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Seeds harbor naturally occurring microbial endophytes that proliferate during seedling development; playing crucial roles in seedling growth, establishment, and protection against fungal pathogens. Resilient actinobacteria of wheat seeds have been explored in this study for their beneficial traits. Ten actinobacteria isolated from the surface-sterilized seeds of wheat variety HD3117 were identified as nine species of Streptomyces and one of Nocardiopsis. Most isolates could grow at 42°C, 5% NaCl, and 10% poly ethylene glycol (PEG); exhibited variable hydrolytic enzyme production for amylase, cellulase, and protease. Few isolates produced indole acetic acid (9.0-18.9 µg ml-1 ) and could solubilize P (11.3-85.2 µg ml-1 ). The isolates were antagonistic against one or more fungal pathogens under test (Fusarium graminearum, Bipolaris sorokiniana, Alternaria sp., and Tilletia indica), of which Streptomyces sampsonii WSA20 inhibited all in dual culture assay. Priming of wheat seeds with the efficient isolate WSA20 led to effective colonization in the root zone and significantly improved germination, shoot and root length in seed germination assay. Significant protection was recorded in microcosm experiment where no symptoms of disease were observed. This study shows the significance of actinobacterial endophytes of wheat seeds in influencing seed germination and seedling growth while protecting from soil-borne pathogens. It is original and suggests that the seed inhabiting efficient actinobacteria may be developed as efficient bioinoculant for sustainable farming system.
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Actinobacteria , Triticum , Triticum/microbiología , Bacterias , Semillas , PlantonesRESUMEN
The key objective of this paper is to study the cyclic codes over mixed alphabets on the structure of FqPQ, where P=Fq[v]⟨v3-α22v⟩ and Q=Fq[u,v]⟨u2-α12,v3-α22v⟩ are nonchain finite rings and αi is in Fq/{0} for i∈{1,2}, where q=pm with m≥1 is a positive integer and p is an odd prime. Moreover, with the applications, we obtain better and new quantum error-correcting (QEC) codes. For another application over the ring P, we obtain several optimal codes with the help of the Gray image of cyclic codes.
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Gamma amino butyric acid (GABA) is a chemical messenger that plays a significant role in muscle relaxation and brain health. Certain lactic acid bacteria (LAB) produce significant levels of GABA and thus act as potential psychobiotic cultures. In the present study, LAB were isolated from non-rhizospheric soil sample of Syzygium cumini (Black plum). A total of 57 LAB were isolated on the basis of their morphological and acid producing characteristic on de Man Rogosa Sharpe (MRS) agar. Only seven isolates were found to produce GABA (0.09-1.13 gL-1) in MRS broth and were identified as Lactococcus. However, L. lactis LP-68 produced highest amount of GABA and was selected for further optimization of culture conditions (pH, temperature and MSG) by response surface methodology (RSM). The optimization resulted in approximately four-fold increase in GABA production (4.11 gL-1). The results indicate that the L. lactis LP-68 can be used as starter culture for production of GABA-enriched functional foods.
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Lactococcus lactis , Prunus domestica , Syzygium , Humanos , Suelo , Ácido gamma-AminobutíricoRESUMEN
Here, different monoclonal antibodies (mAb1, mAb2 and mAb3) of Ebola virus were screened in a real-time and label-free manner using surface plasmon resonance (SPR) to select an appropriate antibody for biosensor applications against a biological warfare agent. For this purpose, a gold SPR chip was modified with 4-mercaptobenzoic acid (4-MBA), and modification was confirmed by FTIR-ATR and EIS. The 4-MBA-modified gold SPR chip was used for immobilization of the recombinant nucleoprotein of Ebola (EBOV-rNP), and the interactions of mAb1, mAb2 and mAb3 were then investigated to determine the best mAb based on the affinity constant (KD), expressed as equilibrium dissociation constant. KD values of 809 nM, 350 pM and 52 pM were found for the interaction of mAb1, mAb2 and mAb3 of Ebola with the immobilized EBOV-rNP, respectively, thus reflecting the high affinity of mAb3. This was confirmed by ELISA results. The thermodynamic parameters (ΔG, ΔH and ΔS) for the interaction between mAb3 and EBOV-rNP were also determined, which revealed that the interaction was spontaneous, endothermic and driven by entropy. The SPR limit of detection of EBOV-rNP with mAb3 was 0.5 pg ml-1, showing mAb3 to be the best high-affinity antibody in our study. This study has opened up new possibilities for SPR screening of different monoclonal antibodies of BWA through the convergence of materials science and optical techniques.
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Ebolavirus/aislamiento & purificación , Fiebre Hemorrágica Ebola/virología , Resonancia por Plasmón de Superficie/métodos , Anticuerpos Monoclonales/química , Anticuerpos Monoclonales/inmunología , Afinidad de Anticuerpos , Benzoatos/química , Ebolavirus/inmunología , Fiebre Hemorrágica Ebola/diagnóstico , Humanos , Compuestos de Sulfhidrilo/química , TermodinámicaRESUMEN
Community-forming traits (CFts) play an important role in the effective colonization of plant-growth-promoting bacterial communities that influence host plants positively by modulating their adaptive functions. In this study, by considering plant-growth-promoting traits (PGPts) and community-forming traits (CFts), three communities were constructed, viz., SM1 (PGPts), SM2 (CFts), and SM3 (PGPts+CFts). Each category isolates were picked up on the basis of their catabolic diversity of different carbon sources. Results revealed a distinctive pattern in the colonization of the communities possessed with CF traits. It was observed that the community with CFts colonized inside the plant in groups or in large aggregations, whereas the community with only PGPts colonized as separate individual and small colonies inside the plant root and leaf. The effect of SM3 in the microcosm experiment was more significant than the uninoculated control by 22.12%, 27.19%, and 9.11% improvement in germination percentage, chlorophyll content, and plant biomass, respectively. The significant difference shown by the microbial community SM3 clearly demonstrates the integrated effect of CFts and PGPts on effective colonization vis-à-vis positive influence on the host plant. Further detailed characterization of the interaction will take this technology ahead in sustainable agriculture.
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Biodiesel is a biological renewable source produced from the conversion of triglycerides to alkyl esters. Palm oil is one of the most used lipid feedstocks for biodiesel production. It becomes necessary to optimize the transesterification reaction parameters to reduce the cost and enhance the quality of biodiesel. This study focuses on the use of homogenous sulfuric acid as a catalyst for the transesterification of palm fatty acids to methyl esters in a batch-scale reactor. A novel examination of transesterification reaction input parameters using the technique for order performance by similarity to ideal solution optimization technique and the effect of these parameters on yield, viscosity, and density of palm biodiesel using 3D surface graphs is investigated in this research. The present optimization approach is implemented to find out the optimum ranking of biodiesel production. From the experimental and numerical simulation, optimum results were observed at the catalyst concentration of 6% (w/w), reaction temperature of 70 °C, the reaction time of 120 min, and alcohol to oil molar ratio of 30:1 at which yield of 95.35%, viscosity of 5.0 cSt, and density of 880 kg/m3 of palm biodiesel were obtained. The different physicochemical properties of produced palm methyl esters are obtained within standards set by international authorities. Selected optimized process parameters can be used for commercial-scale biodiesel production.
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Biocombustibles , Aceites de Plantas , Aceite de Palma , Aceites de Plantas/química , Esterificación , Ésteres/química , Ácidos , Catálisis , EtanolRESUMEN
To combat the shortage of single agro-residue and overcome the problem of seasonal availability, it is beneficial to use mixture of lignocellulosic biomasses. In the present study, efforts were made to use mixed lignocellulosic biomass for production of bioethanol, along with microbial lipids and lactic acid. Upon enzymatic hydrolysis of mixed biomass at varied proportions it was observed that mixture of paddy straw and jute in the ratio 3:1 resulted in best sugar yield (41.50 g/L) at 10% substrate loading. Ethanolic fermentation of mixed substrate hydrolysate by thermotolerant yeast, Saccharomyces cerevisiae JRC6 resulted in 8.39 g/L of ethanol. To maintain sustainability and economic impact, oleaginous yeast (Trichosporon mycotoxinivorans S2) and lactic acid bacteria (Lactobacillus plantarum LP-9) were used for lipid production (14.5 g/L) and lactic acid production (11.08 g/L), respectively. Therefore, this study explored the potential of mixed lignocellulosic biomass to be exploited for production of various value-added products.
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Ácido Láctico , Saccharomyces cerevisiae , Etanol , Fermentación , Lípidos , Biomasa , HidrólisisRESUMEN
Wheat yield can be limited by many biotic and abiotic factors. Heat stress at the grain filling stage is a factor that reduces wheat production tremendously. The potential role of endophytic microorganisms in mitigating plant stress through various biomolecules like enzymes and growth hormones and also by improving plant nutrition has led to a more in-depth exploration of the plant microbiome for such functions. Hence, we devised this study to investigate the abundance and diversity of wheat seed endophytic bacteria (WSEB) from heatS (heat susceptible, GW322) and heatT (heat tolerant, HD3298 and HD3271) varieties by culturable and unculturable approaches. The results evidenced that the culturable diversity was higher in the heatS variety than in the heatT variety and Bacillus was found to be dominant among the 10 different bacterial genera identified. Though the WSEB population was higher in the heatS variety, a greater number of isolates from the heatT variety showed tolerance to higher temperatures (up to 55°C) along with PGP activities such as indole acetic acid (IAA) production and nutrient acquisition. Additionally, the metagenomic analysis of seed microbiota unveiled higher bacterial diversity, with a predominance of the phyla Proteobacteria covering >50% of OTUs, followed by Firmicutes and Actinobacteria. There were considerable variations in the abundance and diversity between heat sensitivity contrasting varieties, where notably more thermophilic bacterial OTUs were observed in the heatT samples, which could be attributed to conferring tolerance against heat stress. Furthermore, exploring the functional characteristics of culturable and unculturable microbiomes would provide more comprehensive information on improving plant growth and productivity for sustainable agriculture.
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Water-diesel emulsion fuel has been found as a prominent alternative fuel by various researchers. Alike this technology, cow-urine (Bos indicus urine) emulsified diesel fuel (GMD emulsion) has been explored in this study. Making of homogeneous and stable emulsion is a crucial aspect in this approach while maintaining the diesel standards. Hence, the applicability of this fuel has been examined based on physicochemical properties for diesel engine application. The stability was assessed by creaming index, droplet size, Pdi, and interfacial tension. The minimum droplet size (278 nm) and 0.282 Pdi was obtained with 5.72 HLB value, 4% (v/v) surfactant, and 10% (v/v) of cow-urine through ultra-sonication. The optimized 10% GMD emulsion was found stable for more than 80 days. Viscosity and density of the fuel got increased with cow-urine and surfactant; however, they found within the standard limits. The physicochemical analysis exhibited the feasibility of the GMD emulsion for diesel engine application.
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Due to growing consumer preference towards natural ingredients in food products, the production of flavors by microbial biotransformation of agrowastes provides an eco-friendly, cost-effective and sustainable pathway for biovanillin production. In the present study, biovanillin was produced by microbial biotransformation of ferulic acid (FA) using Streptomyces sp. ssr-198. The strain was able to grow in glucose medium supplemented with 1 g/L FA and produce 20.91 ± 1.11 mg/L vanillin within 96 h, along with 5.78 ± 0.13 mg/L vanillic acid in 144 h. Estimation of enzymes involved in FA degradation detected maximum feruloyl-CoA synthetase activity (1.21 ± 0.03 U/mg protein) at 96 h and maximum vanillin dehydrogenase activity (0.31 ± 0.008 U/mg protein) at 168 h, with small amounts of ferulic acid esterase activity (0.13 ± 0.002 U/mg protein) in the fermentation medium. Further, the glucose deficient production medium supplemented with 3 g/L of ferulic acid when inoculated with Streptomyces sp. ssr-198 (6% wet weight) produced maximum vanillin (685 ± 20.11 mg/L) within 72 h at 37 °C under agitation (150 rpm) and declined thereafter. Furthermore, in a one-pot experiment, wherein crude ferulic acid esterase (700 IU/g of substrate) from Enterococcus lactis SR1 was added into 10% w/v wheat bran (natural source of ferulic acid) based medium and was inoculated with 1% w/v of Streptomyces sp. ssr-198 resulted in maximum vanillin production (1.02 ± 0.02 mg/g of substrate) within 60 h of incubation. The study provides an insight into synergistic effect of using FAE of E. lactis SR1 and Streptomyces sp. ssr-198 for bioproduction of biovanillin using agro residues. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-021-03006-0.
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The severe outbreak of respiratory coronavirus disease 2019 has increased the significant demand of respiratory mask and its use become ubiquitous worldwide to control this unprecedented respiratory pandemic. The performance of a respiratory mask depends on the efficiency of the filter layer which is mostly made of polypropylene melt blown non-woven (PP-MB-NW). So far, very limited characterization data are available for the PPE-MB-NW in terms to achieve desired particulate filtration efficiency (PFE) against 0.3 µm size, which are imperative in order to facilitate the right selection of PP-MB-NW fabric for the development of mask. In present study, eight different kinds of PP-MB-NW fabrics (Sample A-H) of varied structural morphology are chosen. The different PP-MB-NW were characterized for its pore size and distribution by mercury porosimeter and BET surface area analyzer was explored first time to understand the importance of blind pore in PFE. The PP-MB-NW samples were characterized using scanning electron microscopy so as to know the surface morphology. The filtration efficiency, pressure drop and breathing resistance of various PP-MB-NW fabric samples are investigated in single and double layers combination against the particle size of 0.3, 0.5 and 1 µm. The samples which are having low pore dia, high solid fraction volume, and low air permeability has high filtration efficiency (> 90%) against 0.3 µm particle with high pressure drop (16.3-21.3 mm WC) and breathing resistance (1.42-1.92 mbar) when compared to rest of the samples. This study will pave the way for the judicial selection of right kind of filter layer i.e., PP-MB-NW fabric for the development of mask and it will be greatly helpful in manufacturing of mask in this present pandemic with desired PFE indicating considerable promise for defense against respiratory pandemic.
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Filtros de Aire , COVID-19/prevención & control , Máscaras , Aerosoles/aislamiento & purificación , Filtros de Aire/virología , Diseño de Equipo , Humanos , Máscaras/virología , Tamaño de la Partícula , Polipropilenos/química , SARS-CoV-2/aislamiento & purificación , Textiles/virologíaRESUMEN
Interactions among the plant microbiome and its host are dynamic, both spatially and temporally, leading to beneficial or pathogenic relationships in the rhizosphere, phyllosphere, and endosphere. These interactions range from cellular to molecular and genomic levels, exemplified by many complementing and coevolutionary relationships. The host plants acquire many metabolic and developmental traits such as alteration in their exudation pattern, acquisition of systemic tolerance, and coordination of signaling metabolites to interact with the microbial partners including bacteria, fungi, archaea, protists, and viruses. The microbiome responds by gaining or losing its traits to various molecular signals from the host plants and the environment. Such adaptive traits in the host and microbial partners make way for their coexistence, living together on, around, or inside the plants. The beneficial plant microbiome interactions have been exploited using traditional culturable approaches by isolating microbes with target functions, clearly contributing toward the host plants' growth, fitness, and stress resilience. The new knowledge gained on the unculturable members of the plant microbiome using metagenome research has clearly indicated the predominance of particular phyla/genera with presumptive functions. Practically, the culturable approach gives beneficial microbes in hand for direct use, whereas the unculturable approach gives the perfect theoretical information about the taxonomy and metabolic potential of well-colonized major microbial groups associated with the plants. To capitalize on such beneficial, endemic, and functionally diverse microbiome, the strategic approach of concomitant use of culture-dependent and culture-independent techniques would help in designing novel "biologicals" for various crops. The designed biologicals (or bioinoculants) should ensure the community's persistence due to their genomic and functional abilities. Here, we discuss the current paradigm on plant-microbiome-induced adaptive functions for the host and the strategies for synthesizing novel bioinoculants based on functions or phylum predominance of microbial communities using culturable and unculturable approaches. The effective crop-specific inclusive microbial community bioinoculants may lead to reduction in the cost of cultivation and improvement in soil and plant health for sustainable agriculture.
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Chemical warfare agents (CWAs) have become a pivotal concern for the global community and spurred a wide spectrum of research for the development of new generation protective materials. Herein, a highly effective self-detoxifying filter consisting of in-situ immobilized Zirconium hydroxide [Zr(OH)4] over woven activated carbon fabric [Zr(OH)4@W-ACF] is presented for the removal of CWAs. It was prepared to harness the synergistic effect of high surface area of W-ACF, leads to high dispersion of CWAs and high phosphilicity and reactivity of [Zr(OH)4]. The synthesized materials were characterized by ATR-FTIR, EDX, SEM, TEM, XPS, TGA, and BET surface area analyzer. The kinetics of in-situ degradation of CWAs over Zr(OH)4@W-ACF were studied and found to be following the first-order reaction kinetics. The rate constant was found to be 0.244 min-1 and 2.31 × 10-2 min-1 for sarin and soman, respectively over Zr(OH)4@W-ACF. The potential practical applicability of this work was established by fabricating Zr(OH)4@W-ACF as reactive adsorbent layer for protective suit, and found to be meeting the specified criteria in terms of air permeability, tearing strength and nerve agent permeation as per TOP-08-2-501A:2013 and IS-17380:2020. The degradation products of CWAs were analyzed with NMR and GC-MS. The combined properties of dual functional textile with reactive material are expected to open up new exciting avenues in the field of CWAs protective clothing and thus find diverse application in defence and environmental sector.
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Brucella is alpha-2 Proteobacteria mainly responsible for multi-factorial bacterial zoonotic disease brucellosis with low concentration (10-100 CFU) required to establish the infection. In this study, we developed sandwich ELISA with detection range of 102 to 108 cells mL-1 and limit of detection at 103 cells mL-1 by employing polyclonal rabbit IgG (capture antibody, 10 µg mL-1) and mice IgG (detection antibody, 50 µg mL-1) antibody for its detection. Surface Plasmon Resonance evaluated the interaction of detection antibody with whole cell spiked serum samples at LOD of 102 cells mL-1 along with non co-operative interaction of protein albumin. Further, kinetic evaluation study using detection antibody against cell envelope antigen was performed whereby, Equilibrium Dissociation Constant (KD) and Maximum Binding Capacity (Bmax) were found to be 16.48 pM and 81.67 m° for Brucella abortus S99 and 0.42 pM and 54.50 m° for Brucella melitensis 16 M, respectively. During interference study, sandwich ELISA assay cross-reacted with either of the polyclonal antibody of above Brucella species. Upon validation, no cross-reactivity observed with bacteria-closely related to Brucella. In conclusion, developed semi-quantitative sandwich immunoassay is sensitively rapid in whole cell detection of Brucella and will be useful in development of detection assays from environmental and clinical matrices.
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Anticuerpos Antibacterianos/inmunología , Antígenos Bacterianos/inmunología , Brucella abortus/aislamiento & purificación , Brucella melitensis/aislamiento & purificación , Brucelosis/diagnóstico , Inmunoensayo/normas , Animales , Anticuerpos Antibacterianos/sangre , Brucella abortus/inmunología , Brucella melitensis/inmunología , Brucelosis/inmunología , Brucelosis/microbiología , Femenino , Pruebas Inmunológicas , Ratones , Ratones Endogámicos BALB C , ConejosRESUMEN
Ferulic acid is a known precursor for vanillin production but the significance of agro waste as substrates for its extraction, in combination with microbes is a less explored option. Various lactic acid bacteria were screened for the production of ferulic acid esterase (FAE) and Enterococcus lactis SR1 was found to produce maximum FAE (7.54 ± 0.15 IU/ml) in the synthetic medium under submerged fermentation. To make the process cost effective, various lignocellulosic agroresidues were evaluated for the production of FAE from the bacterium. It was found that wheat bran serves as the best substrate for FAE production (4.18 ± 0.12 IU/ml) from E. lactis SR1. Further, optimization of fermentation conditions for FAE production from E. lactis SR1 using wheat bran as carbon source led to an increase in the enzyme production (7.09 ± 0.21 IU/ml) by 1.5 fold. The FAE produced was used alone or in combination with commercial holocellulase for biological release of FA from the tested agroresidues. The highest release of FA (mg/g) by enzymatic extraction occurred in sugarbeet pulp (2.56), followed by maize bran (1.45), wheat bran (1.39) and rice bran (0.87), when both the enzymes (FAE and holocellulase) were used together. Alkaline extraction and purification of ferulic acid (FA) from these agro residues also showed that sugarbeet pulp contains the highest amount of FA (5.5 mg/g) followed by maize bran (3.0 mg/g), wheat bran (2.8 mg/g) and rice bran (1.9 mg/g), similar to the trend obtained in biological/enzymatic extraction of FA from these residues. Furthermore, the substrates were found to release higher reducing sugars when both commercial holocellulase and FAE were used in combination than by the use of holocellulase alone. Thus, FAEs not only release FA but also enabled hemicellulase and cellulase to release more sugars from plant material.
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Increasing environmental concern, human health and the continuous upgradation in the stringent standards of vehicular emissions have shown much interest in cleaner diesel fuels. Out of various strategies to mitigate the diesel engine emissions, use of water blended diesel in the form of emulsion has grabbed sufficient attention of the fuel research community. Various researches have shown that water-emulsified diesel has sufficient potential to improve the engine performance simultaneously with a significant reduction in the levels of nitrogen oxides (NOx) and particulate matter (PM) emissions. Micro-explosion phenomenon of combustion in emulsion fuel helps to provide efficient and complete combustion which in turn improves brake thermal efficiency. The current study presents a comprehensive review of the usage of water-emulsified diesel fuel in CI engines. Focusing on the performance, combustion, and emission analysis, it also talks in detail about the principle and the chemistry involved in making of a stable and homogeneous water-diesel emulsion compatible for CI engine. The literature survey concludes two crucial points. First, the water-blended diesel emulsion serves as an economical, fuel efficient, and cleaner combustion technology. Second, the optimum blend ratio, emulsifier quantity, and proper process differs in almost all the research papers and hence needed to be standardized.
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Emulsiones/química , Gasolina , Emisiones de Vehículos/análisis , Gasolina/análisis , Óxidos de Nitrógeno/análisis , Material Particulado/análisis , Tensoactivos/química , Agua/químicaRESUMEN
Herein, we demonstrate a nanocomposite material Eu/ZnO/pPy for enhanced performance in photoelectrocatalytic degradation of chemical warfare agent sulphur mustard (SM) at ambient conditions which is growing concern of the Scientific Community amidst the current climate of terrorism. Eu/ZnO/pPy was electrochemically prepared on Au electrode at ambient conditions and was used for electrocatalytic reductive elimination of chloride from SM and results indicated one electron involvement process for the cleavage of the carbon-chloride bond. Surface morphology of Eu/pPy, ZnO/pPy and Eu/ZnO/pPy composites were characterized by SEM and confirmed the formation of the nanoparticles and nanorods on the modified electrode which leads to provide more surface area for the reductive elimination reaction. The elemental composition, functional groups and phase of materials on the modified electrode were deduced using EDX, Raman spectroscopy and XRD, respectively. Eu/ZnO/pPy/Au electrode was utilized for the photoelectrocatalytic degradation of SM as it exhibit excellent electrocatalytic activity and degradation products were analyzed by GC-MS. In the reductive elimination of SM, the following parameters were deduced (i) heterogeneous rate constant (0.127 s-1), (ii) transfer coefficient (0.32) and (iii) number of electron involved (1.0). The enhanced photoelectrocatalytic capability of this nanocomposite could serve as a novel and promising catalyst in defence and environmental applications.
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Sustancias para la Guerra Química/química , Europio/química , Oro/química , Gas Mostaza/química , Nanocompuestos/química , Procesos Fotoquímicos , Pirroles/química , Óxido de Zinc/química , Catálisis , Electrodos , Cromatografía de Gases y Espectrometría de Masas , Irritantes , Nanopartículas/química , Nanotubos/químicaRESUMEN
Surface plasmon resonance (SPR) immunosensor using 4-mercaptobenzoic acid (4-MBA) modified gold (4-MBA/Au) SPR chip was developed first time for the detection of Brucella melitensis (B. melitensis) based on the screening of its complementary DNA target by using two different newly designed DNA probes of IS711 gene. Herein, interaction between DNA probes and target molecule are also investigated and result revealed that the interaction is spontaneous. The kinetics and thermodynamic results derived from the experimental data showed that the interaction between complementary DNA targets and probe 1 is more effective than that of probe 2. Equilibrium dissociation constant (KD) and maximum binding capacity of analyte (Bmax) values for the interaction of complementary DNA target with the immobilized DNA probes were calculated by using kinetic evaluation software, and found to be 15.3 pM (KD) and 81.02m° (Bmax) with probe 1 and 54.9pM and 55.29m° (Bmax), respectively. Moreover, real serum samples analysis were also carried out using immobilized probe 1 and probe 2 with SPR which showed the applicability of this methodology and provides an alternative way for the detection of B. melitensis in less than 10min. This remarkable sensing response of present methodology offer a real time and label free detection of biological warfare agent and provide an opportunity to make miniaturized sensor, indicating considerable promise for diverse environmental, bio-defence, clinical diagnostics, food safety, water and security applications.
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Brucella melitensis/aislamiento & purificación , Brucelosis/microbiología , ADN Bacteriano/análisis , Resonancia por Plasmón de Superficie/métodos , Benzoatos/química , Brucella melitensis/genética , Brucelosis/diagnóstico , Brucelosis/genética , Sondas de ADN/química , Sondas de ADN/genética , ADN Bacteriano/sangre , ADN Bacteriano/genética , Genes Bacterianos , Oro/química , Humanos , Ácidos Nucleicos Inmovilizados/química , Compuestos de Sulfhidrilo/química , TermodinámicaRESUMEN
The cancer profile in the Indian state of Uttarakhand reveals that the breast cancer is the most prevalent type of cancers in females followed by cervical and ovarian type. Literature survey shows that the E6 protein of Human Papilloma Virus-16 (HPV-16) is responsible for causing several forms of cancer in human. Therefore, it is of interest to screen HPV-16 E6 target protein with known natural compounds using computer aided molecular modeling and docking tools. The complete structure of E6 is unknown. Hence, the E6 structure model was constructed using different online servers followed by molecular docking of Colchine, Curcumin, Daphnoretin, Ellipticine and Epigallocatechin-3-gallate; five known natural compounds with best E6 protein model predicted by Phyre2 server. The screening exercise shows that Daphnoretin (with binding free energy of -8.3 kcal/mol), a natural compound derived from Wikstroemia indica has the top binding properties. Thus, it is of interest to consider the compound for further validation.
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Surface plasmon resonance (SPR) screening of monoclonal and polyclonal antibodies of Plasmodium falciparum (MoabPf and PoabPf) for recombinant Histidine rich protein-II antigen (Ag) of Pf (rHRP-II Ag) was conducted in a real-time and label-free manner to select an appropriate antibody (Ab) for biosensor applications. In this study 4-mercaptobenzoic acid (4-MBA) modified gold SPR chip was used for immobilizing the Ag and then Ab was interacted. SEM image showed modification of SPR chip with 4-MBA and EDAX confirmed the presence of 4-MBA on the SPR chip. Equilibrium constant (KD) and maximum binding capacity of analyte (Bmax) values for the interaction of MoabPf or PoabPf with the immobilized rHRP-II Ag were calculated and found to be 0.517 nM and 48.61 m° for MoabPf and 2.288 nM and 46.80 m° for PoabPf, respectively. In addition, thermodynamic parameters such as ΔG, ΔH and ΔS were determined for the interaction between rHRP-II Ag and MoabPf or PoabPf and the values revealed that the interaction is spontaneous, exothermic and driven by entropy. The kinetics and thermodymanic results of this study revealed that the interaction between MoabPf and rHRP-II Ag is more effective than that of PoabPf due to the fact that MoabPf was derived from a single epitope (single clone) whereas the PoabPf was from the mixture of a number of epitopes (polyclones). Finally, SPR methodology was developed for the sensing of malarial antibodies. The limit of detection was found to be 5.6 pg with MoabPf which was found to be the best in our study.