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1.
Lett Appl Microbiol ; 75(5): 1336-1345, 2022 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-35962588

RESUMEN

Biofilm development in Pseudomonas aeruginosa is regulated by its quorum sensing (QS) systems. It has three major QS systems: LasI/R, RhlI/R and PQS/MvfR. Previous studies showed that phenyllactic acid (PLA) binds to RhlR and PqsR and inhibits the Rhl and PQS QS; and eugenol at sublethal concentration inhibits Las and PQS QS systems. Here, we have demonstrated that a combination of sublethal doses of eugenol and PLA enhanced the inhibition of the QS mediated production of the virulence factors and biofilm development of this pathogen. A combination of 50 µmol l-1 eugenol and 0·3 mmol l-1 PLA significantly inhibited the pyocyanin production, protease activity, swarming motility and cytotoxic activities of P. aeruginosa strain PAO1, whereas eugenol and PLA when added individually to PAO1 cultures were less effective in inhibiting its virulence factor expression. Biofilm formation of PAO1 was reduced by 32, 19 and 87% on glass surfaces; and 54, 49 and 93% on catheter surfaces when treated using 50 µmol l-1 eugenol or 0·3 mmol l-1 PLA and their combinations, respectively. The in vitro finding in the reduction of biofilm development was further validated in vivo using a catheter associated medaka fish biofilm model. Our results indicate that a combination of QS inhibitors targeting different QS pathways should be selected while designing therapeutic molecules to achieve maximum QS mediated biofilm inhibition and clinical outcome against P. aeruginosa.


Asunto(s)
Pseudomonas aeruginosa , Factores de Virulencia , Animales , Factores de Virulencia/metabolismo , Piocianina , Eugenol/farmacología , Biopelículas , Percepción de Quorum , Péptido Hidrolasas , Poliésteres , Antibacterianos/farmacología , Proteínas Bacterianas/metabolismo
2.
Cereb Cortex ; 29(12): 5072-5084, 2019 12 17.
Artículo en Inglés | MEDLINE | ID: mdl-30953440

RESUMEN

KCC2 is a brain specific chloride-potassium cotransporter affecting neuronal development including migration and cellular maturation. It modulates chloride homeostasis influencing the switch of GABA from depolarizing to hyperpolarizing, which contributes to the cues that influence the termination of neuronal migration. The expression of KCC2 during migration of interneurons, therefore, correlates with the ability of these cells to respond to GABA as a stop signal. Manipulation of KCC2 in development can affect various aspects of migrating neurons, including the speed. We describe the effect of KCC2 downregulation and inhibition on features of migrating interneurons of normal ferret kits and those treated with methylazoxymethanol acetate, which increases KCC2. Treatment of organotypic cultures with Bisphenol A, an environmental toxin that alters gene expression, also downregulates KCC2 protein. In organotypic slices treated with the KCC2 antagonist VU0240551, chloride imaging shows inhibition of KCC2 via blockade of chloride flux. Time-lapse video imaging of organotypic cultures treated with either drug, shows a significant increase in the average speed, step size, and number of turns made by migrating neurons leaving the ganglionic eminence. Our findings demonstrate the harmful effect of environmental toxins on brain development and potential consequences in the pathogenesis of neurodevelopmental disorders.


Asunto(s)
Movimiento Celular/fisiología , Interneuronas/metabolismo , Neocórtex/embriología , Neurogénesis/fisiología , Simportadores/metabolismo , Contaminantes Ocupacionales del Aire/toxicidad , Animales , Compuestos de Bencidrilo/toxicidad , Movimiento Celular/efectos de los fármacos , Hurones , Interneuronas/efectos de los fármacos , Neocórtex/efectos de los fármacos , Neocórtex/metabolismo , Neurogénesis/efectos de los fármacos , Técnicas de Cultivo de Órganos , Fenoles/toxicidad , Tiazoles/toxicidad , Tioglicolatos/toxicidad , Cotransportadores de K Cl
3.
Parasite Immunol ; 40(7): e12535, 2018 07.
Artículo en Inglés | MEDLINE | ID: mdl-29745990

RESUMEN

Leishmania donovani, the causative parasite of Visceral Leishmaniasis (VL), deviously manipulates host monocytes/macrophages to ensure its survival. Although monocytes/macrophages from patients with VL have demonstrated an impaired oxidative burst and antigen presentation, an unanswered yet pertinent question remains as to whether they are deactivated or alternatively activated. The significantly raised plasma levels of IL-4/IL-13 and IL-10 in VL patients suggested a microenvironment conducive for alternative activation of monocytes/macrophages. Accordingly, the classical markers for IL-4-driven monocytes/macrophages [M(IL-4)] were studied namely intramonocytic CD206+ , circulating CCL22 and CCL17, and were unchanged. Furthermore, the mRNA expression of Kruppel-like factor 4 (KLF4), peroxisome proliferator-activated receptors (PPAR)-γ and arginase-I (ARG-I) in peripheral blood mononuclear cells was unaltered. However, markers for IL-10-driven monocytes/macrophages [M(IL-10)], namely soluble CD163, intramonocytic IL-10, and circulating CXCL13 were significantly increased. Monocytes/macrophages of patients with VL demonstrated an increased expression of markers for M(IL-10), along with the absence of markers for M(IL-4). Taken together, in human VL, manipulation of these IL-10 polarized monocytes-macrophages may pave the way for improved therapeutic outcomes.


Asunto(s)
Interleucina-10/análisis , Leishmania donovani/inmunología , Leishmaniasis Visceral/inmunología , Adulto , Anciano , Femenino , Humanos , Factor 4 Similar a Kruppel , Leishmaniasis Visceral/parasitología , Leucocitos Mononucleares/inmunología , Macrófagos/inmunología , Masculino , Persona de Mediana Edad , Monocitos/inmunología , Estallido Respiratorio
4.
J Appl Microbiol ; 125(5): 1286-1295, 2018 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-29972893

RESUMEN

AIM: The effect of anacardic acid impregnation on catheter surfaces for the prevention of Staphylococcus aureus attachments and biofilm formations were evaluated. METHODS AND RESULTS: Silicon catheter tubes were impregnated using different concentrations of anacardic acids (0·002-0·25%). Anacardic acids are antibacterial phenolic lipids from cashew nut (Anacardium occidentale) shell oil. Anacardic acid-impregnated silicon catheters revealed no significant haemolytic activity and were cytocompatible against fibroblast cell line (L929). Sustained release of anacardic acids was observed for 4 days. Anacardic acid-impregnated silicon catheters efficiently inhibited S. aureus colonization and the biofilm formation on its surface. The in vivo antibiofilm activity of anacardic acid-impregnated catheters was tested in an intraperitoneal catheter-associated medaka fish infection model. Significant reduction in S. aureus colonization on anacardic acid-impregnated catheter tubes was observed. CONCLUSIONS: Our data suggest that anacardic acid-impregnated silicon catheters may help in preventing catheter-related staphylococcal infections. SIGNIFICANCE AND IMPACT OF THE STUDY: This study opens new directions for designing antimicrobial phytochemical-coated surfaces with ideal antibiofilm properties and could be of great interest for biomedical research scientists.


Asunto(s)
Ácidos Anacárdicos/farmacología , Anacardium/química , Antibacterianos/farmacología , Biopelículas/efectos de los fármacos , Staphylococcus aureus/efectos de los fármacos
5.
Appl Opt ; 57(10): AO1-AO2, 2018 Apr 01.
Artículo en Inglés | MEDLINE | ID: mdl-29714265

RESUMEN

The editors of Acousto-Optics 2018 provide an overview of the featured papers in the context of the 13th School of Acousto-Optics and Applications from which the articles were drawn.

6.
Osteoporos Int ; 28(4): 1287-1293, 2017 04.
Artículo en Inglés | MEDLINE | ID: mdl-27921147

RESUMEN

Alendronate therapy has been associated with serious side effects. Altering the alendronate concentration and combining with high-frequency loading as mechanical intervention was explored in this animal study as a treatment for osteoporosis. The bone anabolic potency of high-frequency loading was overruled by the different alendronate dosages applied in the present study. Further exploration of reduced hormonal therapy associated with mechanical interventions in osteoporosis treatment should be sought. INTRODUCTION: The aim of the present study was to investigate the effect of alendronate (ALN) administration at two different dosages, associated or not with high-frequency (HF) loading, on the bone microstructural response. METHODS: Sixty-four female Wistar rats were used, of which 48 were ovariectomized (OVX) and 16 were sham-operated (shOVX). The OVX animals were divided into three groups: two groups were treated with alendronate, at a dosage of 2 mg/kg (ALN(2)) or at a reduced dosage of 1 mg/kg (ALN(1)) three times per week. A third OVX group did not receive pharmaceutical treatment. All four groups were mechanically stimulated via whole body vibration (WBV) at HF (up to 150 Hz) or left untreated (shWBV). ALN and HF were administered for 6 weeks, starting at 10-week post-(sh)OVX. Tibia bone structural parameters were analyzed using ex vivo microcomputed tomography. RESULTS: Trabecular bone loss and structural deterioration resulting from ovariectomy were partially restored by ALN administration, demonstrated by the improvement of trabecular patter factor (Tb.Pf), trabecular separation (Tb.Sp), and structure model index (SMI) of the ALN groups compared to that of the OVX group, regardless of the applied dosage [ALN(2) or ALN(1)] or mechanical loading regime (shWBV or WBV). However, a significant positive effect of the ALN(1) administration on trabecular (decrease of Tb.Sp and SMI) and cortical bone (increase of cortical thickness) microarchitecture compared to that of the OVX status group was observed for both loading regimes was not seen for ALN(2). Furthermore, HF loading resulted in cortical bone changes, with an increased trabeculary area and endocortical perimeter. Finally, the benefits of a combined therapy of ALN with HF loading could not be discerned in the present experimental conditions. CONCLUSIONS: The bone anabolic potency of HF loading was overruled by the ALN dosages applied in the present study. Further altering the ALN dosage combined with robust mechanical stimuli needs to be considered in osteoporosis research and eventually therapy.


Asunto(s)
Alendronato/administración & dosificación , Conservadores de la Densidad Ósea/administración & dosificación , Osteoporosis/terapia , Vibración/uso terapéutico , Alendronato/uso terapéutico , Animales , Densidad Ósea/fisiología , Conservadores de la Densidad Ósea/uso terapéutico , Terapia Combinada , Relación Dosis-Respuesta a Droga , Femenino , Osteoporosis/tratamiento farmacológico , Osteoporosis/fisiopatología , Ovariectomía , Ratas Wistar , Tibia/fisiopatología , Soporte de Peso/fisiología , Microtomografía por Rayos X/métodos
7.
J Environ Manage ; 197: 89-95, 2017 Jul 15.
Artículo en Inglés | MEDLINE | ID: mdl-28334647

RESUMEN

Rice the staple food is a notable intake source of arsenic to the rural population of eastern India through food-chain. A field survey was carried out to study the variation of arsenic load in different parts of rice genotype Shatabdi (most popular genotype of the region) exposed to varying level of arsenic present in the irrigation water and soil. As irrigation is the primary source of arsenic contamination, a study was conducted to assess arsenic load in rice ecosystem under deficit irrigation practices like intermittent ponding (IP), saturation (SAT) and aerobic (AER) imposed during stress allowable stage (16-40 days after transplanting) of the crop (genotype Shatabdi). Present survey showed that arsenic content in water and soil influenced the arsenic load of rice grain. Variation in arsenic among different water and soil samples influenced grain arsenic load to the maximum extent followed by straw. Deviation in root arsenic load due to variation in water and soil arsenic content was lowest. Arsenic concentration of grain is strongly related to the arsenic content of both irrigation water and soil. However, water has 10% higher impact on grain arsenic load over soil. Translocation of arsenic from root to shoot decreased with the increase in arsenic content of water. Imposition of saturated and aerobic environment reduced both yield and grain arsenic load. In contrast under IP a marked decrease in grain arsenic content recorded with insignificant reduction in yield. Deficit irrigation resulted in significant reduction (17.6-25%) in arsenic content of polished rice and the values were lower than that of the toxic level (<0.2 mg kg-1). In contrast the decrease in yield was to the tune of 0.9% under IP regime over CP.


Asunto(s)
Arsénico , Contaminantes del Suelo , Riego Agrícola , Humanos , India , Oryza , Suelo
8.
Ann Oncol ; 27(7): 1291-8, 2016 07.
Artículo en Inglés | MEDLINE | ID: mdl-27117531

RESUMEN

BACKGROUND: In the phase I KEYNOTE-001 study, pembrolizumab demonstrated durable antitumor activity in patients with advanced non-small-cell lung cancer (NSCLC). We sought to characterize the relationship between pembrolizumab dose, exposure, and response to define an effective dose for these patients. PATIENTS AND METHODS: Patients received pembrolizumab 2 mg/kg every 3 weeks (Q3W) (n = 55), 10 mg/kg Q3W (n = 238), or 10 mg/kg Q2W (n = 156). Response (RECIST v1.1) was assessed every 9 weeks. The relationship between the estimated pembrolizumab area under the concentration-time curve at steady state over 6 weeks (AUCss-6weeks) and the longitudinal change in tumor size (sum of longest diameters) was analyzed by regression and non-linear mixed effects modeling. This model was simultaneously fit to all tumor size data, then used to simulate response rates, normalizing the trial data across dose for prognostic covariates (tumor PD-L1 expression and EGFR mutation status). The exposure-safety relationship was assessed by logistic regression of pembrolizumab AUCss-6weeks versus occurrence of adverse events (AEs) of interest based on their immune etiology. RESULTS: Overall response rates were 15% [95% confidence interval (CI) 7%-28%] at 2 mg/kg Q3W, 25% (18%-33%) at 10 mg/kg Q3W, and 21% (95% CI 14%-30%) at 10 mg/kg Q2W. Regression analyses of percentage change from baseline in tumor size versus AUCss-6weeks indicated a flat relationship (regression slope P > 0.05). Simulations showed the exposure-response relationship to be similarly flat, thus indicating that the lowest evaluated dose of 2 mg/kg Q3W to likely be at or near the efficacy plateau. Exposure-safety analysis showed the AE incidence to be similar among the clinically tested doses. CONCLUSIONS: No significant exposure dependency on efficacy or safety was identified for pembrolizumab across doses of 2-10 mg/kg. These results support the use of a 2 mg/kg Q3W dosage in patients with previously treated, advanced NSCLC. CLINICALTRIALSGOV REGISTRY: NCT01295827.


Asunto(s)
Anticuerpos Monoclonales Humanizados/administración & dosificación , Anticuerpos Monoclonales/administración & dosificación , Carcinoma de Pulmón de Células no Pequeñas/tratamiento farmacológico , Pronóstico , Adulto , Anciano , Anticuerpos Monoclonales/efectos adversos , Anticuerpos Monoclonales Humanizados/efectos adversos , Antígeno B7-H1/genética , Carcinoma de Pulmón de Células no Pequeñas/genética , Supervivencia sin Enfermedad , Relación Dosis-Respuesta a Droga , Efectos Colaterales y Reacciones Adversas Relacionados con Medicamentos/patología , Receptores ErbB/genética , Femenino , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Masculino , Persona de Mediana Edad , Estadificación de Neoplasias , Resultado del Tratamiento
10.
Osteoporos Int ; 26(1): 281-90, 2015 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-25164696

RESUMEN

UNLABELLED: High-frequency loading via whole body vibration promotes bone formation and increases bone strength. Whether this translates to positive titanium implant osseointegration in osteoporotic bone was explored in this animal study. An anabolic effect of not only bisphosphonate treatment but also high-frequency loading on implant osseointegration in osteoporotic bone was observed. INTRODUCTION: The present study investigated the impact of high-frequency (HF) loading, applied via whole body vibration (WBV), on titanium implant osseointegration in healthy versus ovariectomy-induced compromised versus pharmacologically treated compromised bone. METHODS: A custom-made Ti implant was inserted into the metaphyseal tibia of 59 rats and left to heal for either 4 or 14 days. Rats were divided into six groups according to their hormonal and mechanical status. WBV, consisting of 10 consecutive frequency steps at an acceleration of 0.3 g, was applied daily for either 4 or 14 days. Tissue samples were processed for quantitative histology at the tibial cortical and medullar level. Data were analyzed by three-way ANOVA and by post hoc pairwise comparisons. RESULTS: The bone healing response at the interface and surrounding titanium implants was negatively influenced by osteoporotic bone conditions, mainly at the trabecular bone level. Furthermore, the administration of bisphosphonates for preventing the ovariectomy-induced impaired peri-implant response was successful. Finally, the effect of HF WBV loading on the peri-implant bone healing was dependent on the bone condition and was anabolic solely in untreated osteoporotic trabecular bone when applied for an extended period of time. CONCLUSIONS: The bone healing response to implant installation is compromised in osteoporotic bone conditions, in particular at the trabecular bone compartment. Meanwhile, not only pharmacological treatment but also mechanical loading via HF WBV can exert a positive effect on implant osseointegration in this specific bone micro-environment. The peri-implant cortical bone, however, seems to be less sensitive to HF WBV loading influences.


Asunto(s)
Implantes Experimentales , Oseointegración/fisiología , Osteoporosis/fisiopatología , Tibia/fisiopatología , Vibración/uso terapéutico , Animales , Femenino , Ovariectomía , Ratas Wistar , Tibia/patología , Tibia/cirugía , Titanio , Soporte de Peso/fisiología
11.
Osteoporos Int ; 26(1): 303-13, 2015 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-25236876

RESUMEN

UNLABELLED: Mechanical loading at high frequency affects bone. Whether this also applies to osteoporotic bone, combined or not with bisphosphonate therapy, was investigated in this animal study through imaging. An anabolic effect of high-frequency loading on osteoporotic bone, however non-synergistic with bisphosphonates, was found, thereby revealing its potential for treatment of osteoporosis. INTRODUCTION: In an effort to elucidate the effect of high-frequency (HF) loading on bone and to optimize its potential for treatment osteoporosis, this study aimed to investigate the effect of HF loading via whole body vibration (WBV), alone or in association with bisphosphonate treatment (alendronate--ALN), on the micro-architecture of ovariectomy (OVX)-induced compromised bone. METHODS: Eighty-four female Wistar rats were ovariectomized (OVX) or sham-operated (shOVX). OVX animals were treated either with ALN (3 days/week at a dose of 2 mg/kg) or with saline solution. Each group (shOVX, OVX, ALN) was further divided into subgroups relative to the loading status (sham-WBV versus WBV) and the duration of experimental period (4 days versus 14 days). (Sham)WBV loading was applied for 10 min/day using 10 consecutive steps of HF loading (130, 135, 140, 145, 150, 130, 135, 140, 145, 150 Hz). Tibial bone structural responses to WBV and/or ALN treatment were analyzed using ex vivo micro-computed tomography. RESULTS: The animal's hormonal status displayed a major impact on the trabecular and cortical bone structural parameters. Furthermore, mechanical treatment with HF WBV increased the cortical thickness and reduced the medullar area in OVX rats. However, OVX trabecular bone was not affected by HF stimuli. Finally, ALN prevented OVX-associated bone loss, but the association of ALN with WBV did not lead to a synergistic bone response in OVX bone. CONCLUSIONS: HF WBV mechanical stimulation displayed an anabolic effect on osteoporotic cortical bone, confirming its therapeutic properties for enhancing compromised bone. Additionally, its association with bisphosphonates' administration did not produce any additive effect on the bone micro-architecture in the present study.


Asunto(s)
Alendronato/uso terapéutico , Conservadores de la Densidad Ósea/uso terapéutico , Osteoporosis/terapia , Vibración/uso terapéutico , Animales , Terapia Combinada , Femenino , Hormonas/sangre , Osteoporosis/tratamiento farmacológico , Osteoporosis/patología , Osteoporosis/fisiopatología , Ovariectomía , Ratas Wistar , Tibia/diagnóstico por imagen , Tibia/patología , Aumento de Peso/fisiología , Soporte de Peso , Microtomografía por Rayos X/métodos
12.
Tissue Antigens ; 84(3): 316-20, 2014 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-24962747

RESUMEN

Asthma is a heterogeneous disease for which a strong genetic basis is firmly established. It is a complex disorder influenced by gene-environment interaction. Human leukocyte antigen (HLA) genes have been shown to be consistently associated with asthma and its related phenotypes in various populations. The aim of this study was to determine the frequency of the selected HLA classes I and II allelic groups in asthmatic and control groups. HLA typing was performed using polymerase chain reaction-sequence-specific typing (PCR-SSP) method. The allele frequency was estimated by direct counting. Frequency of each HLA allelic group was compared between asthmatic group and control group using χ(2) test. P-value was corrected by multiplying with the number of the allelic groups studied. Odds ratio (OR) and its corresponding 95% confidence interval (CI) for each allelic group were calculated using graphpad instat 3.10. The results of this study showed a significantly higher frequency of HLA-DRB1*03 in asthmatics than in controls (11.43% vs 3.64%, OR = 3.78, 95% CI = 1.61-8.85, P = 0.0025, Pcorr < 0.05). Analysis of HLA alleles in low and high total serum immunoglobulin E (IgE) level in asthmatics revealed no significant association. HLA-DRB1*03 may be implicated in the susceptibility to asthma in the pediatric population.


Asunto(s)
Asma/genética , Cadenas HLA-DRB1/genética , Grupos de Población , Asma/inmunología , Niño , Preescolar , Femenino , Frecuencia de los Genes , Interacción Gen-Ambiente , Estudios de Asociación Genética , Predisposición Genética a la Enfermedad , Genotipo , Cadenas beta de HLA-DQ/genética , Prueba de Histocompatibilidad , Humanos , Inmunidad Humoral/genética , Inmunoglobulina E/sangre , India , Masculino , Polimorfismo Genético
13.
Indian J Clin Biochem ; 28(2): 197-200, 2013 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-24426210

RESUMEN

Immunoglobulin (Ig) E has been shown to be a major contributing factor for the development of bronchial hyperresponsiveness in asthma. An elevation in serum IgE levels contributes to asthma and is considered a potent predictor of the development of asthma. The objectives of the present study were to estimate the levels of total serum IgE in asthmatic and healthy control subjects and to investigate the relationship of various demographic and clinical characteristics with the total serum IgE level in asthmatics. We measured the levels of total serum IgE using the ELISA kits (AccuBind, Monobind Inc., USA). The relevant demographic and clinical data were obtained using the questionnaire. The results showed that asthmatic children had significantly elevated level of total serum IgE compared to that of the healthy controls. The levels of total IgE and IL-4 in sera of 44 asthmatic children showed a significant positive correlation. Total serum IgE >150 IU/mL was found to be significantly associated with the age, exposure to cigarette smoke, and raised eosinophil count in asthmatic children. In conclusion, the elevated level of total serum IgE may demonstrate the allergic etiology of asthma in the subjects studied.

14.
SAR QSAR Environ Res ; 33(6): 463-484, 2022 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-35638563

RESUMEN

The quantitative structure-activity relationship (QSAR) modelling of mixtures is not as simple as that for individual chemicals, and it needs additional care to avoid overestimation of the performance. In this research, we have developed a 2D-QSAR model using only 2D interpretable and reproducible descriptors to predict the aquatic toxicity of mixtures of polar and non-polar narcotic substances present in the environment. Partial least squares (PLS) regression has been used to model the response variable (log 1/EC50 against Photobacterium phosphoreum) and the structural features of 84 binary mixtures of polar and nonpolar narcotic toxicants complying with the Organization of Economic Co-operation and Development (OECD) protocols. The model was cross-validated by mixtures-out and compounds-out cross-validation to nullify the developmental bias. The reliability of prediction of the model has been judged by the Prediction Reliability Indicator (PRI) tool using a newly designed set. The new model is robust, reproducible, extremely predictive, easily interpretable, and can be used for reliable prediction of aquatic toxicity of any untested chemical mixtures within the applicability domain. We have additionally used a machine learning-based chemical read-across algorithm in this study to improve the quality of predictions for the toxicity of the mixtures with the modelled descriptors.


Asunto(s)
Relación Estructura-Actividad Cuantitativa , Contaminantes Químicos del Agua , Análisis de los Mínimos Cuadrados , Narcóticos , Reproducibilidad de los Resultados , Contaminantes Químicos del Agua/química , Contaminantes Químicos del Agua/toxicidad
15.
Br J Cancer ; 102(11): 1578-91, 2010 May 25.
Artículo en Inglés | MEDLINE | ID: mdl-20502461

RESUMEN

BACKGROUND: Heat-shock protein 90 (Hsp90) has a crucial role in both the stabilisation and regulation of various proteins, including those related to radioresistance. Inhibition of Hsp90 may therefore provide a strategy for enhancing the radiosensitivity of tumour cells. This study explores the responses of four tumour cell lines (A549, GaMG, HT 1080 and SNB19) to combined treatment with ionising radiation (IR) and two novel inhibitors of Hsp90, NVP-AUY922 and NVP-BEP800. The techniques used included cell and colony counts, expression of Hsp90, Hsp70, Akt, survivin, cleaved caspase 3, p53, cell-cycle progression and associated proteins. DNA damage was analysed by histone gammaH2AX and Comet assays. RESULTS: We found that NVP-AUY922 and NVP-BEP800 enhanced radiosensitivity in all tested cell lines. In contrast, only two cell lines (HT 1080 and GaMG) exhibited an increased rate of apoptosis after drug pretreatment, as revealed by western blot. In all tested cell lines, the expression of histone gammaH2AX, a marker of DNA double-strand breaks, after combined drug-IR treatment was higher and its decay rate was slower than those after each single treatment modality. Drug-IR treatment also resulted in impaired cell-cycle progression, as indicated by S-phase depletion and G2/M arrest. In addition, the cell cycle-associated proteins, Cdk1 and Cdk4, were downregulated after Hsp90 inhibition. INTERPRETATION: These findings show that the novel inhibitors of Hsp90 can radiosensitise tumour cell lines of different entities through destabilisation and depletion of several Hsp90 client proteins, thus causing the depletion of S phase and G2/M arrest, increased DNA damage and repair protraction and, to some extent, apoptosis. The results might have important implications for the radiotherapy of solid tumours.


Asunto(s)
Ciclo Celular/efectos de los fármacos , Daño del ADN/efectos de los fármacos , Reparación del ADN/efectos de los fármacos , Isoxazoles/farmacología , Neoplasias/radioterapia , Pirimidinas/farmacología , Tolerancia a Radiación/efectos de los fármacos , Resorcinoles/farmacología , Benzoquinonas/farmacología , Ciclo Celular/genética , Ciclo Celular/fisiología , Línea Celular Tumoral , Supervivencia Celular , Daño del ADN/genética , Reparación del ADN/genética , Reparación del ADN/efectos de la radiación , ADN de Neoplasias/efectos de los fármacos , ADN de Neoplasias/metabolismo , ADN de Neoplasias/efectos de la radiación , Relación Dosis-Respuesta a Droga , Relación Dosis-Respuesta en la Radiación , Regulación hacia Abajo/efectos de los fármacos , Regulación hacia Abajo/efectos de la radiación , Evaluación Preclínica de Medicamentos , Proteínas HSP90 de Choque Térmico/antagonistas & inhibidores , Humanos , Lactamas Macrocíclicas/farmacología , Neoplasias/genética , Fármacos Sensibilizantes a Radiaciones/farmacología , Regulación hacia Arriba/efectos de los fármacos , Regulación hacia Arriba/efectos de la radiación
16.
J Exp Biol ; 213(Pt 14): 2410-6, 2010 Jul 15.
Artículo en Inglés | MEDLINE | ID: mdl-20581270

RESUMEN

In this study we identify a cGMP-dependent protein kinase (PKG) cascade as a biochemical pathway critical for controlling low-oxygen tolerance in the adult fruit fly, Drosophila melanogaster. Even though adult Drosophila can survive in 0% oxygen (anoxia) environments for hours, air with less than 2% oxygen rapidly induces locomotory failure resulting in an anoxic coma. We use natural genetic variation and an induced mutation in the foraging (for) gene, which encodes a Drosophila PKG, to demonstrate that the onset of anoxic coma is correlated with PKG activity. Flies that have lower PKG activity demonstrate a significant increase in time to the onset of anoxic coma. Further, in vivo pharmacological manipulations reveal that reducing either PKG or protein phosphatase 2A (PP2A) activity increases tolerance of behavior to acute hypoxic conditions. Alternatively, PKG activation and phosphodiesterase (PDE5/6) inhibition significantly reduce the time to the onset of anoxic coma. By manipulating these targets in paired combinations, we characterized a specific PKG cascade, with upstream and downstream components. Further, using genetic variants of PKG expression/activity subjected to chronic anoxia over 6 h, approximately 50% of animals with higher PKG activity survive, while only approximately 25% of those with lower PKG activity survive after a 24 h recovery. Therefore, in this report we describe the PKG pathway and the differential protection of function vs survival in a critically low oxygen environment.


Asunto(s)
Proteínas Quinasas Dependientes de GMP Cíclico/metabolismo , Drosophila melanogaster/fisiología , Oxígeno/metabolismo , Transducción de Señal/fisiología , Animales , GMP Cíclico/antagonistas & inhibidores , GMP Cíclico/metabolismo , Proteínas Quinasas Dependientes de GMP Cíclico/genética , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/enzimología , Activación Enzimática , Inhibidores Enzimáticos/metabolismo , Femenino , Locomoción/fisiología , Masculino , Tasa de Supervivencia
17.
Arch Insect Biochem Physiol ; 74(1): 52-66, 2010 May.
Artículo en Inglés | MEDLINE | ID: mdl-20422718

RESUMEN

Our knowledge of how genes act on the nervous system in response to the environment to generate behavioral plasticity is limited. A number of recent advancements in this area concern food-related behaviors and a specific gene family called foraging (for), which encodes a cGMP-dependent protein kinase (PKG). The desert locust (Schistocerca gregaria) is notorious for its destructive feeding and long-term migratory behavior. Locust phase polyphenism is an extreme example of environmentally induced behavioral plasticity. In response to changes in population density, locusts dramatically alter their behavior, from solitary and relatively sedentary behavior to active aggregation and swarming. Very little is known about the molecular and genetic basis of this striking behavioral phenomenon. Here we initiated studies into the locust for gene by identifying, cloning, and studying expression of the gene in the locust brain. We determined the phylogenetic relationships between the locust PKG and other known PKG proteins in insects. FOR expression was found to be confined to neurons of the anterior midline of the brain, the pars intercerebralis. Our results suggest that differences in PKG enzyme activity are correlated to well-established phase-related behavioral differences. These results lay the groundwork for functional studies of the locust for gene and its possible relations to locust phase polyphenism.


Asunto(s)
Proteínas Quinasas Dependientes de GMP Cíclico/genética , Proteínas Quinasas Dependientes de GMP Cíclico/metabolismo , Conducta Alimentaria/fisiología , Saltamontes/enzimología , Saltamontes/genética , Secuencia de Aminoácidos , Animales , Encéfalo/metabolismo , Proteínas Quinasas Dependientes de GMP Cíclico/química , Femenino , Perfilación de la Expresión Génica , Regulación Enzimológica de la Expresión Génica , Saltamontes/clasificación , Masculino , Datos de Secuencia Molecular , Filogenia , Alineación de Secuencia
18.
Eur Rev Med Pharmacol Sci ; 14(9): 789-94, 2010 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-21061839

RESUMEN

AIM: A total of 625 faecal specimens of diarrheic cases (n-313) and non diarrheic controls (n-312), were screened by RT-PCR to detect Noroviruses in children aged below 5 years in Kolkata, India. MATERIALS AND METHODS: Out of the 313 fecal specimens (cases) screened using CDC primer set, 10 (3.19%) showed amplification in reverse transcription-polymerase chain reaction (RT-PCR) for Norovirus. These included 5 of 260 (1.92%) from hospitalized and 5 of 53 (9.43%) from out patients departament (OPD) cases. RESULTS: Nine (90%) of Norovirus positive cases belonged to genogroup GII and one specimen (10%) was positive for genogroup GI. Among the 312 non diarrheic controls 2 (0.63%) were positive for Norovirus GII. Partial RNA dependent RNA polymerase gene (RdRp) sequences corresponding to the six Norovirus GII positive samples showed homology to the sequences of Djibouti (horn of Africa), Brazil, Italy, Japan and US norovirus strains. CONCLUSION: This study shows the detection of newly emerging Norovirus strains among diarrheic and non diarrheic children in Kolkata.


Asunto(s)
Infecciones por Caliciviridae/epidemiología , Enfermedades Transmisibles Emergentes/epidemiología , Diarrea/epidemiología , Gastroenteritis/epidemiología , Norovirus/genética , Brasil , Infecciones por Caliciviridae/diagnóstico , Infecciones por Caliciviridae/virología , Preescolar , Enfermedades Transmisibles Emergentes/diagnóstico , Enfermedades Transmisibles Emergentes/virología , Diarrea/diagnóstico , Diarrea/virología , Djibouti , Heces/virología , Gastroenteritis/diagnóstico , Gastroenteritis/virología , Genotipo , Humanos , India , Pacientes Internos/estadística & datos numéricos , Italia , Japón , Norovirus/clasificación , Pacientes Ambulatorios/estadística & datos numéricos , Filogenia , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Estados Unidos
19.
Ecotoxicology ; 19(2): 405-18, 2010 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-19830552

RESUMEN

The paper presents the first document to identify the enrichment pattern of acid leachable trace metals (ALTMs) such as Fe, Mn, Cr, Cu, Ni, Pb, Cd, Co, Mo, Ag, As and Ba and their relationship with sediment quality parameters (pH, organic carbon, carbonates and texture) in core sediments (<63 microm particle size) from Indian Sunderban mangrove wetland, formed at the estuarine phase of the river Hugli (Ganges). Textural analysis reveals an overall predominance of mud. The results indicate that the change in pH values causes coagulation and precipitation of ALTMs. Fe and Mn have fairly close distribution patterns of enrichment in surface layers which might be ascribed to early diagnetic processes. The most prominent feature of ALTMs is the enrichment of Fe, Mn, Cr, Cu, Ni, Pb, and Ba in the surface-subsurface layers in the sediment cores, which is mainly attributed to the intense industrial and agricultural activities as well as drainage of untreated domestic sewage to this coastal region. The ALTMs also indicate their association with organic carbon and Fe-Mn oxyhydroxides. The enrichment is well--supported by the correlation, grouping and clustering of ALTMs in statistical analyses. Anthropogenic Factor values indicated ALTMs enrichment for all trace metals due to intense anthropogenic activities. Overall higher values of ALTMs in sediments in comparison to other Indian coastal regions indicate that they are mainly due to the uncontrolled anthropogenic activities in this mangrove estuarine complex. Statistical analyses suggest that five ALTMs (Cu, Pb, As, Mo, Ba) are attached to the organic particles and the clustering of elements separately also indicates that they are from external source. The result of the present study suggests the need for a regular monitoring program which will help to improve the quality of this potential wetland.


Asunto(s)
Ácidos/química , Monitoreo del Ambiente , Sedimentos Geológicos/química , Metales/análisis , Contaminantes Químicos del Agua/análisis , Humedales , Carbono/análisis , Carbonatos/análisis , Concentración de Iones de Hidrógeno , India , Metales/química , Compuestos Orgánicos/análisis , Compuestos Orgánicos/química , Tamaño de la Partícula , Textiles/análisis , Contaminantes Químicos del Agua/química
20.
Arch Environ Contam Toxicol ; 59(1): 49-61, 2010 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-20082069

RESUMEN

The distribution and potential sources of 16 polycyclic aromatic hydrocarbons (PAHs) in sediment cores (<63 microm particle size) of the Sundarban mangrove wetland, northeastern coast of Bay of Bengal (India), were investigated by gas chromatography coupled to mass spectrometry. The total concentrations of 16 PAHs ( summation operator(16)PAHs) ranged from 132 to 2938 ng/g, with a mean of 634 ng/g, and the sum of 10 out of 16 priority PAHs ( summation operator(10)PAH) varied from 123 to 2441 ng/g, with a mean of 555 ng/g, and the 5 carcinogenic PAHs (benzo[b]fluoranthene, benzo[k]fluoranthene, benzo[a]pyrene, indeno[1,2,3-cd]pyrene, and dibenz[a,h]anthracene) accounted for 68-73% of the priority PAHs. Maximum concentrations of the sediment core were obtained at subsoil depth of 12-16 cm. The prevalence of four to six aromatic ring PAHs and cross-plots of specific isomer ratios such as phenanthrene/anthracene, fluoranthene/pyrene, and methylphenanthrenes/phenanthrene suggested the predominance of wood and coal combustion sources, the atmospheric deposition, and surface runoff to be the major transport pathways. A good correlation existed between the benzo[a]pyrene level and the total PAH concentrations, making this compound a potential molecular marker for PAH pollution. Total TEQ (S) (carc) values calculated for samples varied from 6.95 ng/g TEQ (S) (carc) to 119 ng/g TEQ (S) (carc) , with an average of 59 ng/g dry weight TEQ (S) (carc) . The baseline data can be used for regular monitoring, considering the industrial and agricultural growth around this coastal environment.


Asunto(s)
Monitoreo del Ambiente , Sedimentos Geológicos/química , Hidrocarburos Policíclicos Aromáticos/análisis , Contaminantes Químicos del Agua/análisis , Humedales , Avicennia , India , Hidrocarburos Policíclicos Aromáticos/química , Hidrocarburos Policíclicos Aromáticos/toxicidad , Medición de Riesgo , Contaminantes Químicos del Agua/química , Contaminantes Químicos del Agua/toxicidad
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