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1.
World J Microbiol Biotechnol ; 40(1): 2, 2023 Nov 04.
Artigo em Inglês | MEDLINE | ID: mdl-37923802

RESUMO

The stem rot caused by Sclerotium hydrophilum and false smut caused by Ustilaginoidea virens are two of the major production constraints in rice cultivation in India and other countries. Stem rot and false smut can be effectively controlled with synthetic fungicides. However, the indiscriminate use of chemical fungicides may cause development of resistance among the pathogens. In addition to this, synthetic fungicides also exhibit harmful impacts on the environment. Exploiting microbe-based alternatives for managing plant diseases diminishes public concerns about the ill effects of pesticide usage in crops. In this regard, the present study was designed to investigate the potential of native microbial biocontrol agents (BCAs) from rice rhizosphere for the sustainable management of stem rot and false smut diseases in rice. Potential BCAs and pathogens were identified and characterized through morphological, biochemical, and sanger sequencing techniques. Bio-efficacy tests of potential BCAs against stem rot and false smut diseases on rice under glasshouse conditions indicated higher seed vigour index of the treated seeds, significant improvement in the growth of the seedling, increased dry weight, reduction in percentage disease index viz., 70.03% (stem rot) and 69.24% (false smut) over the control plants. Phytohormones indole acetic acid (IAA), abscisic acid (ABA), gibberellic acid (GA), salicylic acid (SA), and zeatin (tZ) were detected and quantified in the four potential BCAs using liquid chromatography- tandem mass spectrometry (LC-MS/MS). Scanning electron microscopy (SEM) studies revealed the endophytic nature of the strains in rice. The study indicated a positive correlation between the diversity and concentration of phytohormones released by the bioagents and enhanced plant growth promotion and disease suppression in rice.


Assuntos
Fungicidas Industriais , Oryza , Reguladores de Crescimento de Plantas , Cromatografia Líquida , Fungicidas Industriais/farmacologia , Espectrometria de Massas em Tandem , Doenças das Plantas/prevenção & controle
2.
Epidemiol Infect ; 143(8): 1599-605, 2015 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-25314901

RESUMO

Dengue fever, a mosquito-borne viral disease, has become a major public health problem with marked expansion in recent decades. Dengue has now become hyperendemic in India with co-circulation of all the four serotypes. Herein, we report an unprecedented outbreak which occurred during August to October 2011 in Odisha, eastern India. This is the first report of a large epidemic in Odisha. Detailed serological and molecular investigation was carried out to identify the aetiology. Almost half of the samples were found to be dengue antigen (NS1) positive. Further molecular assays revealed circulation of mixed dengue serotypes (DENV-2 and DENV-3). Cosmopolitan genotype of DENV-2 and -3 were identified as the aetiology by phylogenetic analysis. Interestingly, a new lineage of DENV-3 within cosmopolitan genotype was incriminated in this outbreak. The emergence of the unprecedented magnitude of the dengue outbreak with the involvement of a novel lineage of DENV in a newer state of India is a major cause for concern. There is an urgent need to monitor phylodynamics of dengue viruses in other endemic areas.


Assuntos
Anticorpos Antivirais/imunologia , Antígenos Virais/imunologia , Vírus da Dengue/imunologia , Epidemias , RNA Viral/genética , Dengue Grave/epidemiologia , Adolescente , Adulto , Dengue/epidemiologia , Dengue/virologia , Vírus da Dengue/genética , Feminino , Genótipo , Humanos , Índia , Masculino , Pessoa de Meia-Idade , Epidemiologia Molecular , Filogenia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sorogrupo , Dengue Grave/virologia , Adulto Jovem
3.
Plant Physiol Biochem ; 206: 108262, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-38091931

RESUMO

We have previously reported a phenomenon of "conditional pathogenesis", in which, a drought-tolerant rhizobacterium, Pseudomonas putida AKMP7, promotes plant growth under well-watered conditions, while, deteriorating plant health under water-stressed conditions, in Arabidopsis thaliana seedlings. To understand the molecular mechanisms behind this phenomenon, we studied the modulation of salicylic acid (SA) biosynthesis as well as SA-responsive gene expression, involved in systemic acquired resistance (SAR), in A. thaliana, by AKMP7, under well-watered and water-stressed conditions. We found that, the plant SA levels were upregulated by AKMP7, both under, well-watered as well as water-stressed conditions. However, the SA signaling gene, Non-expressor of Pathogenesis Related gene 1 (NPR1) and Pathogenesis Related gene 1 (PR1) were upregulated under well-watered conditions and suppressed under water-stress, in AKMP7 inoculated seedlings. To understand the reason for this, we studied the expression of NPR4, a negative regulator of NPR1, and, NPR3, a negative regulator of PR1. We observed that, AKMP7 suppresses NPR1 and, consequently, PR1 genes, by upregulating NPR4 under water stress. To understand the potential role of NPR4 in conditional-pathogenesis, we performed physiological studies with NPR4 knockout mutants of A. thaliana and found that the NPR4 mutants did not exhibit any signs of the characteristic growth retardation caused by AKMP7 inoculation, under water stress. Preliminary studies with the model pathogen, Pseudomonas syringae, indicate that AKMP7 may lead to enhanced disease suppression under well-watered conditions, but not under water-stress. Taken together, our data suggest that, AKMP7 causes conditional pathogenesis by an overall compromise in plant immune response under water-stress.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Pseudomonas putida , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Pseudomonas putida/genética , Pseudomonas putida/metabolismo , Desidratação , Ácido Salicílico/metabolismo , Regulação da Expressão Gênica de Plantas , Doenças das Plantas/microbiologia
4.
Virus Genes ; 46(2): 219-30, 2013 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-23188193

RESUMO

Dengue non-structural protein (NS1) is known to be protective antigen and also has immense application for serodiagnosis. Several serodiagnostic assays available for dengue viral infection are dependent on tissue culture-grown viral proteins. This task is unsafe, laborious, more expensive that makes it unsuitable for routine large-scale production. Although bacterial expression is relatively simple and easy for recombinant protein expression, it is more challenging to make NS1 protein with native structural and immunological features using bacterial expression system. We have successfully developed a method leading to the purification and refolding of recombinant dengue virus type 3 (DENV3) NS1. The gene encoding NS1 was amplified and cloned in pET28a (+) vector. In order to increase the purity of the recombinant NS1, the transgene was engineered to carry 6× Histidine tags at both N and C-terminal ends. The recombinant construct (pETNS1) was transformed into E. coli Rosetta-gami cells and the expression conditions viz IPTG concentration, media type, temperature, and harvest time were optimized. The size of the expressed protein was found to be ~45 kDa and the authenticity of the expressed protein was confirmed using anti-His and anti-NS1 monoclonal antibodies. The NS1 protein was purified under denaturing conditions, to attain the native conformation, NS1 protein was in vitro refolded and dialyzed. The refolded NS1 protein was detected by commercial Immuno chromatographic strip and NS1 specific monoclonal antibodies. IgM antibody capture ELISA was performed using refolded recombinant NS1 protein which recognized the IgM antibodies in dengue-positive samples of acute phase of infection. Our result suggests that rNS1 protein has immense diagnostic potential and can be used in developing point of care diagnostic assays.


Assuntos
Dengue/virologia , Escherichia coli/genética , Expressão Gênica , Proteínas não Estruturais Virais/química , Proteínas não Estruturais Virais/genética , Sequência de Aminoácidos , Dengue/diagnóstico , Vírus da Dengue/química , Vírus da Dengue/genética , Vírus da Dengue/metabolismo , Escherichia coli/metabolismo , Humanos , Dados de Sequência Molecular , Redobramento de Proteína , Alinhamento de Sequência , Proteínas não Estruturais Virais/metabolismo
5.
J Ind Microbiol Biotechnol ; 40(7): 687-95, 2013 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-23608778

RESUMO

Malaria represents the world's greatest public health problem in terms of number of people affected, levels of morbidity and mortality in tropical and subtropical countries. Malaria parasites are members of the Apicomplexa, family of Plasmodiidae. Histidine-rich protein-II secreted by Plasmodium falciparum is known to be a compelling marker in malaria diagnosis and follow-up. In our present study, we have optimized the batch fermentation and downstream process for large scale production of recombinant P. falciparum HRP-II 62 kDa protein for diagnostic application. The culture broth was effectively induced with IPTG twice at different time intervals to sustain induction for a long period. Batch fermentation resulted in a wet weight of 61.34 g/L and dry cell biomass 12.81 g/L. With the improved downstream process, purified recombinant protein had a yield of 304.60 mg/L. The authenticity of the purified recombinant protein was confirmed via western blotting using indigenously developed HRP-II specific monoclonal antibodies and known positive human clinical sera samples. Further, the reactivity of recombinant HRP-II protein was validated using commercially available immuno chromatographic strips. Indirect ELISA using recombinant purified protein recognized the P. falciparum specific antibodies in suspected human sera samples. Our results clearly suggest that the recombinant HRP-II protein produced via batch fermentation has immense potential for routine diagnostic application.


Assuntos
Malária Falciparum/diagnóstico , Plasmodium falciparum , Proteínas/genética , Proteínas/metabolismo , Proteínas Recombinantes/biossíntese , Adolescente , Adulto , Idoso , Anticorpos Monoclonais/imunologia , Técnicas de Cultura Celular por Lotes/métodos , Reatores Biológicos , Western Blotting , Criança , Cromatografia de Afinidade , Ensaio de Imunoadsorção Enzimática , Escherichia coli/citologia , Escherichia coli/genética , Escherichia coli/metabolismo , Feminino , Fermentação , Humanos , Malária Falciparum/parasitologia , Masculino , Pessoa de Meia-Idade , Plasmodium falciparum/genética , Proteínas/imunologia , Proteínas/isolamento & purificação , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/isolamento & purificação
6.
J Ind Microbiol Biotechnol ; 40(2): 245-55, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-23247902

RESUMO

The present Influenza vaccine manufacturing process has posed a clear impediment to initiation of rapid mass vaccination against spreading pandemic influenza. New vaccine strategies are therefore needed that can accelerate the vaccine production. Pichia offers several advantages for rapid and economical bulk production of recombinant proteins and, hence, can be attractive alternative for producing an effective influenza HA based subunit vaccine. The recombinant Pichia harboring the transgene was subjected to fed-batch fermentation at 10 L scale. A simple fermentation and downstream processing strategy is developed for high-yield secretory expression of the recombinant Hemagglutinin protein of pandemic Swine Origin Influenza A virus using Pichia pastoris via fed-batch fermentation. Expression and purification were optimized and the expressed recombinant Hemagglutinin protein was verified by sodium dodecyl sulfate polyacrylamide gel electrophoresis, Western blot and MALDI-TOF analysis. In this paper, we describe a fed-batch fermentation protocol for the secreted production of Swine Influenza A Hemagglutinin protein in the P. pastoris GS115 strain. We have shown that there is a clear relationship between product yield and specific growth rate. The fed-batch fermentation and downstream processing methods optimized in the present study have immense practical application for high-level production of the recombinant H1N1 HA protein in a cost effective way using P. pastoris.


Assuntos
Fermentação , Glicoproteínas de Hemaglutininação de Vírus da Influenza/biossíntese , Vírus da Influenza A Subtipo H1N1/genética , Pichia/metabolismo , Suínos/virologia , Animais , Biomassa , Reatores Biológicos , Eletroforese em Gel de Poliacrilamida , Endotoxinas/análise , Glicoproteínas de Hemaglutininação de Vírus da Influenza/genética , Vírus da Influenza A Subtipo H1N1/metabolismo , Infecções por Orthomyxoviridae/epidemiologia , Infecções por Orthomyxoviridae/virologia , Pandemias , Pichia/genética , Pichia/crescimento & desenvolvimento , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/genética
7.
Indian J Microbiol ; 53(1): 48-55, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24426078

RESUMO

Bacillus anthracis, the causative agent of anthrax is one of the most important biological warfare agents. In this study, surface plasmon resonance (SPR) technology was used for indirect detection of B. anthracis by detecting protective antigen (PA), a common toxin produced by all live B. anthracis bacteria. For development of biosensor, a monoclonal antibody raised against B. anthracis PA was immobilized on carboxymethyldextran modified gold chip and its interaction with PA was characterized in situ by SPR and electrochemical impedance spectroscopy. By using kinetic evaluation software, KD (equilibrium constant) and Bmax (maximum binding capacity of analyte) were found to be 20 fM and 18.74, respectively. The change in Gibb's free energy (∆G = -78.04 kJ/mol) confirmed the spontaneous interaction between antigen and antibody. The assay could detect 12 fM purified PA. When anthrax spores spiked soil samples were enriched, PA produced in the sample containing even a single spore of B. anthracis could be detected by SPR. PA being produced only by the vegetative cells of B. anthracis, confirms indirectly the presence of B. anthracis in the samples. The proposed method can be a very useful tool for screening and confirmation of anthrax suspected environmental samples during a bio-warfare like situation.

8.
Cogn Neurodyn ; 17(4): 985-1008, 2023 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-37522034

RESUMO

Facial Expression Recognition (FER) is the basis for many applications including human-computer interaction and surveillance. While developing such applications, it is imperative to understand human emotions for better interaction with machines. Among many FER models developed so far, Ensemble Stacked Convolution Neural Networks (ES-CNN) showed an empirical impact in improving the performance of FER on static images. However, the existing ES-CNN based FER models trained with features extracted from the entire face, are unable to address the issues of ambient parameters such as pose, illumination, occlusions. To mitigate the problem of reduced performance of ES-CNN on partially occluded faces, a Component based ES-CNN (CES-CNN) is proposed. CES-CNN applies ES-CNN on action units of individual face components such as eyes, eyebrows, nose, cheek, mouth, and glabella as one subnet of the network. Max-Voting based ensemble classifier is used to ensemble the decisions of the subnets in order to obtain the optimized recognition accuracy. The proposed CES-CNN is validated by conducting experiments on benchmark datasets and the performance is compared with the state-of-the-art models. It is observed from the experimental results that the proposed model has a significant enhancement in the recognition accuracy compared to the existing models.

9.
Virus Genes ; 45(3): 440-51, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-22940846

RESUMO

The hemagglutinin (HA) gene of novel Swine Origin Influenza A/California/04/2009 (H1N1) was engineered for expression in Pichia pastoris as a soluble secreted protein. The full length HA-synthetic gene having α secretory tag under the control of AOX1 promoter was integrated into P. pastoris genome through homologous recombination. The resultant Pichia clones having single and multiple copy integrants of the expression cassettes were screened for the expression of full length HA protein in the culture supernatant. In order to completely exploit the expression potential of the P. pastoris expression system, a systematic investigation on the influence of gene copy number on the expression of the recombinant protein was made. A panel of Pichia clones carrying increasing copies of the heterologous gene was selected based on Geneticin resistance and SYBR green-based quantitative real-time PCR approach. Using these strategies, recombinant Pichia transformants carrying up to a maximum of four to six copies of the transgene were identified. After optimising the expression conditions for shaker flask culture, the resultant clones demonstrated that the increase in copy number results in a proportional elevation in the expression level of H1N1HA recombinant protein. Our findings clearly suggest that the gene dosage effect play a vital role in high level expression of the pandemic Influenza HA protein in yeast system.


Assuntos
Dosagem de Genes , Genes Virais , Glicoproteínas de Hemaglutininação de Vírus da Influenza/metabolismo , Vírus da Influenza A Subtipo H1N1/genética , Pichia/metabolismo , Clonagem Molecular , Eletroforese em Gel de Poliacrilamida , Regulação Fúngica da Expressão Gênica , Vetores Genéticos/genética , Vetores Genéticos/metabolismo , Glicoproteínas de Hemaglutininação de Vírus da Influenza/genética , Recombinação Homóloga , Testes de Sensibilidade Microbiana , Organismos Geneticamente Modificados/genética , Organismos Geneticamente Modificados/metabolismo , Pichia/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Transformação Genética , Transgenes
10.
Parasitol Res ; 111(1): 497-501, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22290450

RESUMO

The anti-plasmodial activity of different solvent extracts of Adhatoda vasica (root), Caesalpinia pulcherrima (leaf), Carica papaya (pulp), Erythroxylum monogynum (leaf), Lantana camara (whole plant), Ocimum sanctum (root) and Phyllanthus niruri (whole plant) were studied against Plasmodium falciparum. Of the 35 extracts tested, seven extracts showed good anti-plasmodial activity. Methanol extract of C. pulcherrima showed the lowest IC50 value (10.96 µg/mL) followed by methanol extract of A. vasica (IC(50)=11.1 µg/mL), chloroform extract of O. sanctum (IC(50)=11.47 µg/mL), methanol extract of E. monogynum (IC(50)=12.23 µg/mL), acetone extract of C. pulcherrima (IC(50)=12.49 µg/mL), methanol extract of O. sanctum and acetone extract of A. vasica (IC(50)=14.04 µg/mL). The results of the present study justify the use of these medicinal plants in traditional practice, and also, a further study on the isolation of anti-plasmodial molecules from their active crude extracts is in progress.


Assuntos
Antimaláricos/farmacologia , Extratos Vegetais/farmacologia , Plantas Medicinais/química , Plasmodium falciparum/efeitos dos fármacos , Antimaláricos/isolamento & purificação , Humanos , Concentração Inibidora 50 , Testes de Sensibilidade Parasitária , Extratos Vegetais/isolamento & purificação
11.
J Environ Biol ; 33(3): 673-81, 2012 May.
Artigo em Inglês | MEDLINE | ID: mdl-23029921

RESUMO

The chemical and biological warfare agents are extremely toxic in nature. They act rapidly even in very small quantities and death may occur in minutes. Hence, physical and medical protection must be provided immediately to save life or avoid serious injury. A first aid kit has thus been developed for providing immediate relief from chemical and biological warfare agents (FAKCBW) with the objective of easy detection, personal decontamination, antidote for chemical warfare agents (like nerve agents, sulphur mustard, phosgene, cyanide, radiation exposure and bacterial agents), along with basic medication aid for pain, fever and inflammation. The kit box also includes a user friendly handbook with a simple standard operating procedure. In addition, the kit is rugged to withstand normal jerks, vibration and is water-proof.


Assuntos
Antídotos/uso terapêutico , Armas Biológicas , Substâncias para a Guerra Química , Primeiros Socorros/instrumentação , Intoxicação/tratamento farmacológico , Descontaminação/instrumentação , Lesões por Radiação/prevenção & controle , Fitas Reagentes
12.
Plant Physiol Biochem ; 183: 46-55, 2022 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-35567874

RESUMO

Plant growth-promoting rhizobacteria (PGPR) are beneficial soil bacteria that colonise the rhizosphere and help plants in growth, development, and stress tolerance. While there is a significant body of research elucidating their benefits to plants, studies on the "abnormal" or "unexpected" behavior of these bacteria are almost non-existent. One such study from our laboratory has previously reported a unique situation in which a certain strain of drought and thermo-tolerant PGPR, namely, Pseudomonas putida AKMP7, becomes pathogenic towards Arabidopsis thaliana under drought conditions, but not under normal (well-watered) conditions. In this study, we have probed deeper into this phenomenon of "conditional pathogenesis". We found that, AKMP7 imparts an enhancement in plant growth under well-watered conditions, while, causing a deterioration in plant health under drought conditions. In an attempt to understand the underlying reasons for this phenomenon, we analysed the phytohormones released by Pseudomonas putida AKMP7 using LC-ESI-MS/MS technique. We identified that AKMP7 releases zeatin (a cytokinin), the auxin derivative -indole acetamide and amino acid-conjugates of auxin (indole-3-acetyl-L-alanine, indole-3-acetyl-L-phenylalanine and indole-3-acetyl-L-aspartate) in the growth medium. By treating the plants with commercially obtained forms of these phytohormones, individually or in combination with AKMP7, we identified that zeatin and auxin derivative indole acetamide can play a crucial role in the conditional pathogenesis exhibited by this bacterium on A. thaliana under drought conditions. Our work lays a foundation for further understanding the precise molecular mechanisms involved in this unique phenomenon of conditional/opportunistic pathogenesis.


Assuntos
Arabidopsis , Pseudomonas putida , Acetamidas , Arabidopsis/microbiologia , Ácidos Indolacéticos , Reguladores de Crescimento de Plantas , Plantas , Solo , Microbiologia do Solo , Espectrometria de Massas em Tandem , Zeatina
13.
Virol J ; 8: 524, 2011 Nov 29.
Artigo em Inglês | MEDLINE | ID: mdl-22126628

RESUMO

Currently available vaccines for the pandemic Influenza A (H1N1) 2009 produced in chicken eggs have serious impediments viz limited availability, risk of allergic reactions and the possible selection of sub-populations differing from the naturally occurring virus, whereas the cell culture derived vaccines are time consuming and may not meet the demands of rapid global vaccination required to combat the present/future pandemic. Hemagglutinin (HA) based subunit vaccine for H1N1 requires the HA protein in glycosylated form, which is impossible with the commonly used bacterial expression platform. Additionally, bacterial derived protein requires extensive purification and refolding steps for vaccine applications. For these reasons an alternative heterologous system for rapid, easy and economical production of Hemagglutinin protein in its glycosylated form is required. The HA gene of novel H1N1 A/California/04/2009 was engineered for expression in Pichia pastoris as a soluble secreted protein. The full length HA- synthetic gene having α-secretory tag was integrated into P. pastoris genome through homologous recombination. The resultant Pichia clones having multiple copy integrants of the transgene expressed full length HA protein in the culture supernatant. The Recombinant yeast derived H1N1 HA protein elicited neutralising antibodies both in mice and rabbits. The sera from immunised animals also exhibited Hemagglutination Inhibition (HI) activity. Considering the safety, reliability and also economic potential of Pichia expression platform, our preliminary data indicates the feasibility of using this system as an alternative for large-scale production of recombinant influenza HA protein in the face of influenza pandemic threat.


Assuntos
Anticorpos Neutralizantes/biossíntese , Anticorpos Antivirais/biossíntese , Glicoproteínas de Hemaglutininação de Vírus da Influenza/imunologia , Vírus da Influenza A Subtipo H1N1/imunologia , Vacinas contra Influenza/imunologia , Proteínas Recombinantes de Fusão/imunologia , Animais , Anticorpos Neutralizantes/imunologia , Anticorpos Antivirais/imunologia , Estudos de Viabilidade , Testes de Inibição da Hemaglutinação , Glicoproteínas de Hemaglutininação de Vírus da Influenza/genética , Humanos , Vírus da Influenza A Subtipo H1N1/genética , Vacinas contra Influenza/genética , Masculino , Camundongos , Pichia/genética , Pichia/metabolismo , Coelhos , Proteínas Recombinantes de Fusão/genética , Vacinas Sintéticas/genética , Vacinas Sintéticas/imunologia
14.
Mol Cell Biochem ; 355(1-2): 249-56, 2011 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-21567208

RESUMO

Oxidative stress is triggered by the wound which results in the production of reactive oxygen species (ROS), thereby delaying normal wound repair. Therefore, it is important to reduce the level of ROS to improve healing. A known antioxidant, dehydrozingerone (DHZ) was synthesized and selected for the study. The authors aimed to investigate the wound healing action of topical (100 mg/wound) and systemic (100 mg/kg, p. o.). DHZ on different wound models in normal and dexamethasone (DEX)-suppressed healing. Topical DHZ showed a significant (P < 0.05) rise in tensile strength when compared to control in normal healing. Significant (P < 0.05) wound closure was observed from 3 to 9 days in DHZ oral and gel treated groups. There was a significant (P < 0.05) rise in hydroxyproline content with the DHZ treated groups when compared to control. Systemic DHZ exhibited a significant (P < 0.05) increase in lysyl oxidase (LO) levels of 3.73 ± 0.15 nmol of H(2)O(2) when compared to control. In DEX-suppressed healing, showed good pro-healing activity with respect to the parameters mentioned above. DHZ treatment exhibited a parabolic dose response of ROS inhibition with a plateau effect at 75 µM. There was a steady and constant increase in the % NO inhibition with increasing doses of DHZ. Oral DHZ is effective in accelerating the healing process in both normal and dexamethasone-suppressed wounds. Our study suggests that DHZ (half analog of curcumin) supplementation reduces the steroid-induced delay in wound healing.


Assuntos
Antioxidantes/farmacologia , Dexametasona/farmacologia , Estirenos/farmacologia , Cicatrização/efeitos dos fármacos , Animais , Antioxidantes/metabolismo , Catalase/metabolismo , Linhagem Celular , Cricetinae , DNA/metabolismo , Estabilidade de Medicamentos , Ensaios Enzimáticos , Géis , Glutationa Transferase/metabolismo , Tecido de Granulação/efeitos dos fármacos , Tecido de Granulação/metabolismo , Hidroxiprolina/metabolismo , Lipopolissacarídeos/farmacologia , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Camundongos , Óxido Nítrico/metabolismo , Proteína-Lisina 6-Oxidase/metabolismo , RNA/metabolismo , Ratos , Ratos Wistar , Espécies Reativas de Oxigênio/metabolismo , Pele/efeitos dos fármacos , Pele/enzimologia , Pele/patologia , Superóxido Dismutase/metabolismo , Resistência à Tração/efeitos dos fármacos
15.
J Clin Microbiol ; 46(11): 3759-65, 2008 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-18799699

RESUMO

A disposable amperometric immunosensor was developed for the detection of Plasmodium falciparum histidine-rich protein 2 (PfHRP-2) in the sera of humans with P. falciparum malaria. For this purpose, disposable screen-printed electrodes (SPEs) were modified with multiwall carbon nanotubes (MWCNTs) and Au nanoparticles. The electrodes were characterized by cyclic voltammetry, scanning electron microscopy, and Raman spectroscopy. In order to study the immunosensing performances of modified electrodes, a rabbit anti-PfHRP-2 antibody (as the capturing antibody) was first immobilized on an electrode. Further, the electrode was exposed to a mouse anti-PfHRP-2 antibody from a serum sample (as the revealing antibody), followed by a rabbit anti-mouse immunoglobulin G-alkaline phosphatase conjugate. The immunosensing experiments were performed on bare SPEs, MWCNT-modified SPEs, and Au nanoparticle- and MWCNT-modified SPEs (Nano-Au/MWCNT/SPEs) for the amperometric detection of PfHRP-2 in a solution of 0.1 M diethanolamine buffer, pH 9.8, by applying a potential of 450 mV at the working electrode. Nano-Au/MWCNT/SPEs yielded the highest-level immunosensing performance among the electrodes, with a detection limit of 8 ng/ml. The analytical results of immunosensing experiments with human serum samples were compared with the results of a commercial Paracheck Pf test, as well as the results of microscopy. The specificities, sensitivities, and positive and negative predictive values of the Paracheck Pf and amperometric immunosensors were calculated by taking the microscopy results as the "gold standard." The Paracheck Pf kit exhibited a sensitivity of 79% (detecting 34 of 43 positive samples; 95% confidence interval [CI], 75 to 86%) and a specificity of 81% (correctly identifying 57 of 70 negative samples; 95% CI, 76 to 92%), whereas the developed amperometric immunosensor showed a sensitivity of 96% (detecting 41 of 43 positive samples; 95% CI, 93 to 98%) and a specificity of 94% (correctly identifying 66 of 70 negative samples; 95% CI, 92 to 99%). The developed method is more sensitive and specific than the Paracheck Pf kit.


Assuntos
Antígenos de Protozoários/sangue , Técnicas Biossensoriais/métodos , Testes Imunológicos/métodos , Malária Falciparum/diagnóstico , Proteínas de Protozoários/sangue , Soro/química , Animais , Anticorpos Antiprotozoários , Humanos , Camundongos , Valor Preditivo dos Testes , Coelhos , Sensibilidade e Especificidade
16.
Front Pharmacol ; 8: 868, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29230174

RESUMO

Pelargonidin chloride (PC) is one of the major anthocyanin found in berries, radish and other natural foods. Many natural chemopreventive compounds have been shown to be potent inducers of phase II detoxification genes and its up-regulation is important for oxidative stress related disorders. In the present study, we investigated the effect of PC in ameliorating citrinin (CTN) induced cytotoxicity and oxidative stress. The cytotoxicity of CTN was evaluated by treating HepG2 (Human hepatocellular carcinoma) cells with CTN (0-150 µM) in a dose dependent manner for 24 h, and the IC50 was determined to be 96.16 µM. CTN increased lactate dehydrogenase leakage (59%), elevated reactive oxygen species (2.5-fold), depolarized mitochondrial membrane potential as confirmed by JC-1 monomers and arrested cell cycle at G2/M phase. Further, apoptotic and necrotic analysis revealed significant changes followed by DNA damage. To overcome these toxicological effects, PC was pretreated for 2 h followed by CTN exposure for 24 h. Pretreatment with PC resulted in significant increase in cell viability (84.5%), restored membrane integrity, reactive oxygen species level were maintained and cell cycle phases were normal. PC significantly up-regulated the activity of detoxification enzymes: heme oxygenase 1 (HO-1), glutathione transferase, glutathione peroxidase, superoxide dismutase and quinone reductase. Nrf2 translocation into the nucleus was also observed by immunocytochemistry analysis. These data demonstrate the protective effect of PC against CTN-induced oxidative stress in HepG2 cells and up-regulated the activity of detoxification enzyme levels through Keap1/Nrf2 signaling pathway.

17.
J Vector Borne Dis ; 43(3): 109-14, 2006 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-17024859

RESUMO

BACKGROUND & OBJECTIVES: Plasmodium vivax is geographically widespread and responsible for > 50% of malaria cases in India. Increased drug resistance of the parasite highlights the immediate requirement of early and accurate diagnosis as well as new therapeutics. In view of this, the present study was undertaken to amplify P. vivax (Indian strains) lactate dehydrogenase gene (PvLDH) which has been identified as a good target for antimalarials as well as diagnostics. METHODS: P. vivax infected clinical blood samples were collected from southern part of India and were tested with established diagnostic parameters (ICT, Giemsa staining). Total DNA was extracted from blood samples and subjected to PCR using two sets of primers, one for the amplification of full PvLDH gene (951 bp) and the other for a partial PvLDH gene fragment (422bp), covering a variable antigenic region (140aa) as compared to other plasmodial species. RESULTS & CONCLUSION: PCRs for both the full and partial gene targets were optimised and found to be consistent when tested on several P. vivax positive clinical samples. In addition, full gene PCR was found to specifically detect only P. vivax DNA and could be used as a specific molecular diagnostic tool. These amplified products can be cloned and expressed as a recombinant protein that might be useful for the development and screening of antimalarials as well as for diagnostic purposes.


Assuntos
Genes de Protozoários/genética , L-Lactato Desidrogenase/genética , Plasmodium vivax/genética , Animais , Primers do DNA , Variação Genética , Humanos , Índia , Malária Vivax/diagnóstico , Malária Vivax/parasitologia , Reação em Cadeia da Polimerase , Especificidade da Espécie
18.
Indian J Med Res ; 99: 121-3, 1994 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-8063347

RESUMO

Three isomers of N, N-diethyltolylacetamides were synthesized and evaluated against Aedes aegypti and Culex quinquefasciatus for repellency and with mice for LD50, as an indicator of mammalian toxicity. Of these, N, N-diethyl-p-tolylacetamide (DEPTA) showed comparatively better repellency against C. quinquefasciatus. All the compounds showed higher repellency against C. quinquefasciatus than Ae. aegypti. In addition, N, N-diethyl-m-tolylacetamide (DEMTA) was found to be least toxic to mice.


Assuntos
Acetamidas/farmacologia , Aedes , Culex , Repelentes de Insetos/farmacologia , Acetamidas/síntese química , Acetamidas/toxicidade , Animais , Feminino , Repelentes de Insetos/toxicidade , Dose Letal Mediana , Masculino , Camundongos , Estereoisomerismo
19.
Indian J Med Res ; 103: 84-90, 1996 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-8714144

RESUMO

A malathion resistant colony of C. quinquefasciatus was developed in the laboratory. LC50 and LC90 for larvae were calculated at every generation and the values were 0.3 ppm and 1.13 ppm for first generation and 61.09 ppm, 136.3 ppm for 25th generation respectively. The fold increase in LC50 and LC90 were 2036 and 2726 folds respectively. Cross resistance against propoxur and chlorpyrifos showed 6.64 and 6.52 fold and 600 and 720 fold increase in their LC50 and LC90 values respectively. Triphenyl phosphate (TPP) and piperonyl butoxide (PB) were used as synergists and TPP indicated proportional decrease in LC50 and LC90 values while not much change was observed with PB. No change in biotic potential (larval hatchability, adult emergence and male and female ratio) between susceptible and malathion resistant colonies was observed.


Assuntos
Clorpirifos , Culex , Resistência a Inseticidas , Malation , Propoxur , Animais
20.
Indian J Med Res ; 113: 103-7, 2001 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-11525152

RESUMO

BACKGROUND & OBJECTIVES: The indiscriminate use of insecticides in public health and agriculture has led to the development of resistance to these insecticides in the vector mosquitoes. To understand the development of resistance to synthetic pyrethroids, selection studies on Aedes aegypti were done at Mysore. METHODS: Ae. aegypti collected from the field were subjected to selection experiment with deltamethrin for 16 generations in the laboratory. Cross resistance test was conducted against permethrin and fenvalerate. RESULTS: Tolerance level was found to increase by 333.83 folds in terms of its LC50 values. Cross resistance of this deltamethrin selected line was tested against permethrin and fenvalerate. The results show that the selected line has developed cross resistance as much as 5.19 and 5.92 folds respectively against permethrin and fenvalerate. INTERPRETATION & CONCLUSIONS: The findings show a continuous elevation in tolerance in Ae. aegypti with increase in deltamethrin selection pressure, and development of cross resistance to other insecticides of the same class. The natural or developed tolerance has its implications in the control of these mosquitoes.


Assuntos
Aedes , Resistência a Medicamentos , Inseticidas , Piretrinas , Animais , Nitrilas
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