High resolution melting analysis for the identification of novel mutations in DKC1 and TERT genes in patients with dyskeratosis congenita

Dyskeratosis congenita (DC) is a rare inherited bone-marrow failure syndrome with high clinical heterogeneity. Cells derived from DC patients present short telomeres at early ages, as a result of mutations in genes encoding components of the telomerase complex (DKC1, TERC, TERT, NHP2 and NOP10), or the shelterin complex (TINF2). However, mutations have been identified only in around 50% of the cases, indicating that other genes could be involved in the development of this disease. Indeed, mutations in TCBA1 or chromosome segment C16orf57 have been described recently. We have used HRM technology to perform genetic analysis in the above mentioned genes, in Spanish patients showing both, some clinical features of DC and short telomeres. The mutations have been identified by PCR amplification of DC genes followed by high resolution melting (HRM) and direct DNA sequencing analysis. We have identified seven new families with DC, three with X-linked DC and four with autosomal dominant DC, in which we have found two novel mutations in DKC1 (p.His68Arg and p.Lys390del) and four novel mutations in TERT gene (p.Pro530Leu, p.Arg698Trp, p.Arg971His and p.Arg698Gln). The results show that the use of HRM analysis enables a rapid and inexpensive identification of mutations in dyskeratosis congenita associated genes.     

Repolarization of tumor infiltrating macrophages and increased survival in mouse primary CNS lymphomas after XPO1 and BTK inhibition

Journal of Neuro-Oncology - Patients diagnosed with primary central nervous system lymphoma (PCNSL) often face dismal outcomes due to the limited availability of therapeutic options. PCNSL cells...     

Intensive chemotherapy (high-dose CHOP/ESHAP regimen) followed by autologous stem-cell transplantation in previously untreated patients with peripheral T-cell lymphoma

Aim: To analyze toxicity, response and outcome of a phase IItrial with intensive chemotherapy plus autologous stem-celltransplantation (ASCT) for young patients with peripheral T-celllymphoma (PTCL). Patients and methods: Forty-one patients [30 males and 11 females,median age 47 years] consecutively diagnosed with PTCL receivedthree courses of high-dose cyclophosphamide 2000 mg/m²/day,adriamycin 90 mg/m²/day, vincristine and prednisone alternatingwith three courses of etoposide, cisplatin, cytarabine and prednisone.Responders were submitted to ASCT. Results: Sixty-eight percent of patients received the plannedtreatment. After chemotherapy, 20 patients reached completeresponse (CR), 4 partial response and 17 failed. ASCT was carriedout in 17 of 24 candidates due to lack of mobilization (threecases), toxicity (two), early relapse and patient decision (oneeach). CR rate after treatment was 51%. With a median follow-upof 3.2 years, 5 of 21 CR patients relapsed and 2 died in CRdue to secondary neoplasms. Four-year progression-free survivalwas 30%. Twenty-two patients have died, with a 4-year overallsurvival of 39%. International Prognostic Index was the mainvariable predicting survival. No differences were seen amongthe 24 candidates according to whether or not they underwentASCT. Conclusion: This intensive regimen resulted in moderate CR rate,with manageable toxicity in PTCL. The contribution of ASCT inpreventing relapse is debatable. Novel strategies to increaseCR warrant investigation. Key words: autologous stem-cell transplantation, peripheral T-cell lymphoma, prognosis Received for publication January 7, 2008. Accepted for publication January 9, 2008.     

The direct antiglobulin test in a hospital setting

To evaluate the current use of the DAT in our hospital,we reviewed the charts of all patients who had a DAT performed in our laboratory. The collected data included DAT results and a previously completed laboratory evaluation of suspected hemolytic anemia. Four hundred sixty-three DATs were performed in our laboratory from April 1999 to October 2001. The DAT was negative in 434 (93.7%) cases and positive in 29 (6.3%) cases. A complete laboratory evaluation of suspected hemolytic anemia was seen in 179 (38.7%) cases. The incidence of a positive DAT was higher in the group of patients with > 2 signs of hemolysis (4/34 cases; 11.8%) than in the group of patients with 相似文献   

Osteomas of the paranasal sinuses

A group of 86 patients with osteomas on the bones of the skull and face, 21 suffered from osteomas of the paranasal sinuses. The mean age of the patients was 50 years, with a predominance of male subjects (2/1). The frontal sinus was the frequently involved (57%), followed by the maxillary, ethmoid and the sphenoid sinuses. Diagnosis was made accidentally in 45% of the cases. The most frequent presenting symptom was headache (57%). Simple excision of the osteoma paranasal sinus was possible in 18 cases. We realize a study of the facts, as well as a review of the literature to know the incidence, diagnosis, indications and surgery technics used in these type of osteomas.     

Gene-expression profiling and not immunophenotypic algorithms predicts prognosis in patients with diffuse large B-cell lymphoma treated with immunochemotherapy

Diffuse large B-cell lymphomas (DLBCLs) can be divided into germinal-center B cell-like (GCB) and activated-B cell-like (ABC) subtypes by gene-expression profiling (GEP), with the latter showing a poorer outcome. Although this classification can be mimicked by different immunostaining algorithms, their reliability is the object of controversy. We constructed tissue microarrays with samples of 157 DLBCL patients homogeneously treated with immunochemotherapy to apply the following algorithms: Colomo (MUM1/IRF4, CD10, and BCL6 antigens), Hans (CD10, BCL6, and MUM1/IRF4), Muris (CD10 and MUM1/IRF4 plus BCL2), Choi (GCET1, MUM1/IRF4, CD10, FOXP1, and BCL6), and Tally (CD10, GCET1, MUM1/IRF4, FOXP1, and LMO2). GEP information was available in 62 cases. The proportion of misclassified cases by immunohistochemistry compared with GEP was higher when defining the GCB subset: 41%, 48%, 30%, 60%, and 40% for Colomo, Hans, Muris, Choi, and Tally, respectively. Whereas the GEP groups showed significantly different 5-year progression-free survival (76% vs 31% for GCB and activated DLBCL) and overall survival (80% vs 45%), none of the immunostaining algorithms was able to retain the prognostic impact of the groups (GCB vs non-GCB). In conclusion, stratification based on immunostaining algorithms should be used with caution in guiding therapy, even in clinical trials.