19 research outputs found
Whole exome sequencing identifies genetic variants in inherited thrombocytopenia with secondary qualitative function defects
Inherited thrombocytopenias are a heterogeneous group of disorders characterised
by abnormally low platelet counts which can be associated with abnormal bleeding.
Next generation sequencing has previously been employed in these disorders for the
confirmation of suspected genetic abnormalities, and more recently in the discovery
of novel disease causing genes. However its full potential has not previously been
utilised. Over the past 6 years we have sequenced the exomes from 55 patients,
including 37 index cases and 18 additional family members, all of whom were
recruited to the UK Genotyping and Phenotyping of Platelets study. All patients had
inherited or sustained thrombocytopenia of unknown aetiology with platelet counts
varying from 11-186x109
/L. Of the 51 patients phenotypically tested, 37 (73%), had
an additional secondary qualitative platelet defect. Using whole exome sequencing
analysis we have identified âpathogenicâ or âlikely pathogenicâ variants in 46%
(17/37) of our index patients with thrombocytopenia. In addition, we report variants
of uncertain significance in 12 index cases which include novel candidate genetic
variants in previously unreported genes in four index cases. These results
demonstrate that whole exome sequencing is an efficient method for elucidating
potential pathogenic genetic variants in inherited thrombocytopenia. Whole exome
sequencing also has the added benefit of discovering potentially pathogenic genetic
variants for further study in novel genes not previously implicated in inherited
thrombocytopenia
FamĂlies botĂ niques de plantes medicinals
Facultat de Farmà cia, Universitat de Barcelona. Ensenyament: Grau de Farmà cia, Assignatura: Botà nica FarmacÚutica, Curs: 2013-2014, Coordinadors: Joan Simon, CÚsar Blanché i
Maria Bosch.Els materials que aquĂ es presenten sĂłn els recull de 175 treballs dâuna famĂlia botĂ nica dâinterĂšs medicinal realitzats de manera individual. Els treballs han estat realitzat
per la totalitat dels estudiants dels grups M-2 i M-3 de lâassignatura BotĂ nica FarmacĂšutica
durant els mesos dâabril i maig del curs 2013-14. Tots els treballs sâhan dut a terme a travĂ©s de la plataforma de GoogleDocs i han estat tutoritzats pel professor de lâassignatura i revisats i finalment co-avaluats entre els propis estudiants. Lâobjectiu principal de lâactivitat ha estat fomentar lâaprenentatge autĂČnom i col·laboratiu en BotĂ nica farmacĂšutica
Immunohistochemical analysis of PTEN in endometrial carcinoma: a tissue microarray study with a comparison of four commercial antibodies in correlation with molecular abnormalities
The tumor suppressor gene PTEN/MMAC1 is located on chromosome 10q23.3. Inactivation of PTEN, either by
mutations, deletions, or promoter hypermethylation, has been identified in a wide variety of tumors. Inactivation
of the two alleles of PTEN is required, because it is a tumor suppressor gene. Immunohistochemical staining
may be an effective screening method to demonstrate the absence of the protein in tumors exhibiting PTEN
inactivation. We studied a tissue microarray, constructed from paraffin-embedded blocks of 95 endometrial
carcinomas, 38 of them previously evaluated for alterations in PTEN. We also studied cell blocks obtained from
one PTEN-defective endometrial cancer cell line, after transfection with either a plasmid encoding wild-type
PTEN or the empty vector. The tumor samples were tested with four different anti-PTEN commercial antibodies:
a polyclonal antibody, the monoclonal antibody 28H6, the monoclonal antibody 10P03, and the monoclonal
antibody 6.H2.1. Results were correlated with the presence of abnormalities in PTEN, as well as with the
immunohistochemical expression of phosphorylated AKT. Antibody 28H6 produced a predominant nuclear
staining, while the other three antibodies produced a predominant cytoplasmic staining. There was no
significant correlation between the results obtained with the four antibodies. The monoclonal antibody 6.H2.1
was the only one that exhibited a correlation with the presence of molecular alterations in PTEN, and a
statistically significant association with immunostaining for phosphorylated AKT (r Œ 0.249, P Œ 0.037). The
monoclonal antibody 10P03 exhibited an association with phospho-AKT that did not have statistical
significance. Both 6.H2.1 and 10P03 antibodies stained PTEN-transfected cells, and were negative in the
PTEN-deficient cell line blocks. The polyclonal antibody and the monoclonal antibody 28H6 produced positive
staining in PTEN-deficient cell line blocks, suggesting nonspecific staining. The results indicate that
monoclonal antibody 6.H2.1 may be a suitable alternative for tumors with inactivation of PTEN
Whole exome sequencing identifies genetic variants in inherited thrombocytopenia with secondary qualitative function defects
Inherited thrombocytopenias are a heterogeneous group of disorders characterized by abnormally low platelet counts which can be associated with abnormal bleeding. Next-generation sequencing has previously been employed in these disorders for the confirmation of suspected genetic abnormalities, and more recently in the discovery of novel disease-causing genes. However its full potential has not yet been exploited. Over the past 6 years we have sequenced the exomes from 55 patients, including 37 index cases and 18 additional family members, all of whom were recruited to the UK Genotyping and Phenotyping of Platelets study. All patients had inherited or sustained thrombocytopenia of unknown etiology with platelet counts varying from 11Ă109/L to 186Ă109/L. Of the 51 patients phenotypically tested, 37 (73%), had an additional secondary qualitative platelet defect. Using whole exome sequencing analysis we have identified âpathogenicâ or âlikely pathogenicâ variants in 46% (17/37) of our index patients with thrombocytopenia. In addition, we report variants of uncertain significance in 12 index cases, including novel candidate genetic variants in previously unreported genes in four index cases. These results demonstrate that whole exome sequencing is an efficient method for elucidating potential pathogenic genetic variants in inherited thrombocytopenia. Whole exome sequencing also has the added benefit of discovering potentially pathogenic genetic variants for further study in novel genes not previously implicated in inherited thrombocytopenia
Recommendations for somatic and germline genetic testing of single pheochromocytoma and paraganglioma based on findings from a series of 329 patients
Background Nowadays, 65-80% of pheochromocytoma and paraganglioma (PPGL) cases are explained by germline or somatic mutations in one of 22 genes. Several genetic testing algorithms have been proposed, but they usually exclude sporadic-PPGLs (S-PPGLs) and none include somatic testing. We aimed to genetically characterise S-PPGL cases and propose an evidence-based algorithm for genetic testing, prioritising DNA source. Methods The study included 329 probands fitting three criteria: single PPGL, no syndromic and no PPGL family history. Germline DNA was tested for point mutations in RET and for both point mutation and gross deletions in VHL, the SDH genes, TMEM127, MAX and FH. 99 tumours from patients negative for germline screening were available and tested for RET, VHL, HRAS, EPAS1, MAX and SDHB. Results Germline mutations were found in 46 (14.0%) patients, being more prevalent in paragangliomas (PGLs) (28.7%) than in pheochromocytomas (PCCs) (4.5%) (p= 6.62x10(-10)). Somatic mutations were found in 43% of those tested, being more prevalent in PCCs (48.5%) than in PGLs (32.3%) (p= 0.13). A quarter of S-PPGLs had a somatic mutation, regardless of age at presentation. Head and neck PGLs (HN-PGLs) and thoracic-PGLs (T-PGLs) more commonly had germline mutations (p= 2.0x10(-4) and p= 0.027, respectively). Five of the 29 metastatic cases harboured a somatic mutation, one in HRAS. Conclusions We recommend prioritising testing for germline mutations in patients with HN-PGLs and TPGLs, and for somatic mutations in those with PCC. Biochemical secretion and SDHB-immunohistochemistry should guide genetic screening in abdominal-PGLs. Paediatric and metastatic cases should not be excluded from somatic screening