Biology subjects

Bhatt, S.

Publications and source records attributed to Bhatt, S..

32 records · Page 2Linked to original sources

bioRxiv2022

The cost of sympatry: spatio-temporal patterns in leopard dietary and physiological responses to tiger competition in Rajaji Tiger Reserve, India

1.O_LIApex predators have critical roles in maintaining the structure of ecosystem functioning by controlling intraguild subordinate populations. Such dominant-subordinate interactions involve agonistic interactions including direct (death/displacement) or indirect (physiological and/or health implications) impacts on the subordinates. As these indirect effects are often mediated through physiological processes, it is important to quantify such responses for better understanding of population parameters. C_LIO_LIWe used a well-known sympatric large carnivore intraguild system involving tiger (Panthera tigris) and leopard (Panthera pardus) to understand the dietary and physiological responses under a spatio-temporal gradient of tiger competition pressures in Rajaji Tiger Reserve (RTR), a major source tiger population of the western Terai-Arc Landscape, India between 2015-2020. The park provided a natural experimental set-up for tiger competition with the eastern part (ERTR) having high tiger density and the western (WRTR) part with functionally no competition from tigers. C_LIO_LIWe conducted systematic faecal sampling in the winters of 2015 and 2020 from ERTR and WRTR to assess diet and physiological measures. Analyses of leopard-confirmed faeces suggest a dietary-niche separation as a consequence of tiger competition. In 2020, we found increased occurrence of large-bodied prey species without tiger competition in WRTR. Physiological measures followed the dietary responses where leopards with large-sized prey in diet showed higher fT3M and lower fGCM measures in WRTR. In contrast, ERTR leopards showed lower levels of fT3M as well as fGCM in 2020, possibly due to intense competition from tigers. Overall, these pattens strongly indicate a physiological cost of sympatry where competition with dominant tigers resulted in elevated nutritional stress. C_LIO_LIThe combination of the natural habitat providing unique experimental setup, spatio-temporal sampling strategy and multidisciplinary approaches provide critical conservation perspectives for leopards, particularly in the context of recent increase in tiger numbers across India. We recommend expansion of leopard monitoring and population estimation efforts to buffers, developing appropriate plans for human-leopard conflict mitigation and intensive efforts to understand leopard population dynamics patterns to ensure their persistence during the ongoing Anthropocene. C_LI

physiology↗

bioRxiv2021

BH3 Profiling Identifies Selective BCL-2 Dependence of Adult Early T-cell Progenitor (ETP) Subtype of Acute Lymphoblastic Leukemia (ALL) Patients

Early T-cell precursor (ETP) acute lymphoblastic leukemia (T-ALL) is a high-risk subtype of T-ALL and is associated with poor survival outcomes with chemotherapy. We previously showed that maturation stage of thymocytes distinguishes pro-survival dependencies of pediatric ETP-ALL (BCL-2 dependent) from non-ETP-ALL (BCL-XL dependent). Comparable data in adults are lacking. Our present study focuses on characterizing functional dependencies of adult ETP-ALL and T-ALL on BCL-2 family proteins. Using BH3 profiling on primary tumors, we report that similar to pediatric ALL, adult ETP-ALL is primarily dependent on BCL-2; however, unlike pediatric ALL, adult non-ETP-ALL is co-dependent on both BCL-2 and BCL-XL for survival. By measuring direct mitochondrial permeabilization and cell viability assays, we validated that BH3 profiling predicted on-target cytotoxicity of venetoclax in adult ETP-ALL and navitoclax plus venetoclax in adult T-ALL. These findings provide pre-clinical evidence for venetoclax and navitoclax as potentially efficacious combination therapy for adults with T-ALL. Statement of SignificanceAdults with relapsed T-ALL have poor prognosis and represent an unmet need for novel treatments. We report that adult ETP-ALL is dependent on BCL-2 for survival, similar to pediatric ETP-ALL. However, adult non-ETP-ALL is heterogeneously dependent on both BCL-2 and BCL-XL. We characterize key dependencies on BCL-2 family proteins to direct BH3 mimetics therapy in T-ALL patients.

cancer biology↗

bioRxiv2021

AzuCR RNA modulates carbon metabolism as a dual-function RNA in Escherichia coli

Bacteria have evolved small RNAs (sRNAs) to regulate numerous biological processes and stress responses. While sRNAs generally are considered to be "noncoding", a few have been found to also encode a small protein. Here we describe one such dual-function RNA that modulates carbon utilization in Escherichia coli. The 164 nucleotide RNA was previously shown to encode a 28 amino acid protein (denoted AzuC). We discovered the membrane-associated AzuC protein interacts with GlpD, the aerobic glycerol-3-phosphate dehydrogenase, leading to increased GlpD activity. Overexpression of the RNA encoding AzuC results in a growth defect in glycerol and galactose medium. The defect in galactose medium was still observed for a stop codon mutant derivative, suggesting a potential regulatory role for the RNA. Consistent with this observation, we found that cadA and galE are repressed by base pairing with the RNA (denoted AzuCR). Interestingly, translation of AzuC interferes with the observed repression of cadA and galE by AzuCR and base pairing interferes with AzuC translation, demonstrating that the translation and base pairing functions compete.

microbiology↗

bioRxiv2020

Effects of faecal inorganic contents in accurate measures of stress and nutrition hormone in large felines: implications for physiological assessments in free-ranging animals

Non-invasive stress and nutritional hormones and their interactions are increasingly being used to monitor psychological and nutritional physiology in free-ranging animals at different ecological scales. However, a number of extrinsic and intrinsic factors including hormone-inert dietary materials, inorganic matters etc. are known to affect accurate hormone measures. Here we addressed the impacts of inorganic matter (IOM) on corticosterone and T3 measures in wild tiger (n=193 from Terai Arc landscape, India) and captive lion (n=120 from Sakkarbaug Zoological Garden, Gujarat, India) faeces and evaluated possible corrective measures. The wild tiger samples contained highly variable IOM content (9-98%, mostly with >40% IOM) compared to captive Asiatic lion (17-57%, majority with <40% IOM). We observed significant negative correlation between IOM content and tiger GC (r=-0.48, p=0.000) and T3 (r=-0.60, p=0.000) measures but not in captive lions (r=-0.05, p=0.579). Two corrective measures viz. removing samples with [≥]80% IOM and subsequently expressing concentrations as per gram of organic dry matter (instead of total dry matter) reduced IOM influence on tiger GC and T3 measures without affecting lion GC results. The corrective measures bring out alterations in the tiger T3 results but no changes in GC results. As faecal IOM content is associated with specific behaviours of many carnivore species, our results emphasize the need to reduce IOM-driven hormone data variation for ecologically relevant interpretations for species conservation.

physiology↗

bioRxiv2020

Assessing tiger corridor functionality with landscape genetics and modelling across Terai-Arc Landscape, India

India led the global tiger conservation initiatives since last decade and has doubled its wild tiger population to 2967 (2603-3346). The survival of these growing populations residing inside the continuously shrinking habitats is a major concern, which can only be tackled through focused landscape-scale conservation planning across five major extant Indian tiger landscapes. The Terai-Arc landscape (TAL) is one of the global priority tiger conservation landscapes holding 22% of the countrys wild tigers. We used intensive field-sampling, genetic analyses and GIS modelling to investigate tiger population structure, source-sink dynamics and functionality of the existing corridors across TAL. Genetic analyses with 219 tigers revealed three low, but sigficantly differentiated tiger subpopulations. Overall, we identified Seven source and 10 sink areas in TAL through genetic migrant and gene flow analyses. GIS modelling identified total 19 (10 high, three medium and six low conductance) corridors in this landscape, with 10 being critical to maintain landscape connectivity. We suggest urgent management attention towards 2707 sq. km. non-protected habitat, mitigation measures associated with ongoing linear infrastructure developments and transboundary coordination with Nepal to ensure habitat and genetic connectivity and long-term sustainability of tigers in this globally important landscape.

ecology↗

bioRxiv2020

MAPPING TRENDS IN INSECTICIDE RESISTANCE PHENOTYPES IN AFRICAN MALARIA VECTORS

Mitigating the threat of insecticide resistance in African malaria vector populations requires comprehensive information about where resistance occurs, to what degree, and how this has changed over time. Estimating these trends is complicated by the sparse, heterogeneous distribution of observations of resistance phenotypes in field populations. We use 6423 observations of the prevalence of resistance to the most important vector control insecticides to inform a Bayesian geostatistical ensemble modelling approach, generating fine-scale predictive maps of resistance phenotypes in mosquitoes from the Anopheles gambiae complex across Africa. Our models are informed by a suite of 111 predictor variables describing potential drivers of selection for resistance. Our maps show alarming increases in the prevalence of resistance to pyrethroids and DDT across Sub-Saharan Africa from 2005-2017 as well as substantial spatial variation in resistance trends.

ecology↗

bioRxiv2019

Genetic analyses reveal population structure and recent decline in leopards (Panthera pardus fusca) across Indian subcontinent

Large carnivores maintain the stability and functioning of ecosystems. Currently, many carnivore species face declining population sizes due to natural and anthropogenic pressures. The leopard, Panthera pardus, is probably the most widely distributed and adaptable large carnivore, still persisting in most of its historic range. However, we lack subspecies level data on country or regional scale on population trends, as ecological monitoring approaches are difficult to apply on such wide-ranging species. We used genetic data from leopards sampled across the Indian subcontinent to investigate population structure and patterns of demographic decline. Our genetic analyses revealed four distinct subpopulations corresponding to Western Ghats, Deccan Plateau-Semi Arid, Shivalik and Terai region of north Indian landscapes, each with high genetic variation. Coalescent simulations with 13 microsatellite loci revealed a 75-90% population decline in between 120-200 years ago across India, possibly human induced. Population-specific estimates of genetic decline are in concordance with ecological estimates of local extinction probabilities in four sub-populations obtained from occupancy modelling of historic and current distribution of leopards in India. Our results confirm population decline of a widely distributed, adaptable large carnivore. We re-iterate the relevance of indirect genetic methods for such species, and recommend that detailed, landscape-level ecological studies on leopard populations are critical to future conservation efforts. Our approaches and inference are relevant to other widely distributed, seemingly unaffected carnivores such as the leopard.

genetics↗

bioRxiv2019

Co-localization and confinement of ecto-nucleotidases modulate extracellular adenosine nucleotide pools

1Nucleotides comprise small molecules that perform critical signaling and energetic roles in biological systems. Of these, the concentrations of adenosine and its derivatives, including adenosine tri-, di-, and mono-phosphate are dynamically controlled in the extracellular-space by diphosphohydrolases and ecto-nucleotidases that rapidly degrade such nucleotides. In many instances, the close coupling between cells such as those in synaptic junctions yields tiny extracellular nanodomains, within which the charged nucleotides interact with densely-packed membranes and biomolecules. While the contributions of electrostatic and steric interactions within such nanodomains are known to shape diffusion-limited reaction rates, less is understood about how these factors control the kinetics of sequentially-coupled diphosphohydrolase/nucleotidase-catalyzed reactions. To rank the relative importance of these factors, we utilize reaction-diffusion numerical simulations to systematically probe coupled enzyme activity in narrow junctions. We perform these simulations in nanoscale geometries representative of narrow extracellular compartments, within which we localize sequentially- and spatially-coupled enzymes. These enzymes catalyze the conversion of a representative charged substrate such as adenosine triphosphate (ATP) into substrates with different net charges, such as adenosine monophosphate (AMP) and adenosine (Ado). Our modeling approach considers electrostatic interactions of diffusing, charged substrates with extracellular membranes, and coupled enzymes. With this model, we find that 1) Reaction rates exhibited confinement effects, namely reduced reaction rates relative to bulk, that were most pronounced when the enzyme was close to the pore size and 2) The presence of charge on the pore boundary further tunes reaction rates by controlling the pooling of substrate near the reactive protein akin to ions near trans-membrane proteins. These findings suggest how remarkable reaction efficiencies of coupled enzymatic processes can be supported in charged and spatially-confined volumes of extracellular spaces.

biophysics↗

bioRxiv2019

Tracking progress towards malaria elimination in China: estimates of reproduction numbers and their spatiotemporal variation

China reported zero locally-acquired malaria cases in 2017 and 2018. Understanding the spatio-temporal pattern underlying this decline, especially the relationship between locally-acquired and imported cases, can inform efforts to maintain elimination and prevent re-emergence. This is particularly pertinent in Yunnan province, where the potential for local transmission is highest. Using a geo-located individual-level dataset of cases recorded in Yunnan province between 2011 and 2016, we jointly estimate the case reproduction number, Rc, and the number of unobserved sources of infection. We use these estimates within spatio-temporal geostatistical models to map how transmission varied over time and space, estimate the timeline to elimination and the risk of resurgence. Our estimates suggest that, maintaining current intervention efforts, Yunnan is unlikely to experience sustained local transmission up to 2020. However, even with a mean Rc of 0.005 projected for the year 2019, locally-acquired cases are possible due to high levels of importation.

epidemiology↗

bioRxiv2019

In vitro transposon targeting using a catalytically inactive Cas9

Homology directed genome engineering is limited by transgene size. Although DNA transposons are more efficient with large transgenes, random integrations are potentially mutagenic. Catalytically inactive Cas9 is attractive candidate for targeting a transposase fusion-protein because of its high specificity and affinity for its binding site. Here we demonstrate efficient Cas9 targeting of a mariner transposon. Targeted integrations were tightly constrained at two adjacent TA dinucleotides about 20 bp to one side of the gRNA binding site. Biochemical analysis of the nucleoprotein complexes demonstrated that the transposase and Cas9 moieties of the fusion protein can bind their respective substrates independently. In the presence of the Cas9 target DNA, kinetic analysis revealed a delay between first and second strand cleavage at the transposon end. This step involves a significant conformational change that may be hindered by the properties of the interdomainal linker. Otherwise, the transposase behaved normally and was proficient for integration in vitro and in vivo.

biochemistry↗

bioRxiv2019

Iron nanoparticle-labeled murine mesenchymal stromal cells in an osteoarthritic model persists and demonstrates anti-inflammatory mechanism of action

Osteoarthritis (OA) is characterized by cartilage degradation and chronic joint inflammation. Mesenchymal stem cells (MSCs) have shown promising results in OA, but their mechanism of action is not fully understood. We hypothesize that MSCs polarize macrophages, which are strongly associated with joint inflammation to more homeostatic sub-types. We tracked ferumoxytol (Feraheme, iron oxide nanoparticle)-labeled murine MSCs (Fe-MSCs) in murine OA joints, and quantified changes to homeostatic macrophages.\n\n10-week-old C57BL/6 male mice (n=5/group) were induced to undergo osteoarthritis by destabilization of medical meniscus (DMM) or sham surgery. 3 weeks post-surgery, mice were injected intra-articularly with either fluorescent dye-(DiR) labeled or DiR+ferumoxytol-labeled (DiR+Fe) bone marrow mesenchymal stem cells (MSC, 50x103 MSCs/mouse) or saline (control), to yield 4 groups (n=5 per group for each timepoint [1, 2 and 4weeks]): i) DMM+Saline; ii) DMM+DiR+Fe-MSC; iii) DMM+DiR MSC; iv) SHAM+DiR+Fe-MSC and saline in contralateral knee. 4 weeks after injection, mice were imaged by MRI, and scored for i) OARSI to determine cartilage damage and ii) immunohistochemical changes in CD206, F480 and Prussian Blue/Sca-1 to detect homeostatic macrophages, total macrophages and ferumoxytol-labeled MSCs respectively.\n\nFerumoxytol-labeled MSCs persisted in DMM knee joints at greater levels than in SHAM-MSC knee joints. We observed no difference in OARSI scores between MSC and vehicle groups. Sca-1 and Prussian Blue co-staining confirmed the ferumoxytol label resides in MSCs, although some ferumoxytol label was detected in proximity to MSCs in macrophages, likely due to phagocytosis of apoptotic MSCs, increasing functionality of these macrophages through MSC efferocytosis. We showed decreased MRI synovitis scores in MSC-treated compared to control animals. For the first time we show that MSC-treated OA mice had increased macrophage infiltration (p<0.08) with an increased proportion of CD206+ (homeostatic) macrophages (p<0.01), supporting our hypothesis that MSCs modulate synovial inflammation.

cell biology↗

bioRxiv2019

A practive faeces collection protocol for multidisciplinary research in wildlife science

Faecal samples have become important non-invasive source of information in wildlife biology and ecological research. Despite regular use of faeces, there is no universal protocol available for faeces collection and storage to answer various questions in wildlife biology. We collected 1408 faeces from ten different species using a dry sampling approach, and achieved 94.87% and 86.02% success rate in mitochondrial and nuclear marker amplifications. We also suggest a universal framework to use the same samples for different use. This protocol provides an easy, quick and cheap option to collect non-invasive samples from species living at different environmental conditions to answer multidisciplinary questions in wildlife biology.

ecology↗

bioRxiv2018

Estimating the burden of α-thalassaemia in Thailand using a comprehensive prevalence database for Southeast Asia

Severe forms of -thalassaemia, haemoglobin H disease and haemoglobin Barts hydrops fetalis, are an important public health concern in Southeast Asia. Yet information on the prevalence, genetic diversity and health burden of -thalassaemia in the region remains limited. We compiled a geodatabase of -thalassaemia prevalence and genetic diversity surveys and, using geostatistical modelling methods, generated the first continuous maps of -thalassaemia mutations in Thailand and sub-national estimates of the number of newborns with severe forms in 2020. We also summarised the current evidence-base for -thalassaemia prevalence and diversity for the region. We estimate that 3,595 (95% credible interval 1,717 - 6,199) newborns will be born with severe -thalassaemia in Thailand in 2020, which is considerably higher than previous estimates. Accurate, fine-scale epidemiological data are necessary to guide sustainable national and regional health policies for -thalassaemia control. Our maps and newborn estimates are an important first step towards this aim.\n\nFundingThis work was supported by European Unions Seventh Framework Programme (FP7//2007-2013)/European Research Council [268904 - DIVERSITY]

epidemiology↗

bioRxiv2018

Antimicrobial Efficacy and Safety of a Novel Gas Plasma-Activated Catheter Lock Solution

Antimicrobial lock solutions are important for prevention of microbial colonization and infection of long-term central venous catheters. We investigated the efficacy and safety of a novel antibiotic-free lock solution formed from gas plasma-activated disinfectant (PAD). Using a luminal biofilm model, viable cells of methicillin-resistant Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, and Candida albicans in mature biofilms were reduced by 6 - 8 orders of magnitude with a PAD lock for 60 minutes. Subsequent 24-hour incubation of PAD-treated samples resulted in no detectable regrowth of viable bacteria or fungi. As a comparison, the use of a minocycline/EDTA/ethanol lock solution for 60 minutes led to regrowth of bacteria and fungi, up to 107 - 109 CFU/ml, in 24 hours. The PAD lock solution had minimal impact on human umbilical vein endothelial cell viability, whereas the minocycline/EDTA/ethanol solution elicited cell death in nearly half of human endothelial cells. Additionally, PAD treatment caused little topological change to catheter materials. In conclusion, PAD represents a novel antibiotic-free, non-cytotoxic lock solution that elicits rapid and broad-spectrum eradication of biofilm-laden microbes and which shows promise for the prevention and treatment of intravascular catheter infections.

microbiology↗