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To understand functional duality of the complement system in host defense and lung injury, a more comprehensive view of its localized production in the lung, and the impact of age on complement production are essential. Here, we explored the expression of complement genes through computational analysis of preexisting single cell RNA sequencing data from lung transcriptomes of healthy young (3 months) and old C57BL/6 mice (24 months), and humans. We characterized the distribution of 48 complement genes. Across 28 distinct immune and non-immune cell types in mice, mesothelial cells expressed the greatest number of complement genes (e.g., C1ra, C2, C3), and regulators (e.g., Serping1, Cfh). C5 was abundant in type II alveolar epithelial cells and C1q in interstitial lung macrophages. There were only moderate differences in gene expression between young and old mice. Among 57 human lung cell types, mesothelial cells showed abundant complement expression. A few differences in gene expression (e.g., FCN1, CFI, C6, C7) were also evident between mice and human lung cells. Our findings present a novel perspective on the expression patterns of complement genes in normal lungs. These findings highlight the potential functions of complement in tissue-specific homeostasis and immunity and may foster a mechanistic understanding of its role in lung health and disease.

Coronavirus disease-2019 (COVID-19) provokes a hypercoagulable state with increased incidence of thromboembolism and mortality. Platelets are major effectors of thrombosis and hemostasis. Suitable animal models are needed to better understand COVID-19-associated coagulopathy (CAC) and underlying platelet phenotypes. Here, we assessed K18-hACE2 mice undergoing a standardized SARS-CoV-2 infection protocol to study dynamic platelet responses via mass spectrometry-based proteomics. In total, we found significant changes in >1,200 proteins. Strikingly, protein alterations occurred rapidly by 2 days post-infection (dpi) and preceded outward clinical signs of severe disease. Pathway enrichment analysis of 2dpi platelet proteomes revealed that SARS-CoV-2 infection upregulated complement-coagulation networks (F2, F12, CFH, CD55/CD59), platelet activation-adhesion-degranulation proteins (PF4, SELP, PECAM1, HRG, PLG, vWF), and chemokines (CCL8, CXCL5, CXCL12). When mice started to lose weight at 4dpi, pattern recognition receptor signaling (RIG-I/MDA5, CASP8, MAPK3), and interferon pathways (IFIT1/IFIT3, STAT1) were predominant. Interestingly, SARS-CoV-2 spike protein in the lungs was observed by immunohistochemistry, but in platelets was undetected by proteomics. Similar to patients, K18-hACE2 mice during SARS-CoV-2 infection developed progressive lymphohistiocytic interstitial pneumonia with platelet aggregates in the lungs and kidneys. In conclusion, this model recapitulates activation of coagulation, complement, and interferon responses in circulating platelets, providing valuable insight into platelet pathology during COVID-19.Key PointsSARS-CoV-2-infected humanized ACE2 mice recapitulate platelet reprogramming towards activation-degranulation-aggregation.Complement/coagulation pathways are dominant in platelets at 2 days post-infection (dpi), while interferon signaling is dominant at 4dpi.

The complete mitochondrial genome of Eld's deer ( Rucervus eldii eldii ) has been sequenced, assembled and characterized for the first time using 25 sets of published primers. The Eld's deer, famously known as Sangai or Manipur's brow-antlered deer belongs to the family Cervidae and is critically endangered species in India which needs immediate attention and concern for wildlife conservation. The 16,357 bp long complete mitogenome of R. e. eldii was very similar to most vertebrates in organization with 13 protein-coding genes, 22 transfer RNA, 2 ribosomal RNA and a typical non-coding control region. Its protein-coding region exhibits codon usage common to all mammals. A detailed characterization of the available mitogenomes of Artiodactyla suggested conserved nature of gene organization, codon usage, gene orientation and evolutionary rates of proteins. A detailed characterization of the available mitogenomes of Artiodactyla, suggested conserved nature of gene organization, codon usage, gene orientation and evolutionary rates of proteins. The complete mitogenome and protein-coding genes of R. e. eldii were found to be highly A + T biased. Rate of protein evolution was highest in nad6 and lowest in cox1 . The phylogenetic analyses based on concatenated amino acid and nucleotide sequences of 13 protein-coding genes provided a well-supported broader outline of Cervidae, which agree with the previously described traditional morphological classification. Besides, the well supported phylogeny of artiodactyls mitochondrial genomes helped to resolve relationships between Rucervus eldii eldii (India) and Rucervus eldii hainanus (China) being the other two Eld's deer species remains unresolved i.e. Rucervus eldii thamin (Myanmar) Rucervus eldii siamensis (Thailand). The whole mitogenome sequence of R. e. eldii will significantly benefit systematics, evolutionary studies, biogeography and conservation programs of R. e. eldii and the family Cervidae.

Several reports document the role of tumor necrosis factor alpha (TNF-α) and lipid metabolism in the context of acute inflammation as a causative factor in obesity-associated insulin resistance and as one of the causative parameter of type 2 diabetes mellitus (T2DM). Our aim was to investigate the association between −308G/A and −238G/A polymorphisms located in the promoter region of the TNF-α gene in T2DM in the Indian population with bioinformatics analysis of TNF-α protein networking with an aim to find new target sites for the treatment of T2DM. Demographics of 100 diabetes patients and 100 healthy volunteers were collected in a structured proforma and 3 ml blood samples were obtained from the study group, after approval of Institutional Ethics Committee of the hospital (IEC). The information on clinical parameters was obtained from medical records. Genomic DNA was extracted; PCR–RFLP was performed using TNF-α primers specific to detect the presence of SNPs. Various bioinformatics tools such as STRING software were used to determine its network with other associated genes. The PCR–RFLP studies showed that among the −238G/A types the GG genotype was 87%, GA genotype was 12% and AA genotype was 1%. Almost a similar pattern of results was obtained with TNF-α −308G/A polymorphism. The results obtained were evaluated statistically to determine the significance. By constructing TNF-α protein interaction network we could analyze ontology and hubness of the network to identify the networking of this gene which may influence the functioning of other genes in promoting T2DM. We could identify new targets in T2DM which may function in association with TNF-α. Through hub analysis of TNF-α protein network we have identified three novel proteins RIPK1, BIRC2 and BIRC3 which may contribute to TNF-mediated T2DM pathogenesis. In conclusion, our study indicated that some of the genotypes of TNF-α −308G/A, −238G/A were not significantly associated to type 2 diabetes mellitus, but TNF-α −308G/A polymorphism was reported to be a potent risk factor for diabetes in higher age (>45) groups. Also, the novel hub proteins may serve as new targets against TNF-α T2DM pathogenesis. Keywords: TNF-α, Type 2 diabetes mellitus, Polymorphism, Genotypes, SNPs, Protein interaction network

There is a need to identify novel targets in Acute Lymphoblastic Leukemia (ALL), a hematopoietic cancer affecting children, to improve our understanding of disease biology and that can be used for developing new therapeutics. Hence, the aim of our study was to find new genes as targets using in silico studies; for this we retrieved the top 10% overexpressed genes from Oncomine public domain microarray expression database; 530 overexpressed genes were short-listed from Oncomine database. Then, using prioritization tools such as ENDEAVOUR, DIR and TOPPGene online tools, we found fifty-four genes common to the three prioritization tools which formed our candidate leukemogenic genes for this study. As per the protocol we selected thirty training genes from PubMed. The prioritized and training genes were then used to construct STRING functional association network, which was further analyzed using cytoHubba hub analysis tool to investigate new genes which could form drug targets in leukemia. Analysis of the STRING protein network built from these prioritized and training genes led to identification of two hub genes, SMAD2 and CDK9, which were not implicated in leukemogenesis earlier. Filtering out from several hundred genes in the network we also found MEN1, HDAC1 and LCK genes, which re-emphasized the important role of these genes in leukemogenesis. This is the first report on these five additional signature genes in leukemogenesis. We propose these as new targets for developing novel therapeutics and also as biomarkers in leukemogenesis, which could be important for prognosis and diagnosis.

MDR1, a protein commonly involved in drug transport, has been linked to multi drug resistance and disease progression in cancers such as non-small cell lung cancer. Hence, targeting this protein is essential for improving drug design and preventing adverse drug-drug interactions. The aim of the study was to examine chemotherapeutic drug binding to MDR1 and the interactions therein. We have used Schrodinger suite 2014, to perform homology modelling of human MDR1 based on Mouse MDR1, followed by Induced Fit Docking with Paclitaxel, Docetaxel, Gemcitabine, Carboplatin and Cisplatin drugs. Finally, we evaluated drug binding affinities using Prime/MMGBSA and using these scores we compared the affinities of combination therapies against MDR1. Analysis of the docking results showed Paclitaxel>Docetaxel>Gemcitabine>Carboplatin>Cisplatin as the order of binding affinities, with Paclitaxel having the best docking score. The combination drug binding affinity analysis showed Paclitaxel+Gemcitabine to have the best docking score and hence, efficacy. Through our investigation we have identified the residues Gln 195 and Gln 946 to be more frequently involved in drug binding interactions with MDR1. Our results suggest that, Paclitaxel or combination of Paclitaxel+Gemcitabine could serve as a suitable therapy against MDR1 in NSCLC patients. Thus, our study provides new insight into the possible repurposing of chemotherapeutic drugs in targeting elevated MDR1 levels in NSCLC patients, thereby ensuring better overall outcome. Further our study highlights the use of in silico methodologies in understanding drug binding to protein targets and its relevance to advancing lung cancer therapy.

Atmospheric convection is a natural phenomena associated with heat transport. Convection is strong during daylight periods and rigorous in summer months. Severe ground heating associated with strong winds experienced during these periods. Tropics are considered as the source regions for strong convection. Formation of thunder storm clouds is common during this period. Location of cloud base and its associated dynamics is important to understand the influence of convection on the atmosphere. Lidars are sensitive to Mie scattering and are the suitable instruments for locating clouds in the atmosphere than instruments utilizing the radio frequency spectrum. Thunder storm clouds are composed of hydrometers and strongly scatter the laser light. Recently, a lidar technique was developed at National Atmospheric Research Laboratory (NARL), a Department of Space (DOS) unit, located at Gadanki near Tirupati. The lidar technique employs slant path operation and provides high resolution measurements on cloud base location in real-time. The laser based remote sensing technique allows measurement of atmosphere for every second at 7.5 m range resolution. The high resolution data permits assessment of updrafts at the cloud base. The lidar also provides real-time convective boundary layer height using aerosols as the tracers of atmospheric dynamics. The developed lidar sensor is planned for up-gradation with scanning facility to understand the cloud dynamics in the spatial direction. In this presentation, we present the lidar sensor technology and utilization of its technology for high resolution cloud base measurements during convective conditions over lidar site, Gadanki.

A compact dual polarization lidar (DPL) was designed and developed at National Atmospheric Research Laboratory (NARL) for daytime measurements of the boundary layer aerosol distribution and depolarization properties with very high vertical and temporal resolution. The lidar employs a compact flashlamp pumped Q-switched Nd:YAG laser and operates at 532 nm wavelength. The lidar system uses a stable biaxial configuration between transmitter and receiver units. The receiver utilizes a 150 mm Schmidt Cassegranin telescope for collecting laser returns from the atmosphere. The collected backscattered light is separated into co and cross-polarization signals using a polarization beam splitter cube. A set of mini-PMTs have been used for detection of light from atmosphere during daylight period. A two channel transient recorder system with built-in ADC has been employed for recording the detected light. The entire lidar system is housed in a compact cabinet which can be easily transported for field measurements. During 2014, the lidar system was installed at the Banaras Hindu University (BHU) campus, Varanasi (25.28° N, 82.96° E, 82 m AMSL) and operated for a period of three months in to support the cloud aerosol interaction and precipitation enhancement experiment (CAIPEEX) conducted by Indian Institute of tropical meteorology (IITM). During this campaign period, the lidar measurements were carried out in the vertical direction with spatial resolution of 7.5 m and time sampling of 30s. The lidar measurements revealed the occurrence of boundary layer growth during convective periods and also detected the long-range transport dust layers with significant depolarization. In the present paper, we present the lidar measurements obtained during the campaign period and discuss the observation of transport of dust layer over the experimental site with support of back trajectory analysis and satellite data. The Lidar observations were compared with the available satellite observations also presented here.

α2-adrenergic receptors play a key role in the regulation of sympathetic system, neurotransmitter release, blood pressure and intraocular pressure. Although α2-adrenergic receptors mediate a number of physiological functions in vivo and have great therapeutic potential, the absence of crystal structure of α2-adrenergic receptor subtypes is a major hindrance in the drug design efforts. The therapeutic efficacy of the available drugs is not selective for subtype specificity (α2a, α2b and α2c) leading to unwanted side effects. We used Homology modelling and docking studies to understand and analyze the residues important for agonist and antagonist binding. We have also analyzed binding site volume, and the residue variations which may play important role in ligand binding. We have identified residues through our modelling and docking studies, which would be critical in giving subtype specificity and may help in the development of future subtype-selective drugs.

[Godavarthi S., Jayaraman A. and Gaur A. 2011 Cross-species amplification of human microsatellite markers in pig-tailed and stump-tailed macaques. J. Genet.90, e6–e9. Online only: http://www.ias.ac.in/jgenet/OnlineResources/90/e6.pdf ]

Alterations in cell cycle regulating proteins are a key characteristic in neoplastic proliferation of lymphoblast cells in patients with Acute Lymphoblastic Leukemia (ALL). The aim of our study was to investigate whether the routinely administered ALL chemotherapeutic agents would be able to bind and inhibit the key deregulated cell cycle proteins such as - Cyclins E1, D1, D3, A1 and Cyclin Dependent Kinases (CDK) 2 and 6. We used Schrödinger Glide docking protocol to dock the chemotherapeutic drugs such as Doxorubicin and Daunorubicin and others which are not very common including Clofarabine, Nelarabine and Flavopiridol, to the crystal structures of these proteins. We observed that the drugs were able to bind and interact with cyclins E1 and A1 and CDKs 2 and 6 while their docking to cyclins D1 and D3 were not successful. This binding proved favorable to interact with the G1/S cell cycle phase proteins that were examined in this study and may lead to the interruption of the growth of leukemic cells. Our observations therefore suggest that these drugs could be explored for use as inhibitors for these cell cycle proteins. Further, we have also highlighted residues which could be important in the designing of pharmacophores against these cell cycle proteins. This is the first report in understanding the mechanism of action of the drugs targeting these cell cycle proteins in leukemia through the visualization of drug-target binding and molecular docking using computational methods.