Harnessing microbial phylum-specific molecular markers for assessment of environmental estrogen degradation.

Steroidal estrogens are ubiquitous contaminants that have garnered attention worldwide due to their endocrine-disrupting and carcinogenic activities at sub-nanomolar concentrations. Microbial degradation is one of the main mechanisms through which estrogens can be removed from the environment. Numerous bacteria have been isolated and identified as estrogen degraders; however, little is known about their contribution to environmental estrogen removal. Here, our global metagenomic analysis indicated that estrogen degradation genes are widely distributed among bacteria, especially among aquatic actinobacterial and proteobacterial species. Thus, by using the Rhodococcus sp. strain B50 as the model organism, we identified three actinobacteria-specific estrogen degradation genes, namely aedGHJ, by performing gene disruption experiments and metabolite profile analysis. Among these genes, the product of aedJ was discovered to mediate the conjugation of coenzyme A with a unique actinobacterial C17 estrogenic metabolite, 5-oxo-4-norestrogenic acid. However, proteobacteria were found to exclusively adopt an α-oxoacid ferredoxin oxidoreductase (i.e., the product of edcC) to degrade a proteobacterial C18 estrogenic metabolite, namely 3-oxo-4,5-seco-estrogenic acid. We employed actinobacterial aedJ and proteobacterial edcC as specific biomarkers for quantitative polymerase chain reaction (qPCR) to elucidate the potential of microbes for estrogen biodegradation in contaminated ecosystems. The results indicated that aedJ was more abundant than edcC in most environmental samples. Our results greatly expand the understanding of environmental estrogen degradation. Moreover, our study suggests that qPCR-based functional assays are a simple, cost-effective, and rapid approach for holistically evaluating estrogen biodegradation in the environment.

Endoscopic Hematoma Evacuation for Intracerebral Hemorrhage Under Local Anesthesia: Factors That Affect the Hematoma Removal Rate.

OBJECTIVE: Recent advances in endoscopic surgery have led to more patients being able to undergo endoscopic removal of hypertensive intracerebral hemorrhage (HICH). However, because of the minimal invasiveness, endoscopic HICH removal through a narrow surgical window can result in a low removal rate. The goal of the present study was to investigate the factors that affect the removal rate of HICH evacuation. METHODS: The data from 28 patients with supratentorial HICH who had undergone endoscopic hematoma evacuation were retrospectively analyzed. The inclusion criteria were spontaneous supratentorial HICH with a hematoma volume >30 mL, admission to the hospital within 24 hours of ictus, and a Glasgow coma scale score of ≥4. RESULTS: Of the 28 patients, 9 were women and 19 were men, ranging in age from 41 to 86 years (mean, 60.7 ± 12.7). The hematoma location was the basal ganglia in 25 patients and subcortical in 3 patients. The mean preoperative hematoma volume was 62.4 ± 22.5 mL. The hematoma removal rate was <60% for 11 patients (poor evacuation group) and ≥60% for in 17 patients (good evacuation group). Comparing the 2 groups, chronic renal failure treated with hemodialysis (P = 0.0072, χ2 test), liver cirrhosis (P = 0.023, χ2 test), and surgeon experience with ≥10 cases of endoscopic HICH removal (P = 0.016, χ2 test) were significant factors related to the HICH removal rate. CONCLUSION: To achieve a good removal rate, surgeons should have experience performing the endoscopic procedure. Also, patients with end-stage chronic renal failure or liver cirrhosis should be excluded.

Genomic screening for genes silenced by DNA methylation revealed an association between RASD1 inactivation and dexamethasone resistance in multiple myeloma.

PURPOSE: Epigenetic changes such as DNA methylation play a key role in the development and progression of multiple myeloma. Our aim in the present study was to use genomic screening to identify genes targeted for epigenetic inactivation in multiple myeloma and assess their role in the development of resistance to dexamethasone. EXPERIMENTAL DESIGN: Gene expression was examined using microarray screening, reverse transcription-PCR, and real-time quantitative PCR. DNA methylation was examined using bisulfite PCR, bisulfite sequencing, and bisulfite pyrosequencing in 14 multiple myeloma cell lines, 87 multiple myeloma specimens, and 12 control bone marrow samples. WST-8 assays were used to assess cell viability after treatment with 5-aza-2'-deoxycytidine and/or dexamethasone. RESULTS: Microarray analysis was done to screen for genes up-regulated by 5-aza-2'-deoxycytidine. In RPMI8226 cells, 128 genes were up-regulated, whereas 83 genes were up-regulated in KMS12PE cells. Methylation of 22 genes with CpG islands in their 5' regions, including RASD1, was confirmed. Methylation of RASD1 was associated with its inactivation, which correlated with resistance to dexamethasone. Treating multiple myeloma cells with 5-aza-2'-deoxycytidine restored sensitivity to dexamethasone. Methylation of RASD1 was also detected in a subset of primary multiple myeloma specimens, and the levels of methylation were increased after repeated antitumor treatments. Gene signature analysis revealed various genes to be synergistically induced by treatment with a combination of 5-aza-2'-deoxycytidine plus dexamethasone. CONCLUSION: Our findings indicate that epigenetic inactivation of genes, including RASD1, plays a key role in the development of dexamethasone resistance in multiple myeloma. Moreover, they show the utility of demethylation therapy in cases of advanced multiple myeloma.