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Introduction. Cervicovaginal diversity has been reported as a predictive biomarker for cervical cancer risk. We recently reported the bio-therapeutic potential of vaginal probiotics from healthy Indian women against vaginal pathogens, isolated from the invasive cervical cancer (ICC) patients.Gap Statement. The cervicovaginal microflora from cervical cancer patients has not yet been reported from Indian population.Aim. The present study aimed at comparing the cervicovaginal microbiome between healthy controls (HC) and ICC patients from the Indian population.Methodology. In total, 30 vaginal swabs (15 from HC and 15 from ICC) were subjected to 16S rRNA gene sequencing. Alpha diversity was evaluated by Shannon and Chao1 index; and beta diversity by principle coordinate analysis (PCoA) of weighted and unweighted UniFrac distances. The relative abundance of the microbial taxa was done according to linear discriminant analysis effect size (LEfSe).Results. Predominance of Staphylococcus spp. in ICC and Lactobacillus gasseri in HC groups was observed. Alpha-diversity was found to be higher in ICC as compared to HC but was statistically non-significant. LEfSe analysis revealed Bacteroides fragilis and Escherichia coli as the marker genera in ICC with a marked decrease in Lactobacillus sp. Contrarily, in HC, L. gasseri, L. iners and L. fermentum were found to be abundant.Conclusion. Differences in the vaginal microbiome between healthy and ICC women could help in the early prediction of cervical cancer risk and thus in designing prevention strategies.
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Microbiota , Neoplasias del Cuello Uterino , Humanos , Femenino , Neoplasias del Cuello Uterino/epidemiología , ARN Ribosómico 16S/genética , Vagina , India/epidemiología , Escherichia coliRESUMEN
Abnormal cervicovaginal microbiota play an important role in HPV persistence and progression to cervical cancer. The present study aimed at isolating and identifying potential probiotics from vaginal swabs of healthy women and evaluating their activity against vaginal pathogens isolated from cervical cancer patients. Based on probiotic, acid-bile tolerance and antimicrobial properties, 13 lactic acid bacteria (LAB) from the healthy group were identified by MALDI TOF MS (Matrix Assisted Laser Desorption and Ionisation, Time Of Flight Mass Spectrometry). Among these, four strains, Lactobacillus gasseri P36Mops, Limosilactobacillus fermentum P37Mws, Lactobacillus delbrueckii P31Mcs and Enterococcus faecium P26Mcm, exhibited significant antimicrobial activity against 8 vaginal pathogens (Staphylococcus haemolyticus P41Tcs, Escherichia coli P30Tcs, E. coli P79Bcm, Enterococus faecalis P29Mops, E. faecalis P50Tws, E. faecalis P68Tcb, S. haemolyticus P48Bcb and S. haemolyticus P58Bcb) isolated from precancerous and cervical cancer patients. 16S rRNA sequencing of four potential probiotics revealed congruency with the MALDI-TOF MS identification and phylogenetic analysis showed genetic relationship with previously reported LAB strains. The selected LAB showed strain specific hydrophobicity (35.88-56.70%), auto-aggregation (35.26-61.39%) and antibiotic susceptibility. Interestingly, L. gasseri P36Mops was resistant to five standard antibiotics routinely used against urogenital or vaginal infections. LCMS (Liquid Chromatography Mass Spectrometry) analyses of the CFS (cell-free supernatant) of the four potential probiotics revealed the presence of metabolites such as N-(1-deoxy-1-fructosyl)valine, hygroline, acetoxy-2-hydroxy-16-heptadecen-4-one, avocadyne 4-acetate, avocadyne 2-acetate, taraxinic acid glucosyl ester, 6-hydroxypentadecanedioic acid, with reported antimicrobial activity. The overall data suggest the bio-therapeutic potential of the identified vaginal probiotics against cervical cancer-associated pathogens.
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Lactobacillales , Microbiota , Probióticos , Neoplasias del Cuello Uterino , Antibacterianos/metabolismo , Antibacterianos/farmacología , Escherichia coli/genética , Femenino , Humanos , Filogenia , Probióticos/metabolismo , ARN Ribosómico 16S/genética , ARN Ribosómico 16S/metabolismo , Neoplasias del Cuello Uterino/tratamiento farmacológicoRESUMEN
Nicotine is one of the most easily accessible, commonly abused drugs worldwide and if taken in overdose can cause serious clinical presentation, including cardiac arrhythmias and neurotoxicity mediated through oxidative stress. Its toxicity though rare can cause sudden deaths by cardiovascular arrest, respiratory muscle paralysis, and/or central respiratory failure. Here, we describe a case of intentional fatal ingestion of nicotine sulfate decoction used as a mean for fatal suicidal attempt by a 15-year-old adolescent male who was suffering from childhood-onset depression since about 3 months. He developed drooling of saliva, syncopal attacks, paroxysmal episodes of hematemesis, abdominal pain, signs and symptoms of hypoxia, nonfatal atrial tachycardia, and encephalopathy after ingestion of heavy dose of nicotine-containing concoction; however, he recovered successfully within 24-48 hours without any significant cardiac, respiratory, or neurological deficits (except short-term verbal memory). Authors discussed the details of management and reasons behind the reversible encephalopathy and molecular mechanism of nicotine toxicity. HOW TO CITE THIS ARTICLE: Kamble A, Khairkar P, Kalantri SP, Babhulkar S. Fatal Suicidal Attempt by Deliberate Ingestion of Nicotine-containing Solution in Childhood-onset Depression Mediated through Internet Suicide Guideline: A Case Report. Indian J Crit Care Med 2020;24(8):719-721.
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The intracellular cobalt-type nitrile hydratase was purified from the bacterium Rhodococcuserythropolis. The pure enzyme consisted of two subunits of 29 and 30 kDa. The molecular weight of the native enzyme was estimated to be 65 kDa. At 25 °C the enzyme had a half-life of 25 h. The Michaelis-Menten constants Km and vmax for the enzyme were 0.624 mM and 5.12 µmol/min/mg, respectively, using 3-cyanopyridine as the substrate. The enzyme-containing freely-suspended bacterial cells and the cells immobilized within alginate beads were evaluated for converting the various nitriles to amides. In a packed bed reactor, alginate beads (2 % alginate; 3 mm bead diameter) containing 200 mg/mL of cells, achieved a conversion of >90 % for benzonitrile and 4-cyanopyridine in 38 h (25 °C, pH 7.0) at a feed substrate concentration of 100 mM. The beads could be reused for up to six reaction cycles.
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The cross-linked enzyme aggregate (CLEA) method is used for the dual purpose of combining both the purification and immobilization of enzyme in one step. The present work involved the preparation of a carrier-free, highly active reusable biocatalyst (nitrilase) which encounters least mass-transfer limitations with higher thermal and storage stability. The effect of type of aggregating agent, its concentration as well as that of cross-linking agent was studied. Nitrilase aggregates were prepared using ammonium sulphate (35%) precipitation followed by cross-linking with glutaraldehyde (125 mM) which rendered 70% activity retention. The various cross-linking parameters were optimized in order to increase the activity retention. Stability in terms of temperature, reusability and leaching were also examined. The CLEA preparation showed residual nitrilase activity on repeated use. A highly stable CLEA of nitrilase was finally prepared with maximum activity recovery.
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Aminohidrolasas/metabolismo , Nitrilos/metabolismo , Pseudomonas putida/enzimología , Recombinación Genética , Aminohidrolasas/química , Hidrólisis , Nitrilos/química , Pseudomonas putida/genética , EstereoisomerismoRESUMEN
The reaction conditions for the nicotinamide production by Rhodococcus erythropolis MTCC 1526 have been optimized by statistical experimental design. Application of this approach in the bioprocess can result in rallied product yield, reduced development time, and process variability. In this investigation, response surface methodology and central composite design were employed to predict the levels of variables such as reaction pH (6.5, 7, and 7.5), temperature (15, 20, and 25 degrees C), cell concentration (190, 200, and 210 mg/ml), and substrate concentration (18, 20, and 22 mM) on the production of nicotinamide. A total of 22 experiments were carried out in shake flasks, and a three-dimensional response surface was generated to determine the effect of crucial reaction parameters for the maximum conversion of 3-cynopyridine to nicotinamide. Using this methodology, the optimal values for the reaction conditions were reaction pH 6.85, temperature of 24.8 degrees C, cell concentration of 190.98 mg/ml, and substrate concentration of 21.98 mM. This statistical approach led to the increase of conversion of 3-cynopyridine (93%) as compared to the conversion obtained by one-factor-at-a-time approach (84%).
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Hidroliasas/metabolismo , Niacinamida/biosíntesis , Concentración de Iones de Hidrógeno , Piridinas/metabolismo , Rhodococcus/metabolismo , Propiedades de Superficie , TemperaturaRESUMEN
Culture conditions were optimized for the growth and carbonyl reductase production by a novel yeast strain Candida viswanathii. Response surface methodology was applied for the critical medium components (initial pH, mannitol, yeast extract and calcium chloride) identified earlier by one-factor-at-a-time approach. Central composite design was used for the optimization studies. Using this methodology, the optimal values for the concentration of mannitol, initial pH, yeast extract and calcium chloride were 1.9, 7.5, 1.6 and 4, respectively. This medium was projected to produce, theoretically, growth having an optical density of 1.1 (600 nm) and an enzyme activity of 81.5 U/ml. Using this optimized medium, an experimental growth of 1.1 OD (600 nm) and enzyme activity 80.9 U/ml verified the applied methodology. This approach for medium optimization led to an enhancement of the growth and enzyme activity by 1.3 and 2.3 times higher, respectively, as compared to the unoptimized media.