Biology subjects

Ray, S.

Publications and source records attributed to Ray, S..

At least 109 records · Page 6Linked to original sources

bioRxiv2021

Stimulus-induced narrow-band gamma oscillations in humans can be recorded using open-hardware low-cost EEG amplifier

Stimulus-induced narrow-band gamma oscillations (30-70 Hz) in human electro - encephalograph (EEG) have been linked to attentional and memory mechanisms and are abnormal in mental health conditions such as autism, schizophrenia and Alzheimers Disease. This suggests that gamma oscillations could be valuable both as a research tool and an inexpensive, non-invasive biomarker for disease evaluation. However, since the absolute power in EEG decreases rapidly with increasing frequency following a "1/f" power law, and the gamma band includes line noise frequency, these oscillations are highly susceptible to instrument noise. Previous studies that recorded stimulus-induced gamma oscillations used expensive research-grade EEG amplifiers to address this issue. While low-cost EEG amplifiers have become popular in Brain Computer Interface applications that mainly rely on low-frequency oscillations (< 30 Hz) or steady-state-visually-evoked-potentials, whether they can also be used to measure stimulus-induced gamma oscillations is unknown. We recorded EEG signals using a low-cost, open-source amplifier (OpenBCI) and a traditional, research-grade amplifier (Brain Products GmbH) in male (N = 6) and female (N = 5) subjects (22-29 years) while they viewed full-screen static gratings that are known to induce gamma oscillations. OpenBCI recordings showed gamma response in almost all the subjects who showed a gamma response in Brain Products recordings, and the spectral and temporal profiles of these responses in alpha (8-13 Hz) and gamma bands were highly correlated between OpenBCI and Brain Products recordings. These results suggest that low-cost amplifiers can potentially be used in stimulus induced gamma response detection, making its research, and application in medicine more accessible.

neuroscience↗

bioRxiv2021

A copula based topology preserving graph convolution network for clustering of single-cell RNA-seq data

Annotation of cells in single-cell clustering requires a homogeneous grouping of cell populations. There are various issues in single cell sequencing that effect homogeneous grouping (clustering) of cells, such as small amount of starting RNA, limited per-cell sequenced reads, cell-to-cell variability due to cell-cycle, cellular morphology, and variable reagent concentrations. Moreover, single cell data is susceptible to technical noise, which affects the quality of genes (or features) selected/extracted prior to clustering. Here we introduce sc-CGconv (copula based graph convolution network for single cell clustering), a stepwise robust unsupervised feature extraction and clustering approach that formulates and aggregates cell-cell relationships using copula correlation (Ccor), followed by a graph convolution network based clustering approach. sc-CGconv formulates a cell-cell graph using Ccor that is learned by a graph-based artificial intelligence model, graph convolution network. The learned representation (low dimensional embedding) is utilized for cell clustering. sc-CGconv features the following advantages. a. sc-CGconv works with substantially smaller sample sizes to identify homogeneous clusters. b. sc-CGconv can model the expression co-variability of a large number of genes, thereby outperforming state-of-the-art gene selection/extraction methods for clustering. c. sc-CGconv preserves the cell-to-cell variability within the selected gene set by constructing a cell-cell graph through copula correlation measure. d. sc-CGconv provides a topology-preserving embedding of cells in low dimensional space. The source code and usage information are available at https://github.com/Snehalikalall/CopulaGCN Contact: sumanta.ray@cwi.nl

bioinformatics↗

bioRxiv2021

Spatiotemporal solidification of α-synuclein inside the liquid droplets

Liquid-liquid phase separation (LLPS) and subsequent liquid-to-solid transition is implicated in membraneless organelles formation as well as disease associated protein aggregation. However, how liquid-to-solid transition is initiated inside a liquid droplet remains unclear. Here, using studies at single droplet resolution, we show that liquid-to-solid transition of -synuclein (-Syn) liquid droplets is associated with significant changes in the local microenvironment as well as secondary structure of the protein, which is prominently observed at the center of the liquid droplets. With the ageing of liquid droplets, the "structured" core at the center gradually expands and propagates over entire droplets. Further, during droplet fusion, smaller, homogeneous droplets progressively dissolve and supply proteins to the larger, heterogeneous droplets containing solid-like core at their center. The present study will significantly help to understand the physical mechanism of LLPS and liquid-to-solid transition in biological compartmentalization as well as in protein aggregation associated with human neurodegenerative disorders. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=117 SRC="FIGDIR/small/465113v1_ufig1.gif" ALT="Figure 1"> View larger version (49K): org.highwire.dtl.DTLVardef@2a3406org.highwire.dtl.DTLVardef@bd8148org.highwire.dtl.DTLVardef@172b4eaorg.highwire.dtl.DTLVardef@1c3965f_HPS_FORMAT_FIGEXP M_FIG C_FIG

biophysics↗

bioRxiv2021

Modelling altered signalling of G-protein coupled receptors in inflamed environment to advance drug design

We previously reported the successful design, synthesis and testing of the prototype opioid painkiller NFEPP that does not elicit adverse side effects. Uniquely, this design was based on mathematical modelling of extracellular interactions between G-protein coupled receptors (GPCRs) and ligands, recognizing that GPCRs function differently under pathological versus healthy conditions. We now present a novel stochastic model of GPCR function that includes intracellular dissociation of G-protein subunits and modulation of plasma membrane calcium channels associated with parameters of inflamed and healthy tissue (pH, radicals). The model is validated against in vitro experimental data for NFEPP and fentanyl ligands at different pH values. We found markedly reduced calcium channel inhibition induced by NFEPP at normal pH compared to lower pH, in contrast to the effect of fentanyl, and enhanced constitutive G-protein activation but lower probability of ligand binding with increasing radical concentrations. By means of molecular dynamics simulations, we also assessed qualitative changes of reaction rates due to additional disulfide bridges inside the GPCR binding pocket. The results suggest that, compared to radicals, low pH is a more important determinant of overall GPCR function in an inflamed environment. Future drug design efforts should take this into account.

systems biology↗

bioRxiv2021

Actin capping protein regulates actomyosin contractility to maintain germline architecture in C. elegans

Actin dynamics play an important role in the morphogenesis of cells and tissues, yet the control of actin filament growth takes place at the molecular level. A challenge in the field is to link the molecular function of actin regulators with their physiological function. Here, we report the in vivo role of the actin capping protein CAP-1 in the C. elegans germline. We show that CAP-1 is associated with actomyosin structures in the cortex and rachis, where it keeps the level of contractility in check. A 60% reduction in the level of CAP-1 leads to a 2-fold increase in F-actin and non-muscle myosin II and only a 30% increase in Arp2/3. CAP-1 depletion leads to severe structural defects in the syncytial germline and oocytes, which can be rescued by reducing myosin activity. Thus, we uncover a physiological role for actin capping protein in maintaining C. elegans fertility by regulating the level of actomyosin contractility.

cell biology↗

bioRxiv2021

The potyviral protein 6K1 reduces plant protease activity during Turnip mosaic virus infection

O_LIPotyviral genomes encode just 11 major proteins and multifunctionality is associated to most of these proteins at different stages of virus life cycle. The potyviral protein 6K1 is required for potyvirus replication at the early stages of viral infection and may mediate cell-to-cell movement at later stages. C_LIO_LIOur study demonstrates that the 6K1 protein from Turnip mosaic virus (TuMV) reduces the abundance of transcripts related to jasmonic acid biosynthesis and transcripts that encode cysteine protease inhibitors when expressed in trans in Nicotiana benthamiana relative to controls. Furthermore, 6K1 stability increases when lipoxygenase and cysteine protease activity is inhibited chemically, linking a mechanism to the rapid turnover of 6K1 when expressed in trans. C_LIO_LIUsing transient expression, we show 6K1 is degraded rapidly at early time points in the infection process, whereas at later stages of infection protease activity is reduced and 6K1 becomes more stable, resulting in higher TuMV accumulation in systemic leaves. There was no impact of 6K1 transient expression on TuMV accumulation in local leaves. C_LIO_LITogether, these results suggest a novel function for the TuMV 6K1 protein which has not been reported previously and enhances our understanding of the complex interactions occurring between plants and potyviruses. C_LI

plant biology↗

bioRxiv2021

Co-aggregation and secondary nucleation in the life cycle of human prolactin/galanin functional amyloids

Synergistic-aggregation and cross-seeding by two different amyloid proteins/peptides are well evident in various neurological disorders. However, this phenomenon is not well studied in functional amyloid aggregation. Here, we show Prolactin (PRL) is associated with lactation in mammals and neuropeptide galanin (GAL), which are co-stored in the lactotrophs facilitates the synergic aggregation in the absence of secretory granules helper molecules glycosaminoglycans (GAGS). Interestingly, although each partner possesses homotypic seeding ability, a unidirectional cross-seeding of GAL aggregation can be mediated by PRL seeds. The specificity of co-aggregation by PRL and GAL along with unidirectional cross-seeding suggests tight regulation of functional amyloid formation during co-storage of these hormones in secretory granule biogenesis of female rat lactotrophs. Further mixed fibrils release the constituent functional hormone much faster than the corresponding individual amyloid formed in presence of GAGs, suggesting that co-aggregation of functionally distant hormones might have evolved for efficient storage, synergistic and rapid release of both hormones upon stimulation. The co-aggregation and cross seeding by two different hormones of completely different structures and sequences (PRL and GAL) suggest a novel mechanism of heterologous amyloid formation both in disease and functional amyloids.

biophysics↗

bioRxiv2021

Stimulus-induced narrowband gamma oscillations are test-retest reliable in healthy elderly in human EEG

Visual stimulus-induced narrowband gamma oscillations in electroencephalogram (EEG) recordings have been recently shown to be compromised in subjects with Mild Cognitive Impairment or Alzheimers Disease (AD), suggesting that gamma could be an inexpensive and easily accessible biomarker for early diagnosis of AD. However, to use gamma as a biomarker, its characteristics should remain consistent across multiple recordings, even when separated over long intervals. Previous magnetoencephalography studies in young subjects have reported that gamma power remains consistent over recordings separated by a few weeks to months. Here, we assessed the consistency of slow (20-35 Hz) and fast gamma (36-66 Hz) oscillations induced by static full-field gratings in male (N=20) and female (N=20) elderly subjects (>49 years) in EEG recordings separated by more than a year, and tested the consistency in the magnitude of gamma power, its temporal evolution and spectral profile. Gamma oscillations had distinct spectral and temporal characteristics across subjects, which remained consistent across recordings (average intraclass correlation, ICC of [~]0.7). Alpha oscillations (8-12 Hz) and steady-state-visually-evoked-potentials (SSVEPs) were also found to be reliable. We further tested how EEG features can be used to identify two recordings as belonging to the same versus different subjects and found high classifier performance (area under ROC curve of [~]0.89), with the temporal evolution of slow gamma and spectral profile emerging as the most informative features. These results suggest that EEG gamma oscillations are reliable across recordings and can be used as a clinical biomarker as well as a potential tool for subject identification. Significance statementWe demonstrate the reliability of stimulus-induced gamma oscillations in elderly humans for the first time in EEG. Since gamma has recently been shown to be compromised in patients with Mild Cognitive Impairment or early Alzheimers Disease (AD), together these results mark the first steps towards an EEG based clinical biomarker for early diagnosis of AD. We observed high reliability in the power spectrum, gamma power and its temporal characteristics, within the test-retest period of one year. Alpha and steady-state-visually-evoked potential power were also found to be reliable. These spectral and temporal features could also be used to identify EEG recordings as belonging to the same versus different subjects with high performance, suggesting a potentially key role in subject identification also.

neuroscience↗

bioRxiv2021

Action inhibition revisited: a new method identifiescompromised reactive but intact proactive control inTourette disorder

Tourette disorder (TD) is characterized by tics, which are sudden repetitive involuntary movements or vocalizations. Deficits in inhibitory control in TD patients remain inconclusive from the traditional method of estimating the ability to stop an impending action, which requires careful interpretation of a parameter derived from race model. One possible explanation for these inconsistencies is that race models assumptions are often violated. Here, we used a pair of metrics derived from a recent alternative model to address why stopping performance in TD patients is unaffected by impairments in neural circuitry. These new metrics distinguish between proactive and reactive inhibitory control and estimate them separately. When these metrics were contrasted with healthy controls (HC), we identified robust deficits in reactive control in TD patients, but not in proactive control. The patient population exhibited difficulty in slowing down the speed of movement planning, which they compensated by their intact ability of procrastination. TEASERTourette disorder patients inhibit action by means of strategic postponement to compensate impaired slowness in preparation.

neuroscience↗

bioRxiv2021

Cytotoxic and micronuclei inducing effects of petroleum ether fraction of leaf aqueous extract of Clerodendrum viscosum Vent. in Allium cepa root tip cells

Clerodendrum viscosum is a traditionally used medicinal plant. The present study aimed to analyze a detailed cytotoxic effect of the nonpolar petroleum ether fraction (AQPEF) of leaf aqueous extract of C. viscosum Vent. (LAECV) in Allium cepa root tip cells. The LAECV was fractionated with petroleum ether and tested for A. cepa toxicity at early hours (2 and 4 h treatment) at concentrations 0, 50, 100, 150 and 200 {micro}g mL-1. The highest aberrant cell percentage (10.45{+/-}0.46%) was scored from 150 {micro}g mL-1 followed by100 {micro}g mL-1 (8.75{+/-}0.26%) concentration at 4 h treated samples. The AQPEF treatment induced a significant (p<0.0001) increase in micronuclei frequency; 4.31{+/-}0.33, 5.08{+/-}0.13, 5.05{+/-}0.22 and 3.05{+/-}0.37% respectively at concentrations 50, 100, 150, and 200 {micro}g mL-1. The highest polyploid frequency (20.14{+/-}0.68 %) induced with 100 {micro}g mL-1 of AQPEF at 16 h recovery. 150 {micro}g mL-1 is the most effective concentration of AQPEF to decipher its activity similar to colchicine (150 {micro}g mL-1). In summary, the present study indicates petroleum ether is suitable for extraction of the active phytochemicals of LAECV having cytotoxic effects on A. cepa root tip cells. The AQPEF has colchicine like micronuclei, polyploidy, and mitotic abnormality inducing potentials in A. cepa root apical meristem cells.

pharmacology and toxicology↗

bioRxiv2021

Immunomodulatory Therapy with Glatiramer Acetate Reduces Endoplasmic Reticulum Stress and Mitochondrial Dysfunction in Experimental Autoimmune Encephalomyelitis

Endoplasmic reticulum (ER) stress and mitochondrial dysfunction are found in lesions of multiple sclerosis (MS) and of experimental autoimmune encephalomyelitis (EAE), an animal model of MS, and may contribute to the neuronal loss that underlies permanent impairment. These pathological changes are due to the neuroinflammation that characterizes MS and the strong interplay between the ER and mitochondria. We investigated whether immunomodulatory drug glatiramer acetate (GA) can reduce these changes in the spinal cords of chronic EAE mice by using routine histology, immunostaining, and electron microscopy. EAE spinal cord tissue exhibited increased infiltration/inflammation (upregulation of proinflammatory cytokines), demyelination, mitochondrial dysfunction (increased fission, decreased fusion, and increased biogenesis), ER stress, downregulation of NAD+ dependent pathways, and increased neuronal death. GA reversed these pathological changes, suggesting that immunomodulating therapy can attenuate ER stress, mitochondrial dysfunction, apoptotic cell death, and demyelination in the CNS and increase NAD+ utilizing activities by suppressing neuroinflammation.

neuroscience↗

bioRxiv2021

Generating realistic cell samples for gene selection in scRNA-seq data: A novel generative framework

High dimensional, small sample size (HDSS) scRNA-seq data presents a challenge to the gene selection task in single cell. Conventional gene selection techniques are unstable and less reliable due to the fewer number of available samples which affects cell clustering and annotation. Here, we present an improved version of generative adversarial network (GAN) called LSH-GAN to address this issue by producing new realistic samples and combining this with the original scRNA-seq data. We update the training procedure of the generator of GAN using locality sensitive hashing which speeds up the sample generation, thus maintains the feasibility of applying gene selection procedures in high dimension scRNA-seq data. Experimental results show a significant improvement in the performance of benchmark feature (gene) selection techniques on generated samples of one synthetic and four HDSS scRNA-seq data. Comprehensive simulation study ensures the applicability of the model in the feature (gene) selection domain of HDSS scRNA-seq data. AvailabilityThe corresponding software is available at https://github.com/Snehalikalall/LSH-GAN

bioinformatics↗

bioRxiv2021

Comparative in vitro antioxidant and antibacterial activities of leaf extract fractions of Crimson bottlebrush, Callistemon citrinus (Curtis.) Skeels

ObjectivesThe present study aimed to analyze a comparative in vitro free radical scavenging and antibacterial potentials of leaf aqueous and the successive extract fractions of Callistemon citrinus. MethodsFor in vitro antioxidant activity assessments, 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity, Fe3+ ion reducing antioxidant power (FRAP) assay, and total antioxidant capacity of the extracts were tested. Antibacterial potentials were tested through Agar well diffusion method using both gram-positive and gram-negative bacterial strains. ResultsData indicate the polar successive aqueous fraction (AQF) possesses the highest free radicals scavenging capacity, with lowest EC50 (the required extract concentration to scavenge half of the free radicals) for DPPH and FRAP assays, and contains the highest total phenolics (308.2{+/-}5.9 {micro}g tannic acid equivalent/mg dry extract, DE), flavonoids (516.7{+/-}3.5 {micro}g quercetin equivalent/mg DE), total antioxidant capacity (441.48{+/-}12.8 {micro}g ascorbic acid equivalent/mg DE). Phenolics and flavonoids contents were positively correlated with the in vitro antioxidant activities. The antibacterial study indicates the petroleum ether and chloroform are suitable solvents for extracting antibacterial phytochemicals from C. citrinus leaves that are effective against both gram-positive and negative bacterial strains. ConclusionThe most polar fraction i.e. the successive aqueous extract fraction of C. citrinus leaves exhibited the highest antioxidant activities while the most non-polar petroleum ether extract fraction showed the highest antibacterial potentials thus these extract fractions might have therapeutic importance.

pharmacology and toxicology↗

bioRxiv2021

Appraisal of Cell Cycle Delay and Cytotoxicity Inducing Potential of 3-epicaryoptin in Allium Cepa L.

Diterpenoid 3-epicaryoptin (C26H36O9) is abundant in the leaves of Clerodendrum inerme, a traditionally used medicinal plant, having insect antifeedant activities. Here, we aim to explore the cytogenotoxic effects of compound 3-epicaryoptin in Allium cepa root apical meristem cells. A. cepa roots were treated with 3-epicaryoptin (100, 150 & 200 g mL-1 concentration) and the standard compound colchicine (200 g mL-1 concentration) for 2, 4, 4+16 h (4 h treatment followed by 16 h recovery). Cytogenotoxicity was analysed by studying the root growth retardation (RGR), mitotic index (MI), and cellular aberrations. The result showed statistically significant (p<0.01), concentration-dependent RGR effects of 3-epicaryoptin treatment compared with the negative control. Study of cell frequency in different phases of cell division observed a significant (p<0.001) increase in the metaphase cell percentage (66.2{+/-}0.58 %, 150 g mL-1) and which subsequently caused an increase in the frequency of MI (12.29{+/-}0.34 %, 150 g mL-1) at 4h of 3-epicaryoptin treatment and that was comparable with the colchicine action. The cytological study revealed that the 3-epicaryoptin treatment could induce different types of chromosomal abnormalities such as colchicine like metaphase, vagrant chromosomes, sticky chromosomes, anaphase-bridge, and an increased frequency of micronuclei and polyploid cells. These findings indicate that 3-epicaryoptin is cytogenotoxic, and thus C. inerme should be used with caution in traditional medicine.

pharmacology and toxicology↗

bioRxiv2021

Pupillary aperture is a potential biomarker ofmovement preparation

In response to variable light intensity, the pupils reflexively constrict or dilate to maintain a uniform retinal illumination. The pupillary light reflex (PLR) pathway receives projections from two important areas in primates brain that plan rapid saccadic eye-movement - frontal eye field (FEF) and superior colliculus (SC). The speed with which neurons in these areas increase firing rate to a threshold determines latency of a saccade. Micro-stimulation of FEF/SC neurons below this threshold modulates the magnitude of PLR. Nonetheless, how the saccade latency and pupil dynamics are related remains unknown. Our study shows that the appearance of a bright stimulus evokes pupil constriction at higher rate when the latency of impending saccade to the stimulus is shorter. This inverse relationship between the rate of pupil constriction and the saccade latency is robust irrespective of the reward outcome. In a homeomorphic biomechanical model of pupil, we have projected build-up signal similar to FEF and SC activity to the parasympathetic and sympathetic divisions of the PLR pathway, respectively. Model simulation mimics the observed data to indicate that the FEF and SC activity for eye movement modulates autonomic input to the pupillary muscle plant. A striking similarity between the dynamics of pupil constriction and stochastic rise in neural activity for saccade elicitation suggests that PLR is a potential proxy of movement preparation, and not mere an indicator of attentional orientation. Our study suggests a mechanism of how the retinal luminosity is timely regulated to aid perception by minimizing visual transients due to gaze orientation.

neuroscience↗

bioRxiv2021

Shape analysis of gamma rhythm supports a superlinear inhibitory regime in an inhibition-stabilized network

Visual inspection of stimulus-induced gamma oscillations (30-70 Hz) often reveals a non-sinusoidal shape. Such distortions are a hallmark of non-linear systems and are also observed in mean-field models of gamma oscillations. A thorough characterization of the shape of the gamma cycle can therefore provide additional constraints on the operating regime of such models. However, the gamma waveform has not been quantitatively characterized, partially because the first harmonic of gamma, which arises because of the non-sinusoidal nature of the signal, is typically weak and gets masked due to a broadband increase in power related to spiking. To address this, we recorded local field potential (LFP) from the primary visual cortex (V1) of two awake female macaques while presenting full-field gratings or iso-luminant chromatic hues that produced huge gamma oscillations with prominent peaks at harmonic frequencies in the power spectra. We found that gamma and its first harmonic always maintained a specific phase relationship, resulting in a distinctive shape with a sharp trough and a shallow peak. Interestingly, a Wilson-Cowan (WC) model operating in an inhibition stabilized mode could replicate this shape, but only when the inhibitory population operated in the super-linear regime, as predicted recently. However, another recently developed model of gamma that operates in a linear regime driven by stochastic noise failed to produce salient harmonics or the observed shape. Our results impose additional constraints on models that generate gamma oscillations and their operating regimes.

neuroscience↗

bioRxiv2021

Effect of Leaf Aqueous Extract of Manilkara hexandra on Digestive and Glucose Metabolic Enzymes, and Serum Glucose Level in Labeo rohita Fingerlings

Manilkara hexandra (Roxb.) Dubard (common name Khirni, family: Sapotaceae) is an evergreen traditionally used medicinal plant. The present study aimed to see the effects of leaf aqueous extract of M. hexandra (LAEMH) on digestive and glucose metabolic enzyme action and serum glucose level in rohu, Labeo rohita, fingerlings. Experimental fish were fed a basal diet (Group I), and diets supplemented with LAEMH at 300 mg (Group II) and 600 mg (Group III) kg-1body weight. A significant reduction in serum glucose was noticed in the treated groups when compared with the control group. The reduced amylase activity was noticed in the treated fingerlings in a dose-dependent manner. However, lipase and protease activities didnt differ significantly among the experimental groups. Reduced serum glucose level might be correlated with the decline in the activity of digestive amylase in fish. Group II (3.35{+/-}0.19 U) and Group III (3.49{+/-}0.13 U) were recorded with reduced glucose-6-phosphatase activities than the control group (4.4{+/-}0.39 U). Moreover, the study revealed a decline in fructose-1,6-phosphatase activities in the treated groups in comparison to the control group. The decline in the activities of the metabolic enzymes might be associated with the non-availability of glucose owing to reduced activity of the digestive amylase in the treated groups. In conclusion, the present study established the hypoglycaemic effect of the leaf aqueous extract of M. hexandra in a fish model.

pharmacology and toxicology↗

bioRxiv2020

Deciphering colchicine like actions of clerodin in terms of microtubule destabilization based mitotic abnormalities, G2/M-phase arrest, and plant polyploidy

Clerodin (C24H34O7), a clerodane diterpenoid, is a bitter principle of Clerodendrum viscosum. The present study aimed to decipher colchicine-like actions of clerodin in terms of microtubule destabilization based mitotic abnormalities, G2-M arrest, and plant polyploidy. Purified clerodin showed increased metaphase frequency in Human Peripheral Blood Lymphocytes (HPBLs), Human Embryonic Kidney cells (HEK-293), and Allium cepa root apical meristem cells. Both squashed slide of the onion root tip and flow cytometric analysis of radish protoplast revealed a significantly increased frequency of polyploid cells. Flow cytometric analysis showed an increase in frequencies of G2-M in MCF-7 cells from 6.10 to 16.25% after clerodin (200g/mL) treatment for 24 h. Confocal microscopy imaging of tubulin in clerodin-treated MCF-7 cells revealed microtubule destabilization. Molecular docking and LIGPLOT analysis indicate that clerodin interact in the colchicine binding site, including, single hydrogen bond with Asn 101 of -tubulin. In summary, our experimental data revealed that clerodin has metaphase arresting, microtubule destabilization, and polyploidy inducing ability similar to colchicine. Molecular docking analysis revealed for the first time that clerodin and colchicine interact at the common site of tubulin residue indicating a common mechanism of action. The results also indicate similar cytotoxic potentialities of both clerodin and colchicine even though they belong to different chemical groups. Thus, clerodin may be used in place of colchicine as a plant polyploidy inducing agent in plant breeding programs in Agriculture.

cell biology↗