The small non-coding RNA B11 regulates multiple facets of Mycobacterium abscessus virulence.

Mycobacterium abscessus causes severe disease in patients with cystic fibrosis. Little is known in M. abscessus about the roles of small regulatory RNAs (sRNA) in gene regulation. We show that the sRNA B11 controls gene expression and virulence-associated phenotypes in this pathogen. B11 deletion from the smooth strain ATCC_19977 produced a rough strain, increased pro-inflammatory signaling and virulence in multiple infection models, and increased resistance to antibiotics. Examination of clinical isolate cohorts identified isolates with B11 mutations or reduced expression. We used RNAseq and proteomics to investigate the effects of B11 on gene expression and test the impact of mutations found in clinical isolates. Over 200 genes were differentially expressed in the deletion mutant. Strains with the clinical B11 mutations showed expression trends similar to the deletion mutant, suggesting partial loss of function. Among genes upregulated in the B11 mutant, there was a strong enrichment for genes with B11-complementary sequences in their predicted ribosome binding sites (RBS), consistent with B11 functioning as a negative regulator that represses translation via base-pairing to RBSs. Comparing the proteomes similarly revealed that upregulated proteins were strongly enriched for B11-complementary sequences. Intriguingly, genes upregulated in the absence of B11 included components of the ESX-4 secretion system, critical for M. abscessus virulence. Many of these genes had B11-complementary sequences at their RBSs, which we show is sufficient to mediate repression by B11 through direct binding. Altogether, our data show that B11 acts as a direct negative regulator and mediates (likely indirect) positive regulation with pleiotropic effects on gene expression and clinically important phenotypes in M. abscessus. The presence of hypomorphic B11 mutations in clinical strains is consistent with the idea that lower B11 activity may be advantageous for M. abscessus in some clinical contexts. This is the first report on an sRNA role in M. abscessus.

The benefits of lower ozone due to air pollution emission reductions (2002-2011) in the Eastern United States during extreme heat.

Using the Community Multiscale Air Quality (CMAQ) model and the Benefits Mapping and Analysis Program - Community Edition (BenMAP-CE) tool, we estimate the benefits of anthropogenic emission reductions between 2002 and 2011 in the Eastern United States (US) with respect to surface ozone concentrations and ozone-related health and economic impacts, during a month of extreme heat, July 2011. Based on CMAQ simulations using emissions appropriate for 2002 and 2011, we estimate that emission reductions since 2002 likely prevented 10- 15 ozone exceedance days (using the 2011 maximum 8-hr average ozone standard of 75 ppbv) throughout the Ohio River Valley and 5- 10 ozone exceedance days throughout the Washington, DC - Baltimore, MD metropolitan area during this extremely hot month. CMAQ results were fed into the BenMAP-CE tool to determine the health and health-related economic benefits of anthropogenic emission reductions between 2002 and 2011. We estimate that the concomitant health benefits from the ozone reductions were significant for this anomalous month: 160-800 mortalities (95% confidence interval (CI): 70-1,010) were avoided in July 2011 in the Eastern U.S, saving an estimated $1.3-$6.6 billion (CI: $174 million-$15.5 billion). Additionally, we estimate that emission reductions resulted in 950 (CI: 90-2,350) less hospital admissions from respiratory symptoms, 370 (CI: 180-580) less hospital admissions for pneumonia, 570 (CI: 0-1650) less Emergency Room (ER) visits from asthma symptoms, 922,020 (CI: 469,960-1,370,050) less minor restricted activity days (MRADs), and 430,240 (CI: -280,350-963,190) less symptoms of asthma exacerbation during July 2011.Implications: We estimate the benefits of air pollution emission reductions on surface ozone concentrations and ozone-related impacts on human health and the economy between 2002 and 2011 during an extremely hot month, July 2011, in the eastern United States (US) using the CMAQ and BenMAP-CE models. Results suggest that, during July 2011, emission reductions prevented 10-15 ozone exceedance days in the Ohio River Valley and 5-10 ozone exceedance days in the Mid Atlantic; saved 160-800 lives in the Eastern US, saving $1.3 - $6.5 billion; and resulted in 950 less hospital admissions for respiratory symptoms, 370 less hospital admissions for pneumonia, 570 less Emergency Room visits for asthma symptoms, 922,020 less minor restricted activity days, and 430,240 less symptoms of asthma exacerbation.

An unusual cause of vertigo, tinnitus, and hyperacusis: Vogt-Koyanagi-Harada syndrome.

We describe the case of a 36-year-old woman with a history of vitiligo who presented with an insidious onset of neurologic, vestibular, ocular, and auditory symptoms. She had recently noted the onset of vertigo, tinnitus, and hypersensitivity to sound. Findings on audiometry were within normal limits, although the patient reported some auditory discomfort during the testing. The patient had a history of bilateral uveitis and peripheral neurologic symptoms. She was diagnosed with Vogt-Koyanagi-Harada (VKH) syndrome and started on corticosteroid therapy. Her neurologic, vestibular, ocular, and auditory symptoms resolved. VKH syndrome is an uncommon cause of vertigo and hearing loss, but it should be considered in the differential diagnosis of patients with autoimmunity-related inner ear symptoms.

Gene amplification in esophageal adenocarcinomas and Barrett's with high-grade dysplasia.

PURPOSE: The purpose of this study was to determine the frequency and overall contribution of specific gene amplification events in the formation of Barrett's adenocarcinomas. The relationship of gene amplification to clinical-pathological variables and its potential usefulness as a marker for early cancer detection were also examined. EXPERIMENTAL DESIGN: We used quantitative PCR and Southern blot analysis to screen 87 cases of Barrett's adenocarcinoma for the presence or absence of 13 distinct gene amplification events. Gene amplification was then examined for correlation with other amplification events and clinical variables (survival, stage, nodal involvement, tumor invasion, smoking history, and gender). Additionally, 22 specimens of Barrett's with high-grade dysplasia (HGD) were examined for the presence of gene amplification. RESULTS: One or more amplification events were present in 50 of 87 (57%) adenocarcinomas. The ERBB2 gene was amplified in 19 of 87 (21.8%), CCNE1 in 11 of 87 (12.6%), GATA4 in 9 of 87 (10.3%), KRAS in 9 of 87 (10.3%), EGFR in 7 of 87 (8.0%), CCND1 in 6 of 87 (6.8%), HNF3alpha in 5 of 87 (5.7%), PIK3CA in 5 of 87 (5.7%), C-MYC in 4 of 87 (4.6%), DYRK2 in 2 of 87 (2.3%), and AIB1, AKT1, and IGF1R were amplified in 0 of 87 (0%) of the tumors. CCND1 amplification was found to correlate negatively with survival (P < 0.05). In addition, the ERBB2 amplicon positively correlated (P < 0.05) with GATA4 amplification. Increased copy number of the ERBB2 (1 of 22), GATA4 (1 of 22), KRAS (2 of 22), C-MYC (1 of 22), CCNE1 (2 of 22), and CCND1 (2 of 22) genes was also observed in one or more Barrett's adenocarcinomas with HGD. CONCLUSIONS: The high frequency of gene amplification in esophageal adenocarcinomas and HGD indicates the important role of these events in esophageal adenocarcinoma development. Additionally, these results underscore the possible usefulness of early detection approaches and chemotherapeutic strategies (ErbB2 and cyclin D1) targeted against amplified gene products.