Bacteroidales recruit IL-6-producing intraepithelial lymphocytes in the colon to promote barrier integrity.

Interactions between the microbiota and distal gut are important for the maintenance of a healthy intestinal barrier; dysbiosis of intestinal microbial communities has emerged as a likely contributor to diseases that arise at the level of the mucosa. Intraepithelial lymphocytes (IELs) are positioned within the epithelial barrier, and in the small intestine they function to maintain epithelial homeostasis. We hypothesized that colon IELs promote epithelial barrier function through the expression of cytokines in response to interactions with commensal bacteria. Profiling of bacterial 16S ribosomal RNA revealed that candidate bacteria in the order Bacteroidales are sufficient to promote IEL presence in the colon that in turn produce interleukin-6 (IL-6) in a MyD88 (myeloid differentiation primary response 88)-dependent manner. IEL-derived IL-6 is functionally important in the maintenance of the epithelial barrier as IL-6-/- mice were noted to have increased paracellular permeability, decreased claudin-1 expression, and a thinner mucus gel layer, all of which were reversed by transfer of IL-6+/+ IELs, leading to protection of mice in response to Citrobacter rodentium infection. Therefore, we conclude that microbiota provide a homeostatic role for epithelial barrier function through regulation of IEL-derived IL-6.

Post-Transcriptional Regulation of Hepatic DDAH1 with TNF Blockade Leads to Improved eNOS Function and Reduced Portal Pressure In Cirrhotic Rats.

Portal hypertension (PH) is a major cause of morbidity and mortality in chronic liver disease. Infection and inflammation play a role in potentiating PH and pro-inflammatory cytokines, including TNF, are associated with severity of PH. In this study, cirrhotic bile duct ligated (BDL) rats with PH were treated with Infliximab (IFX, a monoclonal antibody against TNF) and its impact on modulation of vascular tone was assessed. BDL rats had increased TNF and NFkB compared to sham operated rats, and their reduction by IFX was associated with a reduction in portal pressure. IFX treatment also reduced hepatic oxidative stress, and biochemical markers of hepatic inflammation and injury. IFX treatment was associated with an improvement in eNOS activity and increased L-arginine/ADMA ratio and DDAH1 expression. In vitro analysis of HepG2 hepatocytes showed that DDAH1 protein expression is reduced by oxidative stress, and this is in part mediated by post-transcriptional regulation by the 3'UTR. This study supports a role for the DDAH1/ADMA axis on the effect of inflammation and oxidative stress in PH and provides insight for new therapies.

Protein arginine deiminase 4 inhibition is sufficient for the amelioration of collagen-induced arthritis.

Citrullination of joint proteins by the protein arginine deiminase (PAD) family of enzymes is recognized increasingly as a key process in the pathogenesis of rheumatoid arthritis. This present study was undertaken to explore the efficacy of a novel PAD4-selective inhibitor, GSK199, in the murine collagen-induced arthritis model of rheumatoid arthritis. Mice were dosed daily from the time of collagen immunization with GSK199. Efficacy was assessed against a wide range of end-points, including clinical disease scores, joint histology and immunohistochemistry, serum and joint citrulline levels and quantification of synovial autoantibodies using a proteomic array containing joint peptides. Administration of GSK199 at 30 mg/kg led to significant effects on arthritis, assessed both by global clinical disease activity and by histological analyses of synovial inflammation, pannus formation and damage to cartilage and bone. In addition, significant decreases in complement C3 deposition in both synovium and cartilage were observed robustly with GSK199 at 10 mg/kg. Neither the total levels of citrulline measurable in joint and serum, nor levels of circulating collagen antibodies, were affected significantly by treatment with GSK199 at any dose level. In contrast, a subset of serum antibodies reactive against citrullinated and non-citrullinated joint peptides were reduced with GSK199 treatment. These data extend our previous demonstration of efficacy with the pan-PAD inhibitor Cl-amidine and demonstrate robustly that PAD4 inhibition alone is sufficient to block murine arthritis clinical and histopathological end-points.

Ten-year alcohol consumption typologies and trajectories of C-reactive protein, interleukin-6 and interleukin-1 receptor antagonist over the following 12 years: a prospective cohort study.

BACKGROUND: Moderate alcohol consumption is thought to confer cardiometabolic protective effects. Inflammatory pathways are hypothesized to partly underlie this association. OBJECTIVES: The aim of this study was to examine the association between typologies of alcohol consumption and markers of inflammation, and their rate of change over time. METHODS: Data were collected from 8209 participants [69% men; mean age, 50 years (SD 6.1)] of the British Whitehall II study. Alcohol consumption typologies were defined using up to three measures during an approximately 10-year period spanning from 1985 to 1994 as (i) stable nondrinkers, (ii) stable moderate drinkers (referent), (iii) stable heavy drinkers, (iv) nonstable drinkers and (v) former drinkers. C-reactive protein (CRP), interleukin (IL)-6 and IL-1 receptor antagonist (IL-1 RA) were measured up to three times in the following 12 years. RESULTS: Stable moderate drinkers had lower levels of CRP than stable nondrinkers, stable heavy drinkers, former drinkers and nonstable drinkers, but there were no differences in the rate of change in CRP over time between groups. Stable nondrinkers had higher levels of IL-6 as did stable heavy drinkers; rates of change in IL-6 over time were also increased in the latter group. Stable nondrinkers also had higher levels of IL-1 RA. These associations were robust to adjustment for confounding factors. CONCLUSION: Our novel investigation of 10-year drinking typologies shows that stable moderate alcohol consumption is associated with a long-term inflammatory marker profile that is consistent with conferring a reduced risk of developing coronary heart disease.

SWI/SNF chromatin remodeling enzymes in melanocyte differentiation and melanoma.

Epidermal melanocytes are pigment-producing cells derived from the neural crest that protects skin from the damaging effects of solar radiation. Malignant melanoma, a highly aggressive cancer, arises from melanocytes. SWI/SNF enzymes are multiprotein complexes that remodel chromatin structure and have extensive roles in cellular differentiation. Components of the complex have been found to be mutated or lost in several human cancers. This review focuses on studies that implicate SWI/SNF enzymes in melanocyte differentiation and in melanoma.

Feasibility study of FDG PET/CT-derived primary tumour glycolysis as a prognostic indicator of survival in patients with non-small-cell lung cancer.

AIM: To assess the feasibility and prognostic value of measuring total lesion glycolysis of the primary tumour (TLG(primary)) using combined 2-[18F]-fluoro-2-deoxy-d-glucose (FDG) positron-emission tomography/computed tomography (PET/CT) in patients with proven or suspected non-small-cell lung cancer (NSCLC) in the routine diagnostic setting. MATERIALS AND METHODS: At the All wales Research and Diagnostic Positron Emission Tomography Centre in Cardiff (PETIC), in the calendar year 2011, 288 consecutive patients were identified with a single pulmonary mass in whom NSCLC was confirmed or clinically diagnosed following multidisciplinary team review. In a retrospective analysis, for each patient the PET-derived volume of the primary tumour and SUVMEAN was calculated using adaptive thresholds of 40% and 50% of the SUVMAX of the primary tumour. The TLG(primary) (calculated by volume x SUVMEAN) was calculated at these two thresholds and was used to predict survival in a multivariate analysis with TNM (tumour, node, metastasis) stage, age, sex, and SUV(MAX). The primary endpoint was overall survival over a minimum follow-up of at least 7 months. RESULTS: In virtually every case, the primary tumour could be measured using the automated software with minimal use of manual adjustments. In multivariate analysis, TNM clinical stage, log(TLG(primary)) and sex were independent predictors of overall survival. CONCLUSION: Measurements of primary tumour total lesion glycolysis are simple to perform and provide additional prognostic information over and above that provided by TNM staging.

Utility of intraoperative angiography during subaxial foramen transversarium decompression for bow hunter's syndrome.

Bow hunter's syndrome is an uncommon cause of vertebrobasilar insufficiency resulting from rotational compression of the extracranial vertebral artery. While positional compression of any portion of the extracranial vertebral artery has been reported to result in bow hunter's syndrome, the most common site of compression is the V2 segment as it passes through the foramen transversarium of the subaxial cervical spine. A 43-year-old woman presented with increasingly frequent pre-syncopal and syncopal episodes upon leftward head rotation. Pre-operative angiographic studies with the neck rotated to the left demonstrated occlusion of the left vertebral artery by a C4-5 osteophyte arising from the C4 uncinate process. The patient underwent microsurgical decompression of the vertebral artery at C4-5 through a standard anterior transcervical retropharyngeal approach. Selective vertebral artery intraoperative angiography performed with the head passively rotated to the left before and after left vertebral artery decompression showed marked improvement in the luminal diameter and blood flow. The patient's symptoms resolved post-operatively. This case illustrates the second instance of intraoperative angiography used to confirm adequate vertebral artery decompression for bow hunter's syndrome. Intraoperative angiography can be safely used to decrease the extent of vertebral artery decompression in order to minimize the risk of operative complications.

Repair of abdominal aortic aneurysm in heart transplant patients: before or after left ventricular assist device implantation?

Endovascular abdominal aortic aneurysm repair in decompensated heart failure patients requiring ventricular assist device (VAD) placement needs careful consideration of both complex disease states. We present this clinical dilemma and describe our choice of transcatheter aneurysm repair in the face of advanced refractory heart failure following VAD implantation.

Efficient and improved synthesis of Telmisartan.

An efficient synthesis of the angiotensin II receptor antagonist Telmisartan (1) is presented involving a cross coupling of 4-formylphenylboronic acid 10 with 2-(2-bromophenyl)-4,4-dimethyl-2-oxazoline (11) as the key step (90% yield). The benzimidazole moiety 15 was constructed regioselectively via a reductive amination-condensation sequence, replacing the alkylation of the preformed benzimidazole step in the previously published route. This methodology overcomes many of drawbacks associated with previously reported syntheses.

A short and efficient synthesis of valsartan via a Negishi reaction.

An efficient synthesis of the angiotensin-II inhibitor valsartan (Diovan®) is presented. Directed ortho-metalation of 5-phenyl-1-trityl-1H-tetrazole (6) and its Negishi coupling with aryl bromide 5 are the key steps of the synthesis. This method overcomes many of the drawbacks associated with previously reported syntheses.

Optimization of medium constituents for improved chitinase production by Paenibacillus sp. D1 using statistical approach.

AIMS: Statistical optimization of medium components for improved chitinase production by Paenibacillus sp. D1. METHODS AND RESULTS: Urea, K(2)HPO(4), chitin and yeast extract were identified as significant components influencing chitinase production by Paenibacillus sp. D1 using Plackett-Burman method. Response surface methodology (central composite design) was applied for further optimization. The concentrations of medium components for improved chitinase production were as follows (g l(-1)): urea, 0.33; K(2)HPO(4), 1.17; MgSO(4), 0.3; yeast extract, 0.65 and chitin, 3.75. This statistical optimization approach led to the production of 93.2 +/- 0.58 U ml(-1) of chitinase. CONCLUSIONS: The important factors controlling the production of chitinase by Paenibacillus sp. D1 were identified as urea, K(2)HPO(4), chitin and yeast extract. Statistical approach was found to be very effective in optimizing the medium components in manageable number of experimental runs with overall 2.56-fold increase in chitinase production. SIGNIFICANCE AND IMPACT OF THE STUDY: The present investigation provides a report on statistical optimization of medium components for improved chitinase production by Paenibacillus sp. D1. Paenibacillus species are gram-positive, spore-forming bacteria with several PGPR and biocontrol potentials. However, only few reports concerning mycolytic enzyme production especially chitinases are available. Chitinase produced by Paenibacillus sp. D1 represents new source for biotechnological and agricultural use.

How to contain generalized HIV epidemics? A plea for better evidence to displace speculation.

In the worst generalized HIV epidemics in East and Southern Africa, from one-quarter to three-quarters of women aged 15 years can expect to be living with HIV or to have died with AIDS by age 40 years. This disaster continues in the face of massive HIV prevention programmes based on current inexact knowledge of HIV transmission pathways and risks. To stop this disaster, both the public and public health experts need better information about the specific factors that allow HIV to propagate so extensively in countries with generalized epidemics. This knowledge could be acquired by tracing HIV infections to their source - especially tracing HIV infections in women of all ages, and tracing unexplained HIV infections in children with HIV-negative mothers.

Occurrence and detection of AmpC beta lactamases among clinical isolates of E. coli and K. pneumoniae causing UTI.

Presence of Bush class C enzymes in uropathogenic strains of Klebsiella pneumoniae & E. coli resistant to extended spectrum cephalosporins is an emerging threat to clinical therapeutics. These resistant strains result in considerable treatment failure and cannot be detected by routine antibiotic sensitivity screening methods. An effort was therefore made to study AmpC beta lactamase production in E. coli and Klebsiella pneumoniae strains causing UTI. A total of 126 E. coli and 49 K. pneumnoniae strains isolated from urine samples were selected for study out of which AmpC beta lactamase production was seen in 23% E. coli (29 isolates) and 18% K. pneumoniae (49 isolates). The susceptibility of AmpC beta lactamase producers to Imipenem, Nitrofurantoin and Amikacin was found to be 100%, 92% and 80% respectively. Thereby the present study emphasizes the importance of monitoring and control of usage of extended spectrum cephalosporins.

Prevalence of extended spectrum beta-lactamases producing clinical isolates from patients of urinary tract infection in a tertiary care hospital in Delhi.

A total of 250 urinary isolates (188 Escherichia coli and 62 Klebsiella pneumoniae) were studied for ESBL production by double disc approximation test and disc diffusion confirmatory test (NCCLS). ESBL production was found to be 56% in E. coli and 52% in K. pneumoniae. The double disc approximation test showed false ESBL production in five (2.6%) isolates of E. coli and one (1.6%) K. pneumoniae. The susceptibility of ESBL producers to imipenem, amikacin, nitrofurantion was found to be 100%, 86% and 84% respectively. A high degree of co-resistance to co- trimaxazole and norfloxacin was found in strains of ESBL producers. Seventy five per cent of ESBL producers detected were from hospitalized patients admitted in ICU or undergoing surgery.

Effect of aqueous Euphorbia hirta leaf extract on gastrointestinal motility.

The aqueous leaf extract of Euphorbia hirta decreased the gastrointestinal motility in normal rats and decreased the effect of castor oil-induced diarrhoea in mice.

Aminoglycoside resistance in enterococci isolated from paediatric septicaemia in a tertiary care hospital in north India.

BACKGROUND & OBJECTIVES: Enterococci are important nosocomial agents and serious infections caused by them are often treated with a combination of cell wall inhibitor and aminoglycoside. However, the presence of high level aminoglycoside resistance in these isolates makes this treatment combination ineffective. The prevalence of such isolates in a tertiary care set up has important diagnostic and therapeutic implications. The present study was carried out to find out the occurrence of high level aminoglycoside resistant isolates of enterococci in paediatric septicaemia cases in a tertiary care set up in north India. METHODS: Blood of paediatric cases with a clinical diagnosis of septicaemia was cultured to isolate and identify enterococci. Agar screen method was used to detect high level streptomycin and gentamicin resistance in these isolates. Vancomycin susceptibility of these isolates were determined as per the National Committee for Clinical Laboratory Standards (NCCLS) recommendations. RESULTS: Fifty one enterococcal strains were isolated from 21 neonates, nine infants and 21 children with a clinical diagnosis of septicaemia. Sixty eight per cent of these isolates had high level gentamicin resistance and forty three per cent had high level streptomycin resistance. All the isolates with high level streptomycin resistance also had high level gentamicin resistance. More than ninety five per cent of these isolates were sensitive to vancomycin. INTERPRETATION & CONCLUSION: The occurrence of high level gentamicin and high level streptomycin resistance in enterococcal isolates in our set up was high. This would require routine testing of the enterococcal isolates for high level aminoglycoside resistance. Alternative treatment regimes need to be sought, besides prudent use of antibiotics.

Bone mineral status in immigrant Indo-Asian women.

BACKGROUND: Indo-Asian immigrants are known to be at high risk of metabolic bone disease, but the prevalence of osteoporosis in this population is unknown. AIM: To compare the bone mineral at the lumbar spine and femoral neck of Indo-Asian immigrant women with that of age-matched Caucasian women. DESIGN: Retrospective analysis. METHODS: Women of Indo-Asian origin referred for bone density scans in the last five years were identified. The skeletal status of each was compared with an age-matched Caucasian control for bone mineral content (BMC), bone mineral density (BMD) and bone mineral apparent density (BMAD) at the lumbar spine and femoral neck, and hip axis length was measured. RESULTS: At the lumbar spine, Indo-Asians had a significantly lower BMD than Caucasians (0.834 vs. 0.913, p = 0.008), but there was no significant difference when BMAD values were calculated (0.123 vs. 0.122). At the femoral neck, there was no difference in BMD (0.728 vs. 0.712, p = 0.5), and BMAD values were significantly higher among Indo-Asians than Caucasians (0.393 vs. 0.319, p = 0.022). Hip axis length was significantly shorter among Indo-Asian women (10.3 vs. 10.7, p = 0.009). DISCUSSION: Although Indo-Asian women appear to have lower spinal BMD than Caucasians, these differences disappear when BMAD values are calculated. While BMD is an areal density, not taking into account the 'depth' of the bone, BMAD is an estimation of volumetric density. Hence lower BMD values in Asians may be a size-related artefact. Longitudinal studies may be required to evaluate the use of BMD as a marker for fracture risk in this population.