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.

Infección en pacientes neutropénicas sometidas a trasplante autólogo de progenitores hematopoyéticos por cáncer de mama / Infection in patients with neutropenia that undergo an autologous peripheral blood stem cell transplant due to breast cancer

FUNDAMENTOS. La profundidad y duración de la neutropenia y las características de la enfermedad subyacente son factores determinantes en el pronóstico del síndrome febril. Aunque clásicamente se ha considerado al trasplante de progenitores hematopoyéticos (TPH) como un productor de neutropenia de alto riesgo, probablemente la neutropenia observada en el TPH en algunos tumores sólidos podría ser considerada de riesgo intermedio. Evaluamos episodios febriles en pacientes con estas características. MÉTODOS. Analizamos prospectivamente 132 TPH autólogos, obtenidos de sangre periférica, en pacientes con cáncer de mama (1994-1999). Acondicionamiento: STAMP V. Profilaxis antibacteriana: ofloxacino (400 mg/12 h por vía oral). Clasificación del síndrome febril: bacteriemia, microbiológicamente documentado sin bacteriemia, infección clínica y fiebre de origen incierto. RESULTADOS. Presentaron fiebre 122 pacientes (92 por ciento), edad media: 45 años (rango: 27-61). Hubo 32 (26 por ciento) bacteriemias, 13 (11 por ciento) documentaciones microbiológicas sin bacteriemia y 54 (44 por ciento) infecciones clínicas. Mediana de días con neutrófilos <1x109/l: 14 (rango: 11-20). En los 74 pacientes (61 por ciento) que tuvieron factor estimulante de colonias de granulocitos (G-CSF), la media de días para alcanzar 0,5x109/l neutrófilos (7,6) y la media de días ingresado (26) fueron significativamente menores. Hubo foco infeccioso en 80 pacientes (65 por ciento): orofaríngeo en 33 (46 por ciento) y digestivo en 29 (41 por ciento), que fueron los más frecuentes. Se aislaron 48 microorganismos gramnegativos (GN) y 29 grampositivos (GP) (71 por ciento GN resistentes a ofloxacino). Entre 1997-1999 la relación GN/GP fue de 2,3. No hubo muertes relacionadas con la infección. CONCLUSIONES. La excelente evolución de nuestras pacientes permite considerar su neutropenia como de riesgo medio o bajo, lejos de las tasas de mortalidad por infección publicadas en otros tipos de trasplante hematopoyético. El predominio de GN en los últimos años y su escasa sensibilidad a quinolonas debe hacer reconsiderar su uso profiláctico en estas pacientes (AU)