Biogenic synthesis of gold nanoparticle using fractioned cellular components from eukaryotic algae and cyanobacteria

In the present investigation fractioned cellular components like intact pigment bearing thylakoids/chloroplasts, carotenoids, protein, polysaccharides were extracted from the cyanobacterium Anabaena sphaerica and green alga Chlorococcum infusionum. Each of these extracts was used separately in search for efficient reducing agents during gold nanoparticle (GNP) production in pro‐ and eukaryotic algal cell systems. The whole biomass and extracted compounds or cellular structures were exposed in 25 mg L^?1 aqueous hydrogen tetrachloroaurate solutions separately at room temperature. Isolated viable chloroplasts from C. infusionum and thylakoids from A. sphaerica were found to be able to reduce gold ions. The protein extracts of both strains were also able to synthesize GNP at 4°C. Extracted polysaccharides of the two strains responded differently. Polysaccharides from A. sphaerica showed positive response in GNP synthesis, whereas no change was observed for C. infusionum. The carotenoids extracts from both strains acted like an efficient reducing agent. Initially the reducing efficiency of these extracted components was confirmed by the appearance of purple color in biomass or in experimental media. The GNPs, synthesized within the biomass were extracted by sonication with sodium citrate. The UV–vis spectroscopy of extracted purple colored suspensions and media showed the absorption bands at approximately 530–540 nm indicating a strong positive signal of GNP synthesis. Transmission electro n microscopy determined the size and shapes of the particles. The X‐ray diffraction study of the synthesized GNP revealed that the 2θ values appeared at 38.2°, 44.5°, 64.8° and 77.8°. Amongst all, isolated thylakoids and chloroplast showed only spherical GNP production with variable size range at pH 4. Monodisperse GNPs were also synthesized by isolated thylakoids and chloroplast at pH 9. A detailed morphological change of gold treated biomass was revealed employing scanning electron microscopy. The fluorescent property of gold loaded cells was studied by fluorescence microscopy.     

Floral volatiles with colour cues from two cucurbitaceous plants causing attraction of Aulacophora foveicollis

Aulacophora foveicollis Lucas (Coleoptera: Chrysomelidae) is an important phytophagous pest of two cucurbitaceous plants, Momordica cochinchinensis Spreng and Solena amplexicaulis (Lam.) Gandhi. The volatile organic compound profiles from flowers of M. cochinchinensis and S. amplexicaulis were identified and quantified by gas chromatography‐mass spectrometry (GC‐MS) and GC‐flame ionization detector (FID) analyses. Twenty nine and 28 compounds were identified in volatiles of M. cochinchinensis and S. amplexicaulis flowers, respectively. Methyl jasmonate and 3‐octanol were the predominant volatiles of M. cochinchinensis flowers, whereas 1‐octadecanol and 1‐hexanol were most found in the headspace of S. amplexicaulis flowers. Aulacophora foveicollis were more attracted by the flower volatiles of M. cochinchinensis than by those of S. amplexicaulis in a glass Y‐tube olfactometer. A mixture of 1‐heptanol, linalool oxide, 1‐octanol, and nonanal in the proportions present in the headspace of both flower types elicited attraction in the insect. From 25 cm distance, A. foveicollis displayed a preference for artificial flowers of 6.5 cm diameter of S. amplexicaulis flower colour (white) over M. cochinchinensis flower colour (white‐yellow). Finally, a synthetic blend (0.43 μg 1‐heptanol + 1.44 μg linalool oxide + 0.14 μg 1‐octanol + 1.77 μg nonanal dissolved in 25 μl methylene chloride) attracted more beetles when applied in a white artificial flower than when applied in a white‐yellow artificial flower from 40 cm distance. This finding may be helpful in the development of traps for pest management strategies.     

PON-mt-tRNA: a multifactorial probability-based method for classification of mitochondrial tRNA variations

Transfer RNAs (tRNAs) are essential for encoding the transcribed genetic information from DNA into proteins. Variations in the human tRNAs are involved in diverse clinical phenotypes. Interestingly, all pathogenic variations in tRNAs are located in mitochondrial tRNAs (mt-tRNAs). Therefore, it is crucial to identify pathogenic variations in mt-tRNAs for disease diagnosis and proper treatment. We collected mt-tRNA variations using a classification based on evidence from several sources and used the data to develop a multifactorial probability-based prediction method, PON-mt-tRNA, for classification of mt-tRNA single nucleotide substitutions. We integrated a machine learning-based predictor and an evidence-based likelihood ratio for pathogenicity using evidence of segregation, biochemistry and histochemistry to predict the posterior probability of pathogenicity of variants. The accuracy and Matthews correlation coefficient (MCC) of PON-mt-tRNA are 1.00 and 0.99, respectively. In the absence of evidence from segregation, biochemistry and histochemistry, PON-mt-tRNA classifies variations based on the machine learning method with an accuracy and MCC of 0.69 and 0.39, respectively. We classified all possible single nucleotide substitutions in all human mt-tRNAs using PON-mt-tRNA. The variations in the loops are more often tolerated compared to the variations in stems. The anticodon loop contains comparatively more predicted pathogenic variations than the other loops. PON-mt-tRNA is available at http://structure.bmc.lu.se/PON-mt-tRNA/.     

Glucose deprivation induced upregulation of phosphoenolpyruvate carboxykinase modulates virulence in Leishmania donovani

Various physiological stimuli trigger the conversion of noninfective Leishmania donovani promastigotes to the infective form. Here, we present the first evidence of the effect of glucose starvation, on virulence and survival of these parasites. Glucose starvation resulted in a decrease in metabolically active parasites and their proliferation. However, this was reversed by supplementation of gluconeogenic amino acids. Glucose starvation induced metacyclogenesis and enhanced virulence through protein kinase A regulatory subunit (LdPKAR1) mediated autophagy. Glucose starvation driven oxidative stress upregulated the antioxidant machinery, culminating in increased infectivity and greater parasitic load in primary macrophages. Interestingly, phosphoenolpyruvate carboxykinase (LdPEPCK), a gluconeogenic enzyme, exhibited the highest activity under glucose starvation to regulate growth of L. donovani by alternatively utilising amino acids. Deletion of LdPEPCK (Δpepck) decreased virulent traits and parasitic load in primary macrophages but increased autophagosome formation in the mutant parasites. Furthermore, Δpepck parasites failed to activate the Pentose Phosphate Pathway shunt, abrogating NADPH/NADP⁺ homoeostasis, conferring increased susceptibility towards oxidants following glucose starvation. In conclusion, this study showed that L. donovani undertakes metabolic rearrangements via gluconeogenesis under glucose starvation for acquiring virulence and its survival in the hostile environment.     

The cystathionine‐β‐synthase domains on the guanosine 5′’‐monophosphate reductase and inosine 5′‐monophosphate dehydrogenase enzymes from Leishmania regulate enzymatic activity in response to guanylate and adenylate nucleotide levels

The Leishmania guanosine 5′‐monophosphate reductase (GMPR) and inosine 5′‐monophosphate dehydrogenase (IMPDH) are purine metabolic enzymes that function maintaining the cellular adenylate and guanylate nucleotide. Interestingly, both enzymes contain a cystathionine‐β‐synthase domain (CBS). To investigate this metabolic regulation, the Leishmania GMPR was cloned and shown to be sufficient to complement the guaC (GMPR), but not the guaB (IMPDH), mutation in Escherichia coli. Kinetic studies confirmed that the Leishmania GMPR catalyzed a strict NADPH‐dependent reductive deamination of GMP to produce IMP. Addition of GTP or high levels of GMP induced a marked increase in activity without altering the K_m values for the substrates. In contrast, the binding of ATP decreased the GMPR activity and increased the GMP K_m value 10‐fold. These kinetic changes were correlated with changes in the GMPR quaternary structure, induced by the binding of GMP, GTP, or ATP to the GMPR CBS domain. The capacity of these CBS domains to mediate the catalytic activity of the IMPDH and GMPR provides a regulatory mechanism for balancing the intracellular adenylate and guanylate pools.     

Mephebrindole,a synthetic indole analog coordinates the crosstalk between p38MAPK and eIF2α/ATF4/CHOP signalling pathways for induction of apoptosis in human breast carcinoma cells

The efficacy of cancer chemotherapeutics is limited by side effects resulting from narrow therapeutic windows between the anticancer activity of a drug and its cytotoxicity. Thus identification of small molecules that can selectively target cancer cells has gained major interest. Cancer cells under stress utilize the Unfolded protein response (UPR) as an effective cell adaptation mechanism. The purpose of the UPR is to balance the ER folding environment and calcium homeostasis under stress. If ER stress is prolonged, tumor cells undergo apoptosis. In the present study we demonstrated an 3,3′-(Arylmethylene)-bis-1H-indole (AMBI) derivative 3,3′-[(4-Methoxyphenyl) methylene]-bis-(5-bromo-1H-indole), named as Mephebrindole (MPB) as an effective anti-cancer agent in breast cancer cells. MPB disrupted calcium homeostasis in MCF7 cells which triggered ER stress development. Detailed evaluations revealed that mephebrindole by activating p38MAPK also regulated GRP78 and eIF2α/ATF4 downstream to promote apoptosis. Studies extended to in vivo allograft mice models revalidated its anti-carcinogenic property thus highlighting the role of MPB as an improved chemotherapeutic option.     

Polypeptide motions are dominated by peptide group oscillations resulting from dihedral angle correlations between nearest neighbors

To identify basic local backbone motions in unfolded chains, simulations are performed for a variety of peptide systems using three popular force fields and for implicit and explicit solvent models. A dominant "crankshaft-like" motion is found that involves only a localized oscillation of the plane of the peptide group. This motion results in a strong anticorrelated motion of the phi angle of the ith residue (phi(i)) and the psi angle of the residue i - 1 (psi(i-1)) on the 0.1 ps time scale. Only a slight correlation is found between the motions of the two backbone dihedral angles of the same residue. Aside from the special cases of glycine and proline, no correlations are found between backbone dihedral angles that are separated by more than one torsion angle. These short time, correlated motions are found both in equilibrium fluctuations and during the transit process between Ramachandran basins, e.g., from the beta to the alpha region. A residue's complete transit from one Ramachandran basin to another, however, occurs in a manner independent of its neighbors' conformational transitions. These properties appear to be intrinsic because they are robust across different force fields, solvent models, nonbonded interaction routines, and most amino acids.     

Identification of low micromolar dual inhibitors for aldose reductase (ALR2) and poly (ADP-ribose) polymerase (PARP-1) using structure based design approach

Clinical studies have revealed that diabetic retinopathy is a multifactorial disorder. Moreover, studies also suggest that ALR2 and PARP-1 co-occur in retinal cells, making them appropriate targets for the treatment of diabetic retinopathy. To find the dual inhibitors of ALR2 and PARP-1, the structure based design was carried out in parallel for both the target proteins. A series of novel thiazolidine-2,4-dione (TZD) derivatives were therefore rationally designed, synthesized and their in vitro inhibitory activities against ALR2 and PARP-1 were evaluated. The experimental results showed that compounds 5b and 5f, with 2-chloro and 4-fluoro substitutions, showed biochemical activities in micromolar and submicromolar range (IC₅₀ 1.34–5.03 μM) against both the targeted enzymes. The structure-activity relationship elucidated for these novel inhibitors against both the enzymes provide new insight into the binding mode of the inhibitors to the active sites of enzymes. The positive results of the biochemical assay suggest that these compounds may be further optimized and utilized for the treatment of diabetic retinopathy.