Development and evaluation of antimicrobial PVC-grafted polymer for enhanced paint applications.

Demand for antimicrobial paints is increasing globally due to the rising need to control microbial growth and reduce infection risks in various environments. This increased demand underscores the crucial role of advanced antimicrobial coatings in promoting health and safety. In this context, an innovative poly(vinyl chloride) (PVC) grafted polymer with 1-(2-aminoethyl piperazine) (AEP) was prepared and studied in detail. In this study, the prepared polymer was characterized using FTIR and NMR spectroscopy to examine the polymer's chemical structure and employed TGA and DSC for thermal stability analysis. The antimicrobial activity of the grafted polymer was evaluated through the agar diffusion method and showed a significant inhibition zone of 21.6 mm for S. aureus, 16.3 mm for E. coli, 18.3 mm for M. smegmatis, and 20.3 mm for C. albicans at a lowest concentration of 12.5 µg mL-1. To assess surface characteristics, the PVC-g-AEP polymer was mixed with commercial paint and applied to a glass surface. SEM and AFM analysis showed a 5-times increase in porosity while maintaining visual aesthetics. Additionally, the paint displayed excellent stability against water, retaining around 90% of its antimicrobial activity even after 15 washes. This advanced polymer not only exhibits superior antimicrobial properties but also improves paint durability, setting a new benchmark for high-performance antimicrobial coatings and significantly advancing protective paint technology.

Cumulative phylogenetic, sequence and structural analysis of Insulin superfamily proteins provide unique structure-function insights.

The insulin superfamily proteins (ISPs), in particular, insulin, IGFs and relaxin proteins are key modulators of animal physiology. They are known to have evolved from the same ancestral gene and have diverged into proteins with varied sequences and distinct functions, but maintain a similar structural architecture stabilized by highly conserved disulphide bridges. The recent surge of sequence data and the structures of these proteins prompted a need for a comprehensive analysis, which connects the evolution of these sequences (427 sequences) in the light of available functional and structural information including representative complex structures of ISPs with their cognate receptors. This study reveals (a) unusually high sequence conservation of IGFs (>90 % conservation in 184 sequences) and provides a possible structure-based rationale for such high sequence conservation; (b) provides an updated definition of the receptor-binding signature motif of the functionally diverse relaxin family members (c) provides a probable non-canonical C-peptide cleavage site in a few insulin sequences. The high conservation of IGFs appears to represent a classic case of resistance to sequence diversity exerted by physiologically important interactions with multiple partners. We also propose a probable mechanism for C-peptide cleavage in a few distinct insulin sequences and redefine the receptor-binding signature motif of the relaxin family. Lastly, we provide a basis for minimally modified insulin mutants with potential therapeutic application, inspired by concomitant changes observed in other insulin superfamily protein members supported by molecular dynamics simulation.

Synthesis, characterization, and antimicrobial activities of a starch-based polymer.

Due to the rise of nosocomial infections and the increasing threat of antibiotic resistance, new techniques are required to combat bacteria and fungi. Functional antimicrobial biodegradable materials developed from low-cost renewable resources like polysaccharides would enable greater applications in this regard. Our group has developed and characterized a new antimicrobial polymer using commercially available N-ethyl piperazine and starch via simple one-pot method. The prepared antimicrobial polymer was characterized by FTIR and NMR. In addition, the thermal properties of the synthesized antimicrobial polymer were examined through TGA and DSC. The antimicrobial potential of the prepared material was investigated using the bacteria, Staphylococcus aureus, Escherichia coli, and Mycobacterium smegmatis and a fungi Candida albicans. The result indicates that, as the amount of polymer increases, the antimicrobial activity also increases. SA-E-NPz exhibited a zone of inhibition in the range of 8-13 mm, and the MIC was found to be < 0.625 mg against all four microbes. The antimicrobial activity of polymer coated on fabric was also studied. Furthermore, the cytotoxicity studied against human fibroblast cell lines showed that the prepared polymer is non-toxic to the cells. The study concluded that the synthesized polymer shows good antimicrobial activity, is non-toxic to human fibroblast cells, and thus can be used for wound dressing or textile applications.

First Report of Colletotrichum siamense Prihastuti, L. Cai & K.D. Hyde causing anthracnose disease in Santalum album Linn. in India.

Indian sandalwood (Santalum album), valued for its medicinal properties, is an indigenous species of India. Circular or irregular pale yellow lesions surrounded by a purple halo with prominent pinhead sized black fruiting bodies at the centre of the lesion were observed on leaves of sandalwood seedlings in a nursery located in Karnataka, with a disease incidence of 75% (n = 100 investigated plants) during July 2020. The disease prevailed in monsoon followed by winter season (July 2020 - January 2021); summer was less supportive for the disease incidence. As the disease progressed, lesions expanded and merged, causing necrosis of the whole leaf. Isolation of the pathogen involved excision of small sections of diseased tissues from the lesions followed by surface sterilization with 70% ethanol for 30 s and in 1% NaClO for 1 min. Sections were rinsed in sterile distilled water, placed on potato dextrose agar (PDA), and incubated at 25°C for 7 days. Ten isolates of Colletotrichum ssp. were obtained with an isolation frequency = 10/12×100 = 83%. One representative single-spore isolate (CSSA-1) was used for further study. Initially, pure cultures exhibited a white mycelium which later turned gray with time, and had orange conidial masses in a concentric ring pattern with the aggregation of black acervuli at the center of the culture Conidia were single celled, hyaline, and cylindrical having smooth rounded ends and the size ranged from 12.6 to 18.5 µm in length, and 3.5 to 5.6 µm in width (n = 100). The morphological characteristics were in agreement with the species description of fungi in the Colletotrichum gloeosporioides species complex (Weir et al. 2012). To confirm the species designation of the isolate CSSA-1, a multilocus phylogenetic analysis was performed using six genomic loci (Weir et al. 2012; Marin-Felix et al. 2017). The internal transcribed spacer (ITS) region of rDNA and a partial sequence of the beta-tubulin (TUB2), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), chitin synthase 1 (CHS-1), actin (ACT), and glutamine synthetase (GS) genes were amplified using ITS1/ITS4, BT2F/BT4R, GAPDHF/R, 79F/354R, 512F/783R and GSF/GSR primers, respectively. The ITS (OK254122), TUB2 (OL462863), GAPDH (OL462859), CHS-1 (OL462860), actin (OL462861), and glutamine synthetase (OL961822) sequences of representative isolate CSSA-1 showed 99 to 100% identity with sequences MZ148628, MK967339, MN525882, MW192791, MT263504, MF111030, MH370542 and KX578767, respectively to the holotype isolate of Colletotrichum siamense (Prihastuti et al. 2009). The sequences were analysed with representative sequences of Colletotrichum and a multilocus Bayesian inference phylogenetic tree with ITS-GAPDH-ACT-CHS1-GS-TUB2 concatenated data sets (concatenated with Sequence Matrix v.1.8 (Vaidya et al. 2011)) was constructed using Beast version 1.8.4 to confirm the isolate identification (Drummond et al. 2012; Hyde et al. 2014; Weir et al. 2012). Isolate CSSA-1 clustered with C. siamense isolates. To complete Koch's postulates, for the characterized isolate CSSA-1, a pathogenicity test was conducted on 3-month-old sandalwood seedlings by spore spray inoculation. Ten plants were inoculated with a conidial suspension (106 conidia/ml) and control plants inoculated with sterilized water then kept in a glass house at 25°C and >85% relative humidity with a 12-h photoperiod. Humidity was maintained by spraying the plants with water in the morning and evening to enhance the infection. Typical symptoms of anthracnose disease similar to naturally infected leaves were observed, which included circular pale yellow lesions surrounded by a purple halo with prominent pinhead sized black fruiting bodies at the center of the lesion 7 days after inoculation, while the control plants remained unaffected. The pathogen was reisolated from infected leaves and its identity was confirmed as C. siamense based on morphological characteristics. Previously, C. siamense was identified causing disease on chili in India (Gunjan and Shenoy, 2014), but to our knowledge this is the first report of leaf anthracnose caused by C. siamense on Indian sandalwood in India or globally. This study documents crucial information, paving way for epidemiologic studies and design of control strategies to combat this newly emerging disease.

Investigation of effective natural inhibitors for starch hydrolysing enzymes from Simaroubaceae plants by molecular docking analysis and comparison with in-vitro studies.

The present study aims to find the effective natural enzyme inhibitors against alpha-amylase and alpha-glucosidase from the array of compounds identified in plants of the Simaroubaceae family using molecular docking and ADME/Toxicity studies. Among the 218 compounds docked against seven enzymes, buddlenol-A and citrusin-B showed the best binding energies (kcal/mol) of -7.830 and -7.383 against human salivary alpha-amylase and pancreatic alpha-amylase respectively. The other two compounds 9-hydroxycanthin-6-one and bruceolline-B had the best binding energy of -6.461 and -7.576 against N-terminal and C-terminal maltase glucoamylase respectively. Whereas the binding energy of prosopine (-6.499) and fisetinidol (-7.575) was considered as the best against N-terminal and C-terminal sucrase-isomaltase respectively. Picrasidine-X showed the best binding energy (-7.592) against yeast alpha-glucosidase. The study revealed that the seven compounds which showed the best binding energy against respective enzymes are considered as the 'lead hit compounds'. Even though the 'lead hit compounds' are not obeying all the laws of ADMET, the drug-likeness properties of 9-hydroxycanthin-6-one, fisetinidol, picrasidine-X, and prosopine were considerable. Also, kaempferol-3-O-pentoside was the recent compound identified from the Simarouba glauca plant extract found to be one among the top five lead hit compounds against four enzymes. This study provides valuable insight into the direction of developing natural compounds as potential starch hydrolysing enzyme inhibitors for managing type 2 diabetes.

Synthesis and Evaluation of the Insulin-Albumin Conjugate with Prolonged Glycemic Control.

Engineering therapeutic proteins to improve their half-life so as to sustain physiologically relevant extended activity is the need of the hour in biopharmaceutical research. In this study, insulin and bovine serum albumin (BSA) were independently functionalized rationally and were later conjugated to prolong the half-life of insulin. The thiol functionalization of BSA with 2-imminothiolane in the ratio 1:20 yielded an average of 6-8 thiols/BSA, which then reacted with maleimide-functionalized insulin to form an insulin-albumin conjugate. The bioconjugate was purified by size exclusion chromatography, and the increase in size was confirmed by sodium dodecyl-sulfate polyacrylamide gel electrophoresis. Bioconjugation resulted in a multi-fold increase in the hydrodynamic volume of the insulin-albumin conjugate as measured in DLS when compared to BSA. The glucose uptake assay with 3LT3-L1 cell lines was performed, and the mean fluorescence intensity (MFI) of 16.16 observed for the insulin-albumin conjugate was comparable to insulin (19.42). The blood glucose reducing capacity of the insulin-albumin conjugate in streptozotocin induced diabetic male Wistar rats was well maintained up to 72 h when compared to native insulin. Further, a three-fold increase in plasma insulin concentration was observed in bioconjugate treated animals as against insulin treated animals after 24 h of treatment using ELISA. The histological analysis of different organs of the bioconjugate treated rats indicated that it was non-toxic. This study has paved a way for further detailed studies on similar bioconjugates to develop next-generation biotherapeutics for treating diabetes.

Kinetic characterization of purified laccase from Trametes hirsuta: a study on laccase catalyzed biotransformation of 1,4-dioxane.

OBJECTIVE: Laccase is one of the best known biocatalysts which degrade wide varieties of complex molecules that are both non-cyclic and cyclic in structure. The study focused on enzyme kinetics of a purified laccase from Trametes hirsuta L. fungus and its application on biotransformation of a carcinogenic molecule 1,4-dioxane. RESULTS: Laccase was purified from white-rot fungus T. hirsuta L. which showed specific activity of 978.34 U/mg after the purification fold of 54.08. The stable laccase activity (up to 16 h) is shown at 4-6 pH and 20-40 °C temperature range. The purified enzyme exhibited significant stability for 10 metal ions up to 10 mM concentration, except for Fe2+ and Hg2+. The Cu2+ ion induced laccase activity up to 142% higher than the control at 10 mM concentration. The laccase enzyme kinetic parameters Km was 20 ± 5 µM and 400 ± 60 µM, whereas Kcat was 198.29 ± 0.18/s and 80.20 ± 1.59/s for 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) and guaiacol respectively. The cyclic ether 1,4-dioxane (100 ppm) was completely degraded in presence of purified laccase within 2 h of incubation and it was confirmed by HPLC and GC analysis. The oxidation reaction was accelerated by 25, 22, 6 and 19% in presence of 1 mM syringaldehyde, vanillin, ABTS and guaiacol mediators respectively. CONCLUSIONS: In this study, fungal laccase (a natural biocatalyst) based degradation of synthetic chemical 1,4-dioxane was reported for the first time. This method has added advantages over the multiple methods reported earlier being a natural remedy.

Alpha glucosidase inhibition activity of phenolic fraction from Simarouba glauca: An in-vitro, in-silico and kinetic study.

A phenolic rich fraction purified from Simarouba glauca leaves was effective in alpha glucosidase inhibition. The purified fraction named 'fraction-14' had shown significant inhibition of yeast alpha glucosidase enzyme activity (IC50 = 2.4 ± 0.4 µg/mL) when compared to anti-diabetic drug acarbose (IC50 = 2450 ± 24 µg/mL). The purified fraction also had reasonable DPPH (IC50 = 14.4 ± 0.1 µg/mL) and ABTS (IC50 = 7.6 ± 0.5 µg/mL) free radical scavenging activity when compared to the standard ascorbic acid. The LC-MS analysis of bioactive 'fraction-14' revealed four compounds, eclalbasaponin-v (1), cyanidin-3-O-(2'galloyl)-galactoside (2), kaempferol-3-O-glucoside (3) and kaempferol-3-O-pentoside (4) for the first time in S. glauca in this study. The kinetic study of the 'fraction-14' indicates a mixed type of inhibition on the alpha glucosidase enzyme with K i , 6.2 µg/mL. Docking studies showed promising binding energy for the compounds 2 (-7.769 kJ/mol), 3 (-7.04 kJ/mol) and 4 (-7.127 kJ/mol) against yeast alpha glucosidase which was better than acarbose (-6.867 kJ/mol). In conclusion, the phenolic rich fraction from S. glauca possessing good in-vitro antioxidant property and alpha glucosidase enzyme inhibition potential along with mixed inhibition kinetics. Also, better binding energy of compounds (1, 2 & 3) appears to contain potential lead-molecule for antidiabetic therapy.

Scheelite like NaTb(WO4)2 nanoparticles: Green fluorescence and in vitro cell imaging applications.

This study highlights the investigation of the green fluorescence in NaTb(WO4)2 materials (NaTbW Bulk and NaTbW Nano) synthesized via template free hydrothermal method as a function of particle size and morphology. Herein, we demonstrated the biocompatibility and intracellular green fluorescence of NaTbW Nano samples using HeLa cells for cell imaging applications. Powder X-ray diffraction studies showed that the as synthesized NaTbW Bulk and NaTbW Nano crystallize in the Scheelite like tetragonal crystal system with the I41/a space group. The reaction pH and solvent is observed to play a critical role in determining particle size, shape and morphology of these luminescent materials. Furthermore, size dependent optical properties were systematically studied by diffuse reflectance, steady state photoluminescence; time resolved fluorescence lifetime and quantum yield measurements. Both the materials have shown bright green fluorescence upon UV excitation as a function of particle size. Remarkable high quantum yield of NaTbW Bulk indicated its greater luminescence efficiency and the closer CIE coordinates to the commercial green illuminant suggested their potential use in solid state display systems. On the other hand the observed biocompatibility of NaTbW Nano particles towards mammalian cancer HeLa cells, Staphylococcus aureus, Escherichia coli and the intracellular green fluorescence rightly proved its functionality as active bio-probes. Thus, our work summarize the potential use of these Scheelite like NaTb(WO4)2 material for solid state display and bio-imaging applications.

Efficient Synthesis and in Silico Studies of the Benzimidazole Hybrid Scaffold with the Quinolinyloxadiazole Skeleton with Potential α-Glucosidase Inhibitory, Anticoagulant, and Antiplatelet Activities for Type-II Diabetes Mellitus Management and Treating Thrombotic Disorders.

The current study evaluates antidiabetic, anticoagulant, and antiplatelet activity of novel benzimidazole-containing quinolinyl oxadiazoles. These derivatives are synthesized and characterized using spectroscopy (FT-IR, 1H NMR, and mass spectroscopy) and single-crystal X-ray diffraction methods. The inhibitory effects of these compounds were evaluated by the α-glucosidase inhibitory assay and shows the activity in the range of IC50 = 0.66 ± 0.05 to 3.79 ± 0.46 µg/mL. In addition, molecular docking studies revealed that benzimidazole-containing quinolinyl oxadiazoles can correctly dock into the target receptor protein of the human intestinal α-glucosidase, while their bioavailability/drug-likeness was predicted to be acceptable but requires further optimization. On the other hand, compound 8a and 8d showed anticoagulant activity as they enhanced the clotting time from control 180-410 and 180-390 s, respectively, in platelet rich plasma and 230-460 and 230-545 s in platelet poor plasma. Furthermore, only 8a showed antiplatelet activity by inhibiting epinephrine-induced platelet aggregation, and the observed aggregation inhibition was found to be 93.4%. Compounds 8a-f show nontoxic properties because of the non-hydrolyzing properties in the RBC cells. In addition, 8a and 8d show anti-edema and anti-hemorrhagic properties in the experimental mice. These findings reveal that benzimidazole-containing quinolinyl oxadiazoles act as α-glucosidase inhibitors to develop novel therapeutics for treating type-II diabetes mellitus and can act as lead molecules in drug discovery as potential antidiabetic and antithrombotic agents.

Design and regioselective synthesis of trifluoromethylquinolone derivatives as potent antimicrobial agents.

Three series of new trifluoromethyl substituted quinolone derivatives were synthesized (4a-f, 6a-f and 8a-f) from corresponding substituted anilines by multi-step reactions. The regioselective alkylation with different alkyl halides were carried out by approaching two different routes to get the final products in good yield. Newly synthesized compounds were characterized by spectral study and also by C, H, N analyses. Three dimensional structure of 2b and 4b were also confirmed by single crystal X-ray studies. The final compounds (4a-f, 6a-f and 8a-f) were screened for their in-vitro antibacterial and antifungal activity by well plate method (zone of inhibition). The results revealed that, compounds 4a, 6b, 6c and 8e showed significant antibacterial activity as compared to the standard drug Ciprofloxacin. The compound 8a was found to be a potent antifungal agent.