Effectiveness and safety evaluation of thrice weekly double strength vs daily single strength trimethoprim-sulfamethoxazole for prophylaxis of pneumocystis jirovecii pneumonia after kidney transplantation: a two year prospective cohort study.

PURPOSE: It is hard for clinicians to choose the best regimen for pneumocystis jirovecii pneumonia (PJP) prophylaxis. The aim is to evaluate the effectiveness and safety of thrice weekly double strength (TWDS) vs daily single strength (DSS) trimethoprim-sulfamethoxazole (TMP-SMX) for prophylaxis of PJP after kidney transplantation. METHODS: Adult renal transplant recipients (RTRs) who were transplanted between January 1, 2015 and July 1, 2018 were evaluated. A total of 189 RTRs were prescribed PJP prophylactic regimen during the study period (TWDS group: n=98; DSS group: n=91). RESULTS: Morbidity due to PJP infection was significantly higher in TWDS group as compared with DSS group (8.60% vs 1.14%, p= 0.021). There was a significant trend toward higher prevalence of confirmed PJP (log-rank=0.021) in TWDS group. The use of DDS TMP-SMX for prophylaxis after kidney transplantation was associated with a 79% reduction in the incidence of PJP comparing the prophylactic regimen of TWDS. There was no significant difference between the two groups in the overall rate of premature TMP-SMX discontinuation and laboratory indexes. CONCLUSION: Six months of DSS TMP-SMX prophylaxis was more effective than TWDS TMP-SMX regimen with the same safety profile.

The effect of statins on advanced prostate cancer patients with androgen deprivation therapy or abiraterone/enzalutamide: A systematic review and meta-analysis.

WHAT IS KNOWN AND OBJECTIVE: To evaluate the effects of statin use on the treatment outcomes (i.e. overall survival and cancer-specific survival) among advanced prostate cancer (PCa) patients treated with androgen deprivation therapy (ADT) or abiraterone/enzalutamide. METHODS: The original studies, examining the effects of statins on the outcomes (i.e. overall survival and cancer-specific survival) among PCa patients treated with ADT or abiraterone/enzalutamide, were identified through a systematic search by two independent reviewers in the PubMed, Cochrane, Embase, American Society of Clinical Oncology and European Society of Medical Oncology databases. Databases were searched using keywords (abiraterone OR enzalutamide OR androgen deprivation therapy) AND statin. In total, nine eligible studies from 111 references were included for final analysis. RESULTS AND DISCUSSION: Statin use significantly lowered the risk of all-cause mortality (100 709 patients, HR = 0.73, 95%CI = 0.64-0.83, P < .00001) and the risk of cancer-specific mortality (100 343 patients, HR = 0.64, 95% CI = 0.53-0.77, P < .00001) in advanced PCa patients treated with ADT. The sensitivity analysis showed that the results were reliable. However, it could not generate reliable evidence in patients with metastatic castration-resistant prostate cancer (mCRPC) treated with abiraterone/enzalutamide, as relevant studies were limited and had inconsistent results. WHAT IS NEW AND CONCLUSION: The review indicated that the use of statins in combination with ADT was associated with better all-cause survival and cancer-specific survival in patients with advanced PCa. Randomized controlled trials should be conducted to establish efficacy of statins among PCa patients.

Protecting Quantum Dot Fluorescence from Quenching to Achieve a Reliable Automated Multiplex Fluorescence In Situ Hybridization Assay.

Quantum dots (QD) are novel inorganic fluorochromes that are ultra-bright, photo-stable, and available in multiple, highly-resolvable colors. QDs represent an ideal detection material for in situ hybridization (ISH) because they may provide unprecedented resolution and strong signal intensities that are not attainable with traditional fluorophores. Unfortunately, lack of reliability has been an impediment to widespread adoption of QD-based fluorescence in situ hybridization (QD FISH) technology. By optimizing QD-to-target accessibility, we have developed a QD FISH staining procedure that dramatically improves the reliability of an automated ERG/PTEN QD FISH assay (91% 1st pass rate). Here, we report improvements to the assay that protects QD fluorescence from quenching due to trace amounts of heavy metals and minimizes QD background signals. When using this method, highly-consistent staining was observed with the ERG/PTEN QD FISH assay in prostate tissue. Successful staining of several other clinically-relevant genetic markers was also possible. We further demonstrated improved reliability for determining HER2 gene status in breast cancer, identifying anaplastic lymphoma kinase (ALK) gene break-apart in non-small cell lung cancer, and detecting human papillomavirus 16 (HPV16) in cervical intraepithelial neoplasia. The enhanced QD FISH assay allows for examining complicated genetic aberrances without use of enzymatic amplification. Our optimized methods now demonstrate reliability sufficient for QD FISH technology to be a diagnostic tool in a clinical setting.

Studies on BrdU labeling of hematopoietic cells: stem cells and cell lines.

Studies using chronic in vivo BrdU exposure, isolating primitive stem cells, and determining BrdU labeling, indicate that stem cells cycle. BrdU is also incorporated into DNA during damage/repair. DNA, which has incorporated BrdU due to cycle transit is heavier than normal, while the density of DNA with damage/repair incorporation is intermediate. DNA density of purified lineage-rhodamine low (rho(low)) Hoechst low (Ho(low)) stem cells or FDC-P1 cell line cells-was assessed in vitro, after exposure to cytokines and BrdU (cycling model) or cytokines and BrdU with bleomycin to induce strand breaks and hydroxyurea to halt cycle progression (damage/repair model). We determined DNA density using cesium chloride (CsCl) gradients and either fluorometry or dot blot chemiluminesence. DNA from BrdU labeled cycling Lin-rho(lo)Ho(lo) or FDC-P1 cells was heavier than normal DNA, while damage repair DNA had an intermediate density. We then assessed BrdU labeling of Lin-rho(lo)Ho(lo) cells in vivo. We found that 70.9% of lin-rho(lo)Ho(lo) cells labeled at 5 weeks. DNA density of these cells was low, in the damage/repair range, but similar results were obtained with stem cells, which had proliferated in vivo. Dilution of BrdU in in vitro culture of proliferating FDC-P1 cells also resulted in damage/repair density. We conclude that in vitro BrdU labeling models can distinguish between proliferation and damage/repair, but that we cannot obtain high enough in vivo levels to address this issue. All together, while we cannot absolutely exclude damage/repair as contributing to stem cell BrdU labeling, the data indicate that primitive bone marrow stem cells are probably a cycling population.

Cytokine receptor repertoire and cytokine responsiveness of Ho(dull)/Rh(dull) stem cells with differing potentials for G1/S phase progression.

OBJECTIVE: Subsetting of Hoechst 33342 dull (Ho(dull)) hematopoietic stem cells on the basis of rhodamine 123 (Rh) efflux utilizing an improved dual-dye efflux strategy resolves Ho(dull)/Rh(dull) stem cell subsets that differ with regard to their rate of recruitment and progression through the cell cycle upon exposure to cytokines. MATERIALS AND METHODS: Murine bone marrow cells were isolated by negative immunomagnetic selection using lineage-directed antibodies followed by Ho and Rh staining using a dual-dye efflux method. RESULTS: Ho(dull)/Rh(dull) stem cells that efflux Rh more efficiently (R1) exhibit a 4- to 8-hour delay in progression to S phase when stimulated by interleukin-3 (IL-3), IL-6, IL-11, and stem cell factor (SCF) compared to Ho(dull)/Rh(medium) stem cells, which retain low levels of Rh (R2). R1 and R2 cells show a hierarchical entry into S phase upon exposure to any or all of these cytokines. The R1 subset contains proportionately more high proliferative potential colony-forming cells than the R2 subset, but equivalent levels of engraftable stem cells at 3 and 8 weeks after competitive transplantation. Both R1 and R2 cells express c-kit, IL-3R, and IL-11R, whereas IL-6R and c-fms are only expressed by R1 or R2 cells, respectively. Cytokine stimulation of R1 and R2 cells induced cell cycle progression with elevated or induced expression of c-kit, c-fms, IL-2R, and IL-6R. CONCLUSION: These studies indicate that primitive marrow stem cells can be further subsetted by degree of Rh staining to reveal important functional phenotypic differences between cells with different levels of Rh staining.