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One of the principles of green and sustainable chemistry is to use green solvents and one-pot synthetic methods which can eliminate the need for drawn-out separation procedures, purification of intermediate compounds, and the production of waste by-products. Considering these concepts of green and sustainable chemistry, an efficient and novel chemoselective one-pot strategy has been developed for the novel synthesis of 2-Aryl-1,3-azoles via Suzuki and Heck cross-coupling reactions via C-2 arylation of azoles under the discriminate temperature using an ionic liquid as green solvent, promoter and ligand. The chemoselective one-pot synthesis, wide substrate scope, moderate to excellent isolated yield of pure products, mild reaction conditions and recycling/reuse of ionic liquid are highlights of this work.

Summary: Tissue resident macrophages are largely of embryonic (fetal liver) origin and long-lived, while bone marrow–derived macrophages (BMDM) are recruited following an acute perturbation, such as hypoxia in the setting of myocardial ischemia. Prior transcriptome analyses identified BMDM and fetal liver–derived macrophage (FLDM) differences at the RNA expression level. Posttranscriptional regulation determining mRNA stability and translation rate may override transcriptional signals in response to hypoxia. We profiled differentially regulated BMDM and FLDM transcripts in response to hypoxia at the level of mRNA translation. Using a translating ribosome affinity purification (TRAP) assay and RNA-seq, we identified non-overlapping transcripts with increased translation rate in BMDM (Ly6e, vimentin, PF4) and FLDM (Ccl7, Ccl2) after hypoxia. We further identified hypoxia-induced transcripts within these subsets that are regulated by the RNA-binding protein HuR. These findings define translational differences in macrophage subset gene expression programs, highlighting potential therapeutic targets in ischemic myocardium.

Transmit Receive (TR) modules are the hardcore of the radar payload, which has day-night and all weather imaging capability in earth observation satellites having military and civil applications. The TR modules in satellite application will experience critical vibration loads during launch wherein it withstands such loads generated by the launch vehicle.In this paper, Low Temperature Cofired Ceramics (LTCC) based dual channel TR module is analyzed for mechanical performance parameters using finite element method and experimentally tested. Both the channels have been tested independently and the integrated module has been successfully qualified for designed specifications. The analytical calculations have been done to validate the physical parameters such as deflection, stress, natural frequency including thickness of the parts. Further, the optimization of design has been proposed to identify the right material for designing of LTCC substrate lid, mechanical housing, and cover of TR module.The FEM based structural analysis has been carried out with 29,867 nodes and 29,389 elements for LTCC submodule and 342,261 nodes and 960,705 elements for the dual TR module level. It is found through the experimental verification of vibration testing that FEM analysis demonstrated less than 2% errors and provided useful confidence for reliable operation of the radar imaging payload in the orbit.

The formation of functional eggs (oocyte) in ovarian follicles is arguably one of the most important events in early mammalian development since the oocytes provide the bulk genetic and cytoplasmic materials for successful reproduction. While past studies have identified many genes that are critical to normal ovarian development and function, recent studies have highlighted the role of mechanical force in shaping folliculogenesis. In this review, we discuss the underlying mechanobiological principles and the force-generating cellular structures and extracellular matrix that control the various stages of follicle development. We also highlight emerging techniques that allow for the quantification of mechanical interactions and follicular dynamics during development, and propose new directions for future studies in the field. We hope this review will provide a timely and useful framework for future understanding of mechano-signalling pathways in reproductive biology and diseases.

Background: Fully covered intraductal self-expanding metal stents (IDSEMS) have been well described in the management of post-liver transplant (LT) anastomotic strictures (ASs). Their antimigration waists and intraductal nature make them suited for deployment across the biliary anastomosis. Objectives: We conducted a multicentre study to analyse their use and efficacy in the management of AS. Design: This was a retrospective, multicentre observational study across nine tertiary centres in the United Kingdom. Methods: Consecutive patients who underwent endoscopic retrograde cholangiopancreatography with IDSEMS insertion were analysed retrospectively. Recorded variables included patient demographics, procedural characteristics, response to therapy and follow-up data. Results: In all, 162 patients (100 males, 62%) underwent 176 episodes of IDSEMS insertion for AS. Aetiology of liver disease in this cohort included hepatocellular carcinoma ( n = 35, 22%), followed by alcohol-related liver disease ( n = 29, 18%), non-alcoholic steatohepatitis ( n = 20, 12%), primary biliary cholangitis ( n = 15, 9%), acute liver failure ( n = 13, 8%), viral hepatitis ( n = 13, 8%) and autoimmune hepatitis ( n = 12, 7%). Early AS occurred in 25 (15%) cases, delayed in 32 (20%) cases and late in 95 (59%) cases. Age at transplant was 54 years (range, 12–74), and stent duration was 15 weeks (range, 3 days–78 weeks). In total, 131 (81%) had complete resolution of stricture at endoscopic re-evaluation. Stricture recurrence was observed in 13 (10%) cases, with a median of 19 weeks (range, 4–88 weeks) after stent removal. At removal, there were 21 (12%) adverse events, 5 (3%) episodes of cholangitis and 2 (1%) of pancreatitis. In 11 (6%) cases, the removal wires unravelled, and 3 (2%) stents migrated. All were removed endoscopically. Conclusion: IDSEMS appears to be safe and highly efficacious in the management of post-LT AS, with low rates of AS recurrence.

In this experimental study was designed to synthesis the CuO NPs using medicinal plant of Moringa oleifera for eradicate the biofilm formation. In result, the UV-spectrum of the synthesized CuO NPs subjected to the range between 200 and 600 nm. Among these, the prominent absorbance peak at 430 nm indicated the peak was held to the copper oxide nanoparticle (CuO NPs). The particle size of biosynthesized CuO NPs varied from 90 nm to 250 nm with more than 50% distribution appearing between 130 nm and 170 nm. In addition, it was further confirmed by PSA, SEM, TEM and SAED of irregular shape and also crystalline nature. All the synthesis and characterization results were clearly supported to the result and confirmed that the synthesized nanoparticle is CuO NPs. Further, the biosynthesized CuO NPs was performed against biofilm forming K. pneumoniae, S. aureus and A. baumannii. The anti-bacterial activity result was shown with 20 mm, 14 mm and 18 mm zone of inhibition against K. pneumoniae, S. aureus and A. baumannii. The minimum biofilm inhibition concentration result was more evident, that the CuO NPs has excellent anti-biofilm activity at 1000 µg/mL concentration. Furthermore, the decreased survival rate of the CuO NPs was observed at 1000 µg /mL concentration by liquid survival degradation assay. Overall, all the results were clearly confirmed that the CuO NPs has excellent anti-biofilm ability against K. pneumoniae, S. aureus and A. baumannii.

Objective There is a paucity of studies in the literature body evaluating short term outcomes following endoscopic retrograde cholangiopancreatography (ERCP) in patients with inoperable malignant hilar biliary obstruction (MHBO). We aimed to primarily evaluate 30-day mortality in these patients and secondarily, conduct a systematic review of studies reporting 30-day mortality.Design We conducted a retrospective analysis of all patients with inoperable MHBO who underwent ERCP at Leeds Teaching Hospitals NHS Trust between February 2015 and September 2020. Logistic regression models constructed from baseline patient data, the modified Glasgow Prognostic Score (mGPS) and Charlson Comorbidity Index (CCI) were evaluated as predictors of 30-day mortality.Results Eighty-seven patients (49 males) with a mean age of 70.4 years (SD ±12.3) were included. Cholangiocarcinoma was the most common aetiology of MHBO affecting 35/87 (40.2%). Technical success was achieved in 72/87 (82.8%). The 30-day mortality rate was 25.3% (22/87), of which 16 were due to progression of underlying malignant disease. On multivariate analysis, only leucocytosis (OR 4.12, 95% CI 2.70 to 7.41, p=0.02) was an independent predictor of 30-day mortality. Neither mGPS (p=0.47) nor CCI with a cut-off value of ≥7 (p=0.06) were significant predictors of 30-day mortality.Conclusion We demonstrated that 30-day mortality following ERCP for inoperable MHBO remains high despite technical success. Further studies are warranted to identify patients most appropriate for intervention.

Background and Aims. A third of patients with primary biliary cholangitis (PBC) experience poorly understood cognitive symptoms, with a significant impact on quality of life (QOL), and no effective medical treatment. Allopregnanolone, a neurosteroid, is a positive allosteric modulator of gamma-aminobutyricacid-A (GABA-A) receptors, associated with disordered mood, cognition, and memory. This study explored associations between allopregnanolone and a disease-specific QOL scoring system (PBC-40) in PBC patients. Method. Serum allopregnanolone levels were measured in 120 phenotyped PBC patients and 40 age and gender-matched healthy controls. PBC subjects completed the PBC-40 at recruitment. Serum allopregnanolone levels were compared across PBC-40 domains for those with none/mild symptoms versus severe symptoms. Results. There were no overall differences in allopregnanolone levels between healthy controls (median = 0.03 ng/ml (IQR = 0.025)) and PBC patients (0.031 (0.42), p=0.42). Within the PBC cohort, higher allopregnanolone levels were observed in younger patients (r (120) = −0.53, p

Photocatalytic degradation of organic dye by AOP's is an effective method to minimize water pollution. The semiconducting materials have a larger bandgap and difficult to fabricate in the form of 2 D-films. Here we report a simple method for the effective fabrication of undoped and La-doped Bi2O3 thin films by a chemical solution deposition method. The incorporation of lanthanum dopant was successfully done and confirmed with the help of SEM, EDAX, XRD, and other advanced characterization techniques. A brilliant flower-like morphology and incorporation of La ions in Bi2O3 lattice helped the fabricated doped thin film for efficient degradation of carbol fuchsin (CF) organic dye, and Chlorpyrifos (CPS) organic pesticide under 120 min of stimulating light source irradiation. Detailed deposition effect on morphology is discussed. The narrowing of bandgap (2.85 eV) in the doped thin film as compared to undoped Bi2O3 film (3.01 eV) resulted in efficient degradation of CF up to 89% with a rate constant of 0.520 min−1 and CPS to 67% with 0.366 min−1. The scavenger addition confirmed ●OH as the major ROS and, metabolites formed during the degradation are identified by LC-MC analysis of degraded samples of both CF and CPS. The reusability study confirmed the efficiency of films up to multiple catalytic cycles. The work confirmed La-Bi2O3 film was effective for the degradation of moieties from a separate class of organic framework.

A Convenient and green synthesis of 4H-pyran derivatives via one-pot multicomponent reaction of aromatic aldehydes, malononitrile and ethyl acetoacetate using KOH loaded CaO as a catalyst under solvent free condition is reported. The morphology of the catalyst has been characterized by XRD and TEM. This protocol have numerous advantages like lesser reaction time, environment friendly, low cost, easy to separate, and provide higher yield. The synthesized compounds have been characterized and confirmed by different spectroscopic techniques like 1H NMR, 13C NMR, FT- IR, and LC- MS.

Oral drug delivery remains the most common and well tolerated method for drug administration. However, its applicability is often limited due to low drug solubility and stability. One approach to overcome the solubility and stability limitations is the use of amorphous polymeric prodrug formulations, such as poly(β-amino ester) (PBAE). PBAE hydrogels, which are biodegradable and pH responsive, have shown promising results for the controlled release of drugs by improving the stability and increasing the solubility of these drugs. In this work, we have evaluated the potential use of PBAE prodrugs in an oral tablet formulation, studying their sustained drug release potential and storage stability. Curcumin, a low solubility, low stability antioxidant drug was used as a model compound. Poly(curcumin β-amino ester) (PCBAE), a crosslinked amorphous network, was synthesized by a previously published method using a commercial diacrylate and a primary diamine, in combination with acrylate-functionalized curcumin. PCBAE-based tablets were made and exhibited a sustained release for 16 h, following the hydrolytic degradation of PCBAE particles into native curcumin. In addition to the release studies, preliminary storage stability was assessed using standard and accelerated stability conditions. As PCBAE degradation is hydrolysis driven, tablet stability was found to be sensitive to moisture.

Purpose: Despite meticulous techniques, surgical complications continue to be problematic in kidney transplant recipients. Role of routine stenting to reduce complications is controversial. In this study, we compare incidence of early urological complications, lymphoceles, urinary tract infections (UTI) and graft function; with or without double-J stenting. Materials and Methods: All patients who underwent live related donor renal transplantation from February 2014 to February 2016 were included. Transplants prior to February 2015 were without routine stenting; subsequent transplants were with routine stenting. Patients with neurogenic bladder, previously operated bladder and delayed or low urinary output were excluded. Follow-up was for at least three months. Descriptive statistics was performed for all parameters. Chi square test and Fisher's Exact test were used for qualitative variables. For quantitative variables, Mann-Whitney test was used to test median difference and independent samples t-test for mean difference. The p-value ≤0.05 was considered significant. Results: We analysed 74 patients (34 stented and 40 non-stented). There was no difference in the incidence of urinary leak, anastomotic obstruction, lymphoceles or UTI (p>0.4 for all comparisons). However, mean estimated glomerular filtration rate at sixth day, 14th day, one month and two months were 76.1 vs. 61.5 (p=0.025), 72.1 vs. 56.6 (p=0.005), 79.4 vs. 63.1 (p=0.002) and 82.0 vs. 63.3 (p=0.001) in the stented versus non-stented groups. Conclusions: Placement of ureteral stent in renal transplant does not significantly affect the incidence of early urinary complications or UTI. However, graft function is significantly better in stented recipients, at least in the short term.

Purpose: To study clinical presentation, laboratory results, imaging findings and treatment options and outcomes of retroperito-neal fibrosis (RPF). To determine whether it follows the same natural course and response to treatment in the Asian population as in the Western world.Materials and Methods: Medical records of patients diagnosed with RPF on imaging and histopathology between February 2010 and April 2016 were reviewed.Results: Of the 21 patients analyzed, mean age at presentation was 50.81 years. The male to female ratio was 0.9:1. Pain was most common presenting complaint (95.23% cases), almost 85% cases were idiopathic and rests were postradiation induced. The me-dian creatinine level was 1.8 mg/dL. The mean erythrocyte sedimentation rate (ESR) was 53.2 mm/h. Hydronephrosis was present in all patients and 47.6% had atrophic kidneys. Diffuse retroperitoneal mass was present in 61.1%. Ureterolysis with lateralization, omental wrapping or gonadal pedicle wrap was done in 17 cases. Two patients underwent uretero-ureterostomy. One patient un-derwent ileal replacement of ureter, and one ileal conduit. Eighteen patients received concurrent medical treatment, 11 were given tamoxifen, 2 steroids (Prednisolone), and five were given both. Of the 20 patients with follow-up, 70% had complete symptomatic relief; ESR improvement was seen in 77.8%. Follow-up ultrasound showed resolved and decreased hydronephrosis in 20% and 55% respectively. One patient had treatment failure and 17.65% had disease recurrence.Conclusions: RPF is a rare disease with varied presentation and outcomes. The male to female ratio may be equal in Asians and smoking could be lesser contributing factor. More Asian cohort studies are required to support same.

Biochar is a byproduct generated from the hydrothermal liquefaction of biomass, such as corn stover, in an anaerobic environment. This work aims to convert biochar into a value-added product of carbon nanofibrous felt. First, the biochar-containing precursor membrane was prepared from simultaneous electrospinning and electrospraying. After thermal stabilization in air and carbonization in argon, the obtained precursor membrane was converted into a mechanically flexible and robust carbon nanofibrous felt. Electrochemical results revealed that the biochar-derived carbon nanofibrous felt might be a good candidate as a supercapacitor electrode with a good rate capability and high kinetic performance.

Activation of the β2 integrin lymphocyte function-associated antigen-1 (LFA-1) in T cells induces stabilization of proinflammatory AU-rich element (ARE)-bearing mRNAs, by triggering the nuclear-to-cytoplasmic translocation of the mRNA-binding and -stabilizing protein HuR. However, the mechanism by which LFA-1 engagement controls HuR localization is not known. Here, we identify and characterize four key regulators of LFA-1-induced changes in HuR activity: the p38 pathway kinase MK2 and the constitutive nuclear proteins hnRNPs C, H1 and K. LFA-1 engagement results in rapid, sequential activation of p38 and MK2. Post-LFA-1 activation, MK2 inducibly associates with both hnRNPC and HuR, resulting in the dissociation of HuR from hnRNPs C, H1 and K. Freed from the three hnRNPs, HuR translocates from the nucleus to the cytoplasm, and mediates the stabilization of labile cytokine transcripts. Our results suggest that the modulation of T cell cytokine mRNA half-life is an intricate process that is negatively regulated by hnRNPs C, H1 and K and requires MK2 as a critical activator.

In this work, an attempt has been made to validate the quality–productivity improvement framework developed for sponge iron (SI) industry, through its implementation in two different SI plants. The study proved that the developed framework is valid and reliable for coal-based SI industry in India. The same framework may be implemented elsewhere in the world with little modification according to the prevailing environment in that country. The concept can also be expanded to other fields.

Introduction: Low Backache (LBA) is one of the commonest presenting complaint of patients and it is important to evaluate it. Since degenerative changes and neoplastic lesion present initially with LBA, it is important to differentiate the lesions as degenerative, benign or malignant. Once the lesion is differentiated the patient can be managed appropriately. Aim: The aim of the study is application of MRI in the evaluation of lower backache. Materials and Methods: In this cross-sectional study male and female patients with LBA, between the age group 30 years to 70 years were included. Exclusion criteria were history of trauma, surgery, metallic implants. Study was conducted in the Department of Radio-diagnosis of Sree Mookambika Institute of Medical Sciences for a period of 1 year. In patients who had back pain associated with radiculopathy, spinal stenosis, or another specific spinal cause, Magnetic Resonance Imaging (MRI) proved to be essential. Results: This study shows various conditions which causes LBA and the most common cause of LBA was degenerative disc disease, and the most common position of herniation was posterolateral. Most of the disc herniation was seen at L4-L5 level. Conclusion: MRI is an important tool in diagnosing the pathology causing LBA and in guiding the clinician in further management of the patient.

Introduction: We depend on eyesight more than any other of our senses to maneuver through the space around us. In a fraction of a second, our eyes work with our brain to tell us the size, shape, colour, and texture of an object. Our eyes are body’s most highly developed sensory organs. The use of computers and television in the era of information technology has given new heights to the professional success rate and it saves time but on the other hand, it has led to an increase in the number of patients with ocular complaints. Aims: The objective of the study was to study the effect of eye exercise techniques along with kapalbhati pranayama on Visual Reaction Time (VRT). Material & Methods: Total 60 subjects in an age group of 18– 30 were recruited in the study. All the subjects were divided into two equal groups (study group and control group) containing 30 subjects (18 male & 12 female) each. Both the male and female subjects were selected on the basis of their voluntary involvement. Visual reaction time for red and green light was recorded from all 60 subjects before the start of the study. Study group subjects were trained to practice various eye exercise techniques and kapalbhati pranayama for 8 weeks regularly whereas control group were busy with their routine activities. After 8 weeks, visual reaction time was measured for red and green light from all 60 subjects. Statistical Analysis: Data expressed as Mean ± S.D, Student t –test was applied for analysis of data, p value 0.05). Conclusion: The results of our study suggest that simple eye exercises along with pranayama helps in improvement of visual reaction time.

Pegylated interferon (Peg-IFN) in combination with ribavirin is the standard of care in the treatment of chronic hepatitis C (HCV). Peg-IFN is known to have a number of side effects but severe respiratory complications are uncommon. We report two cases, one of Peg-IFN induced interstitial pneumonitis (IP) and the other of bronchiolitis obliterans organising pneumonia (BOOP) in patients with chronic hepatitis C infection. In general, respiratory complications of Peg-IFN are mild and resolve with withdrawal of Peg-IFN. However, as illustrated in our first case fatal interstitial pneumonitis can occur. We present a review of the available literature on Peg-IFN induced lung toxicity. In conclusion, pulmonary toxicity with Peg-IFN is rare but fatality can occur. We highlight the importance of maintaining a high index of suspicion for early diagnosis and prompt treatment, which includes withdrawal of Peg-IFN and consideration of corticosteroid treatment.

Background: Inappropriate or excessive use of topical corticosteroids can lead to cutaneous and systemic adverse effects which occur more commonly with the use of very potent steroids. Monitoring and analysis of the prescription practices of topical steroids can help to achieve rational prescription of these drugs. Aim: The present study was carried out to study and analyze the pattern of prescribing topical corticosteroids among outpatients attending the dermatology clinic in a rural tertiary care and teaching hospital, Ambajogai, Maharashtra. Materials and Methods: A cross-sectional descriptive study was conducted for a duration of two months from August 2011 to September 2011, and 500 prescriptions were randomly collected from the dermatology pharmacy and analyzed. Results: About 66% of the prescriptions contained four to five drugs per prescription. Topical steroids were given in 28.4% of all the prescriptions. In almost all the prescriptions, strength, quantity of the steroid to be used, frequency, site, and duration of application was not mentioned. The chief complaints and diagnoses were not mentioned in about 85% of the prescriptions for topical corticosteroids. About 94.36% of the prescriptions contained very potent steroids. Conclusion: Inadequate prescribing information is a clear characteristic of the dermatological prescriptions containing topical corticosteroids. Doctors should be educated about the importance of giving patients sufficient information regarding the use of steroids. There is a need to revise hospital formulary where low-potency steroids can also be included along with potent ones so that the latter can be avoided in conditions where they are unnecessary.

Recent developments in understanding the role of bile acids (BAs) as signalling molecules in human metabolism and inflammation have opened new avenues in the field of hepatology research. BAs are no longer considered as simple molecules helping in fat digestion but as agents with real therapeutic value in treating complex autoimmune and metabolic liver diseases. BAs and their receptors such as farnesoid X receptor, transmembrane G protein-coupled receptor 5 and peroxisome proliferator-activated receptor have been identified as novel targets for drug development. Some of these novel pharmaceuticals are already in clinical evaluation with the most advanced drugs having reached phase III trials. Chronic liver diseases such as primary biliary cholangitis, primary sclerosing cholangitis and nonalcoholic fatty liver disease, for which there is no or limited pharmacotherapy, are most likely to gain from these developments. In this review we discuss recent and the most relevant basic and clinical research findings related to BAs and their implications for novel therapy for chronic liver diseases.

Posterior reversible encephalopathy syndrome (PRES) is a rare acute neurologic condition, characterized by acute neurological symptoms. The radiological findings are predominantly seen in the posterior cerebral hemispheres and are reversible with appropriate management. We are reporting a case of a 21-year-old woman (primigravida), who presented with loss of vision, head ache and vomiting on 2nd day of post-caesarean section. Caesarean section was done due to fetal distress. On examination, patient appeared drowsy but arousable, had a blood pressure of 140/114 mmHg, rest of the examination was unremarkable. CT and MRI brain was performed to rule out cerebral venous sinus thrombosis and cortical blindness; imaging and clinical findings were suggestive of posterior reversible encephalopathy syndrome (PRES). She was treated successfully with antihypertensives and showed dramatic improvement.

Mycobacterium tuberculosis (Mtb) is known to produce wax esters (WE) when subjected to stress. However, nothing is known about the enzymes involved in biosynthesis of WE and their role in mycobacterial dormancy. We report that two putative Mtb fatty acyl-CoA reductase genes (fcr) expressed in E. coli display catalytic reduction of fatty acyl-CoA to fatty aldehyde and fatty alcohol. Both enzymes (FCR1/Rv3391) and FCR2/Rv1543) showed a requirement for NADPH as the reductant, a preference for oleoyl-CoA over saturated fatty acyl-CoA and were inhibited by thiol-directed reagents. We generated Mtb gene-knockout mutants for each reductase. Metabolic incorporation of( 14)C-oleate into fatty alcohols and WE was severely diminished in the mutants under dormancy-inducing stress conditions that are thought to be encountered by the pathogen in the host. The fatty acyl-CoA reductase activity in cell lysates of the mutants under nitric oxide stress was significantly reduced when compared with the wild type. Complementation restored the lost activity completely in the Δfcr1 mutant and partially in the Δfcr2 mutant. WE synthesis was inhibited in both Δfcr mutants. The Δfcr mutants exhibited faster growth rates, an increased uptake of (14)C-glycerol suggesting increased permeability of the cell wall, increased metabolic activity levels and impaired phenotypic antibiotic tolerance under dormancy-inducing combined multiple stress conditions. Complementation of the mutants did not restore the development of antibiotic tolerance to wild-type levels. Transcript analysis of Δfcr mutants showed upregulation of genes involved in energy generation and transcription, indicating the inability of the mutants to become dormant. Our results indicate that the fcr1 and fcr2 gene products are involved in WE synthesis under in vitro dormancy-inducing conditions and that WE play a critical role in reaching a dormant state. Drugs targeted against the Mtb reductases may inhibit its ability to go into dormancy and therefore increase susceptibility of Mtb to currently used antibiotics thereby enhancing clearance of the pathogen from patients.

Engagement of the β2 integrin, lymphocyte function-associated antigen-1 (LFA-1), results in stabilization of T cell mRNA transcripts containing AU-rich elements (AREs) by inducing rapid nuclear-to-cytosolic translocation of the RNA-stabilizing protein, HuR. However, little is known regarding integrin-induced signaling cascades that affect mRNA catabolism. This study examines the role of the GTPases, Rac 1 and Rac 2, and their downstream effectors, in the LFA-1-induced effects on mRNA.Engagement of LFA-1 to its ligand, ICAM-1, in human peripheral T cells resulted in rapid activation of Rac1 and Rac2. siRNA-mediated knockdown of either Rac1 or Rac2 prevented LFA-1-stimulated stabilization of the labile transcripts encoding IFN-γ and TNF-α, and integrin mediated IFN-γ mRNA stabilization was absent in T cells obtained from Rac2 gene-deleted mice. LFA-1 engagement-induced translocation of HuR and stabilization of TNF- α mRNA was lost in Jurkat cells deficient in the Rac guanine nucleotide exchange factor Vav-1 (J.Vav1). The transfection of J.Vav1 cells with constitutively active Rac1 or Rac2 stabilized a labile β-globin reporter mRNA, in a HuR-dependent manner. Furthermore, LFA-1-mediated mRNA stabilization and HuR translocation in mouse splenic T cells was dependent on the phosphorylation of the mitogen-activated protein kinase kinase, MKK3, and its target MAP kinase p38MAPK, and lost in T cells obtained from MKK3 gene-deleted mice.Collectively, these results demonstrate that LFA-1-induced stabilization of ARE-containing mRNAs in T cells is dependent on HuR, and occurs through the Vav-1, Rac1/2, MKK3 and p38MAPK signaling cascade. This pathway constitutes a molecular switch that enhances immune and pro-inflammatory gene expression in T cells undergoing adhesion at sites of activation and effector function.

BACKGROUND:Mycobacterium tuberculosis (Mtb) becomes dormant and phenotypically drug resistant when it encounters multiple stresses within the host. Inability of currently available drugs to kill latent Mtb is a major impediment to curing and possibly eradicating tuberculosis (TB). Most in vitro dormancy models, using single stress factors, fail to generate a truly dormant Mtb population. An in vitro model that generates truly dormant Mtb cells is needed to elucidate the metabolic requirements that allow Mtb to successfully go through dormancy, identify new drug targets, and to screen drug candidates to discover novel drugs that can kill dormant pathogen. METHODOLOGY/PRINCIPAL FINDINGS:We developed a novel in vitro multiple-stress dormancy model for Mtb by applying combined stresses of low oxygen (5%), high CO(2) (10%), low nutrient (10% Dubos medium) and acidic pH (5.0), conditions Mtb is thought to encounter in the host. Under this condition, Mtb stopped replicating, lost acid-fastness, accumulated triacylglycerol (TG) and wax ester (WE), and concomitantly acquired phenotypic antibiotic-resistance. Putative neutral lipid biosynthetic genes were up-regulated. These genes may serve as potential targets for new antilatency drugs. The triacylglycerol synthase1 (tgs1) deletion mutant, with impaired ability to accumulate TG, exhibited a lesser degree of antibiotic tolerance and complementation restored antibiotic tolerance. Transcriptome analysis with microarray revealed the achievement of dormant state showing repression of energy generation, transcription and translation machineries and induction of stress-responsive genes. We adapted this model for drug screening using the Alamar Blue dye to quantify the antibiotic tolerant dormant cells. CONCLUSIONS/SIGNIFICANCE:The new in vitro multiple stress dormancy model efficiently generates Mtb cells meeting all criteria of dormancy, and this method is adaptable to high-throughput screening for drugs that can kill dormant Mtb. A critical link between storage-lipid accumulation and development of phenotypic drug-resistance in Mtb was established. Storage lipid biosynthetic genes may be appropriate targets for novel drugs that can kill latent Mtb.

Estimation of nitrate accumulation in grapevine plant tissue has a great significance in grape production. In this method, nitrate was extracted in water by using magnesium carbonate and calcium hydroxide in the ratio of 10 : 1 and charcoal by agitating for 15 min, which gives clear, transparent, and rapid extraction by removing pigments, color, and turbidity after filtration. Nitrate content in petiole sample extract was determined by a simple UV-spectrometric screening method at 220 nm wavelength which obeyed Beer's law with linear behavior up to nitrate concentration of 11 ppm. The other species such as nitrite, carbonate, and dissolved organic carbon do not interfere. Spectrophotometric data on nitrate determination by other two methods, i.e., phenoldisulphonic acid and salicylic acid method, are included for comparison. The extraction and nitrate determination methods are innovative, reliable, easy to adapt, reduce time consumption and hazardous waste, moreover, they are suitable for routine laboratories where the sample load is very high., Author(s): Rajendra S. Patil [sup.1] , Vinod S. Nandre [sup.2] , Sagar N. Mhetre [sup.3] , Narayan B. Mhetre [sup.3] , Abhishek P. Sutar [sup.3] , Kishor K. Jadhav [sup.3] [...]