Rapid Detection and Identification of Mucormycetes in Bronchoalveolar Lavage Samples from Immunocompromised Patients with Pulmonary Infiltrates by Use of High-Resolution Melt Analysis

Rapid differential diagnostics of pulmonary infiltrates suspected of invasive fungal disease in an immunocompromised host and early initiation of effective antifungal therapy are crucial for patient outcomes. There are no serological tests available to detect mucormycetes; therefore, PCR-based methods are highly suitable. We validated our previously published PCR followed by high-resolution melt analysis (PCR/HRMA) to detect Rhizopus spp., Rhizomucor pusillus, Lichtheimia corymbifera, and Mucor spp. in bronchoalveolar lavage (BAL) samples from immunocompromised patients who were at risk of invasive fungal disease. All PCR/HRMA-positive samples were retested using novel real-time quantitative PCR (RQ PCR) assays specific to the species identified. In total, between January 2009 and December 2012 we analyzed 99 BAL samples from 86 patients with pulmonary abnormalities using PCR/HRMA. Ninety (91%) BAL samples were negative, and 9 (9%) samples were positive. The sensitivity and specificity of PCR/HRMA were 100% and 93%, respectively. By combining the positive results of PCR/HRMA with positive RQ PCR results, the specificity was raised to 98%. PCR/HRMA, due to its high negative predictive value (99%), represents a fast and reliable tool for routine BAL sample screening for the differential diagnosis of pulmonary infiltrates in immunocompromised patients for the four most clinically important mucormycetes.     

Regulation of dopamine transporter activity by carboxypeptidase E

Background

The dopamine transporter (DAT) plays a critical role in terminating the action of dopamine by rapid reuptake into the presynaptic neuron. Previous studies have revealed that the DAT carboxyl terminus (DAT-CT) can directly interact with other cellular proteins and regulate DAT function and trafficking.

Results

Here, we have identified that carboxypeptidase E (CPE), a prohormone processing exopeptidase and sorting receptor for the regulated secretory pathway, interacts with the DAT-CT and affects DAT function. Mammalian cell lines coexpressing CPE and DAT exhibited increased DAT-mediated dopamine uptake activity compared to cells expressing DAT alone. Moreover, coexpression of an interfering DAT-CT minigene inhibited the effects of CPE on DAT. Functional changes caused by CPE could be attributed to enhanced DAT expression and subsequent increase in DAT cell surface localization, due to decreased DAT degradation. In addition, CPE association could reduce the phosphorylation state of DAT on serine residues, potentially leading to reduced internalization, thus stabilizing plasmalemmal DAT localization.

Conclusion

Taken together, our results reveal a novel role for CPE in the regulation of DAT trafficking and DAT-mediated DA uptake, which may provide a novel target in the treatment of dopamine-governed diseases such as drug addiction and obesity.     

Utilization study of filgrastim (Neutromax) during autologous haematopoietic precursor transplantation for myeloma and lymphoma patients

To describe utilization of a biosimilar product containing filgrastim (Neutromax^®), data of 414 myeloma or lymphoma patients subjected to autologous SCT between 1998 and 2007 were analyzed. Filgrastim was used for mobilization of progenitors (5 days at 300 μg/day) and for the recovery of neutropenia after transplantation (100 μg/day, since day +5). In 2003, the excipient mannitol was replaced by sorbitol. A mean dose of 9.47 × 10⁶ CD34⁺ cells/kg was infused; 100 neutrophils/mm³ required 5-day treatment; 500 neutrophils/mm³, 6 days and 1000 neutrophils/mm³, 7 days. Neutromax^® effect in SCT is similar to reports with other brands. No difference was found between formulations.     

Mycobacterium haemophilum as a Novel Etiology of Cervical Lymphadenitis in an Otherwise Healthy Adult Patient

We describe a case and summarize six additional cases of cervical lymphadenitis in otherwise healthy adults caused by Mycobacterium haemophilum. The organism causes cervicofacial lymphadenitis in healthy children and severe disease in immunocompromised patients but has not been previously reported to cause cervical lymphadenitis in nonimmunocompromised, healthy adults.     

Trends in pemphigus research over 15 years

Background Although pemphigus is a rare autoimmune blistering disease, it attracts the attention of physicians of many disciplines. Objective This study aims to assess the number of articles on pemphigus that have been published over 15 years in dermatology vs. non‐dermatology medical journals, and to evaluate the quality of available evidence. Methods PubMed was searched for articles on pemphigus published between 1 January 1993 to 31 December 2007 using the search word pemphigus. Articles were characterized by publication type and journal type per year. Regression analysis was used to determine the effect of year of publication on number of publications of each type. Results The search yielded 2032 publications on pemphigus during the evaluation period. Sixty‐one per cent were published in dermatology journals. Overall, the number of publications increased linearly with time. Most of this increase was accounted for by publications in non‐dermatology journals. There was an increase in clinical trials over the course of the study period. The number of certain publications with lower quality of evidence, mainly case reports and letters to the editor, increased significantly in the last few years. There was no increase in publications with high quality of evidence. Conclusions The increase on data from non‐dermatology disciplines is a welcome contribution. Nevertheless, high‐quality evidence on pemphigus is still lacking. We trust that the current trend towards evidence‐based dermatology will impact future research on this severe disease.     

Multivariate analyses of prognostic factors associated with hematopoietic recovery in autograft patients with different sources of progenitor cells: A GATMO experience

OBJECTIVE:: To evaluate in a multivariate analysis the prognostic factorsassociated with hematopoietic recovery and the supportive carerequirements after autotransplant of progenitor cells (PC) fromvarious sources: bone marrow (BMPC), BMPC & peripheral blood(PBPC), and PBPC alone. PATIENTS AND METHODS:: A total of 570 patients with hematological malignancies andsolid tumors underwent high-dose therapy followed by autotransplant.PBPC were obtained after mobilization with chemotherapy and/orcytokines. One-hundred five patients received BMPC, 217 receivedBMPC & PBPC and 248 PBPC alone; all of the patients receivedG-CSF or GM-CSF after infusion. RESULTS:: In a multivariate analysis the recovery of neutrophils was adverselyassociated with low numbers of nucleated cells infused (P<0.13),bone marrow progenitor cell source, and diagnosis of multiplemyeloma and acute leukemia (P<0.001). The factors that adverselyaffected platelet recovery were low number of nucleated cellsand diagnosis of multiple myeloma and acute leukemia (P<.001). CONCLUSION:: We conclude that BMPC adversely affect neutrophil recovery whilelow numbers of nucleated cells and diagnosis of multiple myelomaand acute leukemia adversely affect both neutrophil and plateletrecovery. autograft, bone marrow, hematological recovery, peripheral blood, progenitor cells, prognostic factors     

Novel tricyclic poly(ADP-ribose) polymerase-1 inhibitors with potent anticancer chemopotentiating activity: design,synthesis, and X-ray cocrystal structure

A series of novel compounds have been designed that are potent inhibitors of poly(ADP-ribose) polymerase-1 (PARP-1), and the activity and physical properties have been characterized. The new structural classes, 3,4,5,6-tetrahydro-1H-azepino[5,4,3-cd]indol-6-ones and 3,4-dihydropyrrolo[4,3,2-de]isoquinolin-5-(1H)-ones, have conformationally locked benzamide cores that specifically interact with the PARP-1 protein. The compounds have been evaluated with in vitro cellular assays that measure the ability of the PARP-1 inhibitors to enhance the effect of cytotoxic agents against cancer cell lines.     

Estimation of the survival function for Gray's piecewise-constant time-varying coefficients model

Gray's extension of Cox's proportional hazards (PH) model for right-censored survival data allows for a departure from the PH assumption via introduction of time-varying regression coefficients (TVC). For this model estimation of the conditional hazard rate relies on the inclusion of penalized splines. Cubic penalized splines tend to be unstable in the right tail of the distribution and thus quadratic, linear and piecewise-constant penalized splines may be a favourable choice. We derive a survival function estimator for one important member of the class of TVC models--a piecewise-constant time-varying coefficients (PC-TVC) model. Using the first-order Taylor series approximation we also derive an estimate for the variance of the log-transformed and log(-log)-transformed survival function, which in turn leads to estimated confidence limits on the corresponding scales of the survival function. Accuracy in estimating underlying survival times and survival quantiles is assessed for both Cox's and Gray's PC-TVC model using a simulation study featuring scenarios violating the PH assumption. Finally, an example of the estimated survival functions and the corresponding confidence limits derived from Cox's PH and Gray's PC-TVC model, respectively, is presented for a liver transplant data set.