Insights into the binding of thiosemicarbazone derivatives with human serum albumin: spectroscopy and molecular modelling studies

4-[(1Z)-1-(2-carbamothioylhydrazinylidene)ethyl]phenyl acetate [Ace semi],4-[(1Z)-1-(2-carbamothioylhydrazinylidene)ethyl]phenyl propanoate [Pro semi] from the family of thiosemicarbazones derivative has been newly synthesized. It has good anticancer activity as well as antibacterial and it is also less toxic in nature, its binding characteristics are therefore of huge interest for understanding pharmacokinetic mechanism of the drug. The binding of thiosemicarbazone derivative to human serum albumin (HSA) has been investigated by studying its quenching mechanism, binding kinetics and the molecular distance (r) between donor (HSA) and acceptor (thiosemicarbazone derivative) was estimated according to Forster’s theory of non-radiative energy transfer using fluorescence spectroscopy. The binding dynamics has been elaborated using synchronous fluorescence spectroscopy, and the feature of thiosemicarbazone derivative induced structural changes of HSA has been studied by circular dichorism, Fourier transform infrared spectroscopy. Molecular modelling simulations explore the hydrophobic interaction and hydrogen bonding which stabilizes the interaction.     

Biosynthesis of silver nanoparticles using Bacillus subtilis EWP-46 cell-free extract and evaluation of its antibacterial activity

This study highlights the ability of nitrate-reducing Bacillus subtilis EWP-46 cell-free extract used for preparation of silver nanoparticles (AgNPs) by reduction of silver ions into nano silver. The production of AgNPs was optimized with several parameters such as hydrogen ion concentration, temperature, silver ion (Ag⁺ ion) and time. The maximum AgNPs production was achieved at pH 10.0, temperature 60 °C, 1.0 mM Ag⁺ ion and 720 min. The UV–Vis spectrum showed surface plasmon resonance peak at 420 nm, energy-dispersive X-ray spectroscopy (SEM–EDX) spectra showed the presence of element silver in pure form. Atomic force microscopy (AFM) and transmission electron microscopy images illustrated the nanoparticle size, shape, and average particle size ranging from 10 to 20 nm. Fourier transform infrared spectroscopy provided the evidence for the presence of biomolecules responsible for the reduction of silver ion, and X-ray diffraction analysis confirmed that the obtained nanoparticles were in crystalline form. SDS-PAGE was performed to identify the proteins and its molecular mass in the purified nitrate reductase from the cell-free extract. In addition, the minimum inhibitory concentration and minimum bactericidal concentration of AgNPs were investigated against gram-negative (Pseudomonas fluorescens) and gram-positive (Staphylococcus aureus) bacteria.     

Engineering D-Amino Acid Containing Collagen Like Peptide at the Cleavage Site of Clostridium histolyticum Collagenase for Its Inhibition

Collagenase is an important enzyme which plays an important role in degradation of collagen in wound healing, cancer metastasis and even in embryonic development. However, the mechanism of this degradation has not yet been completely understood. In the field of biomedical and protein engineering, the design and development of new peptide based materials is of main concern. In the present work an attempt has been made to study the effect of ^DAla in collagen like peptide (imino-poor region of type I collagen) on the structure and stability of peptide against enzyme hydrolysis. Effect of replacement of ^DAla in the collagen like peptide has been studied using circular dichroic spectroscopy (CD). Our findings suggest that, ^DAla substitution leads to conformational changes in the secondary structure and favours the formation of polyproline II conformation than its L-counterpart in the imino-poor region of collagen like peptides. Change in the chirality of alanine at the cleavage site of collagenase in the imino-poor region inhibits collagenolytic activity. This may find application in design of peptides and peptidomimics for enzyme-substrate interaction, specifically with reference to collagen and other extra cellular matrix proteins.     

Modeling studies on phospholipase A2-inhibitor complexes

Phospholipase A2 (PLA2) is a ubiquitous enzyme that specifically catalyzes hydrolysis of membrane phospholipids to produce lysophospholipids and free fatty acid, namely arachidonic acid, which provides substrate for eicosanoids biosynthesis. Thus, the compounds inhibiting PLA2 have been implicated as potential therapeutic agents in treatment of inflammation related diseases. Plant and marine organisms serve as sources of compounds that act as potential therapeutic agents for treatment of various diseases. The present study reveals the relationship between the structure and function of the medicinally important herbal compounds (acalyphin, chlorogenic acid, stigmasterol, curcumin and tectoridin) and marine compounds (gracilin A and aplysulphurin A). To understand the binding mechanisms of these compounds, molecular modeling studies has been performed with Russell's viper and bovine pancreatic PLA2 as target molecules using molecular operating environment (MOE) software. These compounds show favorable interactions with the amino acid residues at the active site of Russell's viper and bovine pancreatic PLA2, thereby substantiating their proven efficacy as anti-inflammatory compounds and antidotes.     

Rhizoremediation of Cadmium Soil Using a Cadmium-Resistant Plant Growth-Promoting Rhizopseudomonad

Three pseudomonad strains (MKRh1, MKRh3, and MKRh4) isolated from rhizospheres showed a high growth potential in the presence of cadmium, with a minimal inhibitory concentration of 7 mM for cadmium chloride (CdCl₂). Among them, isolate MKRh3 was specifically chosen as the most favorable cadmium-resistant plant growth-promoting rhizobacterium based on its higher 1-aminocyclopropane carboxylic acid deaminase activity, siderophore production, phosphate solubilization, and auxin synthesis and the in vivo growth increment of black gram plants. 16S ribosomal RNA gene sequencing identified MKRh3 as Pseudomonas aeruginosa. The effect of cadmium on black gram plants was studied in soil amended with a gradient of CdCl₂ concentration and the toxicity was evident from stunted growth, poor rooting, and cadmium accumulation. Application of isolate MKRh3 by seed coating overcomes the cadmium toxicity; plants showed lessened cadmium accumulation, extensive rooting, and enhanced plant growth. Further research and development of this promising innate strategy for scale-up to higher-efficiency and large-scale application will be a potent tool to prevent accumulation of cadmium in plants, thereby ensuring food security for humans.     

Isolation and genetic characterization of Toxoplasma gondii from raccoons (Procyon lotor), cats (Felis domesticus), striped skunk (Mephitis mephitis), black bear (Ursus americanus), and cougar (Puma concolor) from Canada

Viable Toxoplasma gondii was isolated by bioassay in mice from tissues of 2 feral cats (Felis domesticus), 2 raccoons (Procyon lotor), a skunk (Mephitis mephitis) trapped in remote locations in Manitoba, Canada, and a black bear (Ursus americanus) from Kuujjuaq, northern Quebec, Canada. Genotyping of these T. gondii isolates using polymorphisms at 10 nuclear markers including SAGI, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and an apicoplast marker Apico revealed 4 genotypes. None of the isolates was clonal archetypal Types I, II, and III found in the United States. These results are in contrast with the Type II genotype that is widespread in domestic animals and humans throughout the United States and Europe. This is the first genotyping of T. gondii isolates from this part of North America.     

Pine cone-mediated green synthesis of silver nanoparticles and their antibacterial activity against agricultural pathogens

The medicinal and physicochemical properties of nanoscale materials are strong functions of the particle size and the materials used in their synthesis. The nanoparticle shape also contributes significantly to their medicinal properties. Several shapes ranging from oval, spherical, rods, to teardrop structures may be obtained by chemical methods. Triangular and hexagonal nanoparticles have been synthesized by using a pine cone extract (PCE). Here, we report the discovery that PCE, when reacted with silver nitrate ions, yields a high percentage of thin, flat, single-crystalline nanohexagonal and nanotriangular silver nanoparticles. The nanohexagonal and nanotriangular nanoparticles appear to grow by a process involving rapid reduction with assembly at room temperature at a high pH. The nanoparticles were characterized by UV–Vis absorption spectroscopy, SEM-EDS, TEM, FTIR, and X-ray diffraction analyses. The anisotropy of the nanoparticle shape results in large near-infrared absorption by the particles. Highly anisotropic particles are applicable in various fields, including agriculture and medicine. The obtained silver nanoparticles (Ag NPs) had significant antibacterial action on both Gram classes of bacteria associated with agriculture. Because the Ag NPs are encapsulated with functional group-rich PCE, they can be easily integrated in various applications.     

Identification and characterization of granule bound starch synthase (GBSS-I) from common buckwheat (Fagopyrum esculentum Moench)

Starch grains present in the endosperm of grains of common buckwheat (Fagopyrum esculentum Moench) show a monomodal distribution with size ranging from 4 to 10 μm. SDS-PAGE analysis of starch granule bound proteins revealed the presence of a single band corresponding to molecular mass of 59.7 kDa. The protein is localized within the central core of the starch grains. Antisera raised against the 59.7 kDa protein cross reacted with the 61 kDa GBSS-I from endosperm starches of maize and the 60 kDa GBSS-I from endosperm starches of rice and wheat, thereby indicating serological homology between the 59.7 kDa buckwheat starch granule bound protein and GBSS-I of wheat, maize and rice. 2D-PAGE of starch granule bound proteins of common buckwheat resolved the fraction into 7 spots with pI ranging from 5.2 to 5.6. N-terminal amino acid sequence for 25 residues of two immunoreactive proteins separated by 2D PAGE showed 94 % homology with N-terminal amino acid sequence of GBSS-I from Hordeum vulgare, Triticum spp. and Phaseolus vulgaris. Even though analysis of the sequence alignment revealed a clear diversification into monocotyledonous and dicotyledonous groups, the protein from buckwheat showed similarities with GBSS-I from both dicots as well as monocots. As is the case with dicots, the sequence of GBSS-I from buckwheat has valine as the 11th residue. GBSS-I from majority of monocots has methionine at this position. The sequence also showed similarities with monocots with valine at P’5 from the N-terminus. GBSS-I from majority of dicots has isoleucine at this position. The significance of these substitution remains to be ascertained.