. The median survival from the time of diagnosis was 45 months in the clonal group, and 40 of 49 deaths were cutaneous T-cell lymphoma (CTCL)-related, whereas 3 deaths in the nonclonal group were unrelated to CTCL (P < 0.01; log-rank test). Multivariate proportional hazards Analysis showed that the absolute Sezary count and lymph node status were independent prognostic variables (P = 0.016 and P = 0.036, respectively). CONCLUSIONS TCR Gene Analysis defines a distinct clinicopathologic group of patients with SS. Clonal patients have a poor prognosis and are likely to die from leukemia/lymphoma, whereas nonclonal patients may have a reactive, inflammatory T-cell disorder. The authors suggest that the definitive diagnostic criteria for patients with SS should include the presence of a clonal TCR Gene rearrangement. Cancer 2001;92:1745–52. © 2001 American Cancer Society.
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T-cell receptor Gene Analysis in the diagnosis of Sézary syndrome.
Journal of the American Academy of Dermatology, 1999Co-Authors: R Russell-jones, S WhittakerAbstract:Diagnosing Sézary syndrome (SS) on clinicopathological grounds alone is far from straightforward, particularly in the early stages of the disease. Atypical lymphocytes may be seen in the peripheral blood of patients with reactive forms of erythroderma, so additional criteria are needed to establish the diagnosis of a T-cell leukemia/lymphoma. A wide variety of confirmatory tests have been proposed in the literature, but there has been no systematic attempt to compare the specificity and sensitivity of these different methods. Recent data indicate that T-cell receptor (TCR) Gene Analysis is the most useful test currently available and that methods based on polymerase chain reaction are more sensitive than Southern blot Analysis. We propose that the diagnostic criteria for SS should include erythroderma, atypical circulating mononuclear cells, and evidence of a clonal T-cell population in the peripheral blood. Clonality can be established with certainty by cytoGenetic or TCR Gene Analysis, but only the latter is sufficiently sensitive to be of value in routine diagnosis. Immunophenotypic data showing an expanded CD4(+)/CD7(-) population, an elevated CD4/CD8 ratio, or restricted V beta expression are not specific to T-cell malignancy and should not be used as a sole diagnostic criteria in SS. Entry criteria for future clinical studies will need to be more rigorous if meaningful comparisons are to be made between different treatment options.
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T-cell receptor Gene Analysis in the diagnosis of Se ́zary syndrome
Journal of the American Academy of Dermatology, 1999Co-Authors: R Russell-jones, S WhittakerAbstract:Abstract Diagnosing Sezary syndrome (SS) on clinicopathological grounds alone is far from straightforward, particularly in the early stages of the disease. Atypical lymphocytes may be seen in the peripheral blood of patients with reactive forms of erythroderma, so additional criteria are needed to establish the diagnosis of a T-cell leukemia/lymphoma. A wide variety of confirmatory tests have been proposed in the literature, but there has been no systematic attempt to compare the specificity and sensitivity of these different methods. Recent data indicate that T-cell receptor (TCR) Gene Analysis is the most useful test currently available and that methods based on polymerase chain reaction are more sensitive than Southern blot Analysis. We propose that the diagnostic criteria for SS should include erythroderma, atypical circulating mononuclear cells, and evidence of a clonal T-cell population in the peripheral blood. Clonality can be established with certainty by cytoGenetic or TCR Gene Analysis, but only the latter is sufficiently sensitive to be of value in routine diagnosis. Immunophenotypic data showing an expanded CD4 + /CD7 – population, an elevated CD4/CD8 ratio, or restricted V beta expression are not specific to T-cell malignancy and should not be used as a sole diagnostic criteria in SS. Entry criteria for future clinical studies will need to be more rigorous if meaningful comparisons are to be made between different treatment options. (J Am Acad Dermatol 1999;41:254-9.)
H Höfler - One of the best experts on this subject based on the ideXlab platform.
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Laser-assisted preparation of single cells from stained histological slides for Gene Analysis.
Histochemistry and cell biology, 1997Co-Authors: I Becker, K F Becker, M H Röhrl, H HöflerAbstract:Individual cells are prepared from histological tissue sections of routinely formalin-fixed and paraffin-embedded tissues using an ultraviolet laser micromanipulator. This technology, in combination with polymerase chain reaction (PCR)-based Gene Analysis, will enable researchers to routinely detect a variety of nucleic acid abnormalities underlying cancer, infection, and Genetic disease with previously unknown sensitivity: at the single cell level. The utility of this technique is demonstrated by PCR amplification and sequencing of the E-cadherin Gene, which codes for a homophilic cell-to-cell adhesion molecule, in early gastric carcinomas of the diffuse type of Lauren's classification. The main characteristics of the laser-assisted microdissection technique are high precision without contamination and easy application. The assignment of individual Gene sequences to single cells will now provide a direct link between molecular biology on the one hand and histology and pathology on the other.
Sandra H. Burnett - One of the best experts on this subject based on the ideXlab platform.
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Phage cluster relationships identified through single Gene Analysis
BMC Genomics, 2013Co-Authors: Kyle C. Smith, Eduardo Castro-nallar, Joshua N. B. Fisher, Donald P. Breakwell, Julianne H. Grose, Sandra H. BurnettAbstract:PhyloGenetic comparison of bacteriophages requires whole genome approaches such as dotplot Analysis, genome pairwise maps, and Gene content Analysis. Currently mycobacteriophages, a highly studied phage group, are categorized into related clusters based on the comparative Analysis of whole genome sequences. With the recent explosion of phage isolation, a simple method for phage cluster prediction would facilitate Analysis of crude or complex samples without whole genome isolation and sequencing. The hypothesis of this study was that mycobacteriophage-cluster prediction is possible using comparison of a single, ubiquitous, semi-conserved Gene. Tape Measure Protein (TMP) was selected to test the hypothesis because it is typically the longest Gene in mycobacteriophage genomes and because regions within the TMP Gene are conserved. A single Gene, TMP, identified the known Mycobacteriophage clusters and subclusters using a Gepard dotplot comparison or a phyloGenetic tree constructed from global alignment and maximum likelihood comparisons. Gepard Analysis of 247 mycobacteriophage TMP sequences appropriately recovered 98.8% of the subcluster assignments that were made by whole-genome comparison. Subcluster-specific primers within TMP allow for PCR determination of the mycobacteriophage subcluster from DNA samples. Using the single-Gene comparison approach for siphovirus coliphages, phage groupings by TMP comparison reflected relationships observed in a whole genome dotplot comparison and confirm the potential utility of this approach to another widely studied group of phages. TMP sequence comparison and PCR results support the hypothesis that a single Gene can be used for distinguishing phage cluster and subcluster assignments. TMP single-Gene Analysis can quickly and accurately aid in mycobacteriophage classification.