On the rise of fear speech in online social media.

Recently, social media platforms are heavily moderated to prevent the spread of online hate speech, which is usually fertile in toxic words and is directed toward an individual or a community. Owing to such heavy moderation, newer and more subtle techniques are being deployed. One of the most striking among these is fear speech. Fear speech, as the name suggests, attempts to incite fear about a target community. Although subtle, it might be highly effective, often pushing communities toward a physical conflict. Therefore, understanding their prevalence in social media is of paramount importance. This article presents a large-scale study to understand the prevalence of 400K fear speech and over 700K hate speech posts collected from Gab.com. Remarkably, users posting a large number of fear speech accrue more followers and occupy more central positions in social networks than users posting a large number of hate speech. They can also reach out to benign users more effectively than hate speech users through replies, reposts, and mentions. This connects to the fact that, unlike hate speech, fear speech has almost zero toxic content, making it look plausible. Moreover, while fear speech topics mostly portray a community as a perpetrator using a (fake) chain of argumentation, hate speech topics hurl direct multitarget insults, thus pointing to why general users could be more gullible to fear speech. Our findings transcend even to other platforms (Twitter and Facebook) and thus necessitate using sophisticated moderation policies and mass awareness to combat fear speech.

Surface Interaction of Ionic Liquids: Stabilization of Polyethylene Terephthalate-Degrading Enzymes in Solution.

The effect of aqueous solutions of selected ionic liquids solutions on Ideonella sakaiensis PETase with bis(2-hydroxyethyl) terephthalate (BHET) substrate were studied by means of molecular dynamics simulations in order to identify the possible effect of ionic liquids on the structure and dynamics of enzymatic Polyethylene terephthalate (PET) hydrolysis. The use of specific ionic liquids can potentially enhance the enzymatic hydrolyses of PET where these ionic liquids are known to partially dissolve PET. The aqueous solution of cholinium phosphate were found to have the smallest effect of the structure of PETase, and its interaction with (BHET) as substrate was comparable to that with the pure water. Thus, the cholinium phosphate was identified as possible candidate as ionic liquid co-solvent to study the enzymatic hydrolyses of PET.

Conserved Dynamic Mechanism of Allosteric Response to L-arg in Divergent Bacterial Arginine Repressors.

Hexameric arginine repressor, ArgR, is the feedback regulator of bacterial L-arginine regulons, and sensor of L-arg that controls transcription of genes for its synthesis and catabolism. Although ArgR function, as well as its secondary, tertiary, and quaternary structures, is essentially the same in E. coli and B. subtilis, the two proteins differ significantly in sequence, including residues implicated in the response to L-arg. Molecular dynamics simulations are used here to evaluate the behavior of intact B. subtilis ArgR with and without L-arg, and are compared with prior MD results for a domain fragment of E. coli ArgR. Relative to its crystal structure, B. subtilis ArgR in absence of L-arg undergoes a large-scale rotational shift of its trimeric subassemblies that is very similar to that observed in the E. coli protein, but the residues driving rotation have distinct secondary and tertiary structural locations, and a key residue that drives rotation in E. coli is missing in B. subtilis. The similarity of trimer rotation despite different driving residues suggests that a rotational shift between trimers is integral to ArgR function. This conclusion is supported by phylogenetic analysis of distant ArgR homologs reported here that indicates at least three major groups characterized by distinct sequence motifs but predicted to undergo a common rotational transition. The dynamic consequences of L-arg binding for transcriptional activation of intact ArgR are evaluated here for the first time in two-microsecond simulations of B. subtilis ArgR. L-arg binding to intact B. subtilis ArgR causes a significant further shift in the angle of rotation between trimers that causes the N-terminal DNA-binding domains lose their interactions with the C-terminal domains, and is likely the first step toward adopting DNA-binding-competent conformations. The results aid interpretation of crystal structures of ArgR and ArgR-DNA complexes.

A new type low-cost, flexible and wearable tertiary nanocomposite sensor for room temperature hydrogen gas sensing.

This paper reports on reduced graphene oxide (rGO), tin oxide (SnO2) and polyvinylidene fluoride (PVDF) tertiary nanocomposite thick film based flexible gas sensor. The nanocomposite of 0.90(PVDF) - 0.10[x(SnO2) - (1 - x)rGO] with different weight percentages (x = 0, 0.15, 0.30, 0.45, 0.6, 0.75, 0.90 and 1) have been prepared by the hot press method. Chromium (Cr) has been deposited on the surface by using E-beam evaporation system, which is used as electrode of the device. Crystal structure, morphology, and electrical characteristics of the device have been explored for the technological application. A correlation between crystallinity, morphology, and electrical properties with these thick films has also been established. The device has been tested at different hydrogen (H2) gas concentration as well as at different response times. A superior response of 0.90(PVDF) - 0.10[0.75(SnO2) - 0.25 rGO] nanocomposite thick film has been observed. Hence, this composition is considered as optimized tertiary nanocomposite for the hydrogen gas sensor application. The sensor response of 49.2 and 71.4% with response time 34 sec and 52 sec for 100 PPM and 1000 PPM H2 gas concentration respectively have been obtained. First time a new kind of low cost and flexible polymer based nanocomposite thick film gas sensor has been explored.

Effect of Co-transfection of Anti-myostatin shRNA Constructs in Caprine Fetal Fibroblast Cells.

Knockdown of myostatin gene (MSTN), transforming growth factor-ß superfamily, and a negative regulator of the skeletal muscle growth, by RNA interference (RNAi), has been reported to increase muscle mass in mammals. The current study was aimed to cotransfect two anti-MSTN short hairpin RNA (shRNA) constructs in caprine fetal fibroblast cells for transient silencing of MSTN gene. In the present investigation, approximately 89% MSTN silencing was achieved in transiently transfected caprine fetal fibroblast cells by cotransfection of two best out of four anti-MSTN shRNA constructs. Simultaneously, we also monitored the induction of IFN responsive genes (IFN), pro-apoptotic gene (caspase3) and anti-apoptotic gene (MCL-1) due to cotransfection of different anti-MSTN shRNA constructs. We observed induction of 0.66-19.12, 1.04-4.14, 0.50-3.43, and 0.42-1.98 for folds IFN-ß, OAS1, caspase3, and MCL-1 genes, respectively (p < 0.05). This RNAi based cotransfection method could provide an alternative strategy of gene knockout and develop stable caprine fetal fibroblast cells. Furthermore, these stable cells can be used as a cell donor for the development of transgenic cloned embryos by somatic cell nuclear transfer (SCNT) technique.

A refined atomic scale model of the Saccharomyces cerevisiae K+-translocation protein Trk1p combined with experimental evidence confirms the role of selectivity filter glycines and other key residues.

Potassium ion (K+) uptake in yeast is mediated mainly by the Trk1/2 proteins that enable cells to survive on external K+ concentration as low as a few µM. Fungal Trks are related to prokaryotic TRK and Ktr and plant HKT K+ transport systems. Overall sequence similarity is very low, thus requiring experimental verification of homology models. Here a refined structural model of the Saccharomyces cerevisiae Trk1 is presented that was obtained by combining homology modeling, molecular dynamics simulation and experimental verification through functional analysis of mutants. Structural models and experimental results showed that glycines within the selectivity filter, conserved among the K-channel/transporter family, are not only important for protein function, but are also required for correct folding/membrane targeting. A conserved aspartic acid in the PA helix (D79) and a lysine in the M2D helix (K1147) were proposed earlier to interact. Our results suggested individual roles of these residues in folding, structural integrity and function. While mutations of D79 completely abolished protein folding, mutations at position 1147 were tolerated to some extent. Intriguingly, a secondary interaction of D79 with R76 could enhance folding/stability of Trk1 and enable a fraction of Trk1[K1147A] to fold. The part of the ion permeation path containing the selectivity filter is shaped similar to that of ion channels. However below the selectivity filter it is obstructed or regulated by a proline containing loop. The presented model could provide the structural basis for addressing the long standing question if Trk1 is a passive or active ion-translocation system.

Binding-competent states for L-arginine in E. coli arginine repressor apoprotein.

Arginine repressor of E. coli is a multifunctional hexameric protein that provides feedback regulation of arginine metabolism upon activation by the negatively cooperative binding of L-arginine. Interpretation of this complex system requires an understanding of the protein's conformational landscape. The ~50 kDa hexameric C-terminal domain was studied by 100 ns molecular dynamics simulations in the presence and absence of the six L-arg ligands that bind at the trimer-trimer interface. A rotational shift between trimers followed by rotational oscillation occurs in the production phase of the simulations only when L-arg is absent. Analysis of the system reveals that the degree of rotation is correlated with the number of hydrogen bonds across the trimer interface. The trajectory presents frames with one or more apparently open binding sites into which one L-arg could be docked successfully in three different instances, indicating that a binding-competent state of the system is occasionally sampled. Simulations of the resulting singly-liganded systems reveal for the first time that the binding of one L-arg results in a holoprotein-like conformational distribution.

Assessment of developmental potential of caprine cloned embryos with ooplasm replenishment under two culture media.

The present study was designed to assess the developmental potential of somatic cell nuclear transfer (SCNT) embryos, with and without replenishment of ooplasm into enucleated oocytes, by culturing separately in two culture media. The enucleated oocytes were replenished with exogenous matured ooplasm under replenished nuclear transfer (RNT) method and compared with conventional nuclear transfer (CNT) method without replenishment. The fusion efficiency in RNT group was found to be significantly higher (P < 0.05) than CNT group (59.39 ± 7.36 vs 45.57 ± 3.68%). The completely fused reconstructed oocytes from both groups were cultured separately in research vitro cleave medium (RVCL) and RVCL-Blast medium. The embryonic development of 2 cell, 4 cell, 8-16 cell and 16-32 cell stages in the RNT group was superior to the CNT group regardless of the culture media used (P < 0.05). The embryonic development of the 8-16 cell, 16-32 cell, morula, and blastocyst stages in the RVCL-Blast medium was significantly higher (P < 0.05) than the RVCL alone for both RNT as well as CNT groups. RNT method with RVCL-Blast produced highly significant (P < 0.01) embryonic development for 8-16 cell and 16-32 cell stage when compared to CNT with RVCL. Conclusively, the combination of RNT with RVCL-Blast culture media enabled an overall increase in the embryonic developmental potential.

Drug target prioritization in Plasmodium falciparum through metabolic network analysis, and inhibitor designing using virtual screening and docking approach.

The genome sequence of Plasmodium falciparum reveals that many metabolic pathways are unique as compared to its human host. Metabolic Network Analysis was carried out to find the essential enzymes critical for the survival of the pathogen. In the present study, choke point and load point analysis was used to locate putative targets. The identified targets were further checked to confirm that no alternate pathway or human homolog exists. Among the top 15 enzymes obtained from this analysis, we have selected P. falciparum orotidine-5'-monophosphate decarboxylase (PfODCase) enzyme as it is sequentially and structurally different from that of humans, for searching novel inhibitors. A five-point 3D pharmacophore was generated for the crystal structure of PfODCase complexes with uridine-5'-monophosphate (U5P). The binding site environment shows three H-bond acceptors, one H-bond donor and one negative ionizable feature. This pharmacophore model was used as a 3D query to perform virtual screening experiments against 2,664,779 standard lead compounds obtained from the freely available ZINC database. Top 10 hits obtained from virtual screening were selected for molecular docking experiments against PfODCase in order to verify their results and to have a better insight into their binding modes. Here, docking of U5P with PfODCase is used as a control. We have identified six compounds, among them, few are U5P analogs and others are novel ones with diverse scaffolds. The key residues: Lys42, Asp20, Lys72, Ser127, Ala184, Gln185 and Arg203 at the main binding pocket of PfODCase are responsible for better stability of diverse ligands. These compounds according to their free energy of binding could serve as potent leads for designing novel inhibitors against malarial ODCase enzyme.