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Das, P.

Publications and source records attributed to Das, P..

At least 73 records · Page 4Linked to original sources

A dynamic generative model can extract interpretable oscillatory components from multichannel neurophysiological recordings

Modern neurophysiological recordings are performed using multichannel sensor arrays that are able to record activity in an increasingly high number of channels numbering in the 100s to 1000s. Often, underlying lower-dimensional patterns of activity are responsible for the observed dynamics, but these representations are difficult to reliably identify using existing methods that attempt to summarize multivariate relationships in a post-hoc manner from univariate analyses, or using current blind source separation methods. While such methods can reveal appealing patterns of activity, determining the number of components to include, assessing their statistical significance, and interpreting them requires extensive manual intervention and subjective judgement in practice. These difficulties with component selection and interpretation occur in large part because these methods lack a generative model for the underlying spatio-temporal dynamics. Here we describe a novel component analysis method anchored by a generative model where each source is described by a bio-physically inspired state space representation. The parameters governing this representation readily capture the oscillatory temporal dynamics of the components, so we refer to it as Oscillation Component Analysis (OCA). These parameters - the oscillatory properties, the component mixing weights at the sensors, and the number of oscillations - all are inferred in a data-driven fashion within a Bayesian framework employing an instance of the expectation maximization algorithm. We analyze high-dimensional electroencephalography and magnetoencephalography recordings from human studies to illustrate the potential utility of this method for neuroscience data. Significance StatementNeuroscience studies often involve simultaneous recordings in a large number of sensors in which a smaller number of dynamic components generate the complex spatio-temporal patterns observed in the data. Current blind source separation techniques produce sub-optimal results and are difficult to interpret because these methods lack an appropriate generative model that can guide both statistical inference and interpretation. Here we describe a novel component analysis method employing a dynamic generative model that can decompose high-dimensional multivariate data into a smaller set of oscillatory components are learned in a data-driven way, with parameters that are immediately interpretable. We show how this method can be applied to neurophysiological recordings with millisecond precision that exhibit oscillatory activity such as electroencephalography and magnetoencephalography.

neuroscience↗

Chimeric identities and reduced stiffness characterise the shoot apex of Arabidopsis stem cell mutants

Stem cell homeostasis in the shoot apical meristem involves a core regulatory feedback loop between the signalling peptide CLAVATA3, produced in stem cells, and the transcription factor WUSCHEL, expressed in the underlying organising centre. clavata mutants display massive meristem overgrowth, which is thought to be caused by stem cell overproliferation, although it is unknown how uncontrolled stem cell divisions lead to this altered morphology. Here we first reveal local buckling defects in mutant meristems, and use analytical models to show how mechanical properties and growth rates may contribute to the phenotype. Indeed, clavata meristems are mechanically more heterogeneous than the wild type, and also display regional growth heterogeneities. Furthermore, stereotypical wild-type meristem organisation is lost in mutants, in which cells simultaneously express distinct fate markers. Finally, cells in mutant meristems are auxin responsive, suggesting that they are functionally different from wild-type stem cells. Thus all benchmarks show that clavata meristem cells are different from wild-type stem cells, suggesting that fasciation is caused by the disruption of a more complex regulatory framework that maintains distinct genetic and functional domains at the shoot apex. Summary statementHeterogeneities in cell mechanics, growth, function and identity contribute to buckling in clavata mutant shoot apices.

plant biology↗

Landing the 'Tiger of Rivers': Understanding Recreational Angling of Mahseers in India using YouTube Videos

Megafish mahseers popularly known as the tiger of rivers, are the dream catch of recreational anglers in India. The present study explored the Recreational Angling (RA) videos of five mahseer species Tor khudree (deccan mahseer), T. putitora (golden mahseer), T. remadevii (humpback mahseer), T. mosal (mosal mahseer) and Neolissochilus hexagonolepis (chocolate mahseer) recorded from India and uploaded on the social media platform YouTube from January 2010 to October 2022. We did not come across any RA videos of T. mosal and T. remadevii on YouTube hence further analyses were carried out on the remaining three focal species. No seasonality was observed in the frequency of RA videos uploaded on YouTube and T. khudree attracted the highest number of views per video. Catch and Release (C&R), an ethical RA practice was noticeably low in the case of N. hexagonolepis. The size of the catch was found to be positively associated with the social engagement received by the RA videos of all the three mahseer species focused. Angler and angling-related remarks and words associated with the emotion trust dominated the comments received by the videos. The results are discussed in light of the trending discourses on developing social media data as a complementary tool for monitoring and managing RA and conserving fish.

ecology↗

Discovery & Evaluation of novel fluorescence molecules for selective recognition of G-quadruplexes structure

Currently, G-quadruplex structure targeting strategies are considered as a promising anticancer approach. In the search of selective and potent G-quadruplex binders, Here we discuss an analysis of a few chroman derivatives ligands: (A) chroman 7-[2-pyrrolo]-pyrrole-[1,2-a]12H pyrrolino[2,3-b]chroman-4-one, and (C) 4-methyl-7-[2-pyrrolo]-pyrrole[1,2-a]12H pyrrolino[2,3-b]chroman-4-one and their respective borondifluoride complexes B and D as a quadruplex targeting compounds which found to stabilize G-quadruplex structure. To investigate the binding characteristics of these molecules with G-quadruplex vs. duplex selectivity, In vitro biophysical studies were performed by steady-state fluorescence, UV-visible titration, fluorescent TO displacement assay, CD thermal melting, circular dichroism spectroscopy, and cellular imaging by employing both telomeric and PRCC G-quadruplex forming sequences. Our investigation shows that these chromam ligands and their complexes are able to selectively bind and stabilize parallel and mixed hybrid topology of G-quadruplex both In vitro and in cellular conditions. A molecular docking study also suggests the binding of these compounds with G-quadruplex conformation. Collectively our study suggests these chroman complexes as a potentially useful fluorescent chemical product for G-quadruplex specific ligands and expands an option for G-quadruplex targeting ligands.

biochemistry↗

Discovery of novel G-quadruplex stabilizing compounds from medicinal plant & Evaluates their cellular toxicity

G-Quadruplexes (G4Q) are higher-order, four-stranded structures that can be formed by repeated guanine tracts in human genomes. In this study, we used a structure-based virtual screening of phytomolecules derived from medicinal plants in order to discover new natural DNA G4Q binders. The top 40 ligands were sorted out based on binding affinity values after completing a docking study on 314 phytomolecule samples with parallel and mixed hybrid structure. Further Thermal melting, circular dichroism, and fluorescence displacement experiment was used as a preliminary screening tool to determine the potential stabilizing properties of {beta}-sitosterol-{beta}-D-glucoside, and Glabrolide. The cytotoxicity experiments were conducted on HEK293T cells and found that both of the tested phyto molecules are non-toxic for up to 150 M concentration. Based on their cytotoxic experiments at the suggested high concentration, these phyto molecules may potentially be employed as G-Quadruplex targets in future research or applications. These results suggest that the plant may be a "lead" in the future for the development of novel therapies for diseases.

biochemistry↗

Exploring G-quadruplex structure in PRCC-TFE3 fusion Oncogene: Plausible use as anti cancer therapy for translocation Renal cell carcinoma (tRCC)

The TFE3 fusion gene, byproduct of Xp11.2 translocation, is the diagnostic marker for translocation renal cell carcinoma (tRCC). Absence of any clinically recognized therapy for tRCC, pressing a need to create novel and efficient therapeutic approaches. Previous studies shown that stabilization of the G-quadruplex structure in oncogenes suppresses their expression machinery. To combat the oncogenesis caused by fusion genes, our objective is to locate and stabilize the G-quadruplex structure within the PRCC-TFE3 fusion gene. Using the Quadruplex- forming G Rich Sequences (QGRS) mapper and the Non-B DNA motif search tool (nBMST) online server, we found putative G-quadruplex forming sequences (PQS) in the PRCC-TFE3 fusion gene. Circular dichroism demonstrating a parallel G-quadruplex in the targeted sequence. Fluorescence and UV-vis spectroscopy results suggest that pyridostatin binds to this newly discovered G-quadruplex. The PCR stop assay, as well as transcriptional or translational inhibition by PQS, revealed that stable G-quadruplex formation affects biological processes. Confocal microscopy of HEK293T cells transfected with the fusion transcript confirmed G- quadruplexes formation in cell. This investigation may shed light on G-quadruplexs functions in fusion genes and may help in the development of therapies specifically targeted against fusion oncogenes, which would enhance the capability of current tRCC therapy approach.

molecular biology↗

Calcineurin inhibition enhances Caenorhabditis elegans lifespan by defecation defects-mediated calorie restriction and nuclear hormone signaling

Calcineurin is a highly conserved calcium/calmodulin-dependent serine/threonine protein phosphatase with diverse functions. Inhibition of calcineurin is known to enhance the lifespan of Caenorhabditis elegans through multiple signaling pathways. Aiming to study the role of calcineurin in regulating innate immunity, we discover that calcineurin is required for the rhythmic defecation motor program (DMP) in C. elegans. Calcineurin inhibition leads to defects in the DMP, resulting in intestinal bloating, rapid colonization of the gut by bacteria, and increased susceptibility to bacterial infection. We demonstrate that intestinal bloating caused by calcineurin inhibition mimics the effects of calorie restriction, resulting in enhanced lifespan. The TFEB ortholog, HLH-30, is required for lifespan extension mediated by calcineurin inhibition. Finally, we show that the nuclear hormone receptor, NHR-8, is upregulated by calcineurin inhibition and is necessary for the increased lifespan. Our studies uncover a role for calcineurin in the C. elegans DMP and provide a new mechanism for calcineurin inhibition-mediated longevity extension.

physiology↗

Small Interactor of PKD2 (SIP), a novel PKD2-related single-pass transmembrane protein, is required for proteolytic processing and ciliary import of Chlamydomonas PKD2

In Chlamydomonas cilia, the ciliopathy-relevant TRP channel PKD2 is spatially compartmentalized into a distal region, in which PKD2 binds the axoneme and extracellular mastigonemes, and a smaller proximal region, in which PKD2 is more mobile and lacks mastigonemes. Here, we show that the two PKD2 regions are established early during cilia regeneration and increase in length as cilia elongate. In abnormally long cilia, only the distal region elongated whereas both regions adjusted in length during cilia shortening. In dikaryon rescue experiments, tagged PKD2 rapidly entered the proximal region of PKD2-deficient cilia whereas assembly of the distal region was hindered, suggesting that axonemal docking of PKD2 requires de novo ciliary assembly. We identified Small Interactor of PKD2 (SIP), a small PKD2-related protein, as a novel component of the PKD2-mastigoneme complex. In sip mutants, stability and proteolytic processing of PKD2 in the cell body were reduced and PKD2-mastigoneme complexes were absent from mutant cilia. Like the pkd2 and mst1 mutants, sip swims with reduced velocity. Cilia of the pkd2 mutant beat with normal frequency and bending pattern but were less efficient in moving cells supporting a passive role of the PKD2-SIP-mastigoneme complexes in increasing the effective surface of Chlamydomonas cilia.

cell biology↗

Changes in Cortical Directional Connectivity during Difficult Listening in Younger and Older Adults

One way to investigate the mechanisms that underlie speech comprehension under difficult listening conditions is via cortical connectivity. The innovative Network Localized Granger Causality (NLGC) framework was applied to magnetoencephalography (MEG) data, obtained from older and younger subjects performing a speech listening task in noisy conditions, in delta and theta frequency bands. Directional connectivity between frontal, temporal, and parietal lobes was analyzed. Both aging- and condition-related changes were found, particularly in theta. In younger adults, as background noise increased, there was a transition from predominantly temporal-to-frontal (bottom-up) connections, to predominantly frontal-to-temporal (top-down). In contrast, older adults showed bidirectional information flow between frontal and temporal cortices even for speech in quiet, not changing substantially with increased noise. Additionally, younger listeners did not show changes in the nature of their cortical links for different listening conditions, whereas older listeners exhibited a switch from predominantly facilitative links to predominantly sharpening, when noise increased. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=169 HEIGHT=200 SRC="FIGDIR/small/541500v1_ufig1.gif" ALT="Figure 1"> View larger version (28K): org.highwire.dtl.DTLVardef@174c132org.highwire.dtl.DTLVardef@37c5a5org.highwire.dtl.DTLVardef@1910e38org.highwire.dtl.DTLVardef@3c94e0_HPS_FORMAT_FIGEXP M_FIG C_FIG HighlightsO_LIDifferent bands show strong differences in directional functional connectivity patterns C_LIO_LIDirectional functional connectivity patterns altered by listening task difficulty C_LIO_LIAging dramatically alters directional functional connectivity patterns in during listening C_LIO_LINature of functional connectivity, additive vs subtractive, depends on age and task C_LI

neuroscience↗

Amorphous calcium phosphate-coated surfaces as a model for bone microenvironment in prostate cancer

Bone metastasis remains one of the biggest challenges in the treatment of prostate cancer, and other solid tumors such as breast, lung, and colon. Modeling a complex microenvironment in-vitro, such as the bone niche, requires interrogation of cell-cell interactions, specific extracellular matrix proteins and a high calcium environment. Here, we present a fast and cost-effective system in which commercially available, non-adhesive, cell culture vessels are coated with amorphous calcium phosphate (ACP) as a surrogate for bone matrix. We further present modified protocols for subculturing cells, as well as nucleic acid and protein collection in high calcium samples. We find that prostate epithelial cell lines show increased adhesion and proliferation when cultured in these surfaces, as well as independence from androgen starvation. We observe gene expression changes on ACP surfaces in early adenocarcinoma cell lines which may reflect alterations relevant to prostate cancer progression. Summary statementTo model the role of calcium in the microenvironment of the metastatic bone niche, we developed a cost-effective way to coat cell culture vessels in bioavailable calcium, and show that it has an effect on prostate cancer cell survival

cancer biology↗

Interferon restores replication fork stability and cell viability in BRCA-defective cells via ISG15

DNA replication and repair defects or genotoxic treatments trigger interferon (IFN)-mediated inflammatory responses. However, whether and how IFN signaling in turn impacts the DNA replication process has remained elusive. Here we show that IFN promotes replication fork stability, cell proliferation and survival in BRCA1/2-defective cancer cells and rescues the lethality of BRCA2-deficient mouse embryonic stem cells. Although IFN activates hundreds of genes, these effects are specifically mediated by the ubiquitin-like modifier ISG15 (IFN-stimulated gene 15). Inactivation of ISG15 or of the enzymes promoting its conjugation, referred as ISGylation, completely suppresses the impact of IFN on the replication process. Depletion of ISG15 significantly reduces cell proliferation rates whereas its upregulation results in increased resistance to the chemotherapeutic drug cisplatin in human BRCA1-mutated triple-negative and mouse BRCA2-deficient breast cancer cells, respectively. Accordingly, cells carrying BRCA1/2 defects consistently show increased ISG15 levels, representing a novel, in-built mechanism of drug resistance linked to BRCAness.

cancer biology↗

SifA SUMOylation governs Salmonella intracellular survival via modulation of lysosomal function

Gastroenteritis causing pathogen Salmonella Typhimurium (S. Tm) during its infection in host cells thrives in a vacuolated compartment, Salmonella Containing Vacuole (SCV), which sequentially acquires host endosomal and lysosomal markers. Long tubular structures, called as Salmonella induced filaments (SIFs), are known to be required for SCVs nutrient acquisition, membrane maintenance and stability. A tightly coordinated interactions involving prominent effector SifA and various host adapters PLEKHM1, PLEKHM2 and Rab GTPases govern SCV integrity and SIF formation. Here, we report for the first time, the functional regulation of SifA is modulated by its SUMOylation at lysine 11. S. Tm expressing lysine 11 mutant SifA (SifAK11R) is defective in intracellular proliferation due to compromised SIF formation and enhanced lysosomal acidification. Furthermore, murine competitive index experiments reveal defective in vivo proliferation and weakened virulence of SifAK11R mutant. Concisely, our results demonstrate that SUMO deficient SifA mutant nearly behaves like a SifA knockout strain which impacts PLEKHM2-M6PR mediated lysosomal acidification pathway. Thus, our results bring forth a novel S. Tm-host crosstalk mechanism involving host mediated effector SUMOylation critical for pathogenicity.

microbiology↗

Discovery of highly potent small molecule pan-coronavirus fusion inhibitors

The unprecedented pandemic of COVID-19, caused by a novel coronavirus, SARS-CoV-2, has led to massive human suffering, death, and economic devastation worldwide. The virus is mutating fast to more transmissible and infectious variants. The Delta variant (B.1.617.2), initially identified in India, and the omicron variant (BA.4 and BA.5) have spread worldwide. In addition, recently alarming antibody evasive SARS-CoV-2 subvariants, BQ and XBB, have been reported. These new variants may pose a substantial challenge to controlling the spread of this virus. Therefore, the continued development of novel drugs having pan-coronavirus inhibition to treat and prevent infection of COVID-19 is urgently needed. These drugs will be critically important in dealing with new pandemics that will emerge in the future. We report the discovery of several highly potent small molecule pan-coronavirus inhibitors. One of which, NBCoV63, showed low nM potency against SARS-CoV-2 (IC50: 55 nM), SARS-CoV (IC50: 59 nM), and MERS-CoV (IC50: 75 nM) in pseudovirus-based assays with excellent selectivity indices (SI: as high as > 900) demonstrating its pan-coronavirus inhibition. NBCoV63 showed equally effective antiviral potency against SARS-CoV-2 mutant (D614G) and several variants of concerns (VOCs) such as B.1.617.2 (Delta), B.1.1.529/BA.1 and BA.4/BA.5 (Omicron) and K417T/E484K/N501Y (Gamma). NBCoV63 also showed similar efficacy profiles to Remdesivir against authentic SARS-CoV-2 (Hong Kong strain) and two of its variants (Delta and Omicron) by plaque reduction in Calu3 cells. Additionally, we show that NBCoV63 inhibits virus-mediated cell-to-cell fusion in a dose-dependent manner. Furthermore, the Absorption, distribution, metabolism, and excretion (ADME) data of NBCoV63 demonstrated drug-like properties.

pharmacology and toxicology↗

High resolution cryo-EM structures of two potently SARS-CoV-2 neutralizing monoclonal antibodies of same donor origin that vary in neutralizing Omicron variants

While vaccines have by large been found to effective against the evolving SARS-CoV-2 variants, the profound and rapid effectivity of monoclonal antibodies (mAbs) in significantly reducing hospitalization to severe disease outcomes have also been demonstrated. In the present study, by high resolution cryo-electron microscopy (cryo-EM), we examined the structural insights of two trimeric spike (S) protein bound mAbs isolated from an Indian convalescent individual infected with ancestral SARS-CoV-2 which we recently reported to potently neutralize SARS-CoV-2 from its ancestral form through highly virulent Delta form however different in their ability to neutralize Omicron variants. Our findings showed binding and conformational heterogeneities of both the mAbs (THSC20.HVTR04 and THSC20.HVTR26) bound to S trimer in its apo and hACE-2 bound forms. Additionally, cryo-EM resolved structure assisted modeling highlighted key residues associated with the ability of these two mAbs to neutralize Omicron variants. Our findings highlighted key interacting features modulating antigen-antibody interacting that can further aid in structure guided antibody engineering to enhance their breadth and potency. HighlightsO_LITwo potent human mAbs obtained from a single donor differ binding to Omicron spikes C_LIO_LIPattern of binding and conformation of these mAbs bound to full length spike differs C_LIO_LIAntibody binding alters the conformational states of S trimer in its apo and hACE-2 bound forms. C_LIO_LICryo-EM structure guided modeling highlighted correlates of interacting residues associated with resistance and sensitivity of BA.1, BA.2, BA.4/BA.5 resistance and sensitivity against these mAbs. C_LI

biophysics↗

Switching state-space modeling of neural signal dynamics

Linear parametric state-space models are a ubiquitous tool for analyzing neural time series data, providing a way to characterize the underlying brain dynamics with much greater statistical efficiency than non-parametric data analysis approaches. However, neural time series data are frequently time-varying, exhibiting rapid changes in dynamics, with transient activity that is often the key feature of interest in the data. Stationary methods can be adapted to time-varying scenarios by employing fixed-duration windows under an assumption of quasi-stationarity. But time-varying dynamics can be explicitly modeled by switching state-space models, i.e., by using a pool of state-space models with different dynamics selected by a probabilistic switching process. Unfortunately, exact solutions for state inference and parameter learning with switching state-space models are intractable. Here we revisit a switching state-space model inference approach first proposed by Ghahramani and Hinton. We provide explicit derivations for solving the inference problem iteratively after applying variational approximation on the joint posterior of the hidden states and the switching process. We introduce a novel initialization procedure using an efficient leave-one-out strategy to compare among candidate models, which significantly improves performance compared to the existing method that relies on deterministic annealing. We then utilize this state-inference solution within a generalized expectation-maximization algorithm to estimate model parameters of the switching process and the linear state-space models with dynamics potentially shared among candidate models. We perform extensive simulations under different settings to benchmark performance against existing switching inference methods and further validate the robustness of our switching inference solution outside the generative switching model class. Finally, we demonstrate the utility of our method for sleep spindle detection in real recordings, showing how switching state-space models can be used to detect and extract transient spindles from human sleep electroencephalograms in an unsupervised manner. Author summaryAn inherent aspect of brain activity is that it changes over time, but existing methods for analyzing neuroscience data typically assume that the underlying activity is strictly stationary, i.e., the properties of that activity do not change over time. One way of handling time-varying data is to break the data into smaller segments that one assumes to be quasi-stationary, but this approach only works if signals vary gradually, and tends to perform poorly when changes are rapid or the target activity is transient in nature. A class of models called linear switching state-space models can explicitly represent time-varying activity, but they pose another set of challenges: exact solutions for such models are intractable, and existing approximate solutions can be highly inaccurate. In this work we present a solution for linear switching state-space models that is able to recover the underlying hidden states and model parameters for time-varying dynamics in a way that is robust to model mis-specification and that outperforms previously proposed methods. We demonstrate the utility of our method by applying it to the problem of sleep spindle detection and show that switching state-space models can automatically detect transient spindle activity from human sleep electroencephalograms.

neuroscience↗

Cut homeodomain transcription factor is a novel regulator of cortical glia morphogenesis and niche maintenance around neural stem cells

Cortex glia in Drosophila central nervous system forms a niche around neural cells for necessary signals to establish cross-talk with their surroundings. These cells grow and expand their thin processes around neural cell bodies. Although essential for the development and function of the nervous system, how these cells make extensive and intricate connected networks remain largely unknown. Here we show that Cut, a homeodomain transcription factor, directly regulates the fate of the cortex glia, impacting NSC homeostasis. Focusing on the thoracic ventral nerve cord (tVNC), we found that Cut is required for normal growth and development of cortex glia and timely increase in DNA content to undergo endomitosis. Knockdown of Cut in cortex glia significantly reduces the growth of cellular processes, the network around NSCs, and their progenys cell bodies. Conversely, overexpression of Cut induces overall growth of the main processes at the expense of side ones. Whereas the Cut knockdown slowdown the timely increase of DNA, Cut overexpression results in a significant increase in nuclear size and volume and a threefold increase in DNA content of cortex glia. Further, we note that constitutively high Cut also interfered with nuclei separation during endomitosis. Since cortex glia form syncytial networks around neural cells, the finding identifies Cut as a novel regulator of glial growth and endomitosis to support a functional nervous system. Article SummaryCut homeodomain transcription factor is crucial for cortex glia growth and the formation of complex cellular processes around neural cells. This regulation ensures a timely increase in DNA content, allowing the cells to enter endomitosis. Constitutively high Cut levels increase the DNA content of these cells to several folds. The finding emphasizes the need to investigate if activated CUX1, the human homolog of Cut, in glioma enhances chromosomal instability and, in conjunction with other mutations, enhances their tumorigenic potential.

developmental biology↗

PointVS: A Machine Learning Scoring Function that Identifies Important Binding Interactions

Over the last few years, many machine learning-based scoring functions for predicting the binding of small molecules to proteins have been developed. Their objective is to approximate the distribution which takes two molecules as input and outputs the energy of their interaction. Only a scoring function that accounts for the interatomic interactions involved in binding can accurately predict binding affinity on unseen molecules. However, many scoring functions make predictions based on dataset biases rather than an understanding of the physics of binding. These scoring functions perform well when tested on similar targets to those in the training set, but fail to generalise to dissimilar targets. To test what a machine learning-based scoring function has learnt, input attribution--a technique for learning which features are important to a model when making a prediction on a particular data point--can be applied. If a model successfully learns something beyond dataset biases, attribution should give insight into the important binding interactions that are taking place. We built a machine learning-based scoring function that aimed to avoid the influence of bias via thorough train and test dataset filtering, and show that it achieves comparable performance on the CASF-2016 benchmark to other leading methods. We then use the CASF-2016 test set to perform attribution, and find that the bonds identified as important by PointVS, unlike those extracted from other scoring functions, have a high correlation with those found by a distance-based interaction profiler. We then show that attribution can be used to extract important binding pharmacophores from a given protein target when supplied with a number of bound structures. We use this information to perform fragment elaboration, and see improvements in docking scores compared to using structural information from a traditional, data-based approach. This not only provides definitive proof that the scoring function has learnt to identify some important binding interactions, but also constitutes the first deep learning-based method for extracting structural information from a target for molecule design.

bioinformatics↗

Ancestral SARS-CoV-2 driven antibody repertoire diversity in an unvaccinated individual correlates with expanded neutralization breadth

Understanding the quality of immune repertoire triggered during natural infection can provide vital clues that form the basis for development of humoral immune response in some individuals capable of broadly neutralizing pan SARS-CoV-2 variants. We assessed the diversity of neutralizing antibody responses developed in an unvaccinated individual infected with ancestral SARS-CoV-2 by examining the ability of the distinct B cell germline-derived monoclonal antibodies (mAbs) in neutralizing known and currently circulating Omicron variants by pseudovirus and authentic virus neutralization assays. The ability of the antibodies developed post vaccination in neutralizing Omicron variants was compared to that obtained at baseline of the same individual and to those obtained from Omicron breakthrough infected individuals by pseudovirus neutralization assay. Broadly SARS-CoV-2 neutralizing mAbs representing unique B cell lineages with non-overlapping epitope specificities isolated from a single donor varied in their ability to neutralize Omicron variants. Plasma antibodies developed post vaccination from this individual demonstrated neutralization of Omicron BA.1, BA.2 and BA.4 with increased magnitude and found to be comparable with those obtained from other vaccinated individuals who were infected with ancestral SARS-CoV-2. Development of B cell repertoire capable of producing antibodies with distinct affinity and specificities for the antigen immediately after infection capable of eliciting broadly neutralizing antibodies offers highest probability in protecting against evolving SARS-CoV-2 variants. ImportanceDevelopment of robust neutralizing antibodies in SARS-CoV-2 convalescent individuals is known, however varies at population level. We isolated monoclonal antibodies from an individual infected with ancestral SARS-CoV-2 in early 2020 that not only varied in their B cell lineage origin but also varied in their capability and potency to neutralize all the known VOC and currently circulating Omicron variants. This indicated establishment of unique lineages that contributed in forming B cell repertoire in this particular individual immediately following infection giving rise to diverse antibody responses that could compensate each other in providing broadly neutralizing polyclonal antibody response. Individuals who were able to produce such potent polyclonal antibody responses after infection have a higher chance of being protected from evolving SARS-CoV-2 variants.

immunology↗