A NANOG-pERK reciprocal regulatory circuit regulates Nanog autoregulation and ERK signaling dynamics.

The self-renewal and differentiation potential of embryonic stem cells (ESCs) is maintained by the regulated expression of core pluripotency factors. Expression levels of the core pluripotency factor Nanog are tightly regulated by a negative feedback autorepression loop. However, it remains unclear how ESCs perceive NANOG levels and execute autorepression. Here, we show that a dose-dependent induction of Fgfbp1 and Fgfr2 by NANOG activates autocrine-mediated ERK signaling in Nanog-high cells to trigger autorepression. pERK recruits NONO to the Nanog locus to repress transcription by preventing POL2 loading. This Nanog autorepression process establishes a self-perpetuating reciprocal NANOG-pERK regulatory circuit. We further demonstrate that this reciprocal regulatory circuit induces pERK heterogeneity and ERK signaling dynamics in pluripotent stem cells. Collectively our data suggest that NANOG induces Fgfr2 and Fgfbp1 to activate ERK signaling in Nanog-high cells to establish a NANOG-pERK reciprocal regulatory circuit. This circuit regulates ERK signaling dynamics and Nanog autoregulation in pluripotent cells.

Cross-population genetic analysis revealed genetic variation and selection in the Ovar-DRB1 gene of Indian sheep breeds.

In sheep, MHC variability is studied widely to explore disease association. The aim of the current study was to explore the genetic diversity of Ovar-DRB diversity across sheep breeds of India. Here, Ovar-DRB1 locus was studied across 20 sheep breeds. DRB1 was amplified (301 bp) and sequenced using a PCR-sequence-based typing approach. Results revealed a high degree of heterozygosity across breeds (mean: 73.99%). Overall mean distance for DRB1 was highest in Sangamneri (0.18) and lowest in Madgyal sheep (0.10). There was a higher rate of transition, across breeds. Further, 39 alleles were isolated in different breeds, out of which 10 were new. To allow easy access and use of the immune-polymorphic database, an online database management system was launched (http://www.mhcdbms.in/). Nucleotide content across breeds for the DRB1 region revealed the richness of GC content (59.26%). Wu-Kabat index revealed vast genetic variation across peptide binding sites (PBS) of DRB1. Residues 6, 66, 69, 52, and 81, were polymorphic showing utility for antigen presentation. All breeds were under positive selection for DRB1 locus (dN > dS). Study revealed the importance of DRB locus diversity for beta chain specifically at PBS across sheep breeds of the Indian subcontinent and presented evidence of positive selection for DRB owing to its evolutionary significance.

Novel fluconazole-resistant zoonotic yeast isolated from mastitis.

A hospital-based cross-sectional study was conducted during 2018-2019 to decipher the prevalence of yeast mastitis. The results indicated a 19.68% prevalence of clinical mastitis in bovines. Among them, 5.51% of samples revealed yeasts constituting 1.09% overall prevalence. Candida albicans was recorded as a significant fungal agent involved in clinical bovine mastitis. We record the association of Kodamaea ohmeri in clinical bovine mastitis. On proteomic and molecular confirmation, K. ohmeri isolates were re-identified from phenotypically identified Candida isolates associated with bovine mastitis. After conventional identification, the yeast isolates were re-identified by MALDI-TOF MS-based proteomic approaches. The D1/D2 domains of 26S-rRNA gene and 5.8S-internal transcribed spacer (ITS) rDNA regions based molecular phylogenetic analysis identified the isolates as K. ohmeri. The isolates were resistant to fluconazole. This study reports the first systemic study of K. ohmeri isolates recovered from bovine clinical mastitis, utilizing conventional, automated, proteomic, and genomic approaches followed by antifungal susceptibility. The findings suggest K. ohmeri as a potent opportunistic emerging pathogen of veterinary and public health concern, need for accurate identification of fungal agents from mycotic mastitis, and use of validated antifungal susceptibility assay because of developing resistance to antimycotic agents. Our findings suggest judicious use of fluconazole and alternative antifungal agents may be considered in case of an outbreak.

Targeting wild-type TP53 using AMG 232 in combination with MAPK inhibition in Metastatic Melanoma; a phase 1 study.

BACKGROUND: Targeting the MDM2-p53 interaction using AMG 232 is synergistic with MAPK inhibitors (MAPKi) in preclinical melanoma models. We postulated that AMG 232 plus MAPKi is safe and more effective than MAPKi alone in TP53-wild type, MAPKi-naïve metastatic melanoma. METHODS: Patients were treated with increasing (120 mg, 180 mg, 240 mg) oral doses of AMG 232 (seven-days-on, 15-days-off, 21-day cycle) plus dabrafenib (D) and trametinib (T) (Arm 1, BRAFV600-mutant) or T alone (Arm 2, BRAFV600-wild type). Patients were treated for seven days with AMG 232 alone before adding T±D. Safety and efficacy were assessed using CTCAE v4.0 and RECIST v1.1 criteria, respectively. Pharmacokinetic (PK) analysis was performed at baseline and steady-state levels for AMG 232. RESULTS: 31 patients were enrolled. Ten and 21 patients were enrolled in Arm 1 and Arm 2, respectively. The most common AMG 232-related adverse events (AEs) were nausea (87%), diarrhea (77%), and fatigue (74%). Seven patients (23%) were withdrawn from the study due to AMG 232-related AEs. Three dose-limiting AEs occurred (Arm 1, 180 mg, nausea; Arm 2, 240 mg, grade 3 pulmonary embolism; Arm 2, 180 mg, grade 4 thrombocytopenia). AMG 232 PK exposures were not altered when AMG 232 was combined with T±D. Objective responses were seen in 8/10 (Arm 1) and 3/20 (Arm 2) evaluable patients. The median progression-free survival for Arm 1 and Arm 2 was 19.0 months-not reached and 2.8 months, respectively. CONCLUSION: The maximum tolerated dose of AMG 232 for both arms was 120 mg. AMG 232 plus T±D exhibited a favorable PK profile. Although objective responses occurred in both arms, adding AMG 232 to T±D did not confer additional clinical benefit.

Building communities of practice through case-based e-learning to prevent and manage TB among people living with HIV-India.

BACKGROUND: Co-management of HIV-TB coinfection remains a challenge globally. Addressing TB among people living with HIV (PLHIV) is a key priority for the Government of India (GoI). In 2016, GoI implemented single-window services to prevent and manage TB in PLHIV. To strengthen HIV-TB service delivery, case-based e-learning was introduced to health care providers at Antiretroviral Therapy centres (ARTc). METHODS: We implemented a hub and spoke model to deliver biweekly, virtual, case-based e-learning at select ARTc (n = 115), from four states of India-Delhi, Uttar Pradesh, Andhra Pradesh and Tamil Nadu. We evaluated feasibility and acceptability of case-based e-learning and its impact on professional satisfaction, self-efficacy, knowledge retention using baseline and completion surveys, session feedback, pre-and post-session assessments. We reviewed routine programmatic data and patient outcomes to assess practices among participating ARTc. RESULTS: Between May 2018 and September 2020, 59 sessions were conducted with mean participation of 55 spokes and 152 participants. For 95% and 88% of sessions ≥ 80% of respondents agreed that topics were clear and relevant to practice, and duration of session was appropriate, respectively. Session participants significantly improved in perceived knowledge, skills and competencies (+ 8.6%; p = 0.025), and technical knowledge (+ 18.3%; p = 0.04) from baseline. Participating ARTc increased TB screening (+ 4.2%, p < 0.0001), TB diagnosis (+ 2.7%, p < 0.0001), ART initiation (+ 4.3%, p < 0.0001) and TB preventive treatment completion (+ 5.2%, p < 0.0001). CONCLUSION: Case-based e-learning is an acceptable and effective modus of capacity building and developing communities of practice to strengthen integrated care. E-learning could address demand for accessible and sustainable continuing professional education to manage complex diseases, and thereby enhance health equity. We recommend expansion of this initiative across the country for management of co-morbidities as well as other communicable and non-communicable diseases to augment the existing capacity building interventions by provide continued learning and routine mentorship through communities of practice.

Consequences of the first and second COVID-19 wave on kidney transplant recipients at a large Indian transplant centre.

BACKGROUND: There is a scarcity of data comparing the consequences of first and second COVID-19 waves on kidney transplant recipients (KTRs) in India. METHODS: We conducted a single-centre retrospective study of 259 KTRs with COVID-19 to compare first wave (March 15-December 31 2020, n = 157) and second wave (April 1-May 31 2021, n = 102). RESULTS: KTRs during second wave were younger (43 vs. 40 years; p-value .04) and also included paediatric patients (0 vs. 5.9%; p-value .003). Symptoms were milder during the second wave (45 vs. 62.7%; p-value .007); COVID-19 positive patients had less frequent cough (32 vs. 13.8%; p-value .001), fever was less frequent (58 vs. 37%; p-value .001), and we observed fewer co-morbidities (11 vs. 20.6%; p-value .04). The percentages of neutrophils (77 vs. 83%; p-value .001) and serum ferritin (439 vs. 688; p-value .0006) were higher during second wave, while lymphocyte counts were reduced (20 vs. 14%; p-value .0001). Hydroxychloroquine (11 vs. 0%; p-value .0001) and tocilizumab (7 vs. 0%; p-value .004) were more frequently prescribed during first wave, while utilization of dexamethasone (6 vs. 27%; p-value .0001) and remdesivir (47 vs. 65%; p-value .03) increased during the second wave. Mucormycosis (1.3 vs. 10%; p-value .01) and ICU admissions (20 vs. 37.2%; p-value .002) were more frequent during second wave. The 28-day mortality rate (9.6 vs. 10%; p-value 1) was not different. CONCLUSIONS: There has been a different clinical spectrum of COVID-19 amongst KTR with similar mortality between the two waves at a large Indian transplant centre.

Prevalence and determinants of workplace violence among nurses in the South-East Asian and Western Pacific Regions: a systematic review and meta-analysis.

BACKGROUND: Workplace violence (WPV) includes verbal abuse, physical violence, bullying or mobbing, assault and sexual harassment. The area has been well researched in the developed and high-income countries among nurses and healthcare professionals, but in the case of the low and upper-middle-income countries, there remains a paucity of comprehensive data on the prevalence of WPV and its contributing factors. AIMS: To estimate the prevalence and determinants of WPV among nurses working in the South-East Asian Region and Western Pacific Regions. DESIGN: Systematic review and Meta-analysis. METHOD: A comprehensive search was done to retrieve articles based on a PRISMA compliant protocol registered in PROSPERO: CRD 42020223605. Study selection, quality assessment and data abstraction were independently done by the team members and discrepancies addressed through mutual consensus. Random-effect meta-analysis, I2 statistics and subgroup analysis were done. RESULTS: The review included 41 studies conducted among 42,222 nurses from 13 countries. The pooled prevalence of WPV, verbal abuse, physical violence, threatening behaviour, physical assault, sexual harassment and bullying/mobbing were 58% (CI 51%-64%), 64% (CI 59%-70%), 23% (CI 14%-34%), 30(CI-11%-52%), 21% (CI 8%-38%), 12% (CI 7%-17%) and 25% (17%-33%), respectively. The various determinants of WPV included attributes related to the patient, nurses and organisation. Moreover, the impact of WPV included physical, psychological repercussions on nurses and the various measures used to manage it. CONCLUSION: Nurses encounter a high level of WPV especially verbal abuse in their workplace settings. Nursing councils and professional nursing organisations should put in their effort towards the formulation and implementation of occupational safety legislation in their respective countries through appropriate political lobbying. RELEVANCE TO CLINICAL PRACTICE: Our review highlights the emerging need to focus on the prevention of WPV among nurses working in lower, middle and upper-middle-income countries. Legislation changes and organisational commitment are vital for ensuring effective policies to combat WPV.

The prevalence of alcohol use disorders using alcohol use disorders identification test (AUDIT) in the Indian setting: - a systematic review and meta-analysis.

There is a wide discrepancy in the epidemiology of alcohol use disorders (AUDs) due to diverse scales and survey approaches. We estimated the prevalence of AUDs by comparing the pooled prevalence based on the alcohol use disorders identification test (AUDIT) Vs. non-AUDIT (all scales other than AUDIT). This review searched the community-based prevalence of AUDs in PubMed, Web of Science, PsycINFO, Scopus, Ovid, and Google Scholar. Articles published during the years from 2000 to 2020 were included. The methodological quality of each study was scored, and data were extracted from the published reports. Pooled prevalence was estimated, and the publication bias was evaluated. Twenty-one studies conducted in different states of India included 73997 community-based respondents, which estimated the overall prevalence of AUDs as 12.5% (95% CI: 9 to 17.3%). The pooled prevalence based on AUDIT was 12.4% (AUDIT ≥8; 95% CI: 8.8 to 17.1%) in which the magnitude of hazardous and harmful alcohol use (8.6%; 95% CI: 5.7 to 12.8%; AUDIT 8-19) was significantly higher than dependent alcohol use (2.3%; 95% CI: 1.1 to 4.8%; AUDIT ≥ 20). The pooled prevalence using the non-AUDIT tool was 14.2(95%; CI: 6-30%). Our findings further reveal that about one in twelve of the population of India have AUDs, and there is a gross variation in the patterns of alcohol use across the country. The high prevalence of AUDs suggests developing a national policy to benefit alcohol use, justifying regional variations.

The perceived social stigma, self-esteem, and its determinants among the health care professionals working in India during COVID 19 pandemic.

BACKGROUND: Health conditions perceived as contagious, dangerous, or incurable are associated with some facets of social stigma. METHODS: A cross-sectional survey was conducted from May 9, 2020 to June 9, 2020, among frontline healthcare workers (HCWs) in India to understand their perceived stigmatizing experiences (SE) and self-esteem during the COVID-19 pandemic. Google forms, an online forms tool, was used to create the survey, and samples were recruited through snowball sampling. Data comprised baseline characteristics of HCWs and their responses to the modified version of the Inventory of Stigmatizing Experiences and the Rosenberg Self-Esteem Scale. RESULTS: Of the 600 participants (mean age: 30.9 ± 6.7 years), 76% comprised of nurses. Most participants were residing in urban areas and working in government sectors in clinical areas. Approximately 66.3% HCWs had at least 1 SE, and 51.7% reported a high impact of stigma (SI) across their various life domains, viz. quality of life, social contacts, self-esteem, and family relations, but 73% had normal self-esteem. The SI was more at the family level than at the individual level. The prevalence of SE (69.5% vs. 56.6%) and psychosocial SI (54.5% vs. 44.1%) was higher among nurses than among doctors. Being a nurse and working in clinical areas were statistically significant (P < 0.05 and < 0.01, respectively) for predicting SE likelihood. CONCLUSION: Although HCWs have their own apprehensions, they do have high self-esteem and continue to deliver professional duties despite their SE. The government should frame guidelines to stop such discrimination and hail the saviors.

Biological, nutritional, and therapeutic significance of Moringa oleifera Lam.

The genus Moringa Adans. comprises 13 species, of which Moringa oleifera Lam. native to India and cultivated across the world owing to its drought and frost resistance habit is widely used in traditional phytomedicine and as rich source of essential nutrients. Wide spectrum of phytochemical ingredients among leaf, flower, fruit, seed, seed oil, bark, and root depend on cultivar, season, and locality. The scientific studies provide insights on the use of M. oleifera with different aqueous, hydroalcoholic, alcoholic, and other organic solvent preparations of different parts for therapeutic activities, that is, antibiocidal, antitumor, antioxidant, anti-inflammatory, cardio-protective, hepato-protective, neuro-protective, tissue-protective, and other biological activities with a high degree of safety. A wide variety of alkaloid and sterol, polyphenols and phenolic acids, fatty acids, flavanoids and flavanol glycosides, glucosinolate and isothiocyanate, terpene, anthocyanins etc. are believed to be responsible for the pragmatic effects. Seeds are used with a view of low-cost biosorbent and coagulant agent for the removal of metals and microbial contamination from waste water. Thus, the present review explores the use of M. oleifera across disciplines for its prominent bioactive ingredients, nutraceutical, therapeutic uses and deals with agricultural, veterinarian, biosorbent, coagulation, biodiesel, and other industrial properties of this "Miracle Tree."

Environmental quality monitoring and impact assessment of solid waste dumpsites in high altitude sub-tropical regions.

Solid Waste Management (SWM) in high altitude regions is critically phased because of the non-availability of suitable facilities for the treatment and handling of large quantities of Municipal Solid Waste (MSW). Open burning practices at hill slopes were noticed which affect the surrounding environment. Hence, it became essential to measure the environmental components around the dumpsites to examine the impacts and suggest new technological solutions. The pollution parameters were monitored in and around the dumpsites, and the data was analysed using statistical tools. The assessment of air quality indicated maximum fine suspended particulate matter (PM2.5) concentration of 206.66 µg/m3 followed by respairable particulate matter (PM10), oxides of nitrogen (NOx) and sulphur dioxide (SO2). Among the gaseous emissions, methane (CH4) concentration was very high (38.53 mg/L) followed by carbon monoxide (CO) concentration (0.96 mg/L). Volatile organic compounds (VOCs) were also detected at few dumpsites with highest observed benzene (C6H6) concentration of 157.53 µg/m3. The soil sample analysis indicated that iron (Fe) concentration dominates followed by manganese (Mn), zinc (Zn), chromium (Cr), copper (Cu), and nickel (Ni). For evaluation of different alternatives for the SWM system, Rapid Impact Assessment Matrix (RIAM) was applied.

Oleander and Datura Poisoning: An Update.

India has a very high incidence of poisoning. While most cases are due to chemicals or drugs or envenomation by venomous creatures, a significant proportion also results from consumption or exposure to toxic plants or plant parts or products. The exact nature of plant poisoning varies from region to region, but certain plants are almost ubiquitous in distribution, and among these, Oleander and Datura are the prime examples. These plants are commonly encountered in almost all parts of India. While one is a wild shrub (Datura) that proliferates in the countryside and by roadsides, and the other (Oleander) is a garden plant that features in many homes. Incidents of poisoning from these plants are therefore not uncommon and may be the result of accidental exposure or deliberate, suicidal ingestion of the toxic parts. An attempt has been made to review the management principles with regard to toxicity of these plants and survey the literature in order to highlight current concepts in the treatment of poisoning resulting from both plants. HOW TO CITE THIS ARTICLE: Pillay VV, Sasidharan A. Oleander and Datura Poisoning: An Update. Indian J Crit Care Med 2019;23(Suppl 4):S250-S255.

FRET Between Riboflavin and 9-Anthraldehyde Based Fluorescent Organic Nanoparticles Possessing Antibacterial Activity.

The aqueous suspension of fluorescent nanoparticles were prepared by using 9-anthradehdye derivative (AH). The nanoparticles (AHNPs) were characterized using DLS-zeta sizer and SEM techniques. The photo physical properties of nanoparticles and precursor were measured and compared using UV-absorption spectroscopy, fluorescence spectroscopy and fluorescence lifetime studies. The significant overlap between fluorescence spectrum of AHNPs and excitation spectrum of Riboflavin (RF) led us to explore Fluorescence Resonance Energy Transfer (FRET) studies between AHNPs and RF in aqueous medium. The mechanism of FRET from AHNPs to RF discussed on spectral observations, thermodynamic parameters and changes produces in fluorescence lifetime in absence and presence of different concentrations of RF to AHNPs. The limit of detection for RF (0.071 µM) is considerably low compared with reported methods. Thus, we explore AHNPs as novel nano probe for quantitative determination of RF in pharmaceutical samples based on FRET study. In addition with this, AHNPs has excellent antibacterial activity than the bulk material for two different bacteria culture viz. E. coli and Bacillus sps. Graphical Abstract 9-anthradehdye based fluorescent nanoparticles (AHNPs) explores as nano probe to detect Riboflavin (RF) in aqueous medium based on Fluorescence Resonance Energy Transfer (FRET) studies. The proposed analytical method successfully applied for quantitative determination of RF in pharmaceutical samples. In addition, with this, AHNPs has excellent antibacterial activity than the bulk material for two different bacteria culture suspension viz. E. coli and Bacillus sps.

Biological, medicinal and toxicological significance of Eucalyptus leaf essential oil: a review.

The genus Eucalyptus L'Heritier comprises about 900 species, of which more than 300 species contain volatile essential oil in their leaves. About 20 species, within these, have a high content of 1,8-cineole (more than 70%), commercially used for the production of essential oils in the pharmaceutical and cosmetic industries. However, Eucalyptus is extensively planted for pulp, plywood and solid wood production, but its leaf aromatic oil has astounding widespread biological activities, including antimicrobial, antiseptic, antioxidant, chemotherapeutic, respiratory and gastrointestinal disorder treatment, wound healing, and insecticidal/insect repellent, herbicidal, acaricidal, nematicidal, and perfumes, soap making and grease remover. In the present review, we have made an attempt to congregate the biological ingredients of leaf essential oil, leaf oil as a natural medicine, and pharmacological and toxicological values of the leaf oil of different Eucalyptus species worldwide. © 2017 Society of Chemical Industry.

Surgical and functional outcomes of cochlear implantation in post-lingual and cross-over patients: First 5-year review of the National Ministry of Health Malaysia cochlear implant programme.

INTRODUCTION: There has been a paradigm shift in the management of acquired sensory neural deafness in the past 30years. This is due to the emergence of implantable hearing devices such as the cochlear implant. The objective of this study is to identify surgical and functional outcomes of post-lingual and cross-over patients implanted with a cochlear implant under the National Ministry of Heath Cochlear Implant (CI) Program between 2009-2013. MATERIALS AND METHODS: We retrospectively reviewed all postlingual and cross-over recipients of cochlear implants under the National Ministry of Heath CI Programme from 2009 to 2013. The outcomes measured were surgical complications and functional outcome. Surgical complications were divided into major and minor complications. Functional outcomes were measured using Categorical Auditory Performances (CAP) scale. RESULTS: A total of 41 post-lingual and 15 cross-over patients were implanted between 2009 and 2013. The age of implantees ranged from 3.6 years to 63.2 years old. There were two major complications (3.6%), one is a case of electrode migration at three months post implantation, and six months post second implantation. Another was a case of device failure at about one-year post implantation. Both patients were reimplanted in the same ear. There was no minor complication. The CAP score for both groups (overall) showed significant improvement with 96.4% achieved CAP score of five and above at 24 months after implantation (p<0.001). The CAP score showed marked improvement at the first 6 months post implantation and continued to improve with time in both groups. CONCLUSION: The Malaysian National Ministry of Health Cochlear implant (CI) Program between 2009-2013 has been a successful programme with good surgical and functional outcomes among the post lingual and cross-over patients.

Winery biomass waste degradation by sequential sonication and mixed fungal enzyme treatments.

To increase the efficiency of winery-derived biomass biodegradation, grape pomace was ultrasonicated for 20min in the presence of 0.25M, 0.5Mand1.0MKOH and 1.0MNaOH. This was followed by treatment with a 1:1 (v/v) mix of crude enzyme preparation derived from Phanerochaete chrysosporium and Trametes versicolor for 18h and a further 18h treatment with a 60:14:4:2 percent ratio combination of enzymes derived from Aspergillus niger: Penicillium chrysogenum: Trichoderma harzianum: P. citrinum, repsectively. Process efficiency was evaluated by its comparison to biological only mixed fungal degradation over 16days. Ultrasonication treatment with 0.5MKOH followed by mixed enzyme treatment yielded the highest lignin degradation of about 13%. Cellulase, ß-glucosidase, xylanase, laccase and lignin peroxidase activities of 77.9, 476, 5,390.5, 66.7 and 29,230.7U/mL, respectively, were observed during biomass degradation. Gas chromatography-mass spectrometry (GC-MS) analysis of the degraded material identified commercially important compounds such as gallic acid, lithocholic acid, glycolic acid and lactic acid which were generated in considerable quantities. Thus, the combination of sonication pre-treatment and enzymatic degradation has the potential to considerably improve the breakdown of agricultural biomass and produce commercially useful compounds in markedly less time (<40h) with respect to biological only degradation (16days).

Comparative analysis of inflammatory gene expression levels in metabolic syndrome & coronary artery disease.

BACKGROUND & OBJECTIVES: Metabolic syndrome (MetS) increases the likelihood of developing coronary artery disease (CAD), and inflammation is involved in the pathogenesis of both these conditions. The present work was conducted to examine the relative expression of 18 key inflammatory genes associated with MetS and incident CAD in a representative group of patients. METHODS: A total of 178 male patients, including 57 with CAD and 121 without CAD, were enrolled in the study. The participants without CAD were characterized for the presence of MetS using modified criteria specific for Asian Indians, which included a lower cut-off for waist circumference (≥90 cm for men). The expression of 18 inflammatory genes was evaluated in peripheral whole blood by quantitative polymerase chain reaction method. RESULTS: Of the 121 participants without CAD, 53 (43.8%) had three or more risk factors (MetS group), 50 (41.3%) had one or two risk factors (non-MetS group), while 18 (14.8%) did not have any risk factors (control group). High nuclear factor-kappa B (NF-κB) expression levels and low interleukin-10 (IL-10) levels were observed in MetS patients. Linear association was seen between NF-κB and vascular endothelial growth factor A (VEGFA) expression and with increase in MetS components. Comparison of gene expression pattern between CAD and MetS revealed significantly higher expression of leukotriene genes - arachidonate 5-lipoxygenase (ALOX5), arachidonate 5-lipoxygenase activating protein (ALOX5 AP), leukotriene A4 hydrolase (LTA4H) and leukotriene C4 synthase (LTC4S), and lower expression of NF-κB, interleukin 1 beta (IL-1ß), monocyte chemoattractant protein-1 (MCP-1/CCL2) and signal transducer and activator of transcription 3 (STAT3) genes in CAD. There was linear increase in expression of LTA4H, LTC4S, IL-8 and VEGFA genes across the four groups, namely from controls, non-MetS, MetS and CAD. INTERPRETATION & CONCLUSIONS: A distinct gene expression pattern was seen in MetS and CAD implying a well-orchestrated inflammatory and immune activity. Specifically, NF-κB might be playing an active role in MetS, allowing further expansion of the inflammatory process with resolution of inflammation in full-blown CAD, wherein other gene players such as leukotrienes may dominate.

Understanding the progression of atherosclerosis through gene profiling and co-expression network analysis in Apob(tm2Sgy)Ldlr(tm1Her) double knockout mice.

The objective of the study was to gain molecular insights into the progression of atherosclerosis in Apob(tm2Sgy)Ldlr(tm1Her) mice, using transcriptome profiles. Weighted gene co network analysis (WGCNA) and time course analysis using limma were used to study disease progression from 0 to 20weeks. Five co-expression modules were identified by WGCNA using the expression values of 2153 genes. Genes associated with autophagy, endoplasmic reticulum stress, inflammation and lipid metabolism were differentially expressed at early stages of atherosclerosis. Time course analysis highlighted activation of inflammatory gene signaling at 4weeks, cell proliferation and calcification at 8weeks, amyloid like structures and oxidative stress at 14weeks and enhanced production of inflammatory cytokines at 20weeks. Our results suggest that maximum gene perturbations occur during early atherosclerosis which could be the danger signals associated with subclinical disease. Understanding these genes and associated pathways can help in improvement of diagnostic and therapeutic targets for atherosclerosis.

Expression of human tissue factor pathway inhibitor on vascular smooth muscle cells inhibits secretion of macrophage migration inhibitory factor and attenuates atherosclerosis in ApoE-/- mice.

BACKGROUND: Tissue factor (TF) and coagulation proteases are involved in promoting atherosclerosis, but the molecular and cellular bases for their involvement are unknown. METHODS AND RESULTS: We generated a new strain (ApX4) of apolipoprotein E-deficient mice expressing a membrane-tethered human tissue factor pathway inhibitor fusion protein on smooth muscle actin-positive cells, including vascular smooth muscle cells (SMCs). ApX4 mice developed little atherosclerosis on either a normal chow or high-fat diet. Lipid levels were similar to those in parental ApoE(-/-) mice, and there was no detectable difference in systemic (circulating) tissue factor pathway inhibitor levels or activity. The small lipid-rich lesions that developed had markedly reduced leukocyte infiltrates, and in contrast to ApoE(-/-) mice, SMCs did not express macrophage migratory inhibitory factor (MIF), including at sites distant from atheromatous lesions. Low levels of circulating MIF in ApX4 mice normalized to levels seen in ApoE(-/-) mice after injection of an inhibitory anti-human tissue factor pathway inhibitor antibody, which also led to MIF expression by tissue factor-positive medial SMCs. MIF production by SMCs in ApoE(-/-) mice in vitro and in vivo was shown to be dependent on tissue factor and protease-activated receptor signaling, which were inhibited in ApX4 mice. CONCLUSIONS: Our data indicate that tissue factor plays a hitherto unreported role in the generation of MIF by SMCs in atherosclerosis-prone ApoE(-/-) mice, inhibition of which significantly prevents the development of atherosclerosis, through inhibition of leukocyte recruitment. These data significantly enhance our understanding of the pathophysiology of this important pathology and suggest new potential translational strategies to prevent atheroma formation.