Targeting alarmin release reverses Sjogren's syndrome phenotype by revitalizing Ca2+ signalling.

BACKGROUND: Primary Sjogren's syndrome (pSS) is a systemic autoimmune disease that is embodied by the loss of salivary gland function and immune cell infiltration, but the mechanism(s) are still unknown. The aim of this study was to understand the mechanisms and identify key factors that leads to the development and progression of pSS. METHODS: Immunohistochemistry staining, FACS analysis and cytokine levels were used to detect immune cells infiltration and activation in salivary glands. RNA sequencing was performed to identify the molecular mechanisms involved in the development of pSS. The function assays include in vivo saliva collection along with calcium imaging and electrophysiology on isolated salivary gland cells in mice models of pSS. Western blotting, real-time PCR, alarmin release, and immunohistochemistry was performed to identify the channels involved in salivary function in pSS. RESULTS: We provide evidence that loss of Ca2+ signaling precedes a decrease in saliva secretion and/or immune cell infiltration in IL14α, a mouse model for pSS. We also showed that Ca2+ homeostasis was mediated by transient receptor potential canonical-1 (TRPC1) channels and inhibition of TRPC1, resulting in the loss of salivary acinar cells, which promoted alarmin release essential for immune cell infiltration/release of pro-inflammatory cytokines. In addition, both IL14α and samples from human pSS patients showed a decrease in TRPC1 expression and increased acinar cell death. Finally, paquinimod treatment in IL14α restored Ca2+ homeostasis that inhibited alarmin release thereby reverting the pSS phenotype. CONCLUSIONS: These results indicate that loss of Ca2+ signaling is one of the initial factors, which induces loss of salivary gland function along with immune infiltration that exaggerates pSS. Importantly, restoration of Ca2+ signaling upon paquinimod treatment reversed the pSS phenotype thereby inhibiting the progressive development of pSS.

Glomerulus Detection Using Segmentation Neural Networks.

Digital pathology is vital for the correct diagnosis of kidney before transplantation or kidney disease identification. One of the key challenges in kidney diagnosis is glomerulus detection in kidney tissue segments. In this study, we propose a deep learning-based method for glomerulus detection from digitized kidney slide segments. The proposed method applies models based on convolutional neural networks to detect image segments containing the glomerulus region. We employ various networks such as ResNets, UNet, LinkNet, and EfficientNet to train the models. In our experiments on a network trained on the NIH HuBMAP kidney whole slide image dataset, the proposed method achieves the highest scores with Dice coefficient of 0.942.

Combating the infodemic: COVID-19 induced fake news recognition in social media networks.

COVID-19 has caused havoc globally due to its transmission pace among the inhabitants and prolific rise in the number of people contracting the disease worldwide. As a result, the number of people seeking information about the epidemic via Internet media has increased. The impact of the hysteria that has prevailed makes people believe and share everything related to illness without questioning its truthfulness. As a result, it has amplified the misinformation spread on social media networks about the disease. Today, there is an immediate need to restrict disseminating false news, even more than ever before. This paper presents an early fusion-based method for combining key features extracted from context-based embeddings such as BERT, XLNet, and ELMo to enhance context and semantic information collection from social media posts and achieve higher accuracy for false news identification. From the observation, we found that the proposed early fusion-based method outperforms models that work on single embeddings. We also conducted detailed studies using several machine learning and deep learning models to classify misinformation on social media platforms relevant to COVID-19. To facilitate our work, we have utilized the dataset of "CONSTRAINT shared task 2021". Our research has shown that language and ensemble models are well adapted to this role, with a 97% accuracy.

Fighting hate speech from bilingual hinglish speaker's perspective, a transformer- and translation-based approach.

Many people have begun to use social media platforms due to the increased use of the Internet over the previous decade. It has a lot of benefits, but it also comes with a lot of risks and drawbacks, such as Hate speech. People in multilingual societies, such as India, frequently mix their native language with English while speaking, so detecting hate content in such bilingual code-mixed data has drawn the larger interest of the research community. The majority of previous work focuses on high-resource language such as English, but very few researchers have concentrated on the mixed bilingual data like Hinglish. In this study, we investigated the performance of transformer models like IndicBERT and multilingual Bidirectional Encoder Representation(mBERT), as well as transfer learning from pre-trained language models like ULMFiT and Bidirectional encoder Representation(BERT), to find hateful content in Hinglish. Also, Transformer-based Interpreter and Feature extraction model on Deep Neural Network (TIF-DNN), is proposed in this work. The experimental results found that our proposed model outperforms existing state-of-art methods for Hate speech identification in Hinglish language with an accuracy of 73%.

Glycolipid Metabolite ß-Glucosylceramide Is a Neutrophil Extracellular Trap-Inducing Ligand of Mincle Released during Bacterial Infection and Inflammation.

Neutrophil extracellular traps (NETs) are implicated in host defense and inflammatory pathologies alike. A wide range of pathogen- and host-derived factors are known to induce NETs, yet the knowledge about specific receptor-ligand interactions in this response is limited. We previously reported that macrophage-inducible C-type lectin (Mincle) regulates NET formation. In this article, we identify glycosphingolipid ß-glucosylceramide (ß-GlcCer) as a specific NET-inducing ligand of Mincle. We found that purified ß-GlcCer induced NETs in mouse primary neutrophils in vitro and in vivo, and this effect was abrogated in Mincle deficiency. Cell-free ß-GlcCer accumulated in the lungs of pneumonic mice, which correlated with pulmonary NET formation in wild-type, but not in Mincle-/-, mice infected intranasally with Klebsiella pneumoniae Although leukocyte infiltration by ß-GlcCer administration in vivo did not require Mincle, NETs induced by this sphingolipid were important for bacterial clearance during Klebsiella infection. Mechanistically, ß-GlcCer did not activate reactive oxygen species formation in neutrophils but required autophagy and glycolysis for NET formation, because ATG4 inhibitor NSC185058, as well as glycolysis inhibitor 2-deoxy-d-glucose, abrogated ß-GlcCer-induced NETs. Forced autophagy activation by tamoxifen could overcome the inhibitory effect of glycolysis blockage on ß-GlcCer-mediated NET formation, suggesting that autophagy activation is sufficient to induce NETs in response to this metabolite in the absence of glycolysis. Finally, ß-GlcCer accumulated in the plasma of patients with systemic inflammatory response syndrome, and its levels correlated with the extent of systemic NET formation in these patients. Overall, our results posit ß-GlcCer as a potent NET-inducing ligand of Mincle with diagnostic and therapeutic potential in inflammatory disease settings.

Comparative study between clinical diagnosis, plain radiography and sonography for the diagnosis of nontraumatic acute abdomen.

Background: Acute abdomen is the most commonly attended surgical emergency. It can be caused by intra-abdominal, extra-abdominal and metabolic causes. A few imaging modalities are at disposal of primary care physicians like plain x-ray and ultrasonography. Materials and Methods: This study has been done to compare the efficacy of clinical diagnosis, plain radiography and sonography in diagnosis of non-traumatic acute abdomen. Every patient under study admitted in the Department of General Surgery underwent thorough clinical evaluation, biochemical investigations, X-rays and sonography. Findings of clinical evaluation, X-Rays and sonography were compared to the final diagnosis found intraoperatively. Results: Clinical diagnosis was made in 47 (94%) out of 50 patients. X-rays were able to diagnose in 20 patients (40%) whereas sonography diagnosed 26 patients (52%). Conclusion: The present study showed that clinical evaluation, x-rays or Ultrasound alone are not sufficient to diagnose cause of non-traumatic acute abdomen in all cases. Clinical evaluation combined with x-rays and ultrasound increases the number and accuracy of pre-operative diagnosis in non-traumatic acute abdomen.

Resolving macrophage polarization through distinct Ca2+ entry channel that maintains intracellular signaling and mitochondrial bioenergetics.

Transformation of naive macrophages into classically (M1) or alternatively (M2) activated macrophages regulates the inflammatory response. Here, we identified that distinct Ca2+ entry channels determine the IFNγ-induced M1 or IL-4-induced M2 transition. Naive or M2 macrophages exhibit a robust Ca2+ entry that was dependent on Orai1 channels, whereas the M1 phenotype showed a non-selective TRPC1 current. Blockade of Ca2+ entry suppresses pNF-κB/pJNK/STAT1 or STAT6 signaling events and consequently lowers cytokine production that is essential for M1 or M2 functions. Of importance, LPS stimulation shifted M2 cells from Orai1 toward TRPC1-mediated Ca2+ entry and TRPC1-/- mice exhibited transcriptional changes that suppress pro-inflammatory cytokines. In contrast, Orai1-/- macrophages showed a decrease in anti-inflammatory cytokines and exhibited a suppression of mitochondrial oxygen consumption rate and inhibited mitochondrial shape transition specifically in the M2 cells. Finally, alterations in TRPC1 or Orai1 expression determine macrophage polarization suggesting a distinct role of Ca2+ channels in modulating macrophage transformation.

Helminth derived factors inhibit neutrophil extracellular trap formation and inflammation in bacterial peritonitis.

Despite their protective antimicrobial function, neutrophil extracellular traps (NETs) have been implicated in propagation of inflammatory responses in several disease conditions including sepsis. Highly diffusible exogenous ROS produced under such inflammatory conditions, can induce exuberant NETs, thus making inhibition of NETs desirable in inflammatory diseases. Here we report that helminth parasite excretory/secretory factors termed as parasitic ligands (PL) inhibit ROS-induced NETs by blocking the activation of nonselective calcium permeable channel Transient Receptor Potential Melastatin 2 (TRPM2). Therapeutic implication of PL mediated blockage of NET formation was tested in preclinical model of septic peritonitis, where PL treatment regulated neutrophil cell death modalities including NET formation and mitigated neutrophil mediated inflammatory response. This translated into improved survival and reduced systemic and local bacterial load in infected mice. Overall, our results posit PL as an important biological regulator of neutrophil functions with implications to a variety of inflammatory diseases including peritonitis.

Experience of setting up of Control room for COVID-19 at NCDC, New Delhi.

Significant public health events of the 21st century include epidemic prone diseases such as severe acute respiratory syndrome (SARS), Middle East respiratory syndrome (MERS), influenza A (H1N1), Ebola virus disease, and coronavirus (SARS-COV-2). Preparedness as well as risk mitigation strategies play an integral role for the success of responses to such health emergencies. An extraordinary cluster of cases of respiratory disease of unknown cause triggered a series of events that constituted a public health risk across the globe through international spread from China and was declared a Public Health Emergency of International Concern (PHEIC) on 30 January, 2020 by the World Health Organization (WHO). To monitor implementation of activities in order to contain the local transmission of COVID-2019 in India, a control room was established at the National Centre for Disease Control (NCDC), New Delhi on 23rd January, 2020 under the Integrated Disease Surveillance Project (IDSP). The main objectives of the control room were to alleviate the concerns and address queries of passengers arriving from the affected countries and also to provide the general public information regarding the measures to be taken as well as the contact details of the respected district health authorities for further necessary action. A total of 183 hunting lines were established at the NCDC, Noida, TB Centre, and the National Health Authority (NHA) Hyderabad and Bengaluru by March 2020. A total of 79,013 calls, 1,04,779 emails, and 1,787 international calls were received w.e.f. 23 January to 30 March, 2020 at the NCDC control room. The NHA Bengaluru and Hyderabad Control room received 3,52,176 calls w.e.f. 15 March to 30 March and TB Noida control room received 55,018 calls w.e.f. 16 March to 30 March, 2020. This prompt action of the center to set up a control room at the NCDC gave the states enough grace period to train their staff and start their individual help lines for addressing people's queries and allay fears.

Detection of African genotype in Hyalomma tick pools during Crimean Congo hemorrhagic fever outbreak, Rajasthan, India, 2019.

Crimean Congo hemorrhagic fever (CCHF) is a zoonotic viral disease presenting with fever and hemorrhagic manifestations in humans. After several outbreaks of CCHF being reported from Gujarat since 2011 till 2019 and from Rajasthan in 2014 and 2015, the present study reports the CCHF outbreak which was recorded from five human cases in three districts Jodhpur, Jaisalmer, and Sirohi of Rajasthan state since August 2019 till November 2019. A high percent of positivity was recorded in livestock animal samples for the CCHFV IgG antibody. CCHF virus (CCHFV) positive human blood samples and Hyalomma tick pool samples were sequenced using next-generation sequencing method. Two different M segment genotypes, encoding glycoprotein precursor, were identified from tick pools in the study: first from Asian and second from African lineage. The L gene (polymerase) and the S gene (nucleocapsid) clustered in the Asian lineage. The present study illustrates the existence of two different CCHFV lineages being circulating within the Hyalomma tick pools in the Rajasthan state, India. We also observed 3.56% amino acid changes between the death and the survived case of CCHFV in the M gene. This report also sets an alarm to enhance human, tick and livestock surveillance in other districts of Rajasthan and nearby states of India. Biosafety measures, barrier nursing along with the availability of personal protective equipment and ribavirin drug will always be a mainstay in preventing nosocomial infection for proper case management.

Therapeutic potential of functionalized siRNA nanoparticles on regression of liver cancer in experimental mice.

Short interfering RNA (siRNA) possesses special ability of silencing specific gene. To increase siRNA stability, transportation and its uptake by tumor cells, effective delivery to the appropriate target cells is a major challenge of siRNA-based therapy. In the present study, an effective, safe and biocompatible survivin siRNA encapsulated, GalNAc decorated PEGylated PLGA nanoconjugates (NCs) viz., GalNAc@PEG@siRNA-PLGA were engineered and their synergistic antitumor efficacy was evaluated for targeted delivery in HCC bearing experimental mice. GalNAc@PEG@siRNA-PLGA NCs were characterized for size, bioavailability, toxicity and biocompatibility. Their antitumor potential was evaluated considering gene silencing, apoptosis, histopathology and survival of treated mice. Exceptional accumulation of hepatocytes, reduction in survivin expression and prominent regression in tumor size confirmed the ASGPR-mediated uptake of ligand-anchored NCs and silencing of survivin gene in a targeted manner. Increased DNA fragmentation and potential modulation of caspase-3, Bax and Bcl-2 factors specified the induction of apoptosis that helped in significant inhibition of HCC progression. The potential synchronous and tumor selective delivery of versatile NCs indicated the effective payloads towards the target site, increased apoptosis in cancer cells and improved survival of treated animals.

M1 Macrophage Polarization Is Dependent on TRPC1-Mediated Calcium Entry.

Macrophage plasticity is essential for innate immunity, but in-depth signaling mechanism(s) regulating their functional phenotypes are ill-defined. Here we report that interferon (IFN) γ priming of naive macrophages induces store-mediated Ca2+ entry and inhibition of Ca2+ entry impairs polarization to M1 inflammatory phenotype. In vitro and in vivo functional analyses revealed ORAI1 to be a primary contributor to basal Ca2+ influx in macrophages, whereas IFNγ-induced Ca2+ influx was mediated by TRPC1. Deficiency of TRPC1 displayed abrogated IFNγ-induced M1 inflammatory mediators in macrophages. In a preclinical model of peritonitis by Klebsiella pneumoniae infection, macrophages showed increased Ca2+ influx, which was TRPC1 dependent. Macrophages from infected TRPC1-/- mice showed inhibited expression of M1-associated signature molecules. Furthermore, in human patients with systemic inflammatory response syndrome, the level of TRPC1 expression in circulating macrophages directly correlated with M1 inflammatory mediators. Overall, TRPC1-mediated Ca2+ influx is essential for the induction/shaping of macrophage polarization to M1 inflammatory phenotype.

Galectin-3 in M2 Macrophages Plays a Protective Role in Resolution of Neuropathology in Brain Parasitic Infection by Regulating Neutrophil Turnover.

Macrophages/microglia with M2-activation phenotype are thought to play important anti-inflammatory and tissue reparative functions in the brain, yet the molecular bases of their functions in the CNS remain to be clearly defined. In a preclinical model of neurocysticercosis using brain infection with a parasite Mesocestoides corti, we previously reported the presence of large numbers of M2 cells in the CNS. In this study using female mice, we report that M2 macrophages in the parasite-infected brain display abundant galectin-3 expression. Disease severity was increased in Galectin-3-/- mice correlating with increased neurological defects, augmented cell death and, importantly, massive accumulation of neutrophils and M2 macrophages in the CNS of these mice. Because neutrophil clearance by efferocytosis is an important function of M2 macrophages, we investigated a possible role of galectin-3 in this process. Indeed, galectin-3-deficient M2 macrophages exhibited a defect in efferocytic clearance of neutrophils in vitro Furthermore, adoptive transfer of M2 macrophages from galectin-3-sufficient WT mice reduced neutrophilia in the CNS and ameliorated disease severity in parasite-infected Galectin-3-/- mice. Together, these results demonstrate, for the first time, a novel role of galectin-3 in M2 macrophage function in neutrophil turnover and resolution of inflammatory pathology in the CNS. This likely will have implications in neurocysticercosis and neuroinflammatory diseases.SIGNIFICANCE STATEMENT Macrophages/microglia with M1-activation phenotype are thought to promote CNS pathology, whereas M2-anti-inflammatory phenotype promote CNS repair. However, the mechanisms regulating M2 cell-protective functions in the CNS microenvironment are undefined. The current study reports that helminth infection of the brain induces an increased expression of galectin-3 in M2 macrophages accumulated in the CNS. Using multiple experimental models in vivo and in vitro, they show that galectin-3 in M2 macrophages functions to clear neutrophils accumulated in the CNS. Importantly, galectin-3 in M2 macrophages plays a central role in the containment of neuropathology and disease severity. These results provide a direct mechanistic evidence of the protective function of M2 macrophages in the CNS.

Ziehl-Neelsen sputum smear microscopy image database: a resource to facilitate automated bacilli detection for tuberculosis diagnosis.

Ziehl-Neelsen stained microscopy is a crucial bacteriological test for tuberculosis detection, but its sensitivity is poor. According to the World Health Organization (WHO) recommendation, 300 viewfields should be analyzed to augment sensitivity, but only a few viewfields are examined due to patient load. Therefore, tuberculosis diagnosis through automated capture of the focused image (autofocusing), stitching of viewfields to form mosaics (autostitching), and automatic bacilli segmentation (grading) can significantly improve the sensitivity. However, the lack of unified datasets impedes the development of robust algorithms in these three domains. Therefore, the Ziehl-Neelsen sputum smear microscopy image database (ZNSM iDB) has been developed, and is freely available. This database contains seven categories of diverse datasets acquired from three different bright-field microscopes. Datasets related to autofocusing, autostitching, and manually segmenting bacilli can be used for developing algorithms, whereas the other four datasets are provided to streamline the sensitivity and specificity. All three categories of datasets were validated using different automated algorithms. As images available in this database have distinctive presentations with high noise and artifacts, this referral resource can also be used for the validation of robust detection algorithms. The ZNSM-iDB also assists for the development of methods in automated microscopy.

Unusual presentation of adult Marfan syndrome as a complex diaphragmatic hiatus hernia.

Marfan syndrome is multisystem connective tissue disorder that primarily involves the skeletal, cardiovascular, and ocular systems. The gastrointestinal complications in Marfan syndrome are rare, with only a few case reports described in the literature. We present a 25-year-old woman who presented with acute abdominal pain for 1 day. The imaging features revealed complex diaphragmatic hiatus hernia with organoaxial gastric volvulus. This is a unique case report about an adult patient with Marfan syndrome who presented with symptomatic paraesophageal hernia and organoaxial gastric volvulus.

Techno-functional differentiation of two vitamin B12 producing Lactobacillus plantarum strains: an elucidation for diverse future use.

An appropriate selection of Lactobacillus strain (probiotic/starter/functional) on the basis of its techno-functional characteristics is required before developing a novel fermented functional food. We compared vitamin B12 (B12, cobalamin) producing Lactobacillus plantarum isolates, BHM10 and BCF20, for functional (vitamin over-production, genomic insight to B12 structural genes, and probiotic attributes) and technological [milks (skim and soy) fermentation and B12 bio-fortification] characteristics. Addition of B12 precursors (5-amonolevulinate and dimethylbenzimidazole) to cobalamin-free fermentation medium increased vitamin production in BHM10, BCF20, and DSM20016 (a positive standard) by 3.4-, 4.4-, and 3.86-folds, respectively. Three important B12 structural genes were detected in L. plantarum species (strains BHM10 and BCF20) by PCR for the first time. The gene sequences were submitted to NCBI GenBank and found phylogenetically closer to respective sequences in B12 producing Lactobacillus reuteri strains. During comparative probiotic testing, BCF20 showed significantly higher (p < 0.05 to p < 0.001) gastrointestinal tolerance and cell surface hydrophobicity (p < 0.05) than BHM10. Moreover, only BCF20 was found positive for BSH activity and also exhibited comparatively better antagonistic potential against potent pathogens. Conversely, high acid and bile susceptible strain BHM10 displayed significantly higher soy milk fermentation and resultant B12 bio-fortification abilities during technological testing. Two B12 quantification techniques, UFLC and competitive immunoassay, confirmed the in vitro and in situ bio-production of bio-available form of B12 after BHM10 fermentation. Conclusively, techno-functional differentiation of two B12 producing strains elucidates their diverse future use; BCF20 either for B12 over-production (in vitro) or as a probiotic candidate, while BHM10 for cobalamin bio-fortification (in situ) in soy milk.

Epigenetic Modulation of Microglial Inflammatory Gene Loci in Helminth-Induced Immune Suppression: Implications for Immune Regulation in Neurocysticercosis.

In neurocysticercosis, parasite-induced immune suppressive effects are thought to play an important role in enabling site-specific inhibition of inflammatory responses to infections. It is axiomatic that microglia-mediated (M1 proinflammatory) response causes central nervous system inflammation; however, the mechanisms by which helminth parasites modulate microglia activation remain poorly understood. Here, we show that microglia display a diminished expression of M1-inflammatory mediators such as tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and nitric oxide synthase 2 (NOS2) in murine neurocysticercosis. Microglia also exhibited a lack of myeloid cell maturation marker major histocompatibility complex (MHC)-II in these parasite-infected brains. Treatment of microglia with helminth soluble/secreted factors (HSFs) in vitro did not induce expression of M1-inflammatory signature molecule NOS2 as well as MHC-II in primary microglia. However, HSF treatment completely inhibited lipopolysaccharide-induced increase in expression of MHC-II, NOS2 and nitric oxide production in these cells. As epigenetic modulation of chromatin states that regulates recruitment of RNA polymerase II (Pol-II) is a key regulatory step in determining gene expression and functional outcome, we next evaluated whether HSF induced modulation of these phenomenon in microglia in vitro. Indeed, HSF downregulated Pol-II recruitment to the promoter region of TNF-α, IL-6, NOS2, MHC-II, and transcription factor CIITA (a regulator of MHC-II expression), by itself. Moreover, HSF suppressed the lipopolysaccharide-induced increase in Pol-II recruitment as well. In addition, HSF exposure reduced the positive histone marks H3K4Me3 and H3K9/14Ac at the promoter of TNF-α, IL-6, NOS2, MHC-II, and CIITA. These studies provide a novel mechanistic insight into helminth-mediated immune suppression in microglia via modulation of epigenetic processes.