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BACKGROUND: Antimicrobial resistance (AMR) is a serious worldwide public health concern that needs immediate action. Probiotics could be a promising alternative for fighting antibiotic resistance, displaying beneficial effects to the host by combating diseases, improving growth, and stimulating the host immune responses against infection. This study was conducted to evaluate the probiotic, antibacterial, and antibiofilm potential of Streptomyces levis strain HFM-2 isolated from the healthy human gut. RESULTS: In vitro antibacterial activity in the cell-free supernatant of S. levis strain HFM-2 was evaluated against different pathogens viz. K. pneumoniae sub sp. pneumoniae, S. aureus, B. subtilis, VRE, S. typhi, S. epidermidis, MRSA, V. cholerae, M. smegmatis, E. coli, P. aeruginosa and E. aerogenes. Further, the ethyl acetate extract from S. levis strain HFM-2 showed strong biofilm inhibition against S. typhi, K. pneumoniae sub sp. pneumoniae, P. aeruginosa and E. coli. Fluorescence microscopy was used to detect biofilm inhibition properties. MIC and MBC values of EtOAc extract were determined at 500 and 1000 µg/mL, respectively. Further, strain HFM-2 showed high tolerance in gastric juice, pancreatin, bile, and at low pH. It exhibited efficient adhesion properties, displaying auto-aggregation (97.0%), hydrophobicity (95.71%, 88.96%, and 81.15% for ethyl acetate, chloroform and xylene, respectively), and showed 89.75%, 86.53%, 83.06% and 76.13% co-aggregation with S. typhi, MRSA, S. pyogenes and E. coli, respectively after 60 min of incubation. The S. levis strain HFM-2 was susceptible to different antibiotics such as tetracycline, streptomycin, kanamycin, ciprofloxacin, erythromycin, linezolid, meropenem, amikacin, gentamycin, clindamycin, moxifloxacin and vancomycin, but resistant to ampicillin and penicillin G. CONCLUSION: The study shows that S. levis strain HFM-2 has significant probiotic properties such as good viability in bile, gastric juice, pancreatin environment, and at low pH; proficient adhesion properties, and antibiotic susceptibility. Further, the EtOAc extract of Streptomyces levis strain HFM-2 has a potent antibiofilm and antibacterial activity against antibacterial-resistant clinical pathogens.
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Antibacterianos , Biofilmes , Testes de Sensibilidade Microbiana , Probióticos , Streptomyces , Biofilmes/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Humanos , Probióticos/farmacologia , Streptomyces/fisiologia , Streptomyces/classificação , Streptomyces/isolamento & purificação , Streptomyces/química , Antibacterianos/farmacologia , Bactérias/efeitos dos fármacos , Bactérias/classificação , Trato Gastrointestinal/microbiologiaRESUMO
BACKGROUND: Gamma-aminobutyric acid (GABA) is a non-protein amino acid with neuroinhibitory, antidiabetic, and antihypertensive properties and is used as a drug for treating anxiety and depression. Some strains of lactobacilli are known to produce GABA and strengthen the gut barrier function which play an important role in ameliorating the effects caused by the pathogen on the gut barrier. The probiotic bacteria are also known to modulate the human fecal microbiota, however, the role of GABA-producing strains on the gut epithelium permeability and gut microbiota is not known. RESULTS: In this study, we report the production of high levels of GABA by potential probiotic bacterium Limosilactobacillus fermentum L18 for the first time. The kinetics of the production of GABA by L18 showed that the maximum production of GABA in the culture supernatant (CS) occurred at 24 h, whereas in fermented milk it took 48 h of fermentation. The effect of L18 on the restoration of lipopolysaccharide (LPS)-disrupted intestinal cell membrane permeability in Caco-2 monolayers showed that it significantly restored the transepithelial electrical resistance (TEER) values, by significantly increasing the levels of junction proteins, occludin and E-cadherin in L18 and LPS-treated Caco-2 cells as compared to only LPS-treated cells. The effect of GABA-secreting L18 on the metataxonome of human stool samples from healthy individuals was investigated by a batch fermentor that mimics the conditions of the human colon. Although, no differences were observed in the α and ß diversities of the L18-treated and untreated samples at 24 h, the relative abundances of bacterial families Lactobacillaceae and Bifidobacteriaceae increased in the L18-treated group, but both decreased in the untreated groups. On the other hand, the relative abundance of Enterobacteriaceae decreased in the L18 samples but it increased in the untreated samples. CONCLUSION: These results indicate that Li. fermentum L18 is a promising GABA-secreting strain that strengthens the gut epithelial barrier by increasing junction protein concentrations and positively modulating the gut microbiota. It has the potential to be used as a psychobiotic or for the production of functional foods for the management of anxiety-related illnesses.
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Microbioma Gastrointestinal , Limosilactobacillus fermentum , Probióticos , Humanos , Células CACO-2 , Lipopolissacarídeos , Função da Barreira Intestinal , Bactérias/metabolismo , Probióticos/uso terapêuticoRESUMO
BACKGROUND: Oxidative stress is responsible for the onset of several chronic and degenerative diseases. Exogenous supply of antioxidants is reported to neutralize the effects of oxidative stress. Several synthetic antioxidants suffer from various side effects which necessitates the exploration of antioxidant compounds from natural sources. Endophytic fungi residing in the plants are gaining the attention of researchers as a source of novel antioxidants. Majority of the research conducted so far on endophytic fungi has been restricted to the members of phylum ascomycota. Basidiomycota, inspite of their immense bioactive potential remain relatively unexploited. This study aimed to assess the ameliorative effects of an endophytic Schizophyllum commune (basidiomycetous fungus) against oxidative stress associated altered antioxidant levels, genotoxicity and cellular damage to different organs in bisphenol A exposed fresh water fish Channa punctatus. RESULTS: Good antioxidant and genoprotective potential was exhibited by S. commune extract in in vitro studies conducted using different antioxidant, DNA damage protection, and cytokinesis blocked micronuclei assays. In vivo studies were performed in fresh water fish Channa punctatus exposed to bisphenol A. A significant decrease in the considered parameters for DNA damage (% micronuclei and comet assay) were recorded in fish treated with S. commune extract on comparison with untreated bisphenol A exposed group. The S. commune extract treated fish also exhibited an increase in the level of antioxidant enzymes viz. catalase, superoxide dismutase and glutathione reductase as well as histoprotective effect on various organs. GC-MS analysis revealed the presence of 3-n-propyl-2,4-pentanedione, n-heptadecanol-1, trans-geranylgeraniol, 3-ethyl-2-pentadecanone, 1-heneicosanol and squalene as some of the compounds in S. commune extract. CONCLUSION: The study highlights the significance of an endophytic basidiomycetous fungus S. commune as a source of antioxidant compounds with possible therapeutic potential.
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Antioxidantes , Schizophyllum , Antioxidantes/farmacologia , Água Doce , Extratos VegetaisRESUMO
BACKGROUND: Increase in the number of infections caused by Gram-negative bacteria in neutropenic cancer patients has prompted the search for novel therapeutic agents having dual anticancer and antimicrobial properties. Bacteriocins are cationic proteins of prokaryotic origin that have emerged as one of the most promising alternative antimicrobial agents with applications as food preservatives and therapeutic agents. Apart from their antimicrobial activities, bacteriocins are also being explored for their anticancer potential. RESULTS: In this study, a broad-spectrum, cell membrane-permeabilizing enterocin with a molecular weight of 65 kDa was purified and characterized from the culture supernatant of vaginal Enterococcus faecium 12a. Enterocin 12a inhibited multidrug-resistant strains of various Gram-negative pathogens such as Salmonella enterica, Shigella flexneri, Vibrio cholerae, Escherichia coli and Gram-positive, Listeria monocytogenes, but had no activities against different strains of gut lactobacilli. The mass spectrometric analysis showed that the enterocin 12a shared partial homology with 4Fe-4S domain-containing redox protein of E. faecalis R712. Further, enterocin 12a selectively inhibited the proliferation of various human cancer cell lines in a dose-dependent manner but not that of normal human peripheral blood mononuclear cells. Enterocin 12a-treated cancer cells showed apoptosis-like morphological changes. CONCLUSION: Enterocin 12a is a novel bacteriocin that has anticancer properties against human cell lines and negligible activity towards non-malignant cells. Therefore, it should be further evaluated for its anticancer potential in animal models.
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Antibacterianos/farmacologia , Anticarcinógenos/farmacologia , Proliferação de Células/efeitos dos fármacos , Enterococcus faecium/química , Apoptose/efeitos dos fármacos , Hidrocarbonetos Aromáticos com Pontes/química , Hidrocarbonetos Aromáticos com Pontes/isolamento & purificação , Hidrocarbonetos Aromáticos com Pontes/farmacologia , Linhagem Celular Tumoral , Permeabilidade da Membrana Celular/efeitos dos fármacos , Enterococcus faecium/metabolismo , Feminino , Humanos , Testes de Sensibilidade Microbiana , Vagina/microbiologiaRESUMO
BACKGROUND: An increasing rate of antibiotic resistance among Gram-negative bacterial pathogens has created an urgent need to discover novel therapeutic agents to combat infectious diseases. Use of bacteriocins as therapeutic agents has immense potential due to their high potency and mode of action different from that of conventional antibiotics. RESULTS: In this study, a novel bacteriocin E20c of molecular weight 6.5 kDa was purified and characterized from the probiotic strain of Enterococcus hirae. E20c had bactericidal activities against several multidrug resistant (MDR) Gram-negative bacterial pathogens. Flow cytometry and scanning electron microscopy studies showed that it killed the Salmonella enterica cells by forming ion-permeable channels in the cell membrane leading to enhanced cell membrane permeability. Further, checkerboard titrations showed that E20c had synergistic interaction with antibiotics such as ampicillin, penicillin, ceftriaxone, and ciprofloxacin against a ciprofloxacin- and penicillin-resistant strain of S. enterica. CONCLUSION: Thus, this study shows the broad spectrum antimicrobial activity of novel enterocin E20c against various MDR pathogens. Further, it highlights the importance of bacteriocins in lowering the minimum inhibitory concentrations of conventional antibiotics when used in combination.
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Antibacterianos/farmacologia , Bacteriocinas/farmacologia , Ciprofloxacina/farmacologia , Salmonella enterica/efeitos dos fármacos , beta-Lactamas/farmacologia , Bacteriocinas/isolamento & purificação , Hidrocarbonetos Aromáticos com Pontes/isolamento & purificação , Hidrocarbonetos Aromáticos com Pontes/farmacologia , Membrana Celular/efeitos dos fármacos , Permeabilidade da Membrana Celular/efeitos dos fármacos , Farmacorresistência Bacteriana Múltipla , Sinergismo Farmacológico , Streptococcus faecium ATCC 9790/química , Streptococcus faecium ATCC 9790/metabolismo , Bactérias Gram-Negativas/efeitos dos fármacos , Testes de Sensibilidade MicrobianaRESUMO
Diabetes is considered as a major health concern worldwide and patients with diabetes are at high risk for infectious diseases. Therefore, α-glucosidase inhibitors possessing antibacterial activity along with the ability to inhibit biofilms would be better therapeutic agents for diabetic patients. In the present study, two fractions (AF1 and AF2) possessing α-glucosidase inhibitory activity were purified from an endophytic fungus Alternaria destruens (AKL-3) isolated from Calotropis gigantea. These were evaluated for their antimicrobial and antibiofilm potential against human pathogens. AF1 exhibited broad spectrum antimicrobial activity against all the tested pathogens. It also significantly inhibited biofilm formation and dispersed the preformed biofilm at sub-optimal concentrations. AF2 possessed lesser activity as compared to AF1. The active compounds were purified using semi preparative HPLC. Some of the active compounds were identified to be phenolic in nature. The active fractions were also determined to be non-mutagenic and non-cytotoxic in safety analysis. The study highlights the role of endophytic fungi as sources of α-glucosidase inhibitors with antimicrobial potential which can have application in management of diabetes.
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Alternaria/química , Anti-Infecciosos/isolamento & purificação , Inibidores de Glicosídeo Hidrolases/farmacologia , Alternaria/isolamento & purificação , Alternaria/metabolismo , Antibacterianos/farmacologia , Anti-Infecciosos/farmacologia , Biofilmes/efeitos dos fármacos , Calotropis , Endófitos/isolamento & purificação , Endófitos/metabolismo , Inibidores de Glicosídeo Hidrolases/metabolismo , Testes de Sensibilidade Microbiana/métodos , alfa-Glucosidases/metabolismoRESUMO
A spectrophotometric method based on diazotization of aniline with triclosan has been developed for the determination of triclosan in water samples. The diazotization process involves two steps: (1) reaction of aniline with sodium nitrite in an acidic medium to form diazonium ion and (2) reaction of diazonium ion with triclosan to form a yellowish-orange azo compound in an alkaline medium. The resulting yellowish-orange product has a maximum absorption at 352 nm which allows the determination of triclosan in aqueous solution in the linear concentration range of 0.1-3.0 µM with R2 = 0.998. The concentration of hydrochloric acid, sodium nitrite, and aniline was optimized for diazotization reaction to achieve good spectrophotometric determination of triclosan. The optimization of experimental conditions for spectrophotometric determination of triclosan in terms of concentration of sodium nitrite, hydrogen chloride and aniline was also carried out by using Box-Behnken design of response surface methodology and results obtained were in agreement with the experimentally optimized values. The proposed method was then successfully applied for analyses of triclosan content in water samples.
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Espectrofotometria/métodos , Triclosan/química , Poluentes Químicos da Água/química , Compostos de Anilina/química , Compostos Azo , Ácido Clorídrico/química , Concentração de Íons de Hidrogênio , Nitrito de Sódio/química , Água/análiseRESUMO
The emergence of antibiotic resistance in pathogens is one of the major health concerns facing mankind as different bacterial strains have developed resistance to antibiotics over the period of time due to overuse and misuse of antibiotics. Besides this, ability to form biofilms is another major factor contributing to antibiotic resistance, which has necessitated the need for exploration for novel and effective compounds with ability to inhibit biofilm formation. Endophytic fungi are reported to exhibit antibacterial and anti-biofilm potential and could serve as a potent source of novel antibacterial compounds. Majority of the bioactivities have been reported from fungi belonging to phylum Ascomycota. Endophytic basidiomycetes, inspite of their profound ability to serve as a source of bioactive compounds have not been exploited extensively. In present study, an attempt was made to assess the antibacterial, anti-biofilm and biofilm dispersion potential of an endophytic basidiomycetous fungus Schizophyllum commune procured from the culture collection of our lab. Ethyl acetate extract of S. commune showed good antibacterial activity against Staphylococcus aureus, Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa, Salmonella enterica and Vibrio cholerae. Minimum inhibitory concentration and minimum bactericidal concentration of the extract were in the range of 1.25-10 mg/ml against the tested bacterial pathogens. The mode of action was determined to be bactericidal which was further confirmed by time kill studies. Good anti-biofilm activity of S. commune extract was recorded against K. pneumoniae and S. enterica, which was further validated by fluorescence microscopy. The present study highlights the importance of endophytic basidiomycetes as source of therapeutic compounds.
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The global prevalence rate of diabetes in 2021 was 6.1% making diabetes one of the top 10 causes of death. Prolonged use of antidiabetic medications is associated with various side effects; therefore, alternative treatment strategies for diabetes need exploration. The antidiabetic properties of Lactiplantibacillus plantarum 2034 was explored both in in vitro and in vivo studies. Secretory metabolites of probiotic L. plantarum 2034 exhibited alpha-glucosidase, alpha-amylase, and lipase inhibitory activities, in vitro. Further, the antidiabetic efficacy of 2034 was evaluated in streptozotocin-nicotinamide-induced diabetic rats. In the therapeutic model, oral administration of L. plantarum resulted in normalization of body weight, fasting blood glucose, total cholesterol (TC), and liver enzymes, and significant (p < 0.05) reduction in insulin and triglyceride (TG) levels. Histological evaluation of pancreas, liver, and kidney showed restoration of normal architecture in probiotic-treated group. Similarly, in a preventive + therapeutic model, 14 days of pre-administration of 2034 in pre, pre + post, and cell-free supernatant resulted in significant reduction in glucose, TG, TC, and liver biochemistry of diabetic rats as compared to untreated diabetic rats. An oral glucose tolerance test showed that the glucose levels normalized within 90 min in all the treated groups. Further, the oxidative stress parameters were also studied that showed that in all the treated groups, the concentration of antioxidant enzymes significantly (p < 0.05) increased as compared to diabetic untreated rats. Thus, administration of L. plantarum 2034 and its metabolites successfully ameliorated hyperglycaemia and hypercholesterolemia in both the models probably due to inhibition of gut enzymes and by increasing the concentration of liver antioxidant enzymes.
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Reactive oxygen species (ROS) cause damage to the DNA producing mutations and formation of tumours such as carcinoma of breast. Tumour cells are known to produce ROS at a greater pace than the non-transformed cells. The increased production of reactive oxygen species causes oxidative stress leading to cell proliferation and hence increased inflammatory conditions. The present study was aimed to investigate the role of oxidative stress in the pathogenesis of breast cancer. Females suffering from breast cancer had significantly decreased Superoxide dismutase (SOD) and reduced glutathione (GSH) levels in comparison to normal females. The compromised antioxidant defence system produces the oxidative stress which in turn creates the inflammatory response shown by concomitant increased adenosine deaminase (ADA) activity in female patients. ADA diminishes the protective molecule adenosine. There were significant variations (p < 0.01) in ADA activity with different clinical stages (stage 1-4) of breast cancer suggesting thereby that estimation of ADA activity can be used as a diagnostic tool to detect the stage of cancer along with cytological studies. Mastectomy was performed and post-operatively serum SOD and ADA activity and plasma GSH levels were estimated. There was a statistically significant increase in activity of SOD and levels of GSH while serum ADA activity decreased significantly, suggesting thereby that oxidative stress is responsible for increased cell proliferation and hence the inflammatory conditions in CA breast that got ameliorated post-operatively.
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Type 2 diabetes mellitus (T2DM) has a heavy disease burden and is one of the leading causes of death worldwide. It is considered to be evolving from a complex and multifactorial metabolic disorder to an inflammatory condition. The strong link between hyperglycemia and oxidative stress has long been established. Oxidative stress leads to the generation of inflammatory mediators and reactive oxygen species, which results in an inflammatory state, which plays a key role in the pathogenesis of diabetic complications. Vitamin D is also known to affect insulin sensitivity. We aimed to correlate the levels of glycated hemoglobin (HbA1c) and vitamin D with oxidative stress. This cross-sectional study included 100 patients with T2DM. Oxidative stress was estimated by lipid peroxidation assay for activity of malondialdehyde (MDA), as well as superoxide dismutase (SOD) levels. HbA1c was estimated using ion-exchange chromatography. Vitamin D was estimated using enzyme-linked immunosorbent assay. The data were analyzed using a t test. The results showed that as the duration of T2DM increased, the levels of SOD and MDA also increased. No significant correlation was found between SOD and MDA with HbA1c (P = 0.995 and 0.195) and vitamin D (P = 0.874 and 0.051), respectively.
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Purpose: To determine the correlation between serum inflammatory and metabolic biomarkers of patients with diabetic retinopathy (DR) and diabetic macular edema (DME). Methods: Serum samples were obtained from 100 diabetic patients. Patients were divided into three groups: group 1 (patients with no DR, n = 27), group 2 (DR with DME, n = 34), and group 3 (DR without DME, n = 39). Serum concentrations of C-reactive protein (CRP) and interleukin-6 (IL-6) were measured by quantitative turbidimetric immunoassay and sandwich chemiluminescence immunoassay, respectively. Metabolic parameters such as glycated hemoglobin (HbA1c), total cholesterol, low-density lipoprotein (LDL), high-density lipoprotein (HDL), triglyceride (TG), serum creatinine, and blood urea were determined by automated analyzer om-360 after standardization. Results: The levels of IL-6 and CRP differed significantly in patients with DR and without DR (P < 0.001 and P = 0.045, respectively). We also found a positive correlation between IL-6 and CRP with the severity of DR. When DR patients with DME were compared to patients without DME, only IL-6 was observed to be significantly elevated (P < 0.001). None of the metabolic markers correlated significantly with DR and DME. Conclusion: Significantly raised levels of serum inflammatory biomarkers can be used to elucidate the significant role of inflammation in the pathogenesis of DR. Therefore, circulating biomarkers can serve as diagnostic and therapeutic predictors for monitoring the onset and progression of DR and DME.
Assuntos
Diabetes Mellitus Tipo 2 , Retinopatia Diabética , Edema Macular , Humanos , Edema Macular/diagnóstico , Edema Macular/etiologia , Retinopatia Diabética/etiologia , Interleucina-6 , Hemoglobinas Glicadas , Proteína C-Reativa , Biomarcadores , Diabetes Mellitus Tipo 2/complicaçõesRESUMO
The growing awareness about the adverse health effects of artificial synthetic preservatives has led to a rapid increase in the demand for safe food preservation techniques and bio preservatives. Thus, in this study, the biopreservatives efficacy of enterocin-producing Enterococcus faecium Smr18 and its enterocin, ESmr18 was evaluated against Salmonella enterica contamination in chicken samples. E. faecium Smr18 is susceptible to the antibiotics penicillin-G, ampicillin, vancomycin, and erythromycin, thereby indicating that it is a nonpathogenic strain. Further, the enterocin ESmr18 was purified and characterised as a 3.8 kDa peptide. It possessed broad spectrum antibacterial activity against both Gram-positive and Gram-negative pathogens including S. enterica serotypes Typhi and Typhimurium. Purified ESmr18 disrupted the cell membrane permeability of the target cell thereby causing rapid efflux of potassium ions from L. monocytogenes and S. enterica. Chicken samples inoculated with S. enterica and packaged in alginate films containing immobilised viable E. faecium resulted in 3 log10 colony forming units (CFU) reduction in the counts of S. enterica after 34 days of storage at 7-8 °C. The crude preparation of ESmr18 also significantly (p < 0.05) reduced the CFU counts of salmonella-inoculated chicken meat model. Purified ESmr18 at the concentration upto 4.98 µg/ml had no cytolytic effect against human red blood cells. Crude preparation of ESmr18 when orally administered in fish did not cause any significant (p < 0.05) change in the biochemical parameters of sera samples. Nonsignificant changes in the parameters of comet and micronucleus assays were observed between the treated and untreated groups of fishes that further indicated the safety profile of the enterocin ESmr18.
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Salmonella enterica serotype Typhi causes chronic enteric fever known as typhoid. Prolonged treatment regimen used for the treatment of typhoid and indiscriminate use of antibiotics has led to the emergence of resistant strains of S. enterica that has further increased the severity of the disease. Therefore, alternative therapeutic agents are urgently required. In this study, probiotic and enterocin-producing bacteria Enterococcus faecium Smr18 was compared for both its prophylactic and therapeutic efficacy in S. enterica infection mouse model. E. faecium Smr18 possessed high tolerance to bile salts and simulated gastric juice, as treatment for 3 and 2 h resulted in 0.5 and 0.23 log10 reduction in the colony forming units, respectively. It exhibited 70% auto aggregation after 24 h of incubation and formed strong biofilms at both pH 5 and 7. Oral administration of E. faecium in BALB/c mice infected with S. enterica significantly (p < 0.05) reduced the mortality of the infected mice and prevented the weight loss in mice. Administration of E. faecium prior to infection inhibited the translocation of S. enterica to liver and spleen, whereas, its administration post-infection completely cleared the pathogen from the organs within 8 days. Further, in both pre- and post-E. faecium-treated infected groups, sera levels of liver enzymes were restored back to normal; whereas the levels of creatinine, urea and antioxidant enzymes were significantly (p < 0.05) reduced compared to the untreated-infected group. E. faecium Smr18 administration significantly increased the sera levels of nitrate by 1.63-fold and 3.22-fold in pre- and post-administration group, respectively. Sera levels of interferon-γ was highest (tenfold) in the untreated-infected group, whereas the levels of interleukin-10 was highest in the post-infection E. faecium-treated group thereby indicating the resolution of infection in the probiotic-treated group, plausibly due to the increased production of reactive nitrogen intermediates.
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Uric acid an endogenous aqueous antioxidant in normal humans is present in much higher concentrations than vitamin C and has been known to cover 2/3(rd) of the free radical scavenging capacity in plasma. In the present study average uric acid levels of patients of rheumatoid arthritis were found to be close to the normal individuals. A unique feature was observed after classifying the patients on the basis of the duration of suffering, the patients having longer duration of disease had least uric acid levels as compared to those suffering from relatively lesser period, similar trend was observed in the ascorbic acid estimations. The decline in uric acid values with progression of the disease was much more than what was observed in case of ascorbic acid suggesting the significant role of uric acid in scavenging of free radicals. Too much lowering of Uric Acid should be checked and vitamin C should be supplemented in diet for maintaining the balance between pro oxidant and antioxidant forces to check pro oxidant insult in rheumatoid arthritis.
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Diarrheal disease caused by Vibrio cholerae is endemic in developing countries including India and is associated with high rate of mortality especially in children. V. cholerae is known to form biofilms on the gut epithelium, and the biofilms once formed are resistant to the action of antibiotics. Therefore agents that prevent the biofilm formation and disperse the preformed biofilms are associated with therapeutic benefits. The use of antibiotics for the treatment of cholera is associated with side effects such as gut dysbiosis due to depletion of gut microflora, and the increasing problem of antibiotic resistance. Thus search for safe alternative therapeutic agents is warranted. Herein, we screened the lactobacilli spp. isolated from the fecal samples of healthy children for their abilities to prevent biofilm formation and to disperse the preformed biofilms of V. cholerae and V. parahaemolyticus by using an in vitro assay. The results showed that the culture supernatant (CS) of all the seven isolates of Lactobacillus spp. used in the study inhibited the biofilm formation of V. cholerae by more than 90%. Neutralization of pH of CS completely abrogated their antimicrobial activities against V. cholera, but had negligible effects on their biofilm inhibitory potential. Further, CS of all the lactobacilli isolates caused the dispersion of preformed V. cholerae biofilms in the range 62-85%; however, pH neutralization of CS reduced the biofilm dispersal potential of the 4 out of 7 isolates by 19-57%. Furthermore, the studies showed that CS of none of the lactobacilii isolates had antimicrobial activity against V. parahaemolyticus, but 5 out of 7 isolates inhibited the formation of its biofilm in the range 62-82%. However, none of the CS dispersed the preformed biofilms of V. parahaemolyticus. The ability of CS to inhibit the adherence of Vibrio spp. to the epithelial cell line was also determined. Thus, we conclude that the biofilm dispersive action of CS of lactobacilli is strain-specific and pH-dependent. As Vibrio is known to form biofilms in the intestinal niche having physiological pH in the range 6-7, the probiotic strains that have dispersive action at high pH may have better therapeutic potential.
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Biofilmes/crescimento & desenvolvimento , Cólera/prevenção & controle , Cólera/terapia , Lactobacillus/metabolismo , Probióticos/uso terapêutico , Vibrio cholerae/crescimento & desenvolvimento , Vibrio parahaemolyticus/crescimento & desenvolvimento , Adolescente , Antibacterianos/farmacologia , Aderência Bacteriana/efeitos dos fármacos , Biofilmes/efeitos dos fármacos , Linhagem Celular , Criança , Pré-Escolar , Cólera/microbiologia , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão , Humanos , Lactobacillus/classificação , Lactobacillus/isolamento & purificaçãoRESUMO
Cancer is the second leading cause of death worldwide and its incidence is expected to grow by almost 70% in the coming 2 decades. Recent microbiome studies in cancer mice models have shown that certain commensal bacteria play protective roles against cancer. Thus, the use of commensal microflora having anticancer activities for the treatment of cancer appears to be an attractive alternative therapeutic strategy. Lactic acid bacteria (LAB) form an integral component of commensal microflora in healthy individuals. As the vaginal ecosystem is enriched in LAB genera, we screened the vaginal LAB microflora of healthy women for their anti-proliferative abilities against various human cancer cell lines. The secreted metabolites of three enterococcal strains, Enterococcus hirae 20c, Enterococcus faecium 12a and L12b, out of 92 LAB isolates selectively inhibited the in vitro proliferation of various human cancer cell lines in a dose-dependent manner but had no activity against normal human peripheral blood monocytes. Further, proteinase K-treatment of the cell-free supernatant (CS) of all the three enterococci abrogated their anti-proliferative abilities, thereby showing the proteinaceous nature of the secreted metabolites in the CS. The microscopic examination of the cell lines showed that CS-treatment induced apoptosis-like morphological changes in the cancer cells. Further, the probiotic characters of the strains were studied, which showed that all the three strains had broad spectrum antimicrobial activities against various Gram-positive and Gram-negative pathogens, including Mycobacterium smegmatis. All the strains tolerated the gastric acidity and bile juice treatments, and had strong adhesive abilities to the colonic epithelial cell line HCT-15. Furthermore, none of the strains had any known secreted virulence factors or harbored virulence genes. This preliminary study highlights an important functional role of the commensal probiotic enterococcal strains E. hirae and E. faecium for the first time by demonstrating their anticancer properties that should be further tested in the in vivo mammalian models.
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A diopside based bioactive system with a nominal composition of xCuO-(45.55-x)CaO-29.44 SiO2-10.28P2O5-14.73 MgO (x=0,1,3 and 5mol%) has been prepared by sol gel technique in the laboratory. X-ray Diffraction, Fourier Transform Infra-Red and Raman Spectroscopy, Field Emission Scanning Electron Microscopy along with Energy Dispersive X-ray Analysis and pH studies have been undertaken on the prepared samples before and after dipping the samples in simulated body fluid. It has been observed that hydroxyapatite layer starts to form with in 24h during immersion in simulated body fluid. Degradation studies have also been employed to check the degradation behavior in Tris-HCl. Dynamic light scattering studies show that particles are mostly agglomerated and have an average size of 356nm. Zeta potential studies have been undertaken to check the surface charge and it has been estimated that samples carry negative charge when dipped in simulated body fluid. Negative surface charge may contribute to attachment and proliferation of osteoblasts. Samples have also shown the antimicrobial properties against the Vibro cholerae and Escherichia coli pathogens. To check the non-toxic nature of the samples, cell cytotoxic and cell culture studies have been undertaken using the MG-63 cell lines. Samples have shown good response with good percentage viability of the cells in the culture media and hence, provides friendly environment to the growth of cells. The particle size, bioactivity, negative values of zeta potential, antimicrobial properties and good cell viability indicate the potential of the synthesized compositions as possible candidates for bone repair applications.
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Cobre/química , Compostos de Fósforo/química , Animais , Materiais Biocompatíveis/química , Linhagem Celular , Sobrevivência Celular , Durapatita/química , HumanosRESUMO
Serum amyloid P-component (SAP), a pentraxin, is known to play an important role in innate immunity to microbial infections; however, nothing is known about it during tuberculosis (TB). Mice intratracheally infected with Mycobacterium tuberculosis Erdman, showed peak SAP levels (442+/-58.2 microg/ml) on day 21, which declined to background levels by day 60. Their serum interleukin-6 levels paralleled SAP levels, whereas, their serum transforming growth factor-beta levels were paradoxical. During the acute phase of infection, the SAP levels positively correlated with the lung mycobacterial load. Purified mouse SAP (1-50 microg/ml) treatment of M. tuberculosis-infected alveolar macrophages (AMs), in vitro, inhibited their intracellular mycobacterial growth; maximum inhibition (1.1 log10 CFU reduction) occurred at 10 microg/ml, and a 4-day treatment appeared optimal. Treatment of AMs with both rabbit anti-mouse SAP polyclonal antibody and mannose-derived simple sugars, separately, blocked the SAP-induced inhibition of mycobacterial growth. The mycobacterial growth inhibition appeared to be nitric oxide (NO)-dependent as NO synthase inhibitors, both aminoguanidine and N(G)-monomethyl-L-arginine, annulled it. Further, SAP treatment of infected AMs induced significant (P<0.05) elaboration of nitrite (72.1+/-8.3 nM/ml), compared to the controls, and these AMs showed augmented expression of inducible NO synthase. This first study demonstrates that during murine TB the SAP levels were increased, and purified mouse SAP inhibited the intra-AM M. tuberculosis growth, in vitro, apparently via NO-dependent mechanism(s). SAP may thus contribute both to the pathogenesis and pulmonary innate immunity in TB.
Assuntos
Macrófagos Alveolares/microbiologia , Mycobacterium tuberculosis/crescimento & desenvolvimento , Mycobacterium tuberculosis/patogenicidade , Componente Amiloide P Sérico/metabolismo , Animais , Células Cultivadas , Contagem de Colônia Microbiana , Modelos Animais de Doenças , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Mycobacterium tuberculosis/efeitos dos fármacos , Óxido Nítrico/biossíntese , Componente Amiloide P Sérico/isolamento & purificação , Componente Amiloide P Sérico/farmacologia , Tuberculose Pulmonar/microbiologia , Regulação para CimaRESUMO
SETTING: Serum amyloid P-component (SAP) plays important roles in host defense during various infectious diseases; however, nothing is known in tuberculosis (TB). OBJECTIVE: To study the SAP response of Mycobacterium tuberculosis H37Rv- and H37Ra-infected mice, and to determine the effect(s) of purified mouse SAP both on their intra-alveolar macrophage (AM) uptake and intra-AM growth in vitro. DESIGN: The SAP levels of mice intratracheally infected with M. tuberculosis H37Rv and H37Ra were determined by ELISA. Mycobacterial AM uptake and intra-AM growth in vitro were determined using fluorescence microscopy and plating, respectively. RESULTS: M. tuberculosis H37Rv-infected mice showed significantly (p < 0.05) increased SAP levels (352.8+/-36.1 microg/ml) with compared mice infected with M. tuberculosis H37Ra (170+/-18.5 microg/ml). During the acute phase of both these infections, enhanced SAP levels correlated with the lung mycobacterial load. In vitro, purified mouse SAP (1-80 microg/ml) inhibited the AM uptake of both the mycobacteria in a concentration-dependent manners to a similar extent; 20 microg/ml SAP appeared optimal. Mycobacterial uptake inhibition was divalent cation- and pH-dependent, and was unaffected both by heat-inactivated and deglycosylated SAP, separately. Curiously, purified mouse SAP (1-80 microg/ml), in a concentration-dependent manner, inhibited the intra-AM growth of both M. tuberculosis H37Rv and H37Ra in vitro; the effect was 0.8 log10 CFUs greater on the latter. Both the mannose-based simple sugars and rabbit anti-mouse SAP polyclonal antibody, separately, annulled the inhibition of mycobacterial growth in vitro. CONCLUSION: This initial study demonstrates that both the SAP response of M. tuberculosis-infected mice, and the SAP-induced intra-AM mycobacterial growth inhibition in vitro were apparently dependent on mycobacterial virulence.