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1.
Curr Microbiol ; 80(1): 20, 2022 Dec 02.
Artigo em Inglês | MEDLINE | ID: mdl-36460801

RESUMO

In the present study, a comprehensive proteomic analysis of Brucella melitensis (B. melitensis) strain ATCC23457 was carried out to investigate proteome alterations in response to in vitro-induced nutrient stress. Our analysis resulted in the identification of 2440 proteins, including 365 hypothetical proteins and 850 potentially secretory proteins representing ~77.8% of the B. melitensis proteome. Utilizing a proteogenomics approach, we provide translational evidence for eight novel putative protein-coding genes and confirmed the coding potential of 31 putatively annotated pseudogenes, thus refining the existing genome annotation. Further, using a label-free quantitative proteomic approach, new insights into the cellular processes governed by nutrient stress, including enrichment of amino acid metabolism (E), transcription (K), energy production and conversion (C), and biogenesis (J) processes were obtained. Pathway analysis revealed the enrichment of survival and homeostasis maintenance pathways, including type IV secretion system, nitrogen metabolism, and urease pathways in response to nutrient limitation. To conclude, our analysis demonstrates the utility of in-depth proteomic analysis in enabling improved annotation of the B. melitensis genome. Further, our results indicate that B. melitensis undergoes metabolic adaptations during nutrient stress similar to other Brucella. sp, and adapts itself for long-term persistence and survival.


Assuntos
Brucella melitensis , Proteômica , Brucella melitensis/genética , Proteoma , Aclimatação , Nutrientes
2.
Proc Natl Acad Sci U S A ; 114(17): 4312-4317, 2017 04 25.
Artigo em Inglês | MEDLINE | ID: mdl-28404733

RESUMO

The anomalous decrease of the viscosity of water with applied pressure has been known for over a century. It occurs concurrently with major structural changes: The second coordination shell around a molecule collapses onto the first shell. Viscosity is thus a macroscopic witness of the progressive breaking of the tetrahedral hydrogen bond network that makes water so peculiar. At low temperature, water at ambient pressure becomes more tetrahedral and the effect of pressure becomes stronger. However, surprisingly, no data are available for the viscosity of supercooled water under pressure, in which dramatic anomalies are expected based on interpolation between ambient pressure data for supercooled water and high pressure data for stable water. Here we report measurements with a time-of-flight viscometer down to [Formula: see text] and up to [Formula: see text], revealing a reduction of viscosity by pressure by as much as 42%. Inspired by a previous attempt [Tanaka H (2000) J Chem Phys 112:799-809], we show that a remarkably simple extension of a two-state model [Holten V, Sengers JV, Anisimov MA (2014) J Phys Chem Ref Data 43:043101], initially developed to reproduce thermodynamic properties, is able to accurately describe dynamic properties (viscosity, self-diffusion coefficient, and rotational correlation time) as well. Our results support the idea that water is a mixture of a high density, "fragile" liquid, and a low density, "strong" liquid, the varying proportion of which explains the anomalies and fragile-to-strong crossover in water.

3.
Malays J Med Sci ; 27(6): 15-26, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33447131

RESUMO

BACKGROUND: Human brucellosis is an important zoonotic disease of public health and often remains neglected owing to lack of sensitive and efficient diagnostic methods. This study evaluates diagnostic utility of in-house designed enzyme-linked immunosorbent assay (ELISA) using whole-cell antigens of Brucella abortus (B. abortus) S19 against the commercially available kits. METHODS: A prospective cohort study involving different populations within the Vidarbha regions of Maharashtra, India was conducted through camps organised from May 2009 to October 2015. A total of 568 serum samples were collected from high-risk people recruited as study cohorts based on inclusion criteria, additional risk factors and clinical symptoms. Samples were evaluated by indirect ELISA using the whole-cell antigens of B. abortus. The results were compared with the commercially available IgG detection ELISA kit to ascertain the specificity and sensitivity of the developed test. RESULTS: Fever, body ache, joint pain, lower back pain, loss of appetite and weight loss were major symptoms associated with the disease. With the cut-off of > 0.8, the positivity of brucellosis infection was at 12.32% (70/568) compared to 9.33% (53/568) as detected by the commercial kit. The in-house developed ELISA method yielded a sensitivity of 87.5% and specificity of 99.18% as compared to the commercial kits (sensitivity -80.30% and specificity -99.6%). DISCUSSION: The B. abortus S19-derived whole-cell protein-based ELISA is rapid and cost-effective and can be used for screening brucellosis infection in lieu of the commercially available ELISA kits.

4.
Indian J Med Res ; 145(6): 833-839, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29067986

RESUMO

BACKGROUND & OBJECTIVES: Central nervous system (CNS) infection caused by Mycobacterium tuberculosis (MTB) is the most severe form of extrapulmonary tuberculosis (EPTB) due to a high level of mortality and morbidity. Limited studies are available on CNS-TB animal model development. The present study describes the development of a murine model of CNS-TB using a clinical strain (C3) isolated from the cerebrospinal fluid (CSF) of CNS-TB patients. METHODS: Groups of mice were infected by the intravenous route with MTB C3 strain isolated from the CSF of CNS-TB patients. Brain and lung tissue were evaluated for bacterial burden, histopathology and surrogate markers of TB infection at 30 and 50 days post-infection. RESULTS: Mice infected intravenously with MTB C3 strains showed progressive development of CNS disease with high bacillary burden in lungs at the initial stage (30 days), which eventually disseminated to the brain at a later stage (50 days). Similarly, high mortality (60%) was associated in mice infected with C3 strain compared to control. INTERPRETATION & CONCLUSIONS: The study showed development of a novel murine model of CNS-TB using the C3 strain of MTB that replicated events of extrapulmonary dissemination. The developed model would be helpful in understanding the pathogenesis of CNS-TB infection for the development of improved therapeutic interventions in future.


Assuntos
Infecções Bacterianas do Sistema Nervoso Central/microbiologia , Modelos Animais de Doenças , Mycobacterium tuberculosis/isolamento & purificação , Tuberculose Pulmonar/microbiologia , Animais , Infecções Bacterianas do Sistema Nervoso Central/líquido cefalorraquidiano , Humanos , Pulmão/microbiologia , Pulmão/patologia , Camundongos , Camundongos Endogâmicos BALB C , Mycobacterium tuberculosis/patogenicidade , Tuberculose Pulmonar/líquido cefalorraquidiano
5.
J Vector Borne Dis ; 54(1): 35-43, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28352044

RESUMO

BACKGROUND & OBJECTIVES: Chikungunya virus (CHIKV) infection has recently witnessed re-emergence, affecting rural areas of India with high morbidity rates. This prospective study was conducted to evaluate seroprevalence and clinical manifestation in targeted villages reporting cases of CHIKV infection. METHODS: A total of 482 patients were recruited from Kalmana and Kothari villages of Ballarpur; Chandrapur district of Maharashtra state, India during CHIKV outbreaks in 2011-12. The serum samples from infected CHIKV patients were simultaneously screened through ELISA for detection of antigen and antibodies (IgM and IgG). Chi-square analysis was used to evaluate differences in seropositivity between age, gender and clinical manifestations of CHIKV. RESULTS: Out of 482 enrolled participants, 197 (41%) males and 285 (59%) females were aged between 5 and 92 yr. The clinical manifestations such as small joint pain (80%), neck stiffness (75%), fever (49%) and large joint pain (47%) were observed amongst CHIKV infected subjects. Mucocutaneous rashes (91%) on knees (71%), feet (56%), fingers and palms (54%) were also observed. Overall, seroprevalence of CHIKV infection was found to be 46% in infected participants during the epidemic period. Among risk factors, ageing and female gender was strongly associated with a raised seroprevalence of CHIKV infection along with symptoms such as rashes, small joints pain and neck stiffness. INTERPRETATION & CONCLUSION: This study reported high seroprevalence rates of CHIKV infection in targeted popula- tions, suggesting its re-emergence in rural India. Proper surveillance is, therefore, necessary to minimize re-emergence and in controlling these impending and sporadic outbreaks.


Assuntos
Febre de Chikungunya/epidemiologia , Febre de Chikungunya/patologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Anticorpos Antivirais/sangue , Antígenos Virais/sangue , Vírus Chikungunya/imunologia , Criança , Pré-Escolar , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Índia/epidemiologia , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , População Rural , Estudos Soroepidemiológicos , Adulto Jovem
6.
Toxicol Ind Health ; 32(3): 485-92, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24193048

RESUMO

Our study aimed to determine the cardiac toxicities of T-2 toxin, a representative mycotoxin that frequently contaminates maize, cereals, and other agricultural products, harvested and stored under damp and cold conditions. Dermal exposure to T-2 toxin caused severe cardiotoxicity in experimental Wistar rats. Electrocardiography studies showed the conduction abnormalities including prolongation of the QT and corrected QT interval, shortening of the PR interval, and tachycardia. Biochemical studies also reported the toxicity of T-2 toxin. T-2 toxin induced acute cardiotoxicity in rats and characterized by significant (p < 0.05) elevation of serum troponin I, creatine kinase (CK) isoenzyme MB, CK isoenzyme NAC, and lactate dehydrogenase as compared to control rats. It is concluded that cardiotoxicity effects of T-2 toxin are thought to be due to direct action on electrocardiac potentials and biochemical changes.


Assuntos
Cardiotoxicidade/patologia , Cardiotoxicidade/fisiopatologia , Toxina T-2/toxicidade , Administração Cutânea , Animais , Pressão Sanguínea/efeitos dos fármacos , Peso Corporal/efeitos dos fármacos , Eletrocardiografia , Coração/efeitos dos fármacos , Coração/fisiopatologia , Masculino , Miocárdio/patologia , Ratos , Ratos Wistar , Toxina T-2/administração & dosagem , Testes de Toxicidade Subcrônica
7.
Environ Sci Technol ; 49(7): 4749-56, 2015 Apr 07.
Artigo em Inglês | MEDLINE | ID: mdl-25738526

RESUMO

Deployment of improved biomass burning cookstoves is recognized as a black carbon (BC) mitigation measure that has the potential to achieve health benefits and climate cobenefits. Yet, few field based studies document BC concentration reductions (and resulting human exposure) resulting from improved stove usage. In this paper, data are presented from 277 real-world cooking sessions collected during two field studies to document the impacts on indoor BC concentrations inside village kitchens as a result of switching from traditional stoves to improved forced draft (FD) stoves. Data collection utilized new low-cost cellphone methods to monitor BC, cooking duration, and fuel consumption. A cross sectional study recorded a reduction of 36% in BC during cooking sessions. An independent paired sample study demonstrated a statistically significant reduction of 40% in 24 h BC concentrations when traditional stoves were replaced with FD stoves. Reductions observed in these field studies differ from emission factor reductions (up to 99%) observed under controlled conditions in laboratory studies. Other nonstove sources (e.g., kerosene lamps, ambient concentrations) likely offset the reductions. Health exposure studies should utilize reductions determined by field measurements inside village kitchens, in conjunction with laboratory data, to assess the health impacts of new cooking technologies.


Assuntos
Poluição do Ar em Ambientes Fechados/estatística & dados numéricos , Culinária/instrumentação , Fuligem/análise , Poluição do Ar em Ambientes Fechados/análise , Biomassa , Carbono/análise , Estudos Transversais , Exposição Ambiental , Utensílios Domésticos , Humanos , Índia , População Rural
8.
J Chem Phys ; 142(1): 014501, 2015 Jan 07.
Artigo em Inglês | MEDLINE | ID: mdl-25573566

RESUMO

A two-channel impedance technique has been used to study the relaxation behavior of 2-ethyl-1-hexanol with polar and non-polar dopants at the few percent concentration level over a wide temperature and frequency range. The non-polar dopants shift both the Debye and the primary structural relaxation time in the same direction, to shorter times for 3-methylpentane and to longer times for squalane, consistent with the relative glass transition temperatures (Tg) of the components. By contrast, polar dopants such as water or methanol modify the α-process towards slower dynamics and increased amplitude, while the Debye process is accelerated and with a decreased amplitude. This effect of adding water to alcohol is explained by water promoting more compact structures with reduced Kirkwood correlation factors. This picture is consistent with a shift in the neutron scattering pre-peak to lower scattering vectors and with simulation work on alcohol-water systems.


Assuntos
Espectroscopia Dielétrica , Hexanóis/química , Difração de Nêutrons , Simulação de Dinâmica Molecular , Água/química
9.
Med J Armed Forces India ; 71(2): 182-5, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25859083

RESUMO

Human papillomavirus (HPV) associated cervical cancer is the leading cause of deaths in India. However, cytological/HPV screening may result in early detection of cervical cancer, resulting in early treatment and reduced mortality. Although reports related to general population is available, data on HPV prevalence among women attending AFMS health care facilities is scarce. Cervical samples were collected for cytological staining by Pap test and molecular detection by PCR, genotyping by HPV specific primers and sequencing. Apart from finding of atypical cells of undetermined significance (ASCUS) in one subject, no evidence of malignancy was observed. A high prevalence of HPV was found in this study group, which was intermediate between previous reports from general population and cervical cancer patients. All the subjects had infection of high risk HPV type16. HPV prevalence was found similar between different age groups. Although, none of the study subjects had malignant changes, but due to high prevalence of high risk HPV infection and other associated risk factors, these subjects might be at an elevated risk of developing cervical cancer. Regular follow-up of these patients who were detected HPV positive are required to screen for cervical malignancy.

10.
Malar J ; 13: 450, 2014 Nov 21.
Artigo em Inglês | MEDLINE | ID: mdl-25416405

RESUMO

BACKGROUND: The R2 repeat region of GLURP has been reported as a good genetic marker for Plasmodium falciparum genotyping. Proper knowledge of the extent and nature of P. falciparum genetic diversity using highly immunogenic R2 repeat region in malaria-endemic areas is a crucial element to understand various aspects related to immunity acquisition and disease pathogenesis. METHODS: Population diversity of P. falciparum GLURP and amino acid sequence repeats in GLURP R2 region was studied in malaria-endemic Assam state, northeast India and compared at an interval of five years during 2005 (Group-A) and 2011 (Group-B). RESULTS: Of the 66 samples, a total of 55 samples showed positive PCR bands for GLURP R2 region and altogether ten types of alleles with size ranging from 501 bp to 1,050 bp (50 bp bin) were observed and coded as genotypes I-X. In Group-A (n = 29), 24 samples were found infected with single, four with double and one with triple P. falciparum genotype, while in Group-B (n = 26), single genotype was found in 23 samples, double in two samples and triple in one sample. Genotype IV showed significant increase (p = 0.002) during 2011 (Group-B). Genotypes I to V were more common in Group-B (62%), however genotypes VI to X were more frequently distributed in Group-A. The expected heterozygosity was found slightly higher in Group-A (HE = 0.87) than Group-B (HE = 0.85), whereas multiplicity of infection (MOI) in Group-A (MOI = 1.21 ± 0.49) and Group-B (MOI = 1.12 ± 0.43) did not display significant variation. The amino acid repeat sequence unit (AAU) DKNEKGQHEIVEVEEILPE (called 'a') was more frequent in the well-conserved part of R2 repeat region. CONCLUSION: The present study is the first extensive study in India which has generated substantial data for understanding the type and distribution of naturally evolved genetic polymorphism at amino acid sequence level in GLURP R2 repeat region in P. falciparum. There was decrease in the PCR amplicon size as well as the number of AAU [amino acid repeat unit] in Group-B displaying the bottleneck effect. The present study described a new type of AAU 'd' which varied from the other previous known AAUs.


Assuntos
Plasmodium falciparum/classificação , Plasmodium falciparum/genética , Polimorfismo Genético , Proteínas de Protozoários/genética , Sequências Repetitivas de Aminoácidos , Adolescente , Adulto , Alelos , Criança , DNA de Protozoário/química , DNA de Protozoário/genética , Feminino , Genótipo , Humanos , Índia , Malária Falciparum/parasitologia , Masculino , Dados de Sequência Molecular , Plasmodium falciparum/isolamento & purificação , Reação em Cadeia da Polimerase , Análise de Sequência de DNA , Adulto Jovem
11.
ScientificWorldJournal ; 2014: 707368, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25383378

RESUMO

Anguillospora longissima, isolated from root as endophytic freshwater hyphomycetes, was evaluated for its bioactivity (antibacterial potential) against five bacterial strains, namely, Gram-positive (Bacillus subtilis MTCC 121) and Gram-negative (Agrobacterium tumefaciens MTCC 609, Escherichia coli MTCC 40, Erwinia chrysanthemum, and Xanthomonas pseudomonas). Antimicrobial activity was assessed by measuring the zone of inhibition with preliminary and secondary antimicrobial assays. The applied fungus was found significant for all tested bacterial strains as showen by their zone of inhibition. In preliminary antimicrobial assay, maximum zone of inhibition was recorded against Gram-negative human pathogenic bacterial strain Escherichia coli (23 mm) followed by Erwinia chrysanthemi (22 mm), Agrobacterium tumefaciens (21 mm), and Xanthomonas phaseoli (21 mm), while minimum zone of inhibition was observed against Bacillus subtilis (20 mm). In secondary antimicrobial assay, the maximum zone of inhibition was recorded against Erwinia chrysanthemi (11 mm) followed by Agrobacterium tumefaciens (10 mm), Xanthomonas phaseoli (10 mm), and Bacillus subtilis (9 mm) and minimum inhibition was found against Escherichia coli (8 mm).


Assuntos
Antibacterianos/farmacologia , Ascomicetos/efeitos dos fármacos , Fungos Mitospóricos/química , Extratos Vegetais/farmacologia , Antibacterianos/química , Bacillus subtilis/efeitos dos fármacos , Erwinia/efeitos dos fármacos , Escherichia coli/efeitos dos fármacos , Humanos , Testes de Sensibilidade Microbiana , Extratos Vegetais/química , Raízes de Plantas/química , Pseudomonas/efeitos dos fármacos
12.
Int J Food Sci Nutr ; 65(7): 856-61, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-24896300

RESUMO

Anemia is a frequent problem in both the primary and secondary health care programs. In contrast, most areas of northeast India are vulnerable to iron toxicity. In the present study, we documented the effect of administration of iron rich water on hemolytic anemia in a Wistar rats' animal model. Hemolytic anemia was induced by phenyl hydrazine through intraperitoneal route and diagnosed by the lowering of blood hemoglobin. After inducing the hemolytic anemia, 24 Wistar rats (n = 6 in four groups) were randomly assigned to 1 mg/l, 5 mg/l, and 10 mg/l ferric oxide iron along with 1 mg/ml ascorbic acid administered through drinking water; a control group was treated with iron-free water. The hematological and biochemical parameters, iron levels in liver, spleen, and kidney were estimated after 30 d of treatment. In the group treated with 5 mg/l iron and ascorbic acid, a significant increase of serum iron and ferritin, and a decrease of TIBC (total iron binding capacity) were observed without changes in other biochemical parameters and histopathological findings. However, in the group treated with 10 mg/l iron and ascorbic acid, hematological changes with significantly higher values for white blood cell count, serum glutamic phospho transaminase, serum glutamic oxaloacetic transaminase, alkaline phosphatase, glucose, splenic, and liver iron content, indicate potential toxicity at this supplementation level. Data suggest that the optimum concentration of iron (5 mg/l) and ascorbic acid solution may improve anemic conditions and may be therapeutically beneficial in the treatment of iron deficiency anemia without any negative impact, while 10 mg/l in drinking water seems to be the threshold for the initiation of toxicity.


Assuntos
Anemia Hemolítica/tratamento farmacológico , Ácido Ascórbico/farmacologia , Água Potável/química , Ferro/farmacologia , Anemia Hemolítica/induzido quimicamente , Animais , Ácido Ascórbico/administração & dosagem , Suplementos Nutricionais , Ferro/administração & dosagem , Ferro/química , Masculino , Fenil-Hidrazinas/toxicidade , Ratos , Ratos Wistar
13.
J Environ Biol ; 35(2): 421-5, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24665772

RESUMO

Pupal stages of Simulium were collected and identified from various breeding habitats of twelve locations in northeastern India. Simulium flies, while attempting to land on human were collected between 07:00-17:00 hrs to understand the biting pattern. Seven species belonging to three sub-genera, Eusimulium (1), Gomphostilbia (1) and Simulium (5) belonging genus Simulium were encountered. Out of total seven species recorded, S. (E) aureohirtum, S. (G) tenuistylum and S. (S) rufibasis were predominant and shared 30.3%, 29.9% and 27.6% of total collection. Stream breeding habitat contributed 47.3% of total catch and was found to be preferred breeding habitat (p < 0.0001). S. (S) christophersi and S. (G) tenuistylum were recorded for the first time from the northeastern region of India. Simuliids biting rhythm showed bimodal pattern and were more active during sunny day (p < 0.0001). Microscopic dissection of simuliids (n = 266) did not incriminate simuliids as vector of onchocerciasis.


Assuntos
Mordeduras e Picadas de Insetos , Onchocerca/isolamento & purificação , Simuliidae/classificação , Simuliidae/fisiologia , Animais , Ecossistema , Interações Hospedeiro-Parasita , Índia , Água
14.
Artigo em Inglês | MEDLINE | ID: mdl-38608221

RESUMO

Objective: In this study we evaluated the utility of Abortus Melitensis Ovis Suis Brucella PCR (AMOS PCR) for the molecular characterization of Brucella species and analyzed the associated risk factors for brucellosis in Central Indian and Meghalayan population. Methods: AMOS PCR was carried out in a total of 160 BSCP-31 PCR-positive DNA samples isolated previously from the blood of Central Indian (n = 90) and Meghalayan cohorts (n = 70). Clinical and associated risk factors recorded earlier were used to establish strain-specific disease outcomes in study cohorts. Results: Brucella melitensis was found to be the dominant strain in both Central Indian and Meghalayan cohorts (57.7% and 54.28%, respectively) followed by Brucella abortus (42.22% and 38.57%). Although rare, brucellosis cases in the Meghalayan population also showed the presence of Brucella suis (7.14%) and Brucella ovis (2.85%). Febrile illness was a major clinical risk factor in both study cohorts, while occupational risk factors like exposure to animals and raw milk consumption were major mediating factors for brucellosis in Central Indian cohorts. On the contrary, meat consumption was found to be significant predisposing factor for brucellosis in Meghalaya. Conclusion: Molecular characterization of Brucella species provides important public health data for mitigation, advocacy, and antimicrobial stewardship.

15.
J Chem Phys ; 139(14): 144503, 2013 Oct 14.
Artigo em Inglês | MEDLINE | ID: mdl-24116631

RESUMO

It is well established that many mono-hydroxy alcohols show an extra relaxation process of the Debye type in addition to the signatures of primary and secondary structural relaxations, which is observed only in dielectric spectroscopy and related techniques. In order to gain further insight into the nature of this Debye peak, we study the linear and nonlinear dielectric behavior of a series of isomeric octyl alcohols and of mixtures of n-propanol with one of the octanols. These samples display systematic variations of the Debye peak intensity and concomitant changes in the Kirkwood correlation factor gK from 0.1 to 4, indicative of different equilibrium constants, K(c∕r), that characterize the populations of non-polar ring and polar open chain structures. For cases where K(c∕r) is not too far from unity, we find that a high electric field shifts K(c∕r) towards more chains, and that the accompanying change in the end-to-end vector of hydrogen-bond connected structures occurs on the Debye time scale. The results suggest that gK is correlated with the spectral separation of the Debye and primary structural peaks, as both features depend on steric hindrance of chain flexibility or bond rotation barriers and on average chain lengths. Based on the complex dynamics of supercooled mono-hydroxy alcohols with three relaxation peaks that cover many orders of magnitude in frequency, it is argued that a frequency dependent gK may be required for assessing the average orientational correlations within hydrogen-bonded structures correctly.

16.
Indian J Med Res ; 137(6): 1180-7, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23852299

RESUMO

BACKGROUND & OBJECTIVES: Clostridium sordellii and its toxins are associated with diseases in animals as well as human. C. sordellii produces two protein toxins (lethal toxin and haemorrhagic toxin). Lethal toxin has gained more importance due its high toxicity. The present study was carried out to develop a sandwich ELISA for detection of lethal toxin of C. sordellii. METHODS: The catalytic domain (1.6 kb) of lethal toxin of C. sordellii was PCR amplified, cloned into pQE30 UA vector and transformed into Escherichia coli SG 13009. Expression conditions were optimized and the recombinant protein was purified under native condition using Ni-NTA affinity chromatography, confirmed by SDS-PAGE and Western blot. Antibody was generated against the purified recombinant protein using Freund's complete and incomplete adjuvants (FCA and FIA) in BALB/c mice and rabbit. A sandwich ELISA was optimized for the detection of lethal toxin. RESULTS: The maximum recombinant protein expression was achieved at 0.5 mM IPTG (isopropylthiogalactoside) induction 4.0 h of post-induction. The polyclonal antibody raised in mice and rabbit showed a titre up to 1:512000. The produced antibody was highly sensitive with the detection limit of 0.3 ng/ml of lethal toxin at 1:4000 dilutions of mice (capturing) and rabbit (revealing) antibody. INTERPRETATION & CONCLUSIONS: An ELISA based detection system was developed for the detection of lethal toxin of C. sordellii. The developed detection system was found to be specific as there was no cross-reactivity with any other clostridial toxins. It will be useful for the detection of lethal toxin of C. sordellii in clinical and environmental samples.


Assuntos
Toxinas Bacterianas/análise , Clostridium sordellii/metabolismo , Ensaio de Imunoadsorção Enzimática/métodos , Animais , Toxinas Bacterianas/imunologia , Domínio Catalítico , Cromatografia de Afinidade , Clonagem Molecular , Infecções por Clostridium/diagnóstico , Infecções por Clostridium/imunologia , Limite de Detecção , Camundongos , Camundongos Endogâmicos BALB C , Reação em Cadeia da Polimerase , Coelhos , Proteínas Recombinantes/química , Fatores de Tempo
17.
Indian J Med Res ; 138(6): 988-94, 2013 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-24521646

RESUMO

BACKGROUND & OBJECTIVE: Japanese encephalitis (JE) outbreaks are common in Assam, northeastern State of India. Information on resistance in known JE vectors in the affected area is important for effective control measures. This study was undertaken to determine the species abundance of JE vectors endemic to Sibsagar district of Assam, and their susceptibility against DDT and deltamethrin. METHODS: Adult mosquitoes were collected using CDC light trap and aspirators from human dwellings from 13 endemic villages falling under three Primary Health Centres. Collected mosquitoes were identified and unfed female mosquitoes were used for DDT and deltamethrin sensitivity bioassay. The bioassay was performed following WHO protocol using standard susceptibility test kit. Knockdown time (KDT) was monitored at every 10 minutes intervals, whereas mortalities were recorded 24 h post-exposure. Vector density and resistance status were mapped using geographic information system (GIS) technique. RESULTS: A total of 7655 mosquitoes were sampled under three genera, i.e. Anopheles, Culex and Mansonia, and nine species, the JE vector Cx. vishnui group (31.78%) was the most predominant species, followed by Ma. uniformis (16.81%) and Ma. indiana (16.45%). All vector species were suspected to be resistant to DDT and sensitive to deltamethrin, except Ma. indiana, which was suspected to deltamethrin resistant. The KDT50 and KDT95 values of vector mosquitoes for DDT were significantly higher as compared to deltamethrin. The probit model used to estimate KDT50 and KDT95 values did not display normal distribution of percentage knockdown with time for all the vectors tested for DDT and deltamethrin, except for Ma. indiana for deltamethrin assay and Cx. gelidus for the DDT assay. INTERPRETATION & CONCLUSION: Differences in insecticide resistance status were observed between insecticides and vector species. The results of this study provided baseline data on insecticide resistance in known JE vectors of Sibsagar, Assam. The maps generated may allow better communication in control operations and comparison of changes in susceptibility status of these vectors over time.


Assuntos
Culicidae/efeitos dos fármacos , Encefalite Japonesa/transmissão , Insetos Vetores/genética , Resistência a Inseticidas/genética , Animais , Culicidae/genética , Culicidae/patogenicidade , DDT/toxicidade , Encefalite Japonesa/genética , Encefalite Japonesa/prevenção & controle , Feminino , Humanos , Índia , Insetos Vetores/efeitos dos fármacos , Nitrilas/toxicidade , Piretrinas/toxicidade , Especificidade da Espécie
19.
Carbohydr Res ; 532: 108917, 2023 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-37572627

RESUMO

Differential scanning calorimeter and broadband dielectric spectroscopy in a broad range of temperatures (150-300 K) were employed to study the d-lyxose aqueous mixture at different hydration levels. Two relaxation processes were observed in all investigated d-lyxose aqueous mixtures. A relaxation process (process-I) usually known as the primary relaxation mode which is accountable for the collective motion of d-lyxose aqueous solution, was observed above the glass transition temperature (Tg). Below Tg, another process designated as process-II was found which is mainly related to the water molecule relaxation inside the d-lyxose matrix. The average relaxation times as a function of temperature and dielectric strengths of both observed relaxation processes (I & II) were analyzed for all hydration levels in d-lyxose. It was identified that the relaxation amplitude of process-II in the d-lyxose aqueous mixture was increased drastically and their activation energies were found to be approximately independent of the content of water above critical concentration, xc = 0.28. This suggests that the dynamical process observed above xc was dominated by the presence of water clusters. In the current aqueous mixture, the critical content of water (xc) is slightly higher as compared to previously reported aqueous mixtures, indicating a more cooperative nature of water molecules with a d-lyxose matrix. Additionally, the Tg of pure water was estimated at 128 ± 5.8 K from the extrapolation of DSC Tg data of the d-lyxose aqueous solution by using the well-known Gordon-Taylor equation. Our current result gives further support to the well-accepted glass transition (Tg) of pure water.


Assuntos
Vitrificação , Água , Água/química , Pentoses , Temperatura , Espectroscopia Dielétrica , Vidro
20.
IEEE Trans Nanobioscience ; 22(2): 365-374, 2023 04.
Artigo em Inglês | MEDLINE | ID: mdl-35853045

RESUMO

Nowadays, Food additives and preservatives have become a hot topic especially formalin, which is a chemical substance used to preserve food. Chronic cancer is caused by swallowing the formalin-contaminated food on a regular basis. As a result, detection of the formalin in food ingredients is a critical need, and this requirement is becoming increasingly important in new terrains. Therefore, a new approach, planar waveguide-based surface plasmon resonance (SPR) sensor is presented in this paper employing silver-chloride materials-black phosphorus structure to detect the formalin. In this study, the optimization of metal thickness and optimal performance of chloride material are analyzed by observing the transmittance power and resonance shift for conventional and chloride-based sensors, respectively. Moreover, the performance parameters, such as sensitivity of 344°/ RIU, quality-factor of [Formula: see text], detection-accuracy of 3.34 and figure-of-merit of 164.74, are observed for the proposed sensor. Furthermore, the performance comparative study is conducted and found that the proposed sensor highlighted the enhanced the SPR sensor and highlighted the performance of FOM. Finally, the electromagnetic filed distributions of the proposed WG-based SPR sensor are shown using the Opti-FDTD software.


Assuntos
Cloretos , Formaldeído , Ressonância de Plasmônio de Superfície , Software , Vibração
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