Miracle-dz/UniFAR-FTUs huggingface dataset:
Expression of the chaplin and rodlin hydrophobic sheath proteins in Streptomyces venezuelae is controlled by σBldN and a cognate anti‐sigma factor, RsbN

The chaplin and rodlin proteins together constitute the major components of the hydrophobic sheath that coats the aerial hyphae and spores in Streptomyces, and mutants lacking the chaplins are unable to erect aerial hyphae and differentiate on minimal media. We have gained insight into the developmental regulation of the chaplin (chp) and rodlin (rdl) genes by exploiting a new model species, Streptomyces venezuelae, which sporulates in liquid culture. Using microarrays, the chaplin and rodlin genes were found to be highly induced during submerged sporulation in a bldN‐dependent manner. Using σBldN ChIP‐chip, we show that this dependence arises because the chaplin and rodlin genes are direct biochemical targets of σBldN. sven3186 (here named rsbN for regulator of sigma BldN), the gene lying immediately downstream of bldN, was also identified as a target of σBldN. Disruption of rsbN causes precocious sporulation and biochemical experiments demonstrate that RsbN functions as a σBldN‐specific anti‐sigma factor.
Exploring breast cancer screening barriers among Barbadian women: a focus group study of mammography in a resource-constrained setting

Breast cancer affects women worldwide. Early detection strategies, notably mammography, aim to reduce mortality from breast cancer. However, mammography is a costly screening tool, generates controversy in terms of its impact and adverse effects, and its uptake remains low among some populations. This qualitative study (12 focus groups with 110 participants) explored experiences with mammography among Barbadian women by investigating how barriers are negotiated in a setting of resource-constrained health care provision without a national screening programme. The study findings indicate that, firstly, Barbadian women have to actively seek understanding of both breast cancer and the mammography process. Women described how, with little public awareness and knowledge, they borrow from available public health information on diabetes and HIV/AIDS to give meaning to mammographic screening. Secondly, many women expressed their fear about mammography and its potential consequences, such as experiencing social stigma and losing a romantic relationship after diagnosis. Thirdly, the cost of screening for women who opted for the more reliable private facilities was discussed, along with the potential cost of health care following a diagnosis and the emotional cost of enduring the societal taboo of breast cancer. If breast cancer screening is to be acceptable for this or similar populations, there should be provision of additional services to ensure better access to free screening or alternative strategies, as well as post-diagnostic social and financial support. The policies to develop these services must also address women’s concerns about screening and breast cancer, and provide comprehensive information to allow informed decisions about screening.
DNA packaging bias and differential expression of gene transfer agent genes within a population during production and release of the Rhodobacter capsulatus gene transfer agent, RcGTA

Rhodobacter capsulatus produces a gene transfer agent (GTA) called RcGTA. RcGTA is a phage‐like particle that packages R. capsulatus DNA and transfers it to other R. capsulatus cells. We quantified the relative frequency of packaging for each gene in the genome by hybridization of DNA from RcGTA particles to an R. capsulatus microarray. All genes were found within the RcGTA particles. However, the genes encoding the RcGTA particle were under‐packaged compared with other regions. Gene transfer bioassays confirmed that the transfer of genes within the RcGTA structural cluster is reduced relative to those of other genes. Single‐cell expression analysis, by flow cytometry analysis of cells containing RcGTA‐reporter gene fusion constructs, demonstrated that RcGTA gene expression is not uniform within a culture. This phenomenon was accentuated when the constructs were placed in a strain lacking a putative lysis gene involved in RcGTA release; a small subpopulation was found to be responsible for ∼ 95% of RcGTA activity. We propose a mechanism whereby high levels of RcGTA gene transcription in the most active RcGTA‐producing cells cause a reduction in their packaging frequency. This subpopulation's role in producing and releasing the RcGTA particles explains the lack of observed cell lysis in cultures.
Effect of Cyp27A1 gene dosage on atherosclerosis development in ApoE‐knockout mice

In humans, sterol 27‐hydroxylase (CYP27A1) deficiency leads to cholesterol deposition in tendons and vasculature. Thus, in addition to its role in bile acid synthesis, where it converts cholesterol to 27‐hydroxycholesterol (27‐OHC), CYP27A1 may also be atheroprotective. Cyp27A1‐deficient (Cyp27A1–/–) mice were crossed with apolipoprotein E (apoE)‐deficient mice. Cyp27A1+/+ /apoE–/– [ApoE‐knockout (KO)], Cyp27A1+/– /apoE–/– heterozygous (het), and Cyp27A1–/– /apoE–/– [double‐knockout (DKO)] mice were challenged with a Western diet (WD) for 3 and 6 mo. ApoE‐KO mice fed a chow diet or a WD were used as the control. The severity of atherosclerosis in DKO mice was reduced 10‐fold. Compared with the control, the DKO mice had no 27‐OHC, total plasma cholesterol and low‐density lipoprotein and very low density lipoprotein (LDL/VLDL) concentrations were reduced 2‐fold, and HDL was elevated 2‐fold. Expression of hepatic CYP7A1, CYP3A, and CYP8B1 were 5‐ to 10‐fold higher. 3‐Hydroxy‐3‐methyl‐glutaryl‐CoA reductase (HMGR) activity increased 4‐fold. Fecal cholesterol was increased. In contrast, het mice fed a WD developed accelerated atherosclerosis and severe skin lesions, possibly because of reduced reverse cholesterol transport due to diminished 27‐OHC production. CYP27A1 activity is involved in the control of cholesterol homeostasis and development of atherosclerosis with a distinct gene dose‐dependent effect. —Zurkinden, L. , Solcà, C. , Vögeli, I. A. , Vogt, B. , Ackermann, D. , Erickson, S. K. , Frey, F. J., Sviridov, D. , Escher, G. Effect of Cyp27A1 gene dosage on atherosclerosis development in ApoE‐knockout mice. FASEB J. 28, 1198–1209 (2014). www.fasebj. org
Cysteine 62 of Bax Is Critical for Its Conformational Activation and Its Proapoptotic Activity in Response to H2O2-induced Apoptosis*

Bax is activated and translocated onto mitochondria to mediate cytochrome c release and apoptosis. The molecular mechanisms of Bax activation during apoptosis remain a subject of debate. We addressed the question of whether reactive oxygen species could directly activate Bax for its subsequent translocation and apoptosis. Using the SW480 human colon adenocarcinoma cell line stably expressing Bax fused to GFP, we showed that H2O2 induces Bax conformational change, mitochondrial translocation, and subsequent oligomerization at mitochondria. We found that H2O2-induced Bax activation is dependent on the conserved cysteine residue 62 of Bax. Mutation of cysteine 62, but not cysteine 126, to serine or alanine abolished its activation by H2O2 but not other death stimuli, both in SW480 and Bax-deficient HCT116 cells, whereas wild type Bax sensitizes these cells to apoptosis. Cysteines of Bax could chemically react with H2O2. Mutation of Bax BH3 domain in the presence of cysteine 62 also abolished Bax proapoptotic activity. We conclude that reactive oxygen species could be a direct signal for Bax activation by reacting with cysteine residues. Our results identify a critical role of cysteine 62 in oxidative stress-induced Bax activation and subsequent apoptosis.
The highly dynamic CRISPR1 system of Streptococcus agalactiae controls the diversity of its mobilome

Clustered regularly interspaced short palindromic repeats (CRISPR) confer immunity against mobile genetic elements (MGEs) in prokaryotes. Streptococcus agalactiae, a leading cause of neonatal infections contains in its genome two CRISPR/Cas systems. We show that type 1‐C CRISPR2 is present in few strains but type 2‐A CRISPR1 is ubiquitous. Comparative sequence analysis of the CRISPR1 spacer content of 351 S. agalactiae strains revealed that it is extremely diverse due to the acquisition of new spacers, spacer duplications and spacer deletions that witness the dynamics of this system. The spacer content profile mirrors the S. agalactiae population structure. Transfer of a conjugative transposon targeted by CRISPR1 selected for spacer rearrangements, suggesting that deletions and duplications pre‐exist in the population. The comparison of protospacers located within MGE or the core genome and protospacer‐associated motif‐shuffling demonstrated that the GG motif is sufficient to discriminate self and non‐self and for spacer selection and integration. Strikingly more than 40% of the 949 different CRISPR1 spacers identified target MGEs found in S. agalactiae genomes. We thus propose that the S. agalactiae type II‐A CRISPR1/Cas system modulates the cohabitation of the species with its mobilome, as such contributing to the diversity of MGEs in the population.
Resolvin D1 primes the resolution process initiated by calorie restriction in obesity‐induced steatohepatitis

Insulin resistance and nonalcoholic steatohepatitis (NASH), characterized by hepatic steatosis combined with inflammation, are major sequelae of obesity. Currently, lifestyle modification (i.e., weight loss) is the first‐line therapy for NASH. However, weight loss resolves steatosis but not inflammation. In this study, we tested the ability of resolvin D1 (RvD1), an anti‐inflammatory and proresolving molecule, to promote the resolution initiated by calorie restriction in obese mice with NASH. Calorie restriction reduced adipose and liver weight (–56 and –13%, respectively; P< 0.001), serum leptin and resistin levels, hepatic steatosis, and insulin resistance. In addition to these, mice receiving RvD1 during the dietary intervention showed increased adiponectin expression at both the mRNA and protein levels and reduced liver macrophage infiltration (–15%, P<0.01). Moreover, RvD1 skewed macrophages from an M1‐ to an M2‐like anti‐inflammatory phenotype, induced a specific hepatic miRNA signature (i.e., miR‐219‐5p and miR‐199a‐5p), and reduced inflammatory adipokine mRNA and protein expression and macrophage innate immune response. In precision‐cut liver slices (PCLSs), which override the influence of circulating factors, RvD1 attenuated hypoxia‐induced mRNA and protein expression of COX‐2, IL‐1β, IL‐6, and CCR7. Of note, RvD1 anti‐inflammatory actions were absent in macrophage‐depleted PCLSs. In summary, RvD1 acts as a facilitator of the hepatic resolution process by reducing the inflammatory component of obesity‐induced NASH. —Rius, B. , Titos, E. , Morán‐Salvador, E. , López‐Vicario, C. , García‐Alonso, V. , González‐Périz, A. , Arroyo, V. , Claria, J. Resolvin D1 primes the resolution process initiated by calorie restriction in obesity‐induced steatohepatitis. FASEB J. 28, 836–848 (2014). www.fasebj. org
Glucocorticoid‐induced TNF receptor family‐related protein ligand regulates the migration of monocytes to the inflamed intestine

Glucocorticoid‐induced TNF receptor family‐related protein (GITR) regulates the function of both T cells and antigen‐presenting cells (APCs), while the function of GITR ligand (GITR‐L) is largely unknown. Here we evaluate the role of GITR‐L, whose expression is restricted to APCs, in the development of enterocolitis. On injecting naive CD4+ T cells, GITR‐L‐/‐Rag‐/‐ mice develop a markedly milder colitis than Rag‐/‐ mice, which correlates with a 50% reduction of Ly6C+CD11b+MHCII+ macrophages in the lamina propria and mesenteric lymph nodes. The same result was observed in αCD40‐induced acute colitis and during peritonitis, suggesting an altered monocyte migration. In line with these observations, the number of nondifferentiated monocytes was approximately 3‐fold higher in the spleen of GITR‐L‐/‐ Rag‐/‐ mice than in Rag‐/‐ mice after αCD40 induction. Consistent with the dynamic change in the formation of an active angiotensin II type 1 receptor (AT1) dimer in GITR‐L‐/‐ splenic monocytes during intestinal inflammation, the migratory capability of splenic monocytes from GITR‐L‐/‐ deficient mice was impaired in an in vitro transwell migration assay. Conversely, αGITR‐L reduces the number of splenic Ly6Chi monocytes, concomitantly with an increase in AT1 dimers. We conclude that GITR‐L regulates the number of proinflammatory macrophages in sites of inflammation by controlling the egress of monocytes from the splenic reservoir. —Liao, G. , van Driel, B. , Magelky, E. , O'Keeffe, M. S. , de Waal Malefyt, R. , Engel, P., Herzog, R. W. , Mizoguchi, E. , Bhan, A. K. , Terhorst, C. Glucocorticoid‐induced TNF receptor family‐related protein ligand regulates the migration of monocytes to the inflamed intestine. FASEB J. 28, 474–484 (2014). www.fasebj. org
CRS–MIS in Candida glabrata: sphingolipids modulate echinocandin–Fks interaction

Infections with the azole‐refractory yeast Candida glabrata are now commonly treated with the echinocandins caspofungin (CSF) or micafungin (MCF). True resistance (> 32‐fold decreased susceptibility) to these lipopeptide inhibitors of cell wall synthesis is rare and strictly associated with mutations in integral membrane proteins Fks1 or Fks2. In contrast, mutants exhibiting 4‐ to 32‐fold CSF reduced susceptibility (CRS) were readily selected in vitro, and surprisingly demonstrated 4‐ to 32‐fold MCF increased susceptibility (MIS). Sequencing and gene deletion demonstrated that CRS–MIS is Fks‐independent. To explore alternative mechanisms, we initially employed Saccharomyces cerevisiae, and observed that CRS was conferred by multiple mutations (fen1Δ, sur4Δ, cka2Δ and tsc10‐ts) disrupting sphingolipid biosynthesis. Following this lead, C. glabrata fen1Δ and cka2Δ deletants were constructed, and shown to exhibit CRS–MIS. Sphingolipid analysis of CRS–MIS laboratory mutants and clinical isolates demonstrated elevated dihydrosphingosine (DHS) and phytosphingosine (PHS) levels, and consistent with this sequencing revealed fen1, sur4, ifa38 and sur2 mutations. Moreover, exogenous DHS or PHS conferred a CRS–MIS phenotype on wild‐type C. glabrata. Exogenous PHS failed, however, to suppress CRS–MIS in a sur2 mutant blocked in conversion of DHS to PHS, implying that accumulation of these intermediates confers CRS–MIS. We conclude that membrane sphingolipids modulate echinocandin–Fks interaction.
Prolonged elevation of galanin and tachykinin expression in mucosal and myenteric enteric nerves in trinitrobenzene sulphonic acid colitis

Abstract Diverticulitis causes recurrent abdominal pain associated with increased mucosal expression of mucosal galanin and substance P (SP). We studied changes in mucosal and myenteric plexus neuropeptides in adult rats using a model of colonic inflammation, trinitrobenzenesulphonic acid colitis. We assessed the effects on the pan‐neuronal markers protein gene product 9.5 (PGP9.5) and neurofilament protein, as well as specific neuropeptides at 1, 2, 3, 4, 6, 8, 10 and 14 weeks. Following the acute injury there was macroscopic resolution of inflammation but minor microscopic abnormalities persisted. Percent area stained of mucosal PGP9.5 fell initially but average levels on days 21 and 28 levels were significantly elevated (P < 0.001), returning to normal by day 42. Percent area staining of PGP9.5 in the muscle rose immediately and remained significantly elevated at 70 days (P < 0.001). SP, neuropeptide K and galanin followed a similar overall pattern. SP to PGP9.5 ratio was significantly increased in the muscle both acutely (days 1–28) and in the long term (days 70 and 98), whereas the galanin to PGP9.5 ratio was significantly increased in the mucosa throughout the study. Low‐grade chronic inflammation after an acute initial insult causes a persistent increase in the expression of galanin in the mucosa and SP in muscle layer.
Serotonergic signalling in the stomach and duodenum of patients with gastroparesis

Abstract Serotonin (5‐HT) is involved in the regulation of motoric and sensory functions of the upper gastrointestinal tract. The aim of the current study was to determine whether serotonergic signalling is altered in patients with idiopathic gastroparesis. Mucosal biopsy specimens were collected from the duodenum, antrum and fundus of 11 patients with idiopathic gastroparesis and 11 healthy controls. Neuroendocrine cells, specifically 5‐HT producing cells, were counted after immunohistochemistry, and non‐neuronal mRNA expression levels of tryptophan hydroxylase (TPH)‐1, 5‐HT transport protein (SERT), 5‐HT3 and 5‐HT4 receptor were quantified by real time RT‐PCR. The number of 5‐HT producing cells was comparable between patients and controls. No difference in expression of TPH‐1 (rate limiting enzyme in 5‐HT biosynthetic pathway) and SERT (responsible for 5‐HT uptake) was found between patients and controls (P > 0.05). In the duodenum, the expression of the 5‐HT3 receptor subunits and the 5‐HT4 receptor was comparable between both groups. However, the 5‐HT4(c) splice variant was expressed more abundantly in healthy controls compared to patients (P = 0.015). This study suggests that the delayed gastric emptying and upper abdominal symptoms in idiopathic gastroparesis do not result from altered mucosal 5‐HT biosynthetic and uptake capacity.
Characterization of myenteric neuropathy in the jejunum of spontaneously diabetic BB‐rats

Abstract Decreased gastric expression and function of neuronal nitric oxide synthase (nNOS) has been proposed as a potential mechanism underlying diabetic gastroparesis. As gastric nNOS expression is vagally controlled, these changes might occur secondarily to vagal neuropathy. In addition, it is unclear whether other inhibitory neurotransmitters are also involved. We used the type 1 diabetic BioBreeding (BB)‐rat model to study jejunal motor control and nNOS expression, which is independent of the vagus. Jejunal segments were used for in vitro contractility studies, and measurement of nNOS expression after 8 or 16 weeks of diabetes compared with age‐ and sex‐matched controls. Unlike electrical field stimulation and acetylcholine (ACh)‐induced contractions, non‐adrenergic non‐cholinergic (NANC) relaxations were significantly reduced in diabetic rats. In contrast to control rats, NANC relaxations in diabetic rats were Nω‐nitro‐l‐arginine methyl ester (l‐NAME) insensitive. Jejunal nNOS expression was significantly decreased in diabetic rats. Both in diabetic and in control animals, l‐NAME resistant relaxations were sensitive to P2‐receptor antagonists. In the jejunum of spontaneously diabetic rats, decreased nitric oxide responsiveness and decreased nNOS protein expression occur while purinergic transmission is unaffected. These findings indicate that nitrergic enteric neuropathy may be a primary dysfunction in diabetes, independent from vagal dysfunction.
Activation of MAPK Kinase 9 Induces Ethylene and Camalexin Biosynthesis and Enhances Sensitivity to Salt Stress in Arabidopsis*

Mitogen-activated protein kinase (MAPK) cascades play important roles in regulating plant growth, development, and responses to various environmental stimuli. We demonstrate that MKK9, an MKK, is an upstream activator of the MPKs MPK3 and MPK6 both in vitro and in planta. Expression of active MKK9 protein in transgenic plants induces the synthesis of ethylene and camalexin through the activation of the endogenous MPK3 and MPK6 kinases. As a consequence, transcription of multiple genes responsible for ethylene biosynthesis, ethylene responses, and camalexin biosynthesis is coordinately up-regulated. The activation of MKK9 inhibits hypocotyl elongation in the etiolated seedlings. MKK9-mediated effects on hypocotyl elongation were blocked by the ethylene biosynthesis inhibitor, aminoethoxyvinylglycine, and ethylene receptor antagonist, Ag+. Expression of active MKK9 protein enhances the sensitivity of transgenic seedlings to salt stress, whereas loss of MKK9 activity reduces salt sensitivity indicating a role for MKK9 in the salt stress response. The results reported here reveal that the MKK9-MPK3/MPK6 cascade participates in the regulation of the biosynthesis of ethylene and camalexin and may be an important axis in the stress responses of Arabidopsis.
NIH research funding and early career physician scientists: continuing challenges in the 21st century

Physician scientists (researchers with either M.D. or M.D.‐Ph.D. degrees) have the unique potential to combine clinical perspectives with scientific insight, and their participation in biomedical research has long been an important topic for policymakers and educators. Given the recent changes in the research environment, an update and extension of earlier studies of this population was needed. Our findings show that physician scientists are less likely to take a major role in biomedical research than they were in the past. The number of physician scientists receiving postdoctoral research training and career development awards is at an all‐time low. Physician scientists today, on average, receive their first major research award (R01 equivalent) at a later age than in the 1980s. The number of first‐time R01‐equivalent awards to physicians is at the same level as it was 30 yr ago, but physicians now represent a smaller percentage of the grant recipients. The long‐term decline in the number of physicians entering research careers was temporarily halted during the period of substantial U.S. National Institutes of Health (NIH) budget growth (1998–2003). These gains are lost, however, in the subsequent years when NIH budgets failed to keep pace with rising costs. —Garrison, H. H. , Deschamps, A. M. NIH research funding and early career physician scientists: continuing challenges in the 21st century. FASEB J. 28, 1049–1058 (2014). www.fasebj. org
A population‐based study of irritable bowel syndrome in a non‐Western population

Abstract Previous studies have found no female predominance in irritable bowel syndrome (IBS) in non‐Western countries. The aim of the study was to assess the prevalence and correlates of Rome II (IBS) in both sexes in Pakistan. A Population‐based survey in a low‐income inner city area using questionnaires to diagnose Rome II IBS and assess distress, disability and stressful life events. Data were collected from 880/938 (93%) randomly selected residents. 13.4% of women and 13.1% men met criteria for Rome II IBS; 34 (3.9%) had diarrhoea‐predominant, 59 (6.7%) had constipation‐predominant IBS and 24 (2.7%) had ‘mixed IBS’. In logistic regression analysis, IBS was associated in men with high income (OR = 1.56; 95% CI: 1.05–2.3) and few years of education (OR = 2.17; 95% CI: 1.2–3.9) and in women with being married (OR = 3.6; 95% CI: 1.1–11.9) and stressful life events score (OR = 1.14; 95% CI: 1.01–1.3). Disability was associated with constipation‐predominant IBS (OR = 1.99; 95% CI: 1.1–3.6), distress (OR = 1.19; 95% CI: 1.14–1.23) and stressful life events (OR = 1.19; 95% CI: 1.1–1.3). Investigations were more likely in men (54%) than in women (27%) (P = 0.003). These findings suggest that the equal sex ratio of IBS in urban Pakistan could result from a close association between marked distress and IBS in men similar to that found in women in western studies.