Unusual structural transition of antimicrobial VP1 peptide

VP1 peptide, an active domain of m-calpain enzyme with antimicrobial activity is found to undergo an unusual conformational transition in trifluoroethanol (TFE) solvent. The nature of, and time dependent variations in, circular dichroism associated with the amide I vibrations, suggest that VP1 undergoes self-aggregation forming anti-parallel β-sheet structure in TFE. Transmission electron micrograph (TEM) images revealed that β-sheet aggregates formed by VP1 possess fibril-like assemblies.     

Interaction between Nbp35 and Cfd1 Proteins of Cytosolic Fe-S Cluster Assembly Reveals a Stable Complex Formation in Entamoeba histolytica

Iron-Sulfur (Fe-S) proteins are involved in many biological functions such as electron transport, photosynthesis, regulation of gene expression and enzymatic activities. Biosynthesis and transfer of Fe-S clusters depend on Fe-S clusters assembly processes such as ISC, SUF, NIF, and CIA systems. Unlike other eukaryotes which possess ISC and CIA systems, amitochondriate Entamoeba histolytica has retained NIF & CIA systems for Fe-S cluster assembly in the cytosol. In the present study, we have elucidated interaction between two proteins of E. histolytica CIA system, Cytosolic Fe-S cluster deficient 1 (Cfd1) protein and Nucleotide binding protein 35 (Nbp35). In-silico analysis showed that structural regions ranging from amino acid residues (P33-K35, G131-V135 and I147-E151) of Nbp35 and (G5-V6, M34-D39 and G46-A52) of Cfd1 are involved in the formation of protein-protein complex. Furthermore, Molecular dynamic (MD) simulations study suggested that hydrophobic forces surpass over hydrophilic forces between Nbp35 and Cfd1 and Van-der-Waal interaction plays crucial role in the formation of stable complex. Both proteins were separately cloned, expressed as recombinant fusion proteins in E. coli and purified to homogeneity by affinity column chromatography. Physical interaction between Nbp35 and Cfd1 proteins was confirmed in vitro by co-purification of recombinant Nbp35 with thrombin digested Cfd1 and in vivo by pull down assay and immunoprecipitation. The insilico, in vitro as well as in vivo results prove a stable interaction between these two proteins, supporting the possibility of its involvement in Fe-S cluster transfer to target apo-proteins through CIA machinery in E. histolytica. Our study indicates that initial synthesis of a Fe-S precursor in mitochondria is not necessary for the formation of Cfd1-Nbp35 complex. Thus, Cfd1 and Nbp35 with the help of cytosolic NifS and NifU proteins can participate in the maturation of non-mitosomal Fe-S proteins without any apparent assistance of mitosomes.     

Design of ant-inspired stochastic control policies for collective transport by robotic swarms

In this paper, we present an approach to designing decentralized robot control policies that mimic certain microscopic and macroscopic behaviors of ants performing collective transport tasks. In prior work, we used a stochastic hybrid system model to characterize the observed team dynamics of ant group retrieval of a rigid load. We have also used macroscopic population dynamic models to design enzyme-inspired stochastic control policies that allocate a robotic swarm around multiple boundaries in a way that is robust to environmental variations. Here, we build on this prior work to synthesize stochastic robot attachment–detachment policies for tasks in which a robotic swarm must achieve non-uniform spatial distributions around multiple loads and transport them at a constant velocity. Three methods are presented for designing robot control policies that replicate the steady-state distributions, transient dynamics, and fluxes between states that we have observed in ant populations during group retrieval. The equilibrium population matching method can be used to achieve a desired transport team composition as quickly as possible; the transient matching method can control the transient population dynamics of the team while driving it to the desired composition; and the rate matching method regulates the rates at which robots join and leave a load during transport. We validate our model predictions in an agent-based simulation, verify that each controller design method produces successful transport of a load at a regulated velocity, and compare the advantages and disadvantages of each method.     

Significance of PELP1 in ER-negative breast cancer metastasis

Breast cancer metastasis is a major clinical problem. The molecular basis of breast cancer progression to metastasis remains poorly understood. PELP1 is an estrogen receptor (ER) coregulator that has been implicated as a proto-oncogene whose expression is deregulated in metastatic breast tumors and whose expression is retained in ER-negative tumors. We examined the mechanism and significance of PELP1-mediated signaling in ER-negative breast cancer progression using two ER-negative model cells (MDA-MB-231 and 4T1 cells) that stably express PELP1-shRNA. These model cells had reduced PELP1 expression (75% of endogenous levels) and exhibited less propensity to proliferate in growth assays in vitro. PELP1 downregulation substantially affected migration of ER-negative cells in Boyden chamber and invasion assays. Using mechanistic studies, we found that PELP1 modulated expression of several genes involved in the epithelial mesenchymal transition (EMT), including MMPs, SNAIL, TWIST, and ZEB. In addition, PELP1 knockdown reduced the in vivo metastatic potential of ER-negative breast cancer cells and significantly reduced lung metastatic nodules in a xenograft assay. These results implicate PELP1 as having a role in ER-negative breast cancer metastasis, reveal novel mechanism of coregulator regulation of metastasis via promoting cell motility/EMT by modulating expression of genes, and suggest PELP1 may be a potential therapeutic target for metastatic ER-negative breast cancer.     

Molecular Markers Reveal Only Two Mud Crab Species of Genus Scylla (Brachyura: Portunidae) in Indian Coastal Waters

The taxonomic ambiguity of the Indian mud crab (genus Scylla de Hann 1833) is still a cause of concern as several papers have been published with misleading identification. This is the first attempt to resolve the taxonomic uncertainty of the mud crab commonly available in Indian coastal waters using molecular genetic markers (ITS-1 and sequencing of COI gene) combined with traditional morphometry. Additionally, we developed a PCR method by which Indian mud crab species can be identified rapidly and effectively. The results clearly indicate that the green morph of the Indian mud crab is Scylla serrata and the brown morph is S. olivacea. The S. serrata commonly mentioned in the literature from India is S. olivacea; the S. tranquebarica noted by many Indian researchers should belong to S. serrata. Caution should be taken when interpreting or implementing the biological, molecular, and aquaculture data in the literature.     

Purification and characterization of an extracellular polysaccharide from haloalkalophilic Bacillus sp. I-450

During the course of investigation of haloalkalophilic bacteria, we screened some heavily polluted soil samples from the mudflats surrounding the city of Inchon, Korea, for their bioflocculant producing ability. Based on the screening, one isolate no. 450 tentatively identified as Bacillus sp. produced an extracellular polysaccharide having flocculation activity. The isolate produced the polysaccharide during the late logarithmic growth phase. The polymer could be recovered from the supernatant of the fermented medium by cold ethanol precipitation and purified by treating with cetylpyridinium chloride (CPC). The polymer was identified as an acidic polysaccharide containing neutral sugars, namely, galactose, fructose, glucose and raffinose, and uronic acids as major and minor components, respectively. The amount of neutral sugars, uronic acid and amino sugars were 52.4, 17.2 and 2.4%, respectively. The molecular weight of the polysaccharide was found to be 2.2×10⁶ Da. The Fourier transform infrared spectrophotometer (FT-IR) revealed typical characteristics of polysaccharides. ¹H NMR spectrum showed that the polymer is a heteroglycan. Thermogravimetric (TGA) analysis indicated the degradation temperature (T_d) at 290 °C. The rheological analysis of the polymer 450 revealed the pseudoplastic property with shear-thinning effect, while the compression test indicated that the polymer had high gel strength, and the S.E.M. studies showed that the polymer has a porous structure with small pore-size distribution indicating the compactness of the polymer.     

Dopamine receptor agonist bromocriptine restrains the follicular development,hatchling success and puberty in Gambusia affinis

The present investigation was conducted to determine the influence of dopamine (DA) receptor agonist bromocriptine (BRO) on reproduction and onset of puberty in the viviparous fish Gambusia affinis. In the first experiment, the mean number of stage I and II follicles (previtellogenic) in 0.8 or 5 mg BRO treated fish did not show significant difference compared to those of experimental controls, whereas the mean number of stage III follicles were significantly lower in 5 mg BRO treated fish compared to experimental controls. However, treatment of 0.8 or 5 mg BRO resulted in significantly lower numbers of stage IV (early vitellogenic) and V (late vitellogenic) follicles compared to those of experimental controls. There was decrease in the percent occurrence of pregnancy and different stages of embryos in BRO treated fish compared with the experimental controls. Concomitant with this, sparsely distributed gonadotropin releasing hormone immunoreactive (GnRH‐ir) fibres were observed in the proximal pars distalis (PPD) region of the pituitary gland in BRO treated fish compared to those of dense accumulations of these fibres in the PPD region of the pituitary gland in experimental controls. In the second experiment, exposure of juveniles (25 DPH) to same doses of BRO for 45 days resulted in complete absence of vitellogenic follicles and presence of few GnRH‐ir fibres in 5 mg BRO treated juvenile in contrast to presence of vitellogenic follicles and dense aggregation of GnRH fibres in treatment controls. Overall, the results of the present investigation suggest that DA affects ovarian follicular and embryonic development and onset of puberty in viviparous species.     

Comparison of controlled pore glass and Kieselguhr-polydimethylacrylamide composite as supports for solid-phase synthesis of 23-residue oligodeoxyribonucleotides in milligram amounts

Two 23-residue oligodeoxyribonucleotides, corresponding to both strands of a DNA duplex at the OR3 site of bacteriophage lambda, have been synthesized in good yields and in milligram quantities by a solid-phase phosphotriester method using two different supports, Kieselguhr-polydimethylacrylamide composite and controlled pore glass. Rapid purification was possible using high-performance liquid chromatography on radial compression ion-exchange columns. The results and utility of the two supports are compared.