Halogenated Secondary Metabolites from Higher Plants: Potent Drug Candidates for Chikungunya Using in silico Approaches.

Chikungunya virus (CHIKV) causes a debilitating fever and joint pain, with no specific antiviral treatment available. Halogenated secondary metabolites from plants are a promising new class of drug candidates against chikungunya, with unique properties that make them effective against the virus. Plants produce these compounds to defend themselves against pests and pathogens, and they are effective against a wide range of viruses, including chikungunya. This study investigated the interactions of halogenated secondary metabolites with nsP2pro, a therapeutic target for CHIKV. A library of sixty-six halogenated plant metabolites screened previously for ADME properties was used. Metabolites without violation of Lipinski's rule were docked with nsP2pro using AutoDock Vina. To find the stability of the pipoxide chlorohydrin-nsP2pro complex, the GROMACS suite was used for MD simulation. The binding free energy of the ligand-protein complex was computed using MMPBSA. Molecular docking studies revealed that halogenated metabolites interact with nsP2pro, suggesting they are possible inhibitors. Pipoxide chlorohydrin showed the greatest affinity to the target. This was further confirmed by the MD simulations, surface accessible area, and MMPBSA studies. Pipoxide chlorohydrin, a halogenated metabolite, was the most potent against nsP2pro in the survey.

Gallate Moiety of Catechin Is Essential for Inhibiting CCL2 Chemokine-Mediated Monocyte Recruitment.

Leukocyte recruitment witnesses an orchestrated complex formation between the chemokines and their molecular partners. CCL2 chemokine that regulates monocyte trafficking is a worthwhile system from the pharmaceutical perspective. In the current study, four major catechins (EC/EGC/ECG/EGCG) were assessed for their inhibitory potential against CCL2-regulated monocyte/macrophage recruitment. Interestingly, catechins with the gallate moiety (ECG/EGCG) could only attenuate the CCL2-induced macrophage migration. These molecules specifically bound to CCL2 on a pocket comprising the N-terminal, ß0-sheets, and ß3-sheets, and the binding affinity of ECGC (Kd = 22 ± 4 µM) is ∼4 times higher than that of the ECG complex (Kd = 85 ± 6 µM). MD simulation analysis evidenced that the molecular specificity/stability of CCL2-catechin complexes is regulated by multiple factors, including stereospecificity, number of hydroxyl groups on the annular ring-B, the positioning of the carbonyl group, and the methylation of the galloyl ring. Further, a significant overlap on the binding surface of CCL2 for EGCG/ECG and receptor interactions as evidenced from NMR data provided the rationale for the observed inhibition of macrophage migration in response to EGCG/ECG binding. In summary, these galloylated epicatechins can be considered as potent protein-protein interaction (PPI) inhibitors that regulate CCL2-directed leukocyte recruitment for resolving inflammatory/immunomodulatory disorders.

Integrated bioinformatics analysis reveals novel key biomarkers in diabetic nephropathy.

Objectives: The underlying molecular mechanisms of diabetic nephropathy have yet not been investigated clearly. In this investigation, we aimed to identify key genes involved in the pathogenesis and prognosis of diabetic nephropathy. Methods: We downloaded next-generation sequencing data set GSE142025 from Gene Expression Omnibus database having 28 diabetic nephropathy samples and nine normal control samples. The differentially expressed genes between diabetic nephropathy and normal control samples were analyzed. Biological function analysis of the differentially expressed genes was enriched by Gene Ontology and REACTOME pathways. Then, we established the protein-protein interaction network, modules, miRNA-differentially expressed gene regulatory network and transcription factor-differentially expressed gene regulatory network. Hub genes were validated by using receiver operating characteristic curve analysis. Results: A total of 549 differentially expressed genes were detected including 275 upregulated and 274 downregulated genes. The biological process analysis of functional enrichment showed that these differentially expressed genes were mainly enriched in cell activation, integral component of plasma membrane, lipid binding, and biological oxidations. Analyzing the protein-protein interaction network, miRNA-differentially expressed gene regulatory network and transcription factor-differentially expressed gene regulatory network, we screened hub genes MDFI, LCK, BTK, IRF4, PRKCB, EGR1, JUN, FOS, ALB, and NR4A1 by the Cytoscape software. The receiver operating characteristic curve analysis confirmed that hub genes were of diagnostic value. Conclusions: Taken above, using integrated bioinformatics analysis, we have identified key genes and pathways in diabetic nephropathy, which could improve our understanding of the cause and underlying molecular events, and these key genes and pathways might be therapeutic targets for diabetic nephropathy.

Deciphering the Potential of Pre and Pro-Vitamin D of Mushrooms against Mpro and PLpro Proteases of COVID-19: An In Silico Approach.

Vitamin D's role in combating the SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2), the virus causing COVID-19, has been established in unveiling viable inhibitors of COVID-19. The current study investigated the role of pre and pro-vitamin D bioactives from edible mushrooms against Mpro and PLpro proteases of SARS-CoV-2 by computational experiments. The bioactives of mushrooms, specifically ergosterol (provitamin D2), 7-dehydrocholesterol (provitamin-D3), 22,23-dihydroergocalciferol (provitamin-D4), cholecalciferol (vitamin-D3), and ergocalciferol (vitamin D2) were screened against Mpro and PLpro. Molecular docking analyses of the generated bioactive protease complexes unravelled the differential docking energies, which ranged from -7.5 kcal/mol to -4.5 kcal/mol. Ergosterol exhibited the lowest binding energy (-7.5 kcal/mol) against Mpro and PLpro (-5.9 kcal/mol). The Molecular Mechanics Poisson-Boltzmann Surface Area (MMPBSA) and MD simulation analyses indicated that the generated complexes were stable, thus affirming the putative binding of the bioactives to viral proteases. Considering the pivotal role of vitamin D bioactives, their direct interactions against SARS-CoV-2 proteases highlight the promising role of bioactives present in mushrooms as potent nutraceuticals against COVID-19.

An organotypic model of high-grade serous ovarian cancer to test the anti-metastatic potential of ROR2 targeted Polyion complex nanoparticles.

High-grade serous ovarian cancer (HGSOC) is the most lethal gynaecological malignancy. Most patients are diagnosed at late stages when the tumour has metastasised throughout the peritoneal cavity. The Wnt receptor ROR2 has been identified as a promising therapeutic target in HGSOC, with limited targeting therapeutic options currently available. Small interfering RNA (siRNA)-based therapeutics hold great potential for inhibiting the function of specific biomarkers, however major challenges remain in efficient delivery and stability. The aim of this study was to investigate the ability of nanoparticles to deliver ROR2 siRNA into HGSOC cells, including platinum resistant models, and estimate the anti-metastatic effect via a 3D organotypic model for ovarian cancer. The nanoparticles were generated by conjugating poly[2-(dimethylamino) ethyl methacrylate] (PDMAEMA) of various chain length to bovine serum albumin (BSA), followed by the condensation of ROR2 siRNA into polyplexes, also termed polyion complex (PIC) nanoparticles. The toxicity and uptake of ROR2 siRNA PIC nanoparticles in two HGSOC cell lines, CaOV3 as well as its cisplatin resistant pair (CaOV3CisR), in addition to primary cells used for the 3D organotypic model were investigated. ROR2 knockdown at both transcriptional and translational levels were evaluated via real-time PCR and western blot analysis, respectively. Following 24 h incubation with the nanoparticles, functional assays were performed including proliferation (IncuCyte S3), transwell migration and 3D co-cultured transwell invasion assays. The PICs nanoparticles exhibited negligible toxicity in the paired CaOV3 cell lines or primary cells. Treating CaOV3 and CaOV3CisR cells with ROR2 siRNA containing PICs nanoparticles significantly inhibited migration and invasion ability. The biocompatible ROR2 siRNA conjugated PICs nanoparticles provide an innovative therapeutic option. ROR2 targeting therapy shows potential in treating HGSOC including platinum resistant forms.

Identification of Key Pathways and Genes in Obesity Using Bioinformatics Analysis and Molecular Docking Studies.

Obesity is an excess accumulation of body fat. Its progression rate has remained high in recent years. Therefore, the aim of this study was to diagnose important differentially expressed genes (DEGs) associated in its development, which may be used as novel biomarkers or potential therapeutic targets for obesity. The gene expression profile of E-MTAB-6728 was downloaded from the database. After screening DEGs in each ArrayExpress dataset, we further used the robust rank aggregation method to diagnose 876 significant DEGs including 438 up regulated and 438 down regulated genes. Functional enrichment analysis was performed. These DEGs were shown to be significantly enriched in different obesity related pathways and GO functions. Then protein-protein interaction network, target genes - miRNA regulatory network and target genes - TF regulatory network were constructed and analyzed. The module analysis was performed based on the whole PPI network. We finally filtered out STAT3, CORO1C, SERPINH1, MVP, ITGB5, PCM1, SIRT1, EEF1G, PTEN and RPS2 hub genes. Hub genes were validated by ICH analysis, receiver operating curve (ROC) analysis and RT-PCR. Finally a molecular docking study was performed to find small drug molecules. The robust DEGs linked with the development of obesity were screened through the expression profile, and integrated bioinformatics analysis was conducted. Our study provides reliable molecular biomarkers for screening and diagnosis, prognosis as well as novel therapeutic targets for obesity.

Hydroxyl Groups on Annular Ring-B Dictate the Affinities of Flavonol-CCL2 Chemokine Binding Interactions.

Owing to the astounding biological properties, dietary plant flavonoids have received considerable attention toward developing unique supplementary food sources to prevent various ailments. Chemokines are chemotactic proteins involved in leukocyte trafficking through their interactions with G-protein-coupled receptors and cell surface glycosaminoglycans (GAGs). CCL2 chemokine, a foremost member of CC chemokines, is associated with the pathogenesis of various inflammatory infirmities, thus making the CCL2-Receptor (CCR2)/GAG axis a potential pharmacological target. The current study is designed to unravel the structural details of CCL2-flavonol interactions. Molecular interactions between flavonols (kaempferol, quercetin, and myricetin) with human/murine CCL2 orthologs and their monomeric/dimeric variants were systematically investigated using a combination of biophysical approaches. Fluorescence studies have unveiled that flavonols interact with CCL2 orthologs specifically but with differential affinities. The dissociation constants (K d) were in the range of 10-5-10-7 µM. The NMR- and computational docking-based outcomes have strongly suggested that the flavonols interact with CCL2, comprising the N-terminal and ß1- and ß3-sheets. It has also been observed that the number of hydroxyl groups on the annular ring-B imposed a significant cumulative effect on the binding affinities of flavonols for CCL2 chemokine. Further, the binding surface of these flavonols to CCL2 orthologs was observed to be extensively overlapped with that of the receptor/GAG-binding surface, thus suggesting attenuation of CCL2-CCR2/GAG interactions in their presence. Considering the pivotal role of CCL2 during monocyte/macrophage trafficking and the immunomodulatory features of these flavonols, their direct interactions highlight the promising role of flavonols as nutraceuticals.

Analysis and visualization of sleep stages based on deep neural networks.

Automatic sleep stage scoring based on deep neural networks has come into focus of sleep researchers and physicians, as a reliable method able to objectively classify sleep stages would save human resources and simplify clinical routines. Due to novel open-source software libraries for machine learning, in combination with enormous recent progress in hardware development, a paradigm shift in the field of sleep research towards automatic diagnostics might be imminent. We argue that modern machine learning techniques are not just a tool to perform automatic sleep stage classification, but are also a creative approach to find hidden properties of sleep physiology. We have already developed and established algorithms to visualize and cluster EEG data, facilitating first assessments on sleep health in terms of sleep-apnea and consequently reduced daytime vigilance. In the following study, we further analyze cortical activity during sleep by determining the probabilities of momentary sleep stages, represented as hypnodensity graphs and then computing vectorial cross-correlations of different EEG channels. We can show that this measure serves to estimate the period length of sleep cycles and thus can help to find disturbances due to pathological conditions.

Determinants of Household Food Insecurity in Rural Areas of the Hilly Region of Kumaun, Uttarakhand, India: A Pilot Study.

The food security status of rural inhabitants of the hilly region of the Uttarakhand State was poorly documented. This study aimed to determine the extent and determinants of household food insecurity in rural-hilly areas of Kumaun, Uttarakhand. A cross-sectional, interview-administered survey was conducted among 155 households. Food insecurity, as measured by Household Food Insecurity Access Scale, was found in a large number of households; 2.6% of households were categorized as severely food-insecure, 53.3% as moderately food-insecure, 7.1% as mildly food-insecure, and 36.8% as food-secure. Multivariate logistic regression analysis revealed that food insecurity was significantly associated with the poverty level of the family, number of children or adolescents in the household, educational level of household head, number of employed family members, and social class (p < 0.05). The food security policies in India must modify focus from food entitlement to include education, income, employment, and household composition. Nutrition-sensitive interventions in agriculture to promote the cultivation of fruits and vegetables in hilly areas. Promotion of poultry farming, goat rearing, freshwater fish farming, and related training will increase the availability of animal food sources. It is an urgent need to design and implement location-specific programs and policies to address food insecurity and food access.

Elucidating the Molecular Interactions of Chemokine CCL2 Orthologs with Flavonoid Baicalin.

An integrated and controlled migration of leukocytes is necessary for the legitimate functioning and maintenance of the immune system. Chemokines and their receptors play a decisive role in regulating the leukocyte migration to the site of inflammation, a phenomena often referred to as chemotaxis. Chemokines and their receptors have become significant targets for therapeutic intervention considering their potential to regulate the immune system. Monocyte chemoattractant protein-1 (MCP-1/CCL2) is a preeminent member of CC chemokine family that facilitates crucial roles by orchestrating the recruitment of monocytes into inflamed tissues. Baicalin (BA), a major bioactive flavonoid, has been reported to attenuate chemokine-regulated leukocyte trafficking. However, no molecular details pertaining to its direct binding to chemokine(s)/receptor(s) are available till date. In the current study, using an array of monomers/dimers of human and murine CCL2 orthologs (hCCL2/mCCL2), we have shown that BA binds to the CCL2 protein specifically with nanomolar affinity (K d = 270 ± 20 nM). NMR-based studies established that BA binds CCL2 in a specific pocket involving the N-terminal, ß1- and ß3-sheets. Docking studies suggested that the residues T16, N17, R18, I20, R24, K49, E50, I51, and C52 are majorly involved in complex formation through a combination of H-bonds and hydrophobic interactions. As the residues R18, R24, and K49 of hCCL2 are crucial determinants of monocyte trafficking through receptor/glycosaminoglycans (GAG) binding in CCL2 human/murine orthologs, we propose that baicalin engaging these residues in complex formation will result in attenuation of CCL2 binding to the receptor/GAGs, thus inhibiting the chemokine-regulated leukocyte trafficking.

Molecular insights into the differential structure-dynamics-stability features of interleukin-8 orthologs: Implications to functional specificity.

Chemokines are a sub-group of chemotactic cytokines that regulate the leukocyte migration by binding to G-protein coupled receptors (GPCRs) and cell surface glycosaminoglycans (GAGs). Interleukin-8 (CXCL8/IL8) is one of the most essential CXC chemokine that has been reported to be involved in various pathophysiological conditions. Structure-function relationships of human IL8 have been studied extensively. However, no such detailed information is available on IL8 orthologs, although they exhibit significant functional divergence. In order to unravel the differential structure-dynamics-stability-function relationship of IL8 orthologs, comparative molecular analysis was performed on canine (laurasians) and human (primates) IL8 proteins using in-silico molecular evolutionary analysis and solution NMR spectroscopy methods. The residue level NMR studies suggested that, although the overall structural architecture of canine IL8 is similar to that of human IL8, systematic differences were observed in their backbone dynamics and low-energy excited states due to amino acid substitutions. Further, these substitutions also resulted in attenuation of stability and heparin binding affinity in the canine IL8 as compared to its human counterpart. Indeed, structural and sequence analysis evidenced for specificity of molecular interactions with cognate receptor (CXCR1) and glycosaminoglycan (heparin), thus providing evidence for a noticeable functional specificity and divergence between the two IL8 orthologs.

Dissecting the differential structural and dynamics features of CCL2 chemokine orthologs.

Chemokines are a sub-group of cytokines that regulate the leukocyte migration. Monocyte chemoattractant protein-1 (MCP/CCL2) is one of the essential CC chemokine that regulates the migration of monocytes into inflamed tissues. It has been observed that the primary sequences of CCL2 orthologs among rodents and primates vary significantly at the C-terminal region. However, no structural details are available for the rodentia family CCL2 proteins. The current study unravelled the structural, dynamics and in-silico functional characteristics of murine CCL2 chemokine using a comprehensive set of NMR spectroscopy techniques and evolutionary approaches. The study unravelled that the N-terminal portion of the murine CCL2 forms a canonical CC chemokine dimer similar to that of human CCL2. However, unlike human CCL2, the murine ortholog exhibits extensive dynamics in the µs-ms timescales. The presence of C-terminal region of the murine CCL2 protein/rodentia family is highly glycosylated, completely disordered, and inhibits the folding of the structured CCL2 regions. Further, it has been observed that the glycosaminoglycan binding surfaces of these orthologs proteins are greatly differed. In a nut shell, this comparative study provided the role of molecular evolution in generating orthologous proteins with differential structural and dynamics characteristics to engage them in specific molecular interactions.

Comparative evaluation of the efficacy of two modes of delivery of Piroxicam (Dolonex(®)) for the management of postendodontic pain: A randomized control trial.

BACKGROUND: Alleviating pain is of utmost importance when treating patients with endodontic pain. AIM: To compare and evaluate the efficacy of two modes of delivery of pretreatment Piroxicam (Dolonex(®), Pfizer) for the management of postendodontic pain. MATERIALS AND METHODS: Sixty-six patients with symptomatic irreversible pulpitis were randomly divided into three groups of 22 subjects Group I - control group, no pharmacological intervention, Group II - patients received pretreatment oral Piroxicam (40 mg), Group III - patients received pretreatment intraligamentary injections totaling 0.4 mL of Piroxicam. Single visit endodontic therapy was performed by a single endodontist. Visual analogue scale was used to record pain before treatment and 4, 8, 12, 24, and 48 h postoperatively. Mann-Whitney U-test and Kruskal-Wallis tests were used to analyze the data. RESULTS: The patients in Groups II and III perceived less postendodontic pain as compared to Group I (P < 0.05), at all the time intervals. At 12, 24, and 48 h, pain experience in patients of Group III was significantly less. CONCLUSIONS: Intraligamentary mode of delivery of Piroxicam was more efficacious.

Influence of Structure, Charge, and Concentration on the Pectin-Calcium-Surfactant Complexes.

Polymer-surfactant complex formation of pectin with different types of surfactants, cationic (cetyltrimethylammonium bromide, CTAB and dodecyl trimethylammonium bromide, DTAB), anionic (sodium dodecyl sulfate, SDS), and neutral (Triton X-100, TX-100), was investigated at room temperature in the presence and absence of cross-linker calcium chloride using light scattering, zeta potential, rheology, and UV-vis spectroscopic measurements where the surfactant concentration was maintained below their critical micellar concentration (CMC). Results indicated that the interaction of cationic surfactant with pectin in the presence and absence of calcium chloride was much stronger compared to anionic and neutral surfactants. The neutral surfactant showed identifiable interaction despite the absence of any charged headgroup, while anionic surfactant showed feeble or very weak interaction with the polymer. The pectin-CTAB or DTAB complex formation was attributed to associative electrostatic and hydrophobic interactions. On comparison between the cationic surfactants, it was found that CTAB interacts strongly with pectin because of its long hydrocarbon chain. The morphology of complexes formed exhibited random coil structures while at higher concentration of surfactant, rod-like or extended random coil structures were noticed. Thus, functional characteristics of the complex could be tuned by varying the type of surfactant (charge and structure) and its concentration. The differential network rigidity (pectin-CTAB versus pectin-DTAB gels) obtained from rheology measurements showed that addition of a very small amount of surfactant (concentration ≪ CMC) was required for enhancing network strength, while the presence of a large amount of surfactant resulted in the formation of fragile gels. No gel formation occurred when the surfactant concentration was close to their CMC values. Considering the importance of pectin in food and pharmaceutical industry, this study is relevant.

Characterization of microstructure, viscoelasticity, heterogeneity and ergodicity in pectin-laponite-CTAB-calcium nanocomposite hydrogels.

In order to customize the viscoelastic properties of pectin gels, it is necessary to work on a composite platform. Herein, the gelation kinetics, and viscoelastic characterization of anionic polysaccharide pectin dispersion prepared in presence of nanoclay laponite are reported using dynamic light scattering and rheology measurements. The ratio Rg/Rh (Rg and Rh are radius of gyration and hydrodynamic radius respectively) determined from light scattering data revealed the presence of random coils of pectin chains inside the gel matrix. When nanoclay laponite was added to the pectin chains solution, two-phase separation was noticed instantaneously. Therefore, the surfactant cetyltrimethylammonium bromide [CTAB] was added to exfoliate the clay platelets in the dispersion, and also in its gel phase. The exfoliating agent cetyltrimethylammonium bromide ([CTAB]≈ cmc/10) helped to enhance the homogeneity and stability of the pectin-clay sols and gels. The storage and loss moduli (G' and G") of the composite gel changed significantly as function of nanoclay laponite content for concentration up to 0.03% (w/v) causing the softening of the gels (gel strength reduced by close to 50%) compared to pectin-calcium gel. However, as the concentration of nanoclay laponite was maintained between 0.01% and 0.03% (w/v), the gel rigidity (G') recovered by 30% (35-45 Pa). The transition from ergodic to non-ergodic state occurred during sol-gel transition owing to the presence of the nanoclay laponite. The gelation time was not too different from the ergodicity breaking time. Thus, the presence of nanoclay laponite in such minute concentration is shown to cause considerable change in the thermo-physical property of the composite gels. This material property modulation will facilitate designing of soft gels having storage modulus continuously varying in the wide range of 10-70 Pa while keeping the gelation temperature mostly unaltered.

Attenuation of the early events of α-synuclein aggregation: a fluorescence correlation spectroscopy and laser scanning microscopy study in the presence of surface-coated Fe3O4 nanoparticles.

The aggregation of α-synuclein (A-syn) has been implicated in the pathogenesis of Parkinson's disease (PD). Although the early events of aggregation and not the matured amyloid fibrils are believed to be responsible for the toxicity, it has been difficult to probe the formation of early oligomers experimentally. We studied the effect of Fe3O4 nanoparticle (NP) in the early stage of aggregation of A-syn using fluorescence correlation spectroscopy (FCS) and laser scanning microscopy. The binding between the monomeric protein and NPs was also studied using FCS at single-molecule resolution. Our data showed that the addition of bare Fe3O4 NPs accelerated the rate of early aggregation, and it did not bind the monomeric A-syn. In contrast, L-lysine (Lys)-coated Fe3O4 NPs showed strong binding with the monomeric A-syn, inhibiting the early events of aggregation. Lys-coated Fe3O4 NPs showed significantly less cell toxicity compared with bare Fe3O4 NPs and can be explored as a possible strategy to develop therapeutic application against PD. To the best of our knowledge, this report is the first example of using a small molecule to attenuate the early (and arguably the most relevant in terms of PD pathogenesis) events of A-syn aggregation.

Coexistence of iso-nonergodic laponite gel and glass in 1-methyl-3-octylimidazolium chloride ionic liquid solutions.

We report unique colloidal gel-glass coexistence in aqueous laponite dispersion (2% w/v) in the presence of 1-methyl-3-octylimidazolium chloride ionic liquid (IL, [C8mim][Cl], concentration 0.01 to 0.05% w/v), where both of the phases had identical nonergodicity and were dynamically interactive. With aging, the nascent heterogeneous dispersion exhibited spontaneous two-phase separation, and the time-dependent relative viscosity followed: η(r) = |ε|(-k) where ε = (t - t(g))/t(g) and t(g) is the time required for the system to get arrested, with k decreasing from 3.13 to 2.54 as the IL concentration was increased from 0 to 0.03% (w/v), implying slowing down of the arrest kinetics. This time was measured from viscosity and rheology studies, revealing the formation of IL-mediated finite size colloidal networks on a time scale of ~4 × 10(3) s, whereas the dispersion developed a large viscosity one decade in time later (~4 × 10(4) s). Homogeneous transparent upper phase was an entropic glass and exhibited substantial storage modulus gain (300-3000 Pa) with an increase in IL concentration (0 to 0.05% (w/v)). The translucent lower gel phase had a much higher storage modulus. Dynamic light scattering measured bimodal relaxation time of concentration fluctuations. The degree of nonergodicity in the two phases was approximately the same, implying laponite-IL cluster exchange across the interface (identical slow-mode diffusivity). In summary, IL-induced first-order phase separation in laponite dispersion produced a homogeneous colloidal gel coexisting with a glass not commonly observed in soft matter systems. This implied that the two phases were dynamically coupled on long time scales, whereas their short-time behavior was distinctively different.

AR-A 014418 Used against GSK3beta Downregulates Expression of hnRNPA1 and SF2/ASF Splicing Factors.

Glioblastoma is one of the most aggressive forms of primary brain tumors of glial cells, including aberrant regulation of glycogen synthase kinase 3 ß (GSK3 ß ) and splicing factors deregulation. Here, we investigate the role of small molecule AR-A014418 and Manzamine A against GSK3 kinase with factual control on splicing regulators. AR-A 014418, 48 hrs posttreatment, caused dose (25-100 µ M) dependent inhibition in U373 and U87 cell viability with also inhibition in activating tyrosine phosphorylation of GSK3alpha (Tyr 279) and beta (Tyr 216). Furthermore, inhibition of GSK3 kinase resulted in significant downregulation of splicing factors (SRSF1, SRSF5, PTPB1, and hnRNP) in U87 cells with downregulation of antiapoptotic genes such as BCL2, BCL-xL, Survivin, MCL1, and BMI1. Similarly, downregulation of splicing factors was also observed in U373 glioma cell after using SiRNA against AKT and GSK3beta kinase. In addition, potential roles of AR-A014418 in downregulation of splicing factors were reflected with decrease in Anxa7 (VA) variant and increase in Anxa7 WT tumor suppressor transcript and protein. The above results suggest that inhibition of GSK3beta kinase activation could be the beneficial strategy to inhibit the occurrence of alternative cancer escape pathway via downregulating the expression of splicing regulators as well as apoptosis.

Effect of unconventional carbon sources on biosurfactant production and its application in bioremediation.

The potential of an alkaliphilic bacterium Klebsiella sp. strain RJ-03, to utilize different unconventional carbon sources for the production of biosurfactant was evaluated. The biosurfactant produced using corn powder, potato peel powder, Madhuca indica and sugarcane bagasse containing medium, exhibited significantly higher viscosity and maximum reduction in surface tension as compared to other substrates. Among several carbon substrates tested, production of biosurfactant was found to be the highest with corn powder (15.40 ± 0.21 g/l) as compared to others. The comparative chemical characterization of purified biosurfactant was done using advance analytical tools such as NMR, FT-IR, SEM, GPC, MALDI TOF-TOF MS, GC-MS, TG and DSC. Analyses indicated variation in the functional groups, monosaccharide composition, molecular mass, thermostability. Higher yield with cheaper raw materials, noteworthy stress tolerance of CP-biosurfactant toward pH and salt as well as compatibility with chemical surfactants and detergents revealed its potential for commercialization and application in bioremediation.

Production and structural characterization of biosurfactant produced by an alkaliphilic bacterium, Klebsiella sp.: evaluation of different carbon sources.

The potential of an alkaliphilic bacterium Klebsiella sp. strain RJ-03, to utilize different carbon sources for the production of an extracellular biosurfactant was evaluated. Among the several carbon substrates tested, production of the crude biosurfactant was found to be the highest with starch (10.1±0.11g/L) followed by sucrose (5.1±0.11g/L), xylose (3.25±0.08g/L), galactose (3.1±0.16g/L) glucose (2.75±0.11g/L) and fructose (2.62±0.07g/L). The crude biosurfactant production was done using starch, sucrose, xylose, galactose and glucose containing medium, that exhibited significantly high viscosity, emulsification activity and maximum reduction in surface tension as compared to those obtained from fructose and maltose. The carbon source has significant effect on the quantity as well as the quality of biosurfactant production. The chemical characteristics of purified biosurfactant was compared by NMR, FT-IR, SEM, GPC, MALDI TOF-TOF MS, GC-MS, TG and DSC analysis, indicating variation in the functional groups, bonds, elements, monosaccharide composition, molecular mass and thermo stability.