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1.
Int J Biol Macromol ; 260(Pt 1): 129450, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38232896

RESUMO

In this study, we fabricated a novel biodegradable functional film using natural polysaccharides by adding jujube seed powder as an active ingredient. Scanning electron microscopy analysis showed agglomerate formation in the film with increasing concentration of seed powder. Fourier transform-infrared spectroscopy study demonstrated an electrostatic interaction between pectin and chitosan. The water solubility and swelling degree significantly decreased from 55.5 to 47.7 % and 66.0 to 41.9 %, respectively, depicting the film's water resistance properties. Higher opacity and lower transmittance value of the film indicated its protective effect towards light-induced oxidation of food. It was observed that the fabricated active film biodegraded to 82.33 % in 6 days. The DPPH radical scavenging activity of 98.02 % was observed for the functional film. The film showed antifungal activity against B. cinerea and P. chrysogenum. The highest zone of inhibition was obtained against food spoiling bacteria B. subtilis followed by S. aureus, P. aeruginosa and E. coli. Genotoxicity studies with the fabricated film showed a mitotic index of 8 % compared to 3 % in the control film. We used the fabricated film to preserve grapefruits, and the result showed that it could preserve grapes for ten days with an increase in antioxidant activity and polyphenolic content.


Assuntos
Quitosana , Frutas , Extratos Vegetais , Ziziphus , Pós/farmacologia , Embalagem de Alimentos , Staphylococcus aureus , Escherichia coli , Quitosana/química , Polissacarídeos/farmacologia , Água/farmacologia , Pectinas/farmacologia
2.
World J Microbiol Biotechnol ; 39(11): 310, 2023 Sep 16.
Artigo em Inglês | MEDLINE | ID: mdl-37715879

RESUMO

Natural antioxidants have become vital to minimize macromolecular damage caused by Reactive Oxygen Species (ROS). This study investigated the antioxidant property of ß-cryptoxanthin (ß-CRX) extracted from Kocuria marina DAGII and its protective effect against macromolecular damages by generating ROS via two models: UV radiation and the Fenton reaction. ß-cryptoxanthin exhibited the highest scavenging activity towards hydrogen peroxide radicals with an IC50 value of 38.30 ± 1.13 µg/ml, favoring the hydrogen atom transfer mechanism. The total antioxidant capacity value of 872.0101 ± 1.84 µg BHT/mg ß-CRX indicated the cumulative ROS scavenging ability of ß-cryptoxanthin. ß-cryptoxanthin could protect against ROS-induced lipid peroxidation, protein oxidation, and DNA damage. The highest lipid peroxidation and protein oxidation inhibition values of ß-cryptoxanthin against ROS were 99.371 ± 0.51% and 78.19 ± 0.15%, respectively. ß-cryptoxanthin also showed a protective effect in maintaining DNA intactness against ROS-mediated DNA damage. Allium cepa test showed the non-genotoxic nature of ß-cryptoxanthin and its protective effect against ROS genotoxic effects. A photo-stability study of ß-cryptoxanthin toward UVA and UVB radiation showed a rapid bleaching result of UVB obeying pseudo-zero order kinetics with an average R2 value of 0.9897 and a higher k value (-6.3 × 10-11 ± 0.2 M/s) than UVA (k value -3.1 × 10-11 ± 0.17 M/s), signifying that UVB is more potent toward photo-degradation. The good SPF value of 23.1737 ± 0.15 showed the UV protection capability of ß-cryptoxanthin. Thus, the present study suggests that ß-cryptoxanthin could be a valuable antioxidant to protect against ROS-induced various macromolecular damages and act as a good UV protectant.


Assuntos
Antioxidantes , beta-Criptoxantina , Antioxidantes/farmacologia , Espécies Reativas de Oxigênio , Dano ao DNA , Hidrogênio
3.
Int J Biol Macromol ; 239: 124281, 2023 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-37001777

RESUMO

Perishable foods like fruits and vegetables, meat, fish, and dairy products have short shelf-life that causes significant postharvest losses, which poses a major challenge for food supply chains. Biopolymers have been extensively studied as sustainable alternatives to synthetic plastics, and pectin is one such biopolymer that has been used for packaging and preservation of foods. Pectin is obtained from abundantly available low-cost sources such as agricultural or food processing wastes and by products. This review is a complete account of pectin extraction from agro-wastes, development of pectin-based composite films and coatings, their characterizations, and their applications in food packaging and preservation. Compared to conventional chemical extraction, supercritical water, ultrasound, and microwave assisted extractions are a few examples of modern and more efficient pectin extraction processes that generate almost no hazardous effluents, and thus, such extraction techniques are more environment friendly. Pectin-based films and coatings can be functionalized with natural active agents such as essential oils and other phytochemicals to improve their moisture barrier, antimicrobial and antioxidant properties. Application of pectin-based active films and coatings effectively improved shelf-life of fresh cut-fruits, vegetables, meat, fish, poultry, milk, and other food perishable products.


Assuntos
Embalagem de Alimentos , Pectinas , Embalagem de Alimentos/métodos , Conservação de Alimentos/métodos , Antioxidantes/farmacologia , Antioxidantes/química , Manipulação de Alimentos
4.
BMC Chem ; 16(1): 42, 2022 Jun 07.
Artigo em Inglês | MEDLINE | ID: mdl-35672858

RESUMO

BACKGROUND: Mangiferin is a C-glycoside xanthone molecule having a wide range of therapeutic properties. Hence, the present study aims to understand the efficacy of mangiferin against colorectal cancer (CRC) and to elucidate the mechanisms of action of mangiferin on colorectal cancer. METHOD: The molecular mechanism of mangiferin against colorectal cancer was studied using Autodock Vina software. Pharmacophore analysis of mangiferin concerning five COX-2 inhibitor drugs was carried out using the PharmaGist server to analyze the possibility of using mangiferin as a COX-2 inhibitor. In vitro analysis of Mangiferin against various cancer cell lines was performed. RESULTS: The molecular mechanism of action of mangiferin against CRC was assessed by docking with multiple target proteins involved in the progression of CRC. Docking studies showed good binding scores (kcal/mol) ranging from - 10.3 to - 6.7. Mangiferin showed a good affinity towards enzymes like COX-2 and LA4H involved in Arachidonic acid (AA) metabolism with a binding score(kcal/mol) of - 10.1 and - 10.3 respectively. The pharmacophore feature assessment of mangiferin was done for COX-2 inhibitor drugs, which further confirmed that mangiferin poses the same pharmacophore feature as that of COX-2 inhibitor drugs. Furthermore, the binding affinity of mangiferin was compared with five COX-2 inhibitor drugs to prove its efficacy as an inhibitor. Mangiferin also had a cytotoxic effect against colorectal cancer (HT 29), cervical cancer (HeLa), and breast cancer (MCF 7) cell lines. The study could establish that Mangiferin might be a promising candidate for the treatment of colorectal cancer. CONCLUSION: In short, these studies exploited the possibility of mangiferin as a lead molecule to develop anticancer/anti-inflammatory drugs for the treatment of CRC.

5.
Plants (Basel) ; 11(3)2022 Feb 06.
Artigo em Inglês | MEDLINE | ID: mdl-35161431

RESUMO

Sulfur is a growth-limiting and secondary macronutrient as well as an indispensable component for several cellular components of crop plants. Over the years various scientists have conducted several experiments on sulfur metabolism based on different aspects of plants. Sulfur metabolism in seeds has immense importance in terms of the different sulfur-containing seed storage proteins, the significance of transporters in seeds, the role of sulfur during the time of seed germination, etc. The present review article is based on an overview of sulfur metabolism in seeds, in respect to source to sink relationships, S transporters present in the seeds, S-regulated seed storage proteins and the importance of sulfur at the time of seed germination. Sulfur is an essential component and a decidable factor for seed yield and the quality of seeds in terms of oil content in oilseeds, storage of qualitative proteins in legumes and has a significant role in carbohydrate metabolism in cereals. In conclusion, a few future perspectives towards a more comprehensive knowledge on S metabolism/mechanism during seed development, storage and germination have also been stated.

6.
Immunol Lett ; 241: 49-54, 2022 01.
Artigo em Inglês | MEDLINE | ID: mdl-34942191

RESUMO

Prohibitin is a highly conserved ubiquitously expressed protein involved in several key cellular functions. Targeting of this protein in the membrane by the virulence polysaccharide, Vi, of human typhoid-causing pathogen, Salmonella enterica serovar Typhi (S. Typhi), results in suppression of IL-2 secretion from T cells activated through the T-cell receptor (TCR). However, the mechanism of this suppression remains unclear. Here, using Vi as a probe, we show that membrane prohibitin associates with the src-tyrosine kinase, p56lck (Lck), and actin in human model T cell line, Jurkat. Activation with anti-CD3 antibody brings about dissociation of this complex, which coincides with downstream ERK activation. The trimolecular complex reappears towards culmination of proximal TCR signaling. Engagement of cells with Vi prevents TCR-triggered activation of Lck and ERK by inhibiting dissociation of the former from prohibitin. These findings suggest a regulatory role for membrane prohibitin in Lck activation and TCR signaling.


Assuntos
Membrana Celular/metabolismo , Complexos Multiproteicos/metabolismo , Proibitinas/metabolismo , Salmonella typhi/patogenicidade , Linfócitos T/fisiologia , Actinas/metabolismo , Humanos , Terapia de Imunossupressão , Células Jurkat , Ativação Linfocitária , Polissacarídeos Bacterianos/imunologia , Ligação Proteica , Receptores de Antígenos de Linfócitos T/metabolismo , Transdução de Sinais , Fatores de Virulência/imunologia
7.
J Clin Invest ; 131(11)2021 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-33822776

RESUMO

Inhibitors of calcineurin phosphatase activity (CNIs) such as cyclosporin A (CsA) are widely used to treat tissue transplant rejection and acute graft-versus-host disease (aGVHD), for which inhibition of gene expression dependent on nuclear factor of activated T cells (NFAT) is the mechanistic paradigm. We recently reported that CNIs inhibit TCR-proximal signaling by preventing calcineurin-mediated dephosphorylation of LckS59, an inhibitory modification, raising the possibility of another mechanism by which CNIs suppress immune responses. Here we used T cells from mice that express LckS59A, which cannot accept a phosphate at residue 59, to initiate aGVHD. Although CsA inhibited NFAT-dependent gene upregulation in allo-aggressive T cells expressing either LckWT or LckS59A, it was ineffective in treating disease when the T cells expressed LckS59A. Two important NFAT-independent T cell functions were found to be CsA-resistant in LckS59A T cells: upregulation of the cytolytic protein perforin in tissue-infiltrating CD8+ T cells and antigen-specific T/DC adhesion and clustering in lymph nodes. These results demonstrate that effective treatment of aGVHD by CsA requires NFAT-independent inhibition of TCR signaling. Given that NFATs are widely expressed and off-target effects are a major limitation in CNI use, it is possible that targeting TCR-associated calcineurin directly may provide effective therapies with less toxicity.


Assuntos
Linfócitos T CD8-Positivos/imunologia , Inibidores de Calcineurina/farmacologia , Ciclosporina/farmacologia , Doença Enxerto-Hospedeiro/tratamento farmacológico , Fatores de Transcrição NFATC/imunologia , Receptores de Antígenos de Linfócitos T/imunologia , Transdução de Sinais/imunologia , Doença Aguda , Animais , Doença Enxerto-Hospedeiro/genética , Doença Enxerto-Hospedeiro/imunologia , Camundongos , Camundongos Knockout , Fatores de Transcrição NFATC/genética , Receptores de Antígenos de Linfócitos T/genética , Transdução de Sinais/genética
8.
Int J Biol Macromol ; 151: 36-46, 2020 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-32070742

RESUMO

Oxidative stress is considered to be the leading cause of many chronic and degenerative diseases leading to death and disability. Antioxidant therapy could be efficient in preventing oxidative stress-induced damages in cells. In the present study the efficacy of mangiferin-chitosan nanoparticles (MCNs) as a therapeutic agent against the oxidative stress, since mangiferin (MGF), a polyphenol protects and attenuates oxidative stress against various diseases. However, the bioactivity of MGF does not result in vivo biological effect owing to its low bioavailability and poor solubility. Mangiferin nanoparticles (NPs) were developed to improve the solubility and bioavailability of mangiferin. Nanoparticles had significant free radical scavenging activity in the cell-free system and had a synergistic action with phase II antioxidant enzymes such as catalases and peroxidases. A nephropathic system was developed to investigate the attenuative role of mangiferin-chitosan nanoparticles against induced oxidative stress, on normal kidney epithelial (NKE) cells. Pre-treatment with the nanoparticles exclusively prevented the induction of cytotoxicity induced by NaF and maintained the level of intracellular antioxidant enzyme in the cells. Nanoparticles had significant lipid peroxidation and protein oxidation inhibition activities. These nanoparticles can be used in food and pharmaceutical industries as a therapeutic agent to prevent the oxidative stress-induced health disorders.


Assuntos
Antioxidantes/química , Antioxidantes/farmacologia , Quitosana/química , Nanopartículas/química , Estresse Oxidativo/efeitos dos fármacos , Xantonas/química , Sobrevivência Celular , Células Cultivadas , Espaço Intracelular/metabolismo , Rim/citologia , Rim/efeitos dos fármacos , Rim/metabolismo , Modelos Biológicos , Oxirredução , Espécies Reativas de Oxigênio/metabolismo , Análise Espectral
9.
Nanotoxicology ; 14(5): 577-594, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-31928284

RESUMO

The successful translation of nanostructure-based bioimaging and/or drug delivery system needs extensive in vitro and in vivo studies on biocompatibility, biodistribution, clearance, and toxicity for its diagnostic applications. Herein, we have investigated the in vitro cyto-hemocompatibility, in vivo biodistribution, clearance, and toxicity in mice after systemic administration of GdF3 nanoparticles loaded PEGylated mesoporous carbon capsule (GdF3-PMCC)-based theranostic system. In vitro cyto-hemocompatibility study showed a very good biocompatibility up to concentration of 500 µg/ml. Biodistribution studies carried out from 1 h to 8 days showed that GdF3-PMCC was found in major organs, such as liver, kidney, spleen, and muscle till 4th day and it was negligible in any tissue after 8th day. The clearance study was carried out for a period of 8 days and it was observed that the urinary system is the main route of excretion of GdF3-PMCC. The tissue toxicity study was done for 15 days and histopathological analysis indicated that the GdF3-PMCC based theranostic system does not have any adverse effect in tissues. Thus, PMCCs are nontoxic and can be applied as theranostic agents in contrast to the other carbon-based systems (PEGylated carbon nanotubes and PEGylated graphene oxide) which showed significant toxicity.


Assuntos
Antineoplásicos/administração & dosagem , Materiais Biocompatíveis/química , Carbono/química , Portadores de Fármacos/química , Compostos de Flúor/administração & dosagem , Gadolínio/química , Nanopartículas/administração & dosagem , Polietilenoglicóis/química , Nanomedicina Teranóstica , Animais , Antineoplásicos/farmacocinética , Antineoplásicos/farmacologia , Antineoplásicos/toxicidade , Apoptose/efeitos dos fármacos , Materiais Biocompatíveis/administração & dosagem , Coagulação Sanguínea/efeitos dos fármacos , Cápsulas , Carbono/administração & dosagem , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Portadores de Fármacos/administração & dosagem , Sistemas de Liberação de Medicamentos , Compostos de Flúor/farmacocinética , Compostos de Flúor/toxicidade , Gadolínio/administração & dosagem , Gadolínio/efeitos adversos , Gadolínio/farmacocinética , Gadolínio/toxicidade , Humanos , Rim/efeitos dos fármacos , Rim/metabolismo , Fígado/efeitos dos fármacos , Fígado/metabolismo , Taxa de Depuração Metabólica , Camundongos , Músculos/efeitos dos fármacos , Músculos/metabolismo , Nanopartículas/química , Polietilenoglicóis/administração & dosagem , Espécies Reativas de Oxigênio , Baço/efeitos dos fármacos , Baço/metabolismo , Distribuição Tecidual
10.
R Soc Open Sci ; 5(7): 172318, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30109058

RESUMO

The dairy industry produces enormous amount of cheese whey containing the major milk nutrients, but this remains unutilized all over the globe. The present study investigates the production of ß-cryptoxanthin (ß-CRX) by Kocuria marina DAGII using cheese whey as substrate. Response surface methodology (RSM) and an artificial neural network (ANN) approach were implemented to obtain the maximum ß-CRX yield. Significant factors, i.e. yeast extract, peptone, cheese whey and initial pH, were the input variables in both the optimizing studies, and ß-CRX yield and biomass were taken as output variables. The ANN topology of 4-9-2 was found to be optimum when trained with a feed-forward back-propagation algorithm. Experimental values of ß-CRX yield (17.14 mg l-1) and biomass (5.35 g l-1) were compared and ANN predicted values (16.99 mg l-1 and 5.33 g l-1, respectively) were found to be more accurate compared with RSM predicted values (16.95 mg l-1 and 5.23 g l-1, respectively). Detailed kinetic analysis of cellular growth, substrate consumption and product formation revealed that growth inhibition took place at substrate concentrations higher than 12% (v/v) of cheese whey. The Han and Levenspiel model was the best fitted substrate inhibition model that described the cell growth in cheese whey with an R2 and MSE of 0.9982% and 0.00477%, respectively. The potential importance of this study lies in the development, optimization and modelling of a suitable cheese whey supplemented medium for increased ß-CRX production.

11.
Proc Natl Acad Sci U S A ; 115(9): 2174-2179, 2018 02 27.
Artigo em Inglês | MEDLINE | ID: mdl-29440413

RESUMO

ZAP-70 is a tyrosine kinase that is essential for initiation of T cell antigen receptor (TCR) signaling. We have found that T cell p38 MAP kinase (MAPK), which is directly phosphorylated and activated by ZAP-70 downstream of the TCR, in turn phosphorylates Thr-293 in the interdomain B region of ZAP-70. Mutant T cells expressing ZAP-70 with an alanine substitution at this residue (ZAP-70T293A) had enhanced TCR proximal signaling and increased effector responses. Lack of ZAP-70T293 phosphorylation increased association of ZAP-70 with the TCR and prolonged the existence of TCR signaling microclusters. These results identify a tight negative feedback loop in which ZAP-70-activated p38 reciprocally phosphorylates ZAP-70 and destabilizes the signaling complex.


Assuntos
Genes Codificadores dos Receptores de Linfócitos T/fisiologia , Proteína-Tirosina Quinase ZAP-70/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Sequência de Aminoácidos , Regulação da Expressão Gênica , Humanos , Células Jurkat , Fosforilação , Transdução de Sinais , Proteína-Tirosina Quinase ZAP-70/genética , Proteínas Quinases p38 Ativadas por Mitógeno/genética
12.
Eng Life Sci ; 17(7): 739-748, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32624819

RESUMO

Various natural polymers with hydrophilic properties have been used to form hydrogels for the encapsulation and delivery of nutrients and drugs in food and pharmaceutical industries. Among them, chitosan (ChiHG)- and alginate (AlgHG)- based hydrogels have been extensively explored for delivery of several nutraceuticals in recent years. Release of natural canthaxanthin (CX) obtained from Dietzia maris NITD (accession number: HM151403) has been investigated with emphasis on biomedical applications. Significant changes (P < 0.05) in degree of swelling ( % ) and moisture content (% dry basis) were found after encapsulation of bacterial canthaxanthin (BCX), but the gel content remained unchanged. BCX encapsulation efficiency was calculated to be 55.92% and 60.45% in ChiHG and AlgHG, respectively. A noticeable change in heat of fusion ( Δ H m ) d melting point ( T m ) was recorded in ChiHG and AlgHG after BCX encapsulation. Swelling and BCX release from gel matrix was performed under two different pH (1.2 and 7.4). The results showed that swelling of hydrogel and BCX release was facilitated at higher pH (7.4) than acidic pH (1.2). With regard to the release kinetics data, it was found that BCX is released from both ChiHG and AlgHG in a non - Fickian diffusion transport method. In addition, antioxidant activity of BCX encapsulated hydrogels was found significantly higher (P < 0.001) in terms of DPPH, ABTS, nitrite, hydroxyl radical scavenging and reducing power assay. These results indicated that BCX can be successfully encapsulated into a polymeric hydrogel to obtain a dynamic biomaterial that may be used in drug delivery applications in future.

13.
Nat Immunol ; 18(2): 196-204, 2017 02.
Artigo em Inglês | MEDLINE | ID: mdl-27941787

RESUMO

Calcineurin is a phosphatase whose primary targets in T cells are NFAT transcription factors, and inhibition of calcineurin activity by treatment with cyclosporin A (CsA) or FK506 is a cornerstone of immunosuppressive therapies. Here we found that calcineurin was recruited to the T cell antigen receptor (TCR) signaling complex, where it reversed inhibitory phosphorylation of the tyrosine kinase Lck on Ser59 (LckS59). Loss of calcineurin activity impaired phosphorylation of Tyr493 of the tyrosine kinase ZAP-70 (ZAP-70Y493), as well as some downstream pathways in a manner consistent with signaling in cells expressing LckS59A (Lck that cannot be phosphorylated) or LckS59E (a phosphomimetic mutant). Notably, CsA inhibited integrin-LFA-1-dependent and NFAT-independent adhesion of T cells to the intercellular adhesion molecule ICAM-1, with little effect on cells expressing mutant Lck. These results provide new understanding of how widely used immunosuppressive drugs interfere with essential processes in the immune response.


Assuntos
Calcineurina/metabolismo , Proteína Tirosina Quinase p56(lck) Linfócito-Específica/metabolismo , Receptores de Antígenos de Linfócitos T/metabolismo , Linfócitos T/imunologia , Proteína-Tirosina Quinase ZAP-70/metabolismo , Animais , Adesão Celular/efeitos dos fármacos , Ciclosporina/farmacologia , Humanos , Imunossupressores/farmacologia , Molécula 1 de Adesão Intercelular/metabolismo , Células Jurkat , Ativação Linfocitária/efeitos dos fármacos , Antígeno-1 Associado à Função Linfocitária/metabolismo , Proteína Tirosina Quinase p56(lck) Linfócito-Específica/genética , Camundongos , Camundongos Transgênicos , Fatores de Transcrição NFATC/genética , Fatores de Transcrição NFATC/metabolismo , Fosforilação/efeitos dos fármacos , Fosforilação/genética , Ligação Proteica , Transdução de Sinais , Linfócitos T/efeitos dos fármacos , Tacrolimo/farmacologia
14.
Bioprocess Biosyst Eng ; 40(1): 99-113, 2017 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-27628580

RESUMO

In the present investigation, growth kinetics of Kocuria marina DAGII during batch production of ß-Cryptoxanthin (ß-CRX) was studied by considering the effect of glucose and maltose as a single and binary substrate. The importance of mixed substrate over single substrate has been emphasised in the present study. Different mathematical models namely, the Logistic model for cell growth, the Logistic mass balance equation for substrate consumption and the Luedeking-Piret model for ß-CRX production were successfully implemented. Model-based analyses for the single substrate experiments suggested that the concentrations of glucose and maltose higher than 7.5 and 10.0 g/L, respectively, inhibited the growth and ß-CRX production by K. marina DAGII. The Han and Levenspiel model and the Luong product inhibition model accurately described the cell growth in glucose and maltose substrate systems with a R 2 value of 0.9989 and 0.9998, respectively. The effect of glucose and maltose as binary substrate was further investigated. The binary substrate kinetics was well described using the sum-kinetics with interaction parameters model. The results of production kinetics revealed that the presence of binary substrate in the cultivation medium increased the biomass and ß-CRX yield significantly. This study is a first time detailed investigation on kinetic behaviours of K. marina DAGII during ß-CRX production. The parameters obtained in the study might be helpful for developing strategies for commercial production of ß-CRX by K. marina DAGII.


Assuntos
beta-Criptoxantina/biossíntese , Micrococcaceae/crescimento & desenvolvimento , Modelos Biológicos , Cinética
15.
Biotechnol Res Int ; 2014: 293434, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24864214

RESUMO

Xenorhabdus indica KB-3, a well-known protease producer, was isolated from its entomopathogenic nematode symbiont Steinernema thermophilum. Since medium constituents are critical to the protease production, the chemical components of the selected medium (soya casein digest broth) were optimized by rotatable central composite design (RCCD) using response surface methodology (RSM). The effects of all five chemical components (considered as independent variables), namely tryptone, soya peptone, dextrose, NaCl, and dipotassium phosphate, on protease production (dependent variable) were studied, and it was found that tryptone and dextrose had maximum influence on protease production. The protease production was increased significantly by 66.31% under optimal medium conditions (tryptone-5.71, soya peptone-4.9, dextrose-1.45, NaCl-6.08, and dipotassium phosphate-0.47 in g/L). To best of knowledge, there are no reports on optimization of medium component for protease production by X. indica KB-3 using RSM and their application in fibrinolysis. This study will be useful for industrial processes for production of protease enzyme from X. indica KB-3 for its application in the field of agriculture and medicine.

16.
J Infect Dis ; 210(1): 79-88, 2014 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-24470505

RESUMO

T cells are critical to immunity against pathogenic Salmonella including Salmonella Typhi which causes systemic infection, typhoid, in humans. The strategies that this pathogen employs to keep T-cell mediated immune responses in check during establishment of systemic infection are not completely understood. Here, we show that the virulence polysaccharide Vi, which distinguishes S. Typhi from localized gastroenteritis-producing nontyphoidal Salmonella serovars, is a potent inhibitor of T-cell activation. Vi released by S. Typhi interacts with the membrane prohibitin complex and inhibits IL-2 secretion from T cells stimulated through the T-cell receptor (TCR) but does not affect PMA-activated interleukin 2 (IL-2) secretion. Treatment with Vi suppresses early activation events including TCR down-regulation, actin polymerization, and phosphorylation of ERK. Coadministration of Vi with anti-CD3 Ab reduces secretion of IL-2 and interferon γ in mice. Our findings reveal a mechanism by which S. Typhi may target T-cell immunity during establishment of typhoid.


Assuntos
Interações Hospedeiro-Patógeno , Interleucina-2/antagonistas & inibidores , Ativação Linfocitária/efeitos dos fármacos , Polissacarídeos Bacterianos/metabolismo , Proteínas Repressoras/antagonistas & inibidores , Salmonella typhi/imunologia , Linfócitos T/efeitos dos fármacos , Animais , Membrana Celular/efeitos dos fármacos , Membrana Celular/imunologia , Células Cultivadas , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Proibitinas , Linfócitos T/imunologia
17.
3 Biotech ; 4(2): 107-120, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-28324442

RESUMO

Mangiferin, a bioactive compound having potent nutraceutical, strong antioxidant and pharmacological significance has been extracted using microwave-assisted extraction (MAE) technique from Curcuma amada, commonly known as mango ginger. The extraction solvent ethanol is eco-friendly, nontoxic and reduces the risk of environmental hazards. The influence of several independent variables such as microwave power, ethanol concentration, extraction (irradiation) time and pre-leaching time has been studied under varying conditions using one-factor-at-a-time analysis to obtain an optimal extraction ratio. The maximum mangiferin content of 1.1156 mg/g is obtained at microwave power of 550 W and extraction time of 50 s with 80 % ethanol as a solvent and pre-leaching time of 20 min. The results indicate that microwave power and ethanol concentration have the most significant effect on the yield of mangiferin content. The presence of mangiferin in final Curcuma amada extract is confirmed through high-performance liquid chromatography and the functional groups are identified through Fourier transform infrared spectroscopy analyses using standard mangiferin. The experimental profiles are fitted into a two-parameter modified first-order kinetic model and a three-parameter modified logistic model and checked using the goodness-of-fit criterion. The Curcuma amada retained its antioxidant activity after MAE treatment and the antioxidant activity of mangiferin obtained after extraction using DPPH free radical scavenging assay is studied.

18.
J Microbiol Biotechnol ; 23(11): 1536-43, 2013 Nov 28.
Artigo em Inglês | MEDLINE | ID: mdl-23928843

RESUMO

Proteases produced by Xenorhabdus are known to play a significant role in virulence leading to insect mortality. The present study was undertaken to purify and characterize protease from Xenorhabdus indica, an endosymbiont of nematode Steinernema thermophilum, and to decipher its role in insect mortality and its efficacy to control Helicoverpa armigera. A set of 10 strains of Xenorhabdus isolated from different regions of India were screened for protease activity on the basis of zone of clearing on gelatin agar plates. One potent strain of Xenorhabdus indica was selected for the production of protease, and the highest production (1,552 U/ml) was observed at 15-18 h of incubation at 28°C in soya casein digest broth. The extracellular protease was purified from culture supernatant using ammonium sulfate precipitation and ion-exchange chromatography. The enzyme was further characterized by SDS-PAGE and zymography, which confirmed the purity of the protein and its molecular mass was found to be ~52 kDa. Further MALDI-TOF/TOF analysis and effect of metal chelating agent 1,10-phenanthrolin study revealed the nature of the purified protease as a secreted alkaline metalloprotease. The bioefficacy of the purified protease was also tested against cotton bollworm (Helicoverpa armigera) and resulted in 67.9 ± 0.64% mortality within one week. This purified protease has the potential to be developed as a natural insecticidal agent against a broad range of agriculturally important insects.


Assuntos
Inseticidas/isolamento & purificação , Inseticidas/metabolismo , Lepidópteros/efeitos dos fármacos , Metaloproteases/isolamento & purificação , Metaloproteases/metabolismo , Xenorhabdus/enzimologia , Animais , Bioensaio , Precipitação Química , Cromatografia por Troca Iônica , Meios de Cultura/química , Eletroforese em Gel de Poliacrilamida , Índia , Lepidópteros/fisiologia , Metaloproteases/química , Peso Molecular , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Análise de Sobrevida , Xenorhabdus/crescimento & desenvolvimento , Xenorhabdus/isolamento & purificação
19.
3 Biotech ; 2(1): 27-30, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-22582154

RESUMO

Mangiferin present in Curcuma amada was extracted with the help of microwave assisted extraction (MAE). The extraction solvent used was ethanol, which is eco-friendly and reduced the risk of environmental hazard. The mangiferin content was found to increase until 500 W, but decreased as the microwave power was increased further. A similar threshold was also obtained for microwave irradiation time. Following a mathematical analysis, an optimal mangiferin yield of 41 µg/mL was obtained from an extraction time of 15.32 s for a microwave power of 500 W.

20.
Bioprocess Biosyst Eng ; 35(8): 1375-88, 2012 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-22451081

RESUMO

Dietzia maris NIT-D, a canthaxanthin producer, was isolated during routine screening of pigment-producing bacteria. Response surface methodology was applied for statistical designing of process parameters for biomass and canthaxanthin production. The effects of four process parameters (considered as independent variables), namely temperature (10-30 °C), pH (4.75-5.75), shaker speed (75-135 rpm) and percentage inoculum (0.5-2.5 %) on the biomass and canthaxanthin yield (considered as dependent variables) were studied. As much as 122 mg L(-1) of canthaxanthin was obtained when Dietzia maris NIT-D was incubated for 120 h at 25 °C and 120 rpm, initial pH and percentage inoculum being 5.5 and 2 % respectively. The pigment yield is the highest reported till date, with Dietzia maris as the test organism. The maximum biomass yield was 7.39 g L(-1) under optimized process parameters. The predicted values were also verified by validation experiments in 5-day fermentation. Different mathematical models were used to describe growth and production, considering the effect of glucose in batch mode. The kinetic constants were calculated by fitting the experimental data to the models. Cell growth was inhibited beyond a glucose concentration of 15 g L(-1). Andrews' model gave the best fit with a R (2) value of 0.9993. During the exponential growth phase, the specific growth rate was found to remain fairly constant with respect to time. There was no inhibitory effect due to intracellular product accumulation for all concentrations of glucose. This observation is the first of its kind, as previous studies have reported that increasing accumulation of intracellular carotenoid exerts greater degree of inhibition on growth.


Assuntos
Cantaxantina/biossíntese , Corynebacterium/crescimento & desenvolvimento , Corynebacterium/metabolismo , Modelos Biológicos , Concentração de Íons de Hidrogênio
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