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This study determined the anti-listerial activity of indigenous probiotics from traditional fermented foods of Western Himalaya against meat borne Listera monocytogens isolates from Himachal Pradesh. One hundred samples of meat and meat products like chicken (n = 25), chevon (goat meat, n = 20), fish (n = 20) and pork (n = 30) were collected and were analyzed for the presence of Listeria spp. by recommended culture and biochemical methods. L. monocytogens isolates were confirmed by PCR targeting the virulence gene hlyA (haemolysin A) and by16S rRNA sequencing. Anti-listerial activity of probiotic bacteria isolated from indigenous fermented foods of Himachal Pradesh was determined by well diffusion method using Lactobacillus rhamnosus GG (ATCC 53103) as the reference strain. Five percent of tested samples were found positive for L. monocytogens with incidence of 8.0% in chicken (2/25), 10.0% in fish (2/20) and 4.0% in chevon meat (1/25). None of the tested pork samples were found contaminated with L. monocytogenes. Among 11 indigenous probiotics used in this study, highest antagonistic activity was exhibited by Lactobacillus plantarum (ADF 10) and Enterococcus faecium (ADF1) which was equivalent to the reference strain.
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BACKGROUND: Probiotics are believed to have properties that lower the risk of colon cancer. However, the mechanisms by which they exert their beneficial effects are relatively unknown. AIM: To assess the impact of probiotics in preventing induction of colon carcinogenesis in rats. METHODS: The rats were divided into six groups viz., normal control, Lactobacillus plantarum (AdF10)-treated, Lactobacillus rhamnosus GG (LGG)-treated, 1,2-dimethylhydrazine (DMH)-treated, L. plantarum (AdF10) + DMH-treated and L. rhamnosus GG (LGG) + DMH-treated. Both the probiotics were supplemented daily at a dose of 2 × 1010 cells per day. DMH at a dose of 30 mg/kg body weight was administered subcutaneously twice a week for the first 4 weeks and then once every week for a duration of 16 weeks. Glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), glutathione peroxidase (GPx), glutathione-S-transferase (GST) and catalase as protein expression of genes involved in apoptosis were assessed during DMH-induced colon carcinogenesis in rats. RESULTS: DMH treatment decreased the activity of GSH, GPx, GST, SOD and catalase. However, AdF10 and LGG supplementation to DMH-treated rats significantly increased the activity of these enzymes. Further, DMH treatment revealed alterations in the protein expressions of various genes involved in the p53-mediated apoptotic pathway such as p53, p21, Bcl-2, Bax, caspase-9 and caspase-3, which, however, were shifted towards normal control levels upon simultaneous supplementation with probiotics. CONCLUSION: The present study suggests that probiotics can provide protection against oxidative stress and apoptotic-related protein disregulation during experimentally induced colon carcinogenesis.
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1,2-Dimetilidrazina , Carcinógenos , Neoplasias do Colo/prevenção & controle , Probióticos/uso terapêutico , Animais , Apoptose , Neoplasias do Colo/etiologia , Modelos Animais de Doenças , Feminino , Lactobacillus plantarum , Lacticaseibacillus rhamnosus , Estresse Oxidativo , Ratos , Ratos Sprague-DawleyRESUMO
Post traumatic urethral injury is uncommon in children. The management of this condition is dependent on the severity of injury. Initial suprapubic cystostomy with delayed repair is the conventional treatment. Successful reconstruction of urethral injury may be followed by urethral stricture, incontinence, impotence, and retrograde ejaculation. Successful repair of post traumatic urethral injury followed by secondary incontinence in children has not been well addressed in literature. We report the management of one such child, with satisfactory outcome with implantation of a new model of single piece artificial urinary sphincter in the bulbar urethra by perineal approach.
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Fluorescent probes based on semiconducting polymer nanoparticles (NPs) such as polyaniline (PANI) usually require external fluorophore doping to provide fluorescence function. Direct use of PANI-based NPs for bioimaging applications has been limited by PANI's weak blue fluorescence and aggregation-induced quenching in physiological medium. In this report, we developed a facile solid-state synthesis method to produce fluorescent polyaniline nanoparticles (FPNs) that are not only water-soluble but also exhibit high intensity and pH-sensitive red fluorescence. The FPNs showed high photoluminescence quantum yield (PLQY) of 19.3 % at physiological pH, which makes FPNs ideal for application as fluorescent nanoprobes in bioimaging. Moreover, we performed an in-depth study of photoluminescence dependence on pH and the phenomena of exciton-polaron quenching at low pH was highlighted. We also found that the ratio of emission intensity at 600 nm and 650 nm increased from 0.04 to 1.65 as pH was raised from 2.6 to 11.8, which could find its application in ratiometric pH sensing. FPNs exhibited excellent biocompatibility with >85 % cell viability for fibroblasts NIH/3 T3 and prostate cancer 22RV1 cells even at concentrations as high as 1000 µg/mL. In addition, fluorescence microscopy demonstrated concentration-dependent red fluorescence in the cytoplasm owing to the cellular uptake of FPNs in prostate cancer cells.
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Nanopartículas , Neoplasias da Próstata , Compostos de Anilina , Corantes Fluorescentes , Humanos , Concentração de Íons de Hidrogênio , Masculino , Imagem ÓpticaRESUMO
In this study, we examined the relationship between the endothelial selectins (P-selectin and E-selectin) and whether they are critical for alpha4-integrin-dependent leukocyte recruitment in inflamed (late phase response), cremasteric postcapillary venules. Animals were systemically sensitized and 2 wk later challenged intrascrotally with chicken ovalbumin. Leukocyte rolling flux, adhesion, and emigration were assessed at baseline and 4 and 8 h postantigen challenge. There was a significant increase in leukocyte rolling flux, adhesion, and emigration in sensitized and challenged mice at both 4 and 8 h. At 8 h, the increase in leukocyte rolling flux was approximately 50% inhibitable by an anti-alpha4-integrin antibody, 98% inhibitable by fucoidin (a selectin-binding carbohydrate), and 100% inhibitable by an anti-P-selectin antibody. P-selectin-deficient animals displayed no leukocyte rolling or adhesion at 8 h after challenge. However, at 8 h there were many emigrated leukocytes in the perivascular space suggesting P-selectin-independent rolling at an earlier time point. Indeed, at 4 h postantigen challenge in P-selectin-deficient mice, there was increased leukocyte rolling, adhesion, and emigration. The rolling in the P-selectin-deficient mice at 4 h was largely alpha4-integrin dependent. However, there was an essential E-selectin-dependent component inasmuch as an anti-E-selectin antibody completely reversed the rolling, and in E-selectin and P-selectin double deficient mice rolling, adhesion and emigration were completely absent. These results illustrate that P-selectin underlies all of the antigen-induced rolling with a brief transient contribution from E-selectin in the P-selectin-deficient animals. Finally, the antigen-induced alpha4-integrin-mediated leukocyte recruitment is entirely dependent upon endothelial selectins.
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Antígenos CD/fisiologia , Hipersensibilidade Imediata/imunologia , Inflamação/imunologia , Selectina L/fisiologia , Selectina-P/fisiologia , Animais , Adesão Celular , Galinhas , Cruzamentos Genéticos , Hipersensibilidade Imediata/fisiopatologia , Imunização , Inflamação/fisiopatologia , Integrina alfa4 , Selectina L/genética , Leucócitos/imunologia , Leucócitos/fisiologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos , Camundongos Knockout , Ovalbumina/imunologia , Selectina-P/genética , Fatores de TempoRESUMO
Organic photovoltaic (OPV) cells suffer from low charge carrier mobilities of polymers, which renders it important to achieve complete optical absorption in active layers thinner than optical absorption length. Active layers conformally deposited on light-trapping, microscale textured, grating-type surfaces is one possible approach to achieve this objective. In this report, we analyze the design of such grating-type OPV cells using finite element method simulations. The energy dissipation of electromagnetic field in the active layer is studied as a function of active layer thickness, and pitch and height of the underlying textures. The superiority of textured geometry in terms of light trapping is clearly demonstrated by the simulation results. We observe 40% increase in photonic absorption in 150 nm thick active layer, for textures with 2 microm pitch and 1.5 microm height.
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The objective of this study is to describe the time of onset of contractions in twin pregnancies that result in delivery, so as to investigate whether there is a diurnal influence. A UK maternity department database was used to identify twin deliveries over a 5-year period, and the time of onset of labour was retrieved from these records. Two hundred and eleven women with spontaneous onset of labour in twin pregnancies were studied. A significant diurnal rhythm in the timing of contractions was noted, with 45% of deliveries occurring in labour that commenced between midnight and 08.00 hours. This periodicity is similar for twin pregnancies that end in preterm (24-36 weeks of gestation; χ² = 17.2; P < 0.01) or term deliveries (37-40 weeks of gestation; χ² = 13; P < 0.05). The periodicity of onset of labour in multiple pregnancies demonstrates a similar rhythm to singleton pregnancies, where labour most commonly begins between midnight and 08.00 hours.
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Ritmo Circadiano , Início do Trabalho de Parto/fisiologia , Trabalho de Parto Prematuro/fisiopatologia , Gravidez Múltipla/fisiologia , Feminino , Humanos , Gravidez , Fatores de Tempo , GêmeosRESUMO
The present study was designed to investigate the effects of a selective COX-2 inhibitor, etoricoxib in rats on the hematological and toxicity parameters in colon and kidney at two different doses of the drug, one within the therapeutic anti-inflammatory range as based on the reported ED50 value (Eto-1) while the other at ten times higher (Eto-2), relative to the toxicity studies which have not been reported so far. The results showed that the control and the drug treated animals achieved similar linear growth rate and also showed no major alterations in the histological parameters in the liver and kidney tissue. The animals treated with lower dose of etoricoxib showed an overall decrease in total leukocytes counts as well as in the number of neutrophils, lymphocytes, monocytes and eosinophills while the higher dose of the drug produced a highly significant increase in all the cell counts. However, the drug treatment at both the dose level produced significant fall in the activities of alkaline phosphatase, sucrase, lactase and maltase in the kidney but increased the activity of alkaline phosphatase in colon. The treatment of etoricoxib did not produce any change in the nitric oxide and citrulline levels in kidney while an increase was noted in the colonic tissue. It was concluded that etoricoxib is a relatively safe drug at its anti-inflammatory ED50 dose in rats when the hematological parameters and the structural and functional characteristics of kidney and colonic tissues were studied.
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Anti-Inflamatórios não Esteroides/farmacologia , Colo/efeitos dos fármacos , Colo/enzimologia , Inibidores de Ciclo-Oxigenase/farmacologia , Rim/efeitos dos fármacos , Rim/enzimologia , Leucócitos/efeitos dos fármacos , Piridinas/farmacologia , Sulfonas/farmacologia , Animais , Anti-Inflamatórios não Esteroides/administração & dosagem , Inibidores de Ciclo-Oxigenase/administração & dosagem , Etoricoxib , Masculino , Piridinas/administração & dosagem , Ratos , Ratos Wistar , Sulfonas/administração & dosagemRESUMO
A few mixed ligand transition metal carbodithioate complexes of the general formula [M(4-MPipzcdt)x(phen)y]Y (M = Mn(II), Co(II), Zn(II); 4-MPipzcdt = 4-methylpiperazine-1-carbodithioate; phen = 1,10-phenanthroline; x = 1 and y = 2 when Y = Cl; x = 2 and y = 1 when Y = nil) were synthesized and screened for their antimicrobial activity against Candida albicans, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Enterococcus faecalis by disk diffusion method. All the complexes exhibited prominent antimicrobial activity against tested pathogenic strains with the MIC values in the range <8-512 gmL(-1). The complexes [Mn(4-MPipzcdt)2(phen)] and [Co(4-MPipzcdt)(phen)2]Cl inhibited the growth of Candida albicans at a concentration as low as 8 µgmL(-1). The complexes were also evaluated for their toxicity towards human transformed rhabdomyosarcoma cells (RD cells). Moderate cell viability of the RD cells was exhibited against the metal complexes.
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The present study was designed to evaluate the effects of three non-steroidal anti-inflammatory drugs (NSAIDs) with varying cycloxygenase selectivities on the small intestinal biochemical composition, function and histology during 1, 2-dimethylhydrazine (DMH) administration. Sprague Dawley male rats were divided into five different groups viz: Group 1 (control, vehicle treated), Group 2 (DMH-treated, 30 mg/kg body weight/week in 1 mM EDTA-saline, subcutaneously), Group 3 (DMH + aspirin-60 mg/kg body weight), Group 4 (DMH + celecoxib-6 mg/kg body weight), Group 5 (DMH + etoricoxib-0.64 mg/kg body weight). After six weeks of treatment, brush border membrane was isolated from the jejunum segment of all the groups and changes in the associated enzymes such as sucrase, lactase, maltase, alkaline phosphatase, membrane lipid composition, fluorescence polarizations of diphenylhexatriene, pyrene excimer formation, histological changes and surface characteristics were studied. The results indicated a significant alteration in the enzyme activity as well as changes in the structure and function of the intestine in the presence of the pro-carcinogen, DMH, which suggests the possible chemopreventive efficacy of NSAIDs against the intestinal cancer.
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1,2-Dimetilidrazina/farmacologia , Anti-Inflamatórios não Esteroides/farmacologia , Aspirina/farmacologia , Carcinógenos/farmacologia , Inibidores de Ciclo-Oxigenase/farmacologia , Mucosa Intestinal/efeitos dos fármacos , Intestino Delgado/efeitos dos fármacos , Pirazóis/farmacologia , Piridinas/farmacologia , Sulfonamidas/farmacologia , Sulfonas/farmacologia , 1,2-Dimetilidrazina/administração & dosagem , Experimentação Animal , Animais , Anti-Inflamatórios não Esteroides/administração & dosagem , Aspirina/administração & dosagem , Peso Corporal , Carcinógenos/administração & dosagem , Celecoxib , Inibidores de Ciclo-Oxigenase/administração & dosagem , Etoricoxib , Polarização de Fluorescência , Neoplasias Intestinais/prevenção & controle , Intestino Delgado/enzimologia , Intestino Delgado/metabolismo , Intestino Delgado/fisiologia , Intestino Delgado/ultraestrutura , Masculino , Lipídeos de Membrana/metabolismo , Microscopia Eletrônica de Varredura , Pirazóis/administração & dosagem , Piridinas/administração & dosagem , Ratos , Ratos Sprague-Dawley , Sulfonamidas/administração & dosagem , Sulfonas/administração & dosagem , Fatores de TempoRESUMO
Oxidative stress has been implicated in brain ageing and in age-related neurodegenerative disorders. Since N-acetylcysteine (NAC) has recently been shown to prevent oxidative damage in ageing brain, we have examined the effects of this thiolic antioxidant on the age associated oxidative stress related parameters in rat brain regions. The lipid peroxide formation, reduced glutathione (GSH) content along with the activities of superoxide dismutase (SOD) and catalase were determined in the cerebral cortex and cerebellum brain regions of the young (4 months) and older (14 months) female rats. The lipid peroxidation was observed to be increased in the cerebral cortex regions accompanied by simultaneous decrease in the GSH content in both the regions of older rats. The SOD activity was reduced in both the regions while catalase was reduced only in cerebellum region of the older rats. Following NAC supplementation (160 mg/kg. b. wt./ day), lipid peroxidation was observed to be reduced which was accompanied by enhanced GSH levels, along with enhanced SOD and catalase in both the brain regions of older age rats. Further, in the younger age rats the NAC treatment resulted in the decrease of lipid peroxidation in both the regions that was accompanied by the increase catalase activity in cerebral cortex region along with increase in GSH content and SOD in cerebellum regions. Our result suggests that the normal brain ageing is associated with the decrease in antioxidative defense status and the supplementation of thiol antioxidants like NAC may prove helpful in managing the age related brain disorders characterized by compromised antioxidative defense systems.
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Acetilcisteína/farmacologia , Envelhecimento/efeitos dos fármacos , Envelhecimento/metabolismo , Cerebelo/efeitos dos fármacos , Cerebelo/metabolismo , Córtex Cerebral/efeitos dos fármacos , Córtex Cerebral/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Animais , Feminino , Ratos , Ratos WistarRESUMO
ABSTRACT The anticancer efficacy of two different classes of NSAIDs, the nonspecific cyclooxygenase (COX) inhibitor aspirin and the specific COX-2 inhibitor celecoxib, was examined at their therapeutic anti-inflammatory doses during 1,2-dimethylhydrazine (DMH)-induced colon carcinogenesis in a rat model. Eight to 10-week-old male rats of Sprague strain were divided into four groups. While group 1 served as control and received the vehicle of the drugs, groups 2, 3, and 4 were administered freshly prepared DMH in 1 mM EDTA saline (pH 7.0) (30 mg/kg body weight/week, subcutaneously). Groups 3 and 4 were also given a daily treatment of aspirin (60 mg/kg body weight, orally) and celecoxib (6 mg/kg body weight, orally), respectively, both prepared in carboxy-methyl cellulose. Animals were sacrificed at the end of 12 weeks and colons from different groups were subjected to macroscopic and histopathological studies, enzymatic activities of superoxide dismutase (SOD) and catalase (CAT), and determination of lipid peroxide level. The maximum number of raised mucosal lesions in proximal, middle, and distal regions of the colon was found in the DMH group alone, and the lowest number was found in the celecoxib-treated DMH group. Histological studies also showed the highest occurrence of dysplastic aberrant crypt foci (ACF) associated with enlarged lymphoid follicles in all the three portions of colon (i.e., proximal, middle, and distal). The aspirin-administered DMH group had lesser ACF in the proximal and middle portions and no ACF in the distal region. The celecoxib-administered DMH group showed no ACF in the middle region of the rat colon. DMH treatment induced lipid peroxidation and inhibited the activities of SOD and CAT. Both the aspirin- and celecoxib-treated DMH groups showed a marked lowering of the lipid peroxide level along with a significant enhancement of CAT activity when compared with the DMH-treated group. The results show that celecoxib was found to be more effective in reducing the ACF occurrence and aggregates of lymphoid tissue than the nonselective COX inhibitor aspirin, and suggests a possible chemoprevention modality in colon cancer. This may have important implications as COX-2 selective drugs at anti-inflammatory doses are better tolerated clinically than standard NSAIDs, thus making them potentially better chemopreventive agents in colon cancer.
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The microbial dynamics expressed in terms of culturable microbial populations i.e. bacteria, fungi, actinomycetes and Azotobacter were measured after 33 years of continuous application of mineral fertilizers and amendments to an acid alfisol. The bacterial, fungal and Azotobacter populations were maximum in plots treated with mineral fertilizers and FYM (100%NPK+FYM) while actinomycetes population was maximum in mineral fertilizes and lime treated plots (100%NPK+Lime). The bacterial population decreased and fungal population increased with increasing levels of NPK i.e. from 50% to 150%NPK. Bacillus species of bacteria and Gliocladium, Aspergillus and Rhizopus species of fungi were the main dominating culturable microorganisms in all the treatments. The FYM and lime amended plots sustained crop productivity and microbial populations at higher levels than rest of the mineral fertilizer treatments. The nitrogenous fertilizers alone had the most deleterious effect on crop productivity and the biological soil environment.
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In the present study, different transcripts of Trichoderma harzianum ThHP-3 were evaluated for their response against four fungal pathogens Fusarium oxysporum, Colletotrichum capsici, Colletotrichum truncatum and Gloesercospora sorghi using RT-qPCR. The time course study of T. harzianum transcripts related to signal transduction, lytic enzymes, secondary metabolites and various transporters revealed variation in expression against four fungal pathogens. In a broader term, the transcripts were upregulated at various time intervals but the optimum expression of cyp3, abc, nrp, tga1, pmk, ech42 and glh20 varied with respect to host fungi. Additionally, the expression of transcripts related to transporters/cytochromes was also observed against Fusarium oxysporum after 96h whereas transcripts related to secondary metabolites and lytic enzymes showed significant difference in expression against Colletotrichum spp. from 72 to 96h. This is first study on transcriptomic response of T. harzianum against pathogenic fungi which shows their host specific response.
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Fusarium/fisiologia , Especificidade de Hospedeiro , Controle Biológico de Vetores , Plantas/microbiologia , Trichoderma/fisiologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Trichoderma/genéticaRESUMO
In the present study the effects of two cycloxygenase-2 (COX-2) selective inhibitors, celecoxib and nimesulide as compared to a non-selective COX inhibitor, aspirin was studied in the rat intestine. Female Wistar rats weighing between 150-175 g were divided into four groups having 8 animals each as follows: Group 1(Control), Group 2- Aspirin (40 mg/kg), Group 3- Nimesulide (10 mg/kg) and Group 4- Celecoxib (10 mg/kg). After 35 days of treatment the animals were sacrificed, intestine removed and the effects on the antioxidant defense system, membrane composition and functions along with the membrane specific enzymes were studied in different regions of the intestine. The study showed a significant increase in the lipid peroxide levels as TBA-reactive substance as well as the conjugated dienes, except for celecoxib treated group which showed a decrease. Significant decrease was also observed in the level of reduced glutathione (GSH), superoxide dismutase (SOD), glutathione-s-transferase and catalase activities for aspirin and nimesulide group while Celecoxib caused an increase in glutathione reductase (GR). Aspirin and nimesulide exhibited an increase in the brush border membrane (BBM) bound enzyme activities like sucrase, lactase, maltase and alkaline phosphatase in the small intestine while celecoxib showed decrease in lactase, maltase and alkaline phosphatase. The phospholipid content increased only for aspirin treated group while cholesterol decreased in all the treatment groups. Also celecoxib treatment brought about an increase in glycolipid content. The membrane fluidity was studied by the rotational diffusion of 1, 6, diphenyl, 1, 3, 5 hexatriene (DPH) incorporated in the membrane and the fluorescence polarization (p), fluorescence anisotropy(r), anisotropy parameter [r0/r-1](-1) and order parameter [S2 = (4/3r - 0.1)/r0] were recorded. No significant change in the fluorescence parameters were observed in the BBM and the liposomes made from the BBM lipids for the treatment groups. These results indicate that celecoxib may be accepted as a safer drug in terms of overall gastro-intestinal toxicity as compared to the aspirin and nimesulide.
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Anti-Inflamatórios não Esteroides/farmacologia , Mucosa Intestinal/efeitos dos fármacos , Animais , Antioxidantes/fisiologia , Aspirina/farmacologia , Celecoxib , Inibidores de Ciclo-Oxigenase/farmacologia , Feminino , Mucosa Intestinal/imunologia , Mucosa Intestinal/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Pirazóis/farmacologia , Ratos , Ratos Wistar , Segurança , Sulfonamidas/farmacologiaRESUMO
Monensin, a carboxylic ionophore, is well known for Na(+)/H(+) exchanger activity across biological membranes. It is also used in the poultry industry for its useful effects as a food additive. The present study has been designed to investigate the effects of monensin on some oxidative stress-related parameters in rat testis. Monensin was administered intratesticularly (5 mug/testis) to both testes by a single dose to Wistar rats for different time periods. After the completion of the respective treatments, various parameters reflecting the antioxidant defense system of the tissue were monitored and marked changes were found in the activities of various enzymes as well as in the levels of reduced glutathione and lipid peroxidation. After 1, 2, 3, and 4 days of monensin treatment, the activity of superoxide dismutase was found to be unaltered. However, after 2 days of monensin treatment, glutathione-S-transferase and catalase showed inhibition in their activities along with the depletion of glutathione (reduced) accompanied by a marked increase in lipid peroxidation. The increase in lipid peroxidation was noticeable even after 1 day of monensin administration. The inhibition in glutathione-S-transferase and glutathione peroxidase activities was also observed along with an increase in lipid peroxidation at the end of the 3-day posttreatment period, while, the 4-day posttreatment schedule caused an increase in the activity of glutathione reductase and glutathione peroxidase that was also accompanied by an inhibition of catalase. The findings of the present study are indicative of the potential of monensin in testicular tissue in contraceptive intervention.
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Twenty-three Saccharomyces cerevisiae strains isolated from different fermented foods of Western Himalayas have been studied for strain level and functional diversity in our department. Among these 23 strains, 10 S. cerevisiae strains on the basis of variation in their brewing traits were selected to study their organoleptic effect at gene level by targeting ATF1 gene, which is responsible for ester synthesis during fermentation. Significant variation was observed in ATF1 gene sequences, suggesting differences in aroma and flavor of their brewing products. Apple is a predominant fruit in Himachal Pradesh and apple cider is one of the most popular drinks all around the world hence, it was chosen for sensory evaluation of six selected yeast strains. Organoleptic studies and sensory analysis suggested Sc21 and Sc01 as best indigenous strains for soft and hard cider, respectively, indicating their potential in enriching the local products with enhanced quality.
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In the present study, endochitinase of T. harzianum isolate-ThHP3 induced against mycelium of F. oxysporum was cloned, sequenced and characterized. The complete nucleotide sequence contained an ORF of 1293bp corresponding to 430 amino acids with 46kDa molecular weight and theoretical pI 5.59. The precursor protein contained 22 amino acids long signal peptide at N terminus. The domain architecture of endochitinase showed low complexity regions, presence of 1W9P domain specific to cyclopentapeptide and lack of carbohydrate binding modules. The ligand binding site of ech46 endochitinase was constituted by 10 amino acids. The cDNA encoding ech46 endochitinase was ligated into pET28a vector and transformed to E. coli BL21. The predicted molecular weight of recombinant endochitinase without signal peptide was 49.4kDa with a theoretical pI 6.67. SDS-PAGE analysis of purified 6xHis tagged protein showed a single band of 49kDa. The refolded enzyme was active under acidic conditions with a temperature and pH optima of 50°C and 4. Km and Vmax for recombinant endochitinase using 4-pNP-(GlcNAc)3 were 315.2±0.36µM and 0.140±0.08µMmin-1, respectively and the calculated kcat was 6.44min-1. The RT-qPCR revealed induction of ech46 by phytopathogenic fungi.
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Quitinases/genética , Proteínas Fúngicas/genética , Trichoderma/enzimologia , Sequência de Aminoácidos , Sequência de Bases , Quitinases/química , Quitinases/isolamento & purificação , Clonagem Molecular , Eletroforese em Gel de Poliacrilamida , Éxons/genética , Proteínas Fúngicas/química , Proteínas Fúngicas/isolamento & purificação , Concentração de Íons de Hidrogênio , Íntrons/genética , Íons , Metais/farmacologia , Modelos Moleculares , Peso Molecular , Filogenia , Domínios Proteicos , Reação em Cadeia da Polimerase em Tempo Real , Proteínas Recombinantes/metabolismo , Homologia Estrutural de Proteína , TemperaturaRESUMO
BACKGROUND: Direct experimental evidence suggests that total enteral nutrition (TEN) reduces septic morbidity compared with bowel rest and total parenteral nutrition (TPN) and that mucosal support and maintenance of gut barrier function is a key mechanism. This effect is supported indirectly by clinical studies, but this question has not previously been investigated directly in the postoperative patient. This study examined the hypothesis that early enteral feeding after major upper gastrointestinal surgery may modulate gut barrier function and decrease the risk of major infective complications compared with bowel rest and parenteral nutrition. METHODS: A randomized clinical trial of 67 patients (TPN = 34; TEN = 33) fed postoperatively for 7 days was performed. Thirty-day major morbidity and mortality were monitored. Intestinal permeability was measured using the lactulose/mannitol test preoperatively and on postoperative days 1 and 7. Systemic anti-endotoxin core immunoglobulin G and M antibodies and serum albumin and C-reactive protein were quantified at these time points. RESULTS: No clinical benefit was observed in patients fed enterally compared with the parenterally fed group. Intestinal permeability was increased on the 1st postoperative day in association with evidence of endotoxin exposure. By day 7, enteral feeding compared with parenteral feeding had failed to significantly influence any of the gut barrier or systemic parameters. CONCLUSIONS: This randomized controlled trial of TEN vs TPN after major upper gastrointestinal surgery failed to show a clinical benefit for the enteral route. Moreover, enteral nutrition did not modulate gut barrier function postoperatively.
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Fenômenos Fisiológicos do Sistema Digestório , Procedimentos Cirúrgicos do Sistema Digestório , Nutrição Enteral , Nutrição Parenteral Total , Anticorpos/sangue , Distinções e Prêmios , Proteína C-Reativa/metabolismo , Endotoxinas/imunologia , Neoplasias Esofágicas/cirurgia , Humanos , Neoplasias Pancreáticas/cirurgia , Permeabilidade , Pesquisadores , Albumina Sérica/metabolismo , Neoplasias Gástricas/cirurgia , Resultado do TratamentoRESUMO
Lead (Pb) is known to disrupt the pro-oxidant/antioxidant balance of tissues, which leads to biochemical and physiological dysfunction. Oxidative stress is considered a possible molecular mechanism involved in Pb neurotoxicity. Considering the vulnerability of the brain to oxidative stress under Pb neurotoxicity, this study investigated the effects of exposure of the thiol antioxidant N-acetylcysteine (NAC) on lead-induced oxidative damage and lipid peroxidation in brain regions of the rat. Wister strain rats were exposed to lead in the form of lead acetate (20 mg/kg body wt/d) for a period of 2 wk and the effects of NAC on lead-induced neurotoxicity in rat brain regions were assessed by postadministration of NAC (160 mg/kg body wt/d) for a period of 3 wk. The lipid peroxidation byproduct, malondialdehyde (MDA) increased following lead exposure in both of the regions, and the antioxidant capacities of the cell in terms of the activity of antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT) was diminished. Following NAC treatment, lead-induced lipid peroxidation decreased and antioxidant enzyme activities improved, with CAT showing enhancement in the cerebral region only and SOD showing enhancements in the cerebellar region. Our result suggests that thiol-antioxidant supplementation following Pb exposure might enhance the reductive status of brain regions by arresting the lipid peroxidative damage in brain regions.