Molecular insights into clinical trials for immune checkpoint inhibitors in colorectal cancer: Unravelling challenges and future directions.

Colorectal cancer (CRC) is a complex disease with diverse etiologies and clinical outcomes. Despite considerable progress in development of CRC therapeutics, challenges remain regarding the diagnosis and management of advanced stage metastatic CRC (mCRC). In particular, the five-year survival rate is very low since mCRC is currently rarely curable. Over the past decade, cancer treatment has significantly improved with the introduction of cancer immunotherapies, specifically immune checkpoint inhibitors. Therapies aimed at blocking immune checkpoints such as PD-1, PD-L1, and CTLA-4 target inhibitory pathways of the immune system, and thereby enhance anti-tumor immunity. These therapies thus have shown promising results in many clinical trials alone or in combination. The efficacy and safety of immunotherapy, either alone or in combination with CRC, have been investigated in several clinical trials. Clinical trials, including KEYNOTE-164 and CheckMate 142, have led to Food and Drug Administration approval of the PD-1 inhibitors pembrolizumab and nivolumab, respectively, for the treatment of patients with unresectable or metastatic microsatellite instability-high or deficient mismatch repair CRC. Unfortunately, these drugs benefit only a small percentage of patients, with the benefits of immunotherapy remaining elusive for the vast majority of CRC patients. To this end, primary and secondary resistance to immunotherapy remains a significant issue, and further research is necessary to optimize the use of immunotherapy in CRC and identify biomarkers to predict the response. This review provides a comprehensive overview of the clinical trials involving immune checkpoint inhibitors in CRC. The underlying rationale, challenges faced, and potential future steps to improve the prognosis and enhance the likelihood of successful trials in this field are discussed.

NLRP3/GSDMD mediated pyroptosis induces lung inflammation susceptibility in diesel exhaust exposed mouse strains.

BACKGROUND: Genetic diversity among species influences the disease severity outcomes linked to air pollution. However, the mechanism responsible for this variability remain elusive and needs further investigation. OBJECTIVE: To investigate the genetic factors and pathways linked with differential susceptibility in mouse strains associated with diesel exhaust exposure. METHODS: C57BL/6 and Balb/c mice were exposed to diesel exhaust (DE) for 5 days/week for 30 min/day for 8 weeks. Body weight of mice was recorded every week and airway hyperresponsiveness towards DE exposure was recorded after 24 h of last exposure. Mice were euthanised to collect BALF, blood, lung tissues for immunobiochemical assays, structural integrity and genetic studies. RESULTS: C57BL/6 mice showed significantly decreased body weight in comparison to Balb/c mice (p < 0.05). Both mouse strains showed lung resistance and damage to elastance upon DE exposure compared to respective controls (p < 0.05) with more pronounced effects in C57BL/6 mice. Lung histology showed increase in bronchiolar infiltration and damage to the wall in C57BL/6 mice (p < 0.05). DE exposure upregulated pro-inflammatory and Th2 cytokine levels in C57BL/6 in comparison to Balb/c mice. C57BL/6 mice showed increase in Caspase-1 and ASC expression confirming activation of downstream pathway. This showed significant activation of inflammasome pathway in C57BL/6 mice with ∼2-fold increase in NLRP3 and elevated IL-1ß expression. Gasdermin-D levels were increased in C57BL/6 mice demonstrating induction of pyroptosis that corroborated with IL-1ß secretion (p < 0.05). Genetic variability among both species was confirmed with sanger's sequencing suggesting presence of SNPs in 3'UTRs of IL-1ß gene influencing expression between mouse strains. CONCLUSIONS: C57BL/6 mice exhibited increased susceptibility to diesel exhaust in contrast to Balb/c mice via activation of NLRP3-related pyroptosis. Differential susceptibility between strains may be attributed via SNPs in the 3'UTRs of the IL-1ß gene.

Forward genetic screen using a gene-breaking trap approach identifies a novel role of grin2bb-associated RNA transcript (grin2bbART) in zebrafish heart function.

LncRNA-based control affects cardiac pathophysiologies like myocardial infarction, coronary artery disease, hypertrophy, and myotonic muscular dystrophy. This study used a gene-break transposon (GBT) to screen zebrafish (Danio rerio) for insertional mutagenesis. We identified three insertional mutants where the GBT captured a cardiac gene. One of the adult viable GBT mutants had bradycardia (heart arrhythmia) and enlarged cardiac chambers or hypertrophy; we named it "bigheart." Bigheart mutant insertion maps to grin2bb or N-methyl D-aspartate receptor (NMDAR2B) gene intron 2 in reverse orientation. Rapid amplification of adjacent cDNA ends analysis suggested a new insertion site transcript in the intron 2 of grin2bb. Analysis of the RNA sequencing of wild-type zebrafish heart chambers revealed a possible new transcript at the insertion site. As this putative lncRNA transcript satisfies the canonical signatures, we called this transcript grin2bb associated RNA transcript (grin2bbART). Using in situ hybridization, we confirmed localized grin2bbART expression in the heart, central nervous system, and muscles in the developing embryos and wild-type adult zebrafish atrium and bulbus arteriosus. The bigheart mutant had reduced Grin2bbART expression. We showed that bigheart gene trap insertion excision reversed cardiac-specific arrhythmia and atrial hypertrophy and restored grin2bbART expression. Morpholino-mediated antisense downregulation of grin2bbART in wild-type zebrafish embryos mimicked bigheart mutants; this suggests grin2bbART is linked to bigheart. Cardiovascular tissues use Grin2bb as a calcium-permeable ion channel. Calcium imaging experiments performed on bigheart mutants indicated calcium mishandling in the heart. The bigheart cardiac transcriptome showed differential expression of calcium homeostasis, cardiac remodeling, and contraction genes. Western blot analysis highlighted Camk2d1 and Hdac1 overexpression. We propose that altered calcium activity due to disruption of grin2bbART, a putative lncRNA in bigheart, altered the Camk2d-Hdac pathway, causing heart arrhythmia and hypertrophy in zebrafish.

Resveratrol mitigates miR-212-3p mediated progression of diesel exhaust-induced pulmonary fibrosis by regulating SIRT1/FoxO3.

BACKGROUND: Diesel exhaust (DE) exposure contributes to the progression of chronic respiratory diseases and is associated with dysregulation of microRNA expression. The present study aims to investigate the involvement of miRNAs and target genes in DE-induced lung fibrosis. METHODS: C57BL/6 mice were divided into three groups. Group 1 mice were exposed to filtered air (Control). Group 2 mice were exposed to DE for 30 min per day, 5 days per week, for 8 weeks (DE). Group 3 mice received DE exposure along with resveratrol on alternate days for the last 2 weeks (DE + RES). Mice were sacrificed to isolate RNA from lung tissue for miRNA microarray profiling. Bronchoalveolar lavage fluid and lung tissues were collected for cell count and biochemical analysis. RESULTS: DE exposure resulted in differential expression of 28 miRNAs with fold change >2 (p < 0.05). The upregulated miR-212-3p was selected for further analysis. Consensus analysis revealed enrichment of SIRT1 in the FoxO pathway, along with a co-annotation of reduced body weight (p < 0.05). A549 cells transfected with a miR-212-3p inhibitor showed a dose-dependent increase in SIRT1 expression, indicating SIRT1 as a direct target. Treatment with resveratrol restored SIRT1 and miR-212-3p expression and led to a reduction in inflammatory cytokines (p < 0.05). The modulation of SIRT1 correlated negatively with macrophage infiltration, confirming its role in regulating cellular infiltration and lung inflammation. Fibronectin, alpha-SMA, and collagen levels were significantly decreased in DE + RES compared to DE group suggesting modulation of cellular functions and resolution of lung fibrosis. Furthermore, a significant decrease in FoxO3a and TGF-ß gene expressions was observed upon resveratrol administration thereby downregulating pro-fibrotic pathway. CONCLUSIONS: The present study demonstrates resveratrol treatment stabilizes SIRT1 gene expression by attenuating miR-212-3p in DE-exposed mice, leading to downregulation of TGF-ß and FoxO3a expressions. The study highlights the therapeutic role of resveratrol in the treatment of DE-induced pulmonary fibrosis.

Selection for insecticide resistance can promote Plasmodium falciparum infection in Anopheles.

Insecticide resistance is under strong selective pressure in Anopheles mosquitoes due to widespread usage of insecticides in vector control strategies. Resistance mechanisms likely cause changes that profoundly affect mosquito physiology, yet it remains poorly understood how selective pressures imposed by insecticides may alter the ability of the mosquito to host and transmit a Plasmodium infection. From pyrethroid-resistant field-derived Anopheles gambiae s.l. mosquitoes, we established resistant (RES) and susceptible (SUS) colonies by either selection for, or loss of insecticide resistance. We show increased oocyst intensity and growth rate as well as increased sporozoite prevalence and intensity in RES compared to SUS females infected with Plasmodium falciparum. The increase in infection intensity in RES females was not associated with the presence of the kdrL1014F mutation and was not impacted by inhibition of Cytochrome P450s. The lipid transporter lipophorin (Lp), which was upregulated in RES compared to SUS, was at least partly implicated in the increased intensity of P. falciparum but not directly involved in the insecticide resistance phenotype. Interestingly, we observed that although P. falciparum infections were not affected when RES females were exposed to permethrin, these females had decreased lipid abundance in the fat body following exposure, pointing to a possible role for lipid mobilization in response to damage caused by insecticide challenge. The finding that selection for insecticide resistance can increase P. falciparum infection intensities and growth rate reinforces the need to assess the overall impact on malaria transmission dynamics caused by selective pressures mosquitoes experience during repeated insecticide challenge.

Diesel exhaust exposure impairs recovery of lung epithelial and cellular damage in murine model.

Studies have investigated the relationship between diesel exhaust (DE) exposure and lung health, highlighting the potential for DE to induce pulmonary inflammation and oxidative stress. However, the resolution of inflammation upon withdrawal of DE exposure needs further investigation. Therefore, resolution of diesel exhaust-induced lung damage was studied in the murine model. Mice (6 weeks) were divided into three groups. Group 1 (control) mice were exposed to filtered air, Group 2 (DE) mice were exposed to DE (5.1 ± 0.7 mg/m3) & Group 3 (DE-FA) mice were exposed to DE followed by filtered air exposure. Airway hyper-responsiveness was recorded after 24 h of the last exposure. BALF and lung samples were collected for cytokine estimation, immunobiological assays, and western blot analysis. DE exposure showed an increase in lung resistance thereby causing alteration in lung function parameters (p < 0.05) which was restored in the DE-FA group. BALF analysis showed a significant increase in total cell count and protein content in DE with no resolution in DE-FA groups (p < 0.05). Lung histology showed no reduction in the bronchiolar thickness and damage in the DE-FA group suggesting irreversible lung damage (p < 0.05). The significant increase in inflammatory cytokine levels, and collagen deposition showed persistent inflammatory phase and lung damage in the DE-FA group(p < 0.05). ZO-1 was significantly decreased in both test groups indicating disintegrated lung epithelium where in claudin-5 expression showed increased lung permeability. A significant increase in neutrophil elastase activity and decreased expression of, Elafin, resulted in lung epithelial damage in the DE-FA group. Lung injury marker alpha1-antitrypsin was increased in DE-FA groups indicating an immune defense mechanism against neutrophil elastase. The study showed that DE exposure causes persistent lung damage via neutrophil elastase-associated disruption of the epithelial barrier integrity and membrane dysfunction.

In silico tools to assess the potential allergenicity of shiitake mushrooms (Lentinula edodes).

BACKGROUND: Computational tools may have an edge over conventional methods for the preliminary evaluation of food allergenicity. In this study, the allergenic potential of Lentinula edodes was evaluated and validated using in silico tools. RESULTS: The potential cross-reactivity of mushroom proteins with fungal allergens was determined using sequence alignment - the Fast Alignment (FASTA) and Basic Local Alignment Search Tool (BLAST) algorithm. Eight L. edodes proteins were cross-reactive with allergens from fungal origin, showing 52%-89% sequence identity using FASTA algorithm-based alignment. The BLAST data were corroborated by percentage identity and query coverage. Physico-chemical property-based allergenicity was deciphered by AlgPred, Allermatch, and AllergenFP software, which predicted six out of eight proteins as potential allergens. Sequence alignment showed 66%-86% conservancy between mushroom protein and known fungal allergens. Secondary structure and amino acid composition supported structural affinity between query and fungal proteins. Three-dimensional structures of five mushroom proteins were generated, quality assessed, and superimposed with fungal allergens, suggesting possible allergenicity of mushroom proteins. An enzyme-linked immunosorbent assay (ELISA) demonstrated immunoglobulin E (IgE) binding in 13 out of 21 food-hypersensitive patients' sera. CONCLUSION: In silico tools provide preliminary indications about the potential allergenicity and cross-reactivity of mushroom proteins. This approach may be used for the prelusive allergenicity assessment of allergen sources. © 2022 Society of Chemical Industry.

Study the association between spirometry based functional grading and six minute walk distance in chronic respiratory disease patients at a rural tertiary care centre of India.

Introduction: Six-min walk test (6MWT) is easy to use, the least expensive, and a quick measure of physical function and it reflects the capacity to perform our day-to-day activities hence quality of life can be assessed with 6MWT. This study was planned to assess the role of 6MWT in chronic respiratory disease patients and its association with spirometry-based functional grading at a rural tertiary care center of northern India. Materials and Methods: This was a hospital-based cross-sectional study done between December 2019 and July 2021. In this study, 110 patients were included as per inclusion and exclusion criteria. 6MWT and spirometry were conducted as per the American Thoracic Society/European Research Society recommendation using Spiropalm 6MWT and the association between 6MWT and spirometry was assessed. Results: A total of 110 chronic respiratory disease patients were included in the study. There were 69 (63%) males while 41 (37%) were females. Among study participants, chronic obstructive pulmonary disease patients were the most common 48 (43.6%) patients, followed by asthma 28 (25.5%), posttuberculosis sequelae patients 22 (20%), interstitial lung disease 9 (8.2%), and bronchiectasis 3 (2.7%) patients were found. There was a significant positive correlation of 6-min walk distance (6MWD) and % predicted 6MWD with spirometric parameters, forced expiratory volume in 1 s (FEV1), forced vital capacity (FVC) and there was a significant positive correlation of 6MWD with FEV1% (predicted) also. 6MWD and % predicted 6MWD negatively correlated with FEV1/FVC and association between 6MWD and FEV1/FVC was not statistically significant and between % predicted 6MWD and FEV1/FVC, it was found statistically significant. Conclusion: The 6MWD traveled by chronic respiratory disease patients was significantly lower than the predicted 6MWD and 6MWD correlated with spirometric variables well. Therefore, it can conclude that 6MWT is a useful alternative of spirometry in the management of chronic respiratory disease patients in resource-limited settings.

Résumé Introduction: Le test de marche de six minutes (6MWT) est facile à utiliser, le moins coûteux et constitue une mesure rapide de la fonction physique. Il reflète la capacité à effectuer nos activités quotidiennes et la qualité de vie peut donc être évaluée à l'aide du test de marche de six minutes. Cette étude avait pour but d'évaluer le rôle du6MWT chez les patients atteints de maladies respiratoires chroniques et son association avec le classement fonctionnel basé sur la spirométrie dans un centre de soins tertiaires rural du nord de l'Inde. Matériels et méthodes: Il s'agit d'une étude transversale en milieu hospitalier réalisée entre décembre 2019 et juillet 2021. Dans cette étude, 110 patients ont été inclus selon les critères d'inclusion et d'exclusion. Le 6MWT et la spirométrie ont été effectués conformément aux recommandations de l'American Thoracic Society et de l'European Research Society. Thoracic Society/European Research Society en utilisant le Spiropalm 6MWT et l'association entre le 6MWT et la spirométrie a été évaluée. Résultats: Au total, 110 patients atteints de maladies respiratoires chroniques ont été inclus dans l'étude. Il y avait 69 hommes (63 %) et 41 femmes (37 %).41 (37 %) étaient des femmes. Parmi les participants à l'étude, les patients atteints de bronchopneumopathie chronique obstructive étaient les plus nombreux (48 (43,6 %)), suivis de l'asthme (28 (25,5 %)) suivis par l'asthme 28 (25,5%), les séquelles de la tuberculose 22 (20%), la pneumopathie interstitielle 9 (8,2%) et la bronchectasie 3 (2,7%).3 (2,7 %). Il existe une corrélation positive significative entre la distance de marche de 6 minutes (6MWD) et le % prédit de la 6MWD avec les paramètres spirométriques, l'expiration forcée et le taux de mortalité.avec les paramètres spirométriques, le volume expiratoire forcé en 1 s (VEMS), la capacité vitale forcée (CVF) et le volume de l'air expiré.) et la capacité vitale forcée (CVF), et il existe une corrélation positive significative entre le 6MWD et le VEMS.entre le 6MWD et le VEMS(prédit). Le 6MWD et le % prédit du 6MWD étaient négativement corrélés avec le VEMS / CVF et l'association entre le 6MWD et le % prédit du VEMS./CVF et l'association entre le6MWD et le VEMS/n'était pas statistiquement significative et entre le % prédit du 6MWD et le VEMS/CVF, elle s'est avérée statistiquement significative. Conclusion: Le 6MWD parcouru par les patients atteints de maladies respiratoires chroniques était significativement plus bas que le 6MWD prédit et le 6MWDétait bien corrélé avec les variables spirométriques. On peut donc conclure que le 6MWT est une alternative utile à la spirométrie dans la prise en charge des patients atteints de maladies respiratoires chroniques dans les pays à ressources limitées.des patients souffrant de maladies respiratoires chroniques dans des contextes où les ressources sont limitées. Mots-clés: distance de marche de 6 minutes, test de marche de 6 minutes, maladies respiratoires chroniques, spirométrie.

Prognostic Role of Catecholamine in Moderate-to-Severe Traumatic Brain Injury: A Prospective Observational Cohort Study.

Objective Traumatic brain injury leads to the activation of sympathetic nervous system and elevation in serum catecholamine levels. The aim of this study was to determine whether catecholamine level obtained within 24 hours of traumatic brain injury provides a reliable prognostic marker for outcome. Materials and Methods This study was a prospective observational cohort study on 36 moderate-to-severe traumatic brain injury. Plasma epinephrine (E), norepinephrine (NE), and dopamine (DA) levels were measured by using computed tomography enzyme-linked immunosorbent assay test and compared with Glasgow coma scale (GCS) that was obtained concurrently. Neurological outcome was determined by GCS at day 7 of treatment and by Glasgow outcome scale at mean follow-up of 9.73 ± 2.26 months. Results Patients with GCS 3 to 4 had markedly increase in baseline mean E (771.5 ± 126.0), NE (2,225.0 ± 215.4), and DA (590.2 ± 38.8) levels as compared with control, while patients with better GCS (11-12) had mildly elevated levels. Patients with GCS 5 to 10 had intermediate values. Cases with markedly elevated baseline E, NE, and DA level were either died or remained in poor GCS (3 or 4) at day 7 of treatment and remained in persistent vegetative state at mean follow-up of 9.73 ± 2.26 months. Cases with only mildly elevated E, NE, and DA level were improved to better GCS on treatment and had good recovery on follow-up. Conclusion These data indicate that a markedly elevated catecholamine level was an excellent endogenous and readily quantifiable marker that appears to reflect the extent of brain injury and predict the likelihood of recovery.

Hospital-Based Retrospective Cross-sectional Study to Analyse the Causes of Maternal Deaths at a Tertiary Health Care Facility.

Context: Maternal mortality is considered a key health indicator of Maternal and Child Health. Considering the fact that complications are preventable and most of them are modifiable, the study has been planned to analyse maternal deaths in order to suggest recommendations for preventing it. There are various delays according to the three-delay model at primary and secondary level; therefore, interventions are needed at those levels to prevent maternal deaths. Aims: To determine the various direct and indirect causes of maternal deaths, analyse the association of medical and social factors with maternal deaths and ^to determine the predictors of maternal deaths. Settings and Design: Hospital-based retrospective cross-sectional study of all the maternal deaths occurring in the last 4 years at a tertiary health care facility. Methods and Material: Data were collected from the Facility Based Maternal Death Review forms. Statistical Analysis Used: Data were entered and analysed by IBM SPSS version 25.0 software. Results: For maternal deaths, direct obstetric causes were responsible in 128 (74.4%) and indirect causes in 45 (26.2%) cases followed by unspecified causes in 78 (45.3%) and 1 (0.6%) coincidental cause. Statistically significant associations were observed between maternal death and period of gestation, mode of delivery and outcome of delivery (P = 0.12, P = 0.04 and P < 0.001, respectively). Conclusions: The health professionals of primary and secondary level should be well equipped to diagnose the complications and to manage it as early as possible. Thus, maternal mortality rates can be decreased to significant level.

Metabolic Syndrome and Its Correlates Among Female Chronic Obstructive Pulmonary Disease Patients at a Rural Tertiary Health Care Center in Northern India.

BACKGROUND: Chronic obstructive pulmonary disease (COPD) is a lung disease that is thought to result from chronic inflammation that may affect other organ systems. Similarly, metabolic syndrome includes central obesity, hypertriglyceridemia, low high-density lipoprotein cholesterol (HDL-C), hyperglycemia, and hypertension. The prevalence of metabolic syndrome and its associated factors among female COPD patients in northern India needs to be evaluated. AIM AND OBJECTIVES: To find the prevalence of metabolic syndrome and its correlates among female chronic obstructive pulmonary disease patients at a rural tertiary health care center in northern India. MATERIALS AND METHODS: A cross-sectional study was conducted between January 2019 and June 2020 at a rural tertiary health care center in northern India. The female patients who presented with symptoms of COPD and fulfilled the inclusion criteria were included and classified by Global Initiative for Chronic Obstructive Lung Disease (GOLD 2020) guideline while the clinical diagnosis of metabolic syndrome was made according to National Cholesterol Education Program: Adult Treatment Panel III (NCEP: ATP III) criteria. RESULTS: A total of 210 female COPD patients were included, the mean age of patients who had metabolic syndrome was 63.38±10.54 years. Metabolic syndrome was diagnosed in 60.48% of patients. There was a significant difference between female COPD patients with and without metabolic syndrome regarding body weight, BMI (body mass index), waist circumference, systolic blood pressure (SBP) and diastolic blood pressure (DBP), blood sugar, serum triglyceride, serum HDL-C. Whereas no significant difference was found between patients with and without metabolic syndrome group regarding smoking exposure, biomass fuel exposure, duration of biomass fuel exposure, mMRC (modified Medical Research Council) grading of breathlessness, GOLD grading of airflow limitation, route and duration of corticosteroid used. In our study, we also found a significant association between the severity of airflow limitation of COPD with the duration of biomass fuel exposure and BMI. Also, there was a significant association between biomass fuel exposure and the presence of cough in female COPD patients. CONCLUSION: Metabolic syndrome is a prevalent entity in female COPD patients among the northern Indian population. Body weight, BMI, waist circumference, SBP, DBP, fasting blood sugar, serum triglyceride, and serum HDL-C have a significant impact on developing metabolic syndrome in female COPD patients. Duration of biomass fuel exposure and BMI also have a significant impact on the severity of airflow limitation in female COPD patients. So early detection and treatment of parameters of metabolic syndrome are important to reduce complications.

Prevalence of high-risk pregnancy among pregnant women enrolled under Pradhan Mantri Surakshit Matritva Abhiyan in government health facilities of district Etawah, Uttar Pradesh: A cross-sectional study.

Background: According to the Sample Registration System report, India has reduced the maternal mortality rate from 130 per 100,000 live births in 2014-2016 to 113 per 100,000 live births in 2016-2018. The main purpose of antenatal care is to identify "high-risk" cases as early as possible from a large group of antenatal mothers and provide them skilled and appropriate care. Objective: To determine the prevalence of high-risk pregnancy (HRP) in pregnant females availing services under pradhan mantri surakshit matritva abhiyan (PMSMA) and to assess awareness of pregnant mothers about services provided under PMSMA in district Etawah of Uttar Pradesh. Material and Methods: Community-based cross-sectional study was carried out among 400 female beneficiaries who were registered under the PMSMA scheme and delivered their baby at any government health facility during one year of study period. Results: It was observed that from all the antenatal women visiting the community health center for HRP day under the PMSMA scheme, 162 (40.5%) were categorized as HRPs and 238 (59.6%) of them were nonhigh-risk pregnancies. A statistically significant association was observed (P-value = 0.005 at 95% CI) between the difference in the proportion of HRPs and the educational status of the pregnant mothers. Out of 400 beneficiaries, 167 (41.75%) were aware of the PMSMA scheme. Conclusion: Regular antenatal care (ANC) check-ups, early identification of HRP, health education, and timely screening are needed to reduce maternal mortality.

Choline chloride attenuates the allergic airway disease by inhibiting the lysophosphatidylcholine induced response in mouse model.

Aims: Allergic airway disease manifestation is induced by lysophosphatidylcholine (LPC) through CD1d-restricted Natural killer T (NKT) cells. Choline chloride (ChCl) and LPC both have the "choline" moiety in their structure and this may interplay the effect in allergic airway disease pathway. Main methods: To test the hypothesis, mice were sensitized with cockroach extract (CE); challenged with CE or exposed to LPC and were given ChCl 1hr later. Key findings: A significant increase in Airway hyperresponsiveness (AHR), total and differential cell count, Th2 cytokines, 8-isoprostanes level in bronchoalveolar lavage fluid (BALF) and inflammation score based on lung histology were observed on challenge with CE or exposure to LPC (p â€‹< â€‹0.05) indicating LPC induced airway disease manifestation in mice. These parameters were reduced significantly after administering mice with ChCl (p â€‹< â€‹0.05). The inflammatory parameters were significantly increased in LPC exposed mice, not sensitized with CE, which were significantly decreased when mice were administered with ChCl demonstrating its role in the inhibition of LPC induced allergic airway disease manifestation. Docking of CD1d with LPC and ChCl indicated the competitive inhibition of LPC induced effect by ChCl. This was validated in vivo in the form of decreased CD1d-restricted NKT cells in BALF and lung of the immunized mice on ChCl administration. There was no effect of ChCl administration on CD1d expression in BALF and lung cells. Significance: This study shows that ChCl attenuates the allergic response by inhibiting the LPC induced- NKT cell mediated AHR, inflammation and oxidative stress by competitive inhibition to LPC in binding to CD1d.

Using an antimalarial in mosquitoes overcomes Anopheles and Plasmodium resistance to malaria control strategies.

The spread of insecticide resistance in Anopheles mosquitoes and drug resistance in Plasmodium parasites is contributing to a global resurgence of malaria, making the generation of control tools that can overcome these roadblocks an urgent public health priority. We recently showed that the transmission of Plasmodium falciparum parasites can be efficiently blocked when exposing Anopheles gambiae females to antimalarials deposited on a treated surface, with no negative consequences on major components of mosquito fitness. Here, we demonstrate this approach can overcome the hurdles of insecticide resistance in mosquitoes and drug resistant in parasites. We show that the transmission-blocking efficacy of mosquito-targeted antimalarials is maintained when field-derived, insecticide resistant Anopheles are exposed to the potent cytochrome b inhibitor atovaquone, demonstrating that this drug escapes insecticide resistance mechanisms that could potentially interfere with its function. Moreover, this approach prevents transmission of field-derived, artemisinin resistant P. falciparum parasites (Kelch13 C580Y mutant), proving that this strategy could be used to prevent the spread of parasite mutations that induce resistance to front-line antimalarials. Atovaquone is also highly effective at limiting parasite development when ingested by mosquitoes in sugar solutions, including in ongoing infections. These data support the use of mosquito-targeted antimalarials as a promising tool to complement and extend the efficacy of current malaria control interventions.

Diesel exhaust exposure in mice induces pulmonary fibrosis by TGF-ß/Smad3 signaling pathway.

BACKGROUND: Epidemiological studies suggest increased risk of lung cancer associated with diesel exhaust (DE) exposure. However, DE-induced lung fibrosis may lead to cancer and needs investigation. OBJECTIVES: To study the mechanism involved in the initiation of DE- induced lung fibrosis. METHODS: C57BL/6 mice were exposed to DE for 30 min/day for 5 days/weeks for 8 weeks. Pulmonary function test was performed to measure lung function. Mice were euthanized to collect BALF, blood, and lung tissue. BALF was used for cell count and cytokine analysis. Lung tissue slides were stained to examine structural integrity. RNA from lung tissue was used for RT-PCR. Immunoblots were performed to study fibrosis and EMT pathway. RESULTS: Mice exposed to DE increase lung resistance and tissue elastance with decrease in inspiratory capacity (p < 0.05) suggesting lung function impairment. BALF showed significantly increased macrophages, neutrophils and monocytes (p < 0.01). Additionally, there was an increase in inflammation and alveolar wall thickening in lungs (p < 0.01) correlates with cellular infiltration. Macrophages had black soot deposition in lung tissue of DE exposed mice. Lung section staining revealed increase in mucus producing goblet cells for clearance of soot in lung. DE exposed lung showed increased collagen deposition and hydroxyproline residue (p < 0.01). Repetitive exposure of DE in mice lead to tissue remodeling in lung, demonstrated by fibrotic foci and smooth muscles. A significant increase in α-SMA and fibronectin (p < 0.05) in lung indicate progression of pulmonary fibrosis. TGF-ß/Smad3 signaling was activated with increase in P-smad3 expression in DE exposed mice. Decreased expression of E-cadherin and increased vimentin (p < 0.05) in lungs of DE exposed mice indicate epithelial to mesenchymal transition. CONCLUSION: DE exposure to mice induced lung injury and pulmonary fibrosis thereby remodeling tissue. The study demonstrates TGF-ß/SMAD3 pathway involvement with an activation of EMT in DE exposed mice.

Effect of 0.5% povidone-iodine on the nasopharyngeal and oropharyngeal viral loads in patients with COVID-19: A double-blind placebo-controlled randomized clinical trial.

Background and Objective: The povidone-iodine (PvP-I) nasal antiseptic has been shown to completely inactivate the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in vitro at variable concentrations. This study was performed to investigate the effect of 0.5% PvP-I nasal drops and oral gargles on the nasopharyngeal and oropharyngeal viral loads in SARS-CoV-2-positive patients. Methods: This was a double-blind, placebo-controlled, randomized clinical trial among patients aged ≥18 years with reverse-transcriptase polymerase chain reaction confirmed in the mild to moderate category of SARS-CoV-2 infection. A total of 32 patients were randomly assigned to receive either freshly prepared 0.5% PvP-I solution or distilled water in the form of supervised self-administered 4-5 nasal drops, followed by 20 ml for gargling for at least 30 seconds. The main outcome measure was the mean change in viral titer and Ct values in the nasopharyngeal and oropharyngeal samples at baseline, 5 minutes, and 3 hours post intervention. Results: The mean change in viral titers across the time duration for the test group when compared with the control group was not statistically significant (P = 0.109). However, the mean change in Ct value was found to be borderline statistically significant (P = 0.042). Noticeable differences were noted among the mean viral titers and Ct values in the intervention group when plotted against the time of testing as compared to the control group. PvP-I solution at 0.5% dilution was well tolerated, and no evident side effects were reported. Conclusions: This study shows that 0.5% PvP-I has an effect on reducing nasopharyngeal and oropharyngeal viral loads in COVID-19 patients. This can be of substantial aid for the primary care physicians, especially for the practitioners in remote and resource poor areas.

Evaluation of Cognitive Functions in Traumatic Brain Injury Patients Using Mini Mental State Examination and Clock Drawing Test.

BACKGROUND: Traumatic brain injuries (TBIs) are the leading cause of morbidity, mortality, disability, and socioeconomic losses globally, but of more concern, in India and other developing countries. The Mini mental state examination (MMSE) and clock drawing test (CDT) are the two mostly adapted methods for cognitive impairment screening. Therefore, it is necessary to establish a robust evaluation system exclusively for post-TBI cognitive impairment. MATERIALS AND METHODS: One hundred and thirty-four cases treated previously at the health facility for TBIs were evaluated for cognitive functions during the follow-up period ranging from 3 weeks to 6 months in the out-patient department. All cases underwent mini-mental score examination (MMSE) and CDT to assess their cognitive performances. The data were analyzed statistically using Chi-square and ANOVA tests of significance. RESULTS: Statistically significant association (P < 0.001) between the cognitive status of patients on the basis of overall MMSE score and the site of brain injury was observed. It was noted that 76 (56.7%) of the cases had cognitive impairment (MMSE score <24) with majority 44 (32.8%) patients having frontal lobe injuries, followed by 14 (10.1%) having brain injuries in the temporal lobe. On the other hand, using CDT score, it was observed that 102 (76.1%) of the cases had cognitive impairment (CDT score <5) with the majority 49 (36.6%) cases having frontal lobe injury followed by 19 (14.2%) having brain injury in the parietal lobe. CONCLUSION: The CDT was able to access cognitive function disruption in those patients, in whom the mini-mental score examination was not able to assess the same, and this difference in detection capabilities of both the tests was statistically found significant.

Impact of COVID-19 pandemic on maternal and child health services in Uttar Pradesh, India.

BACKGROUND: In the wake of the Covid-19 Pandemic, parts of the public health system at increased risk of reduced efficiency include healthcare services for women and children. This in turn could reverse all the progress achieved over the years in reducing maternal and child mortality. In this study, an attempt has been made to assess the indirect effect of the pandemic on maternal and child health services in public health facilities. METHODS: Data pertaining to maternal and child health services being provided under specific Government programmes, were collected from public health facilities of District Sant Kabir Nagar in Uttar Pradesh, India. Comparative analysis of the data from the pandemic phase with data from the year 2019 was done to determine the impact on services. RESULTS: Reduced coverage across all maternal and child health interventions was observed in the study. There was an overall decrease of 2.26 % in number of institutional deliveries. Antenatal care services were the worst affected with 22.91% decline. Immunization services were also dramatically decreased by more than 20%. CONCLUSION: The response of the public healthcare delivery system to the Covid-19 Pandemic is negatively affecting both the provision and utilization of maternal and child healthcare services. It is deterrent to the progress achieved in maternal and child health parameters over the years. Better response strategies should be put in place to minimize lag in service deliwvery.

Anti-ulcerogenic effect of methanolic extract of Elaeagnus conferta Roxb. seeds in Wistar rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Elaeagnus conferta Roxb. (Elaeagnaceae) is a subtropical shrub mainly native to India, Vietnam, Malaysia and South China, whose various parts are used for treatment of diabetes, gastric ulcers, pain, oxidative stress and pulmonary disorders. Though the other parts of the plant have been reported for their ethnic use i.e. fruits as astringent locally and for cancer systemically, leaves for body pain and flowers for pain in chest and the seeds are mentioned as edible, there is no report per se on the medicinal use of seeds. Based on the fact that seeds of closely resembling species i.e. Elaeagnus rhamnoides has demonstrated significant anti-gastroulcerative property, the probability of the seeds of E. conferta possessing similar activity seemed quite significant. AIM OF THE STUDY: Phytochemical investigation and assessment of pharmacological mechanism(s) involved in anti-ulcer effect of methanolic extract of the seeds of E. conferta. MATERIALS AND METHODS: Bioactive phytoconstituents were isolated by column chromatography. These were identified by spectroscopic techniques including infrared (IR) spectroscopy, nuclear magnetic resonance (NMR) and mass spectrometry. Methanolic extract (MEC) of the seeds was prepared by cold maceration and its anti-ulcerogenic potential was evaluated using indomethacin (50 mg/kg) and water immersion stress models in male rats. The animals were pre-treated with different doses of MEC (400 and 800 mg/kg) and the therapeutic effect was compared with standard drug i.e. ranitidine (RANT; 50 mg/kg). The ameliorative effects of MEC were investigated on gastric juice pH, total acidity, free acidity and ulcer index. The assays of malionaldehyde (MDA), catalase (CAT), superoxide dismutase (SOD), glutathione (GSH) and pro-inflammatory cytokines i.e. interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) were carried out to find out the possible mechanism(s) of protection. Further, histopathological changes were also studied. RESULTS: Chromatography studies and further confirmation by spectroscopic techniques revealed the presence of four different compounds in MEC i.e oleic acid (1), stearic acid (2), ascorbic acid (3) and quercetin (4). MEC exhibited anti-ulcerogenic effect in dose dependent manner which may be attributed to suppression of pro-inflammatory cytokines (IL-6, TNF-α) and MDA (112.7%), and up-regulation of protective factors such as CAT (90.48%), SOD (92.77%) and GSH (90.01%). Ulcer inhibition, reduction in total and free acidity and increase in gastric juice pH were observed in MEC treated rats as compared to disease control animals. Histopathological findings confirmed decreased cell infiltration, less epithelial cell damage and regeneration of gastric mucosa in dose dependent manner. CONCLUSIONS: The anti-ulcer effect of MEC may be attributed to its ability to scavenge free radicals and anti-inflammatory property via suppression of TNF-α and IL-6, thus offers a complete and holistic approach for management of peptic ulcer.

A deep learning-based COVID-19 automatic diagnostic framework using chest X-ray images.

The lethal novel coronavirus disease 2019 (COVID-19) pandemic is affecting the health of the global population severely, and a huge number of people may have to be screened in the future. There is a need for effective and reliable systems that perform automatic detection and mass screening of COVID-19 as a quick alternative diagnostic option to control its spread. A robust deep learning-based system is proposed to detect the COVID-19 using chest X-ray images. Infected patient's chest X-ray images reveal numerous opacities (denser, confluent, and more profuse) in comparison to healthy lungs images which are used by a deep learning algorithm to generate a model to facilitate an accurate diagnostics for multi-class classification (COVID vs. normal vs. bacterial pneumonia vs. viral pneumonia) and binary classification (COVID-19 vs. non-COVID). COVID-19 positive images have been used for training and model performance assessment from several hospitals of India and also from countries like Australia, Belgium, Canada, China, Egypt, Germany, Iran, Israel, Italy, Korea, Spain, Taiwan, USA, and Vietnam. The data were divided into training, validation and test sets. The average test accuracy of 97.11 ± 2.71% was achieved for multi-class (COVID vs. normal vs. pneumonia) and 99.81% for binary classification (COVID-19 vs. non-COVID). The proposed model performs rapid disease detection in 0.137 s per image in a system equipped with a GPU and can reduce the workload of radiologists by classifying thousands of images on a single click to generate a probabilistic report in real-time.