Panel Sequencing for Clinically Oriented Variant Screening and Copy Number Detection in Chronic Lymphocytic Leukemia Patients.

According to current guidelines, in chronic lymphocytic leukemia (CLL), only the TP53 molecular status must be evaluated prior to every treatment's initiation. However, additional heterogeneous genetic events are known to confer a proliferative advantage to the tumor clone and are associated with progression and treatment failure in CLL patients. Here, we describe the implementation of a comprehensive targeted sequencing solution that is suitable for routine clinical practice and allows for the detection of the most common somatic single-nucleotide and copy number variants in genes relevant to CLL. We demonstrate that this cost-effective strategy achieves variant detection with high accuracy, specificity, and sensitivity. Furthermore, we identify somatic variants and copy number variations in genes with prognostic and/or predictive value, according to the most recent literature, and the tool provides evidence about subclonal events. This next-generation sequencing (NGS) capture-based target assay is an improvement on current approaches in defining molecular prognostic and/or predictive variables in CLL patients.

Astroglial distribution and sexual differences in neural metabolism in mammillary bodies.

The sexual differences in cerebral nuclei are produced by the organizational and the activational function of gonadal hormones. The different performances by male and female rats in memory tasks requiring use of the mammillary bodies (MBs), could be due to structural and functional sexual dimorphic differences. Our work quantifies the number of glial fibrillary acidic protein immunoreactive (GFAP-IR) astrocytes, and neuronal metabolic activity measured by the cytochrome oxidase (CO) histochemistry in the MBs in rats of both sexes. We find that there is no difference in astroglial number in the medial mammillary nucleus (MMN) and in the lateral mammillary nucleus (LMN) of males, females in estrus and diestrus adult rats. However, we do find statistically significant differences between the sexes in the neuronal oxidative metabolism influenced by the estrous cycle. We, therefore, conclude that there are functional and not structural sex differences in the MBs.

Quality control of bacterial contamination in autologous peripheral blood stem cells for transplantation.

BACKGROUND AND OBJECTIVES: Microbiological follow-up is part of quality control of peripheral blood stem cell (PBSC) manipulation. DESIGN AND METHODS: We prospectively studied microbiological cultures performed in 865 consecutive untreated autologous PBSC harvests from 348 patients. Our aim was to know the rate of microbiological contamination, the optimum moment to evaluate the sample and the clinical significance of the positive findings. RESULTS: Fifty-nine of the 852 samples (6.9%) yielded a positive culture after PBSC collection (sample 1) and 62 samples also yielded positive results before cryopreservation (7.2%) (sample 2). At the time of the analysis, a total of 520 aphereses had been infused and the number of positive cultures after thawing (sample 3) and after washing (sample 4; 82 aphereses) was 5.4% and 2.3%, respectively. Most of the positive cultures were due to coagulase-negative staphylococci (48 isolates). After thawing 15 coagulase-negative staphylococci and 2 enterococci isolates were recovered. Comparison between samples using a marginal homogeneity test showed no differences in the rate of contamination observed at the different sampling points. INTERPRETATION AND CONCLUSIONS: Positive microbiological findings in collected PBSC are not due to contamination within the laboratory. Cryopreservation using DMSO does not eradicate bacteria and manipulation does not seem to affect results. To simplify the procedure it would be possible to eliminate the microbiological controls performed immediately before cryopreservation.