Optimizing CaCl2-mediated transformation of Pseudomonas aeruginosa SDK-6 with pJN105 using OFAT: A novel and efficient cloning approach.

Cloning and expression of a gene in the desired host is required for optimum production in recombinant strains. The present research is the first attempt to optimize the physiological conditions for the transformation of Pseudomonas aeruginosa SDK-6 with pJN105. Different factors, such as inoculum size, incubation period, heat shock temperature, and heat shock time were optimized using one factor at a time (OFAT) followed by the selection of transformants using gentamicin resistance marker. The maximum number of transformants (2.002 ± 0.077 × 105 cfu/ µg of plasmid DNA) were reported with 0.5% (v/v) inoculum, an incubation period of 3 h, and heat shock treatment at 50 °C for 1 min. An overall 12-fold increase in transformation efficiency was observed. The presence of a 6055 bp band on agarose gel confirmed the transformation of Pseudomonas aeruginosa with the vector pJN105.

Construction of Pseudomonas aeruginosa SDK-6 with synthetic lipase gene cassette and optimization of different parameters using response surface methodology for over-expression of recombinant lipase.

Lipases are industrially important enzymes having vast applications in various fields. Cloning and expression of lipase enzyme-encoding genes in suitable host lead to their widespread use in different fields. The present study represents the first attempt towards the expression of the synthetic lipase gene in Pseudomonas aeruginosa. An alkalophilic lipase gene (GenBank accession number: NP_388152) from Bacillus subtilis was synthetically designed and introduced in the pJN105 vector and subsequently cloned in Pseudomonas aeruginosa SDK-6. Agarose gel electrophoresis confirmed the transformation of SDK-6, exhibiting a band difference of ~ 700 bp between native and recombinant pJN105. Further amplification of cloned lipase gene was confirmed using PCR amplification with Lip 1 and Lip 2 primers respectively, followed by restriction analysis. Approximately 15-fold increase in lipase production was observed in recombinant Pseudomonas as compared to the native strain. One factor at a time (OFAT) analysis revealed L-arabinose, inoculum size (0.5%; v/v), and agitation (120 rpm) as significant factors affecting the over-expression of lipase enzyme. Optimization of enzyme induction conditions by central composite design (CCD) led to 1.60-fold increase in the production of lipase at 0.65% (w/v) inducer concentration, OD600-1.075 before induction and 35 °C post induction temperature with overall lipase production of 50.50 IU/mL. Statistical validation of observed value via ANOVA showed an F-value of 138.70 at p < 0.01 with R2 of 0.9921.

An insight into microbial sources, classification, and industrial applications of xylanases: A rapid review.

Endo 1,4-ß-d-xylanases (EC3.2.1.8) are one of the key lignocellulose hydrolyzing enzymes. Xylan, which is present in copious amounts on earth, forms the primary substrate of endo-xylanases, which can unchain the constituent monosaccharides linked via ß-1,4-glycosidic bonds from the xylan backbone. Researchers have shown keen interest in the xylanases belonging to glycoside hydrolase families 10 and 11, whereas those placed in other glycoside hydrolase families are yet to be investigated. Various microbes such as bacteria and fungi harbor these enzymes for the metabolism of their lignocellulose fibers. These microbes can be used as miniature biofactories of xylanase enzymes for a plethora of environmentally benign applications in pulp and paper industry, biofuel production, and for improving the quality of food in bread baking and fruit juice industry. This review highlights the potential of microbes in production of xylanase for industrial biotechnology.

Pectinases as promising green biocatalysts having broad-spectrum applications: Recent trends, scope, and relevance.

Pectinases are a collection of multiple enzymes that have a common substrate, that is, pectin. They can act on different parts of pectin due to the structural heterogeneity of pectin. Therefore, they have been placed in different groups, such as protopectinases, polygalacturonases, polymethylesterases, pectin lyases, and pectate lyases. They are naturally present both in multicellular organisms such as higher plants and in unicellular organisms such as microbes. In past decade, it has been witnessed that chemical and mechanical methods employed in industrial processes have led to environmental hazards and serious health disorders, thus increasing the search for eco-friendly approaches with minimal health risks. Hence, microbial enzymes have been extensively used as safer alternative for these environmentally unsafe methods. Among these microbial enzymes, pectinases hold great significance and is one of the principal enzymes that have been used commercially. It is predominantly used as a green biocatalyst for fruit, fiber, oil, textile, beverage, pulp, and paper industry. Thus, this review focuses on the structure of pectin, microbial sources of pectin, and principle industrial applications of pectinases.

QEEG Predictors of Treatment Response in Major Depressive Disorder- A Replication Study from Northwest India.

Background: Predicting treatment response with antidepressant is a challenging task for clinicians and researchers. An important limitation of an antidepressant trial is the increased time spent before an adequacy of trial can be decided. Quantitative Electroencephalography has shown some evidence in identifying early changes seen with antidepressants. No data has been reported from Indian population on its predictive capabilities. Aim: To examine whether early changes in frontal and prefrontal theta value in QEEG could predict antidepressant treatment response. Methods: Structured clinical assessments were conducted at baseline and after one week in a sample of treatment-seeking adults with major depressive disorder (n = 50). Patients were started on SSRI (Escitalopram, fluoxetine, paroxetine or sertraline) and followed for 8 weeks. QEEG recordings were carried out at baseline and week 1 and its parameters (relative theta power and cordance) were assessed to identify its predictive value for treatment response. Treatment response was assessed using Hamilton depression rating scale with 50% reduction after 8 weeks being considered as response. Results: Mean age of the sample was 39 ± 10 years and majority of them were females (64%). A significant reduction was found in relative frontal theta value (p = 0.021) from baseline to one week in responders. However, linear regression revealed that this change could not predict the treatment response (p = 0.37). Conclusions: QEEG changes are observed in initial phase of antidepressant treatment but these changes can't predict the treatment response.

In vitro Antimicrobial Activity of Cucumis L. and Momordica L. against Human Pathogens.

Plants are the rich source of compounds having antimicrobial properties against human pathogens. The present study has been carried out to evaluate the antimicrobial potential of Cucumis melo var. agrestis (morphotype I), Cucumis melo var. agrestis (morphotype II), Cucumis melo var. momordica L., Cucumis melo L., Momordica balsamina L., Momordica charantia L., Momordica dioica L. against Staphylococcus aureus, Pseudomonas flourescens, Bacillus coagulans, and Klebsiella pneumoniae. Crude extract of Cucumis L. and Momordica L. species were prepared with methanol, acetone and water for the determination of antimicrobial properties. Maximum yield was reported in methanol extract while minimum in acetone for all plant species. The maximum zone of inhibition of about 32.3 ± 0.57 mm was found against Staphylococcus aureus in Cucumis melo L., 21.3 ± 0.57 mm for Pseudomonas flourescens in Cucumis melo var. agrestis (morphotype II), 17 ± 0 mm for Klebsiella pneumoniae in Momordica balsamina L., and 23.3 ± 0.57 mm for Bacillus coagulans in Cucumis melo var. agrestis (morphotype II) extracts, respectively. The most active antimicrobial plants species were reported to be Cucumis melo var. agrestis (morphotype I), Cucumis melo L. and Momordica charantia L. having antimicrobial activities against all tested microorganisms.

Screening and Characterization of Next-Generation Biofuels Producing Bacterial Strains.

Production of fuels from renewable resources is of utmost importance due to fast depletion of fossil resources and related environmental issues. The present study explored the intrinsic capability of microbial strains to produce alka(e)nes, the next-generation biofuel, thus to reduce the dependence upon current petroleum fuels. Eight bacterial strains, namely, SDK-1, SDK-2, SDK-6, SDK-7, SDK-8, SDK-9, SDK-10, and SDK-11 were isolated from sludge and soil samples collected from different sources using lauric acid as a substrate with a potential to produce alka(e)nes. Production of different medium- and long-chain alka(e)nes by these isolates was confirmed via gas chromatography-mass spectrometer (GC-MS) analysis. SDK-1 (7.2%), SDK-2 (3.72%), and SDK-6 (3.52%) produced significant proportion of medium-chain hydrocarbons as compared to SDK-10 and control with no production. These isolates may be further investigated for production of these alternative sources of energy. In contrary, maximum fraction of long-chain hydrocarbons is produced in SDK-8 (75.28%) followed by SDK-9 (61.51%). Similarly more than 50% of the total hydrocarbons produced in SDK-8 constitute fossil mimic hydrocarbons while only 10.78% fractions were found in SDK-10. Since these fractions resemble different hydrocarbons obtained from crude oil, hence may be explored for their wide applications in different fields. Biochemical characterization and sequencing of the 16S rRNA gene revealed the homology of SDK-1, SDK-2 and SDK-6 with Pseudomonas aeruginosa, SDK-7 and SDK-9 with Enterobacter cloacae, SDK-8 with Klebsiella pnuemoniae, SDK-10 with Enterobacter hormaechei and SDK-11 with Pseudomonas nitroreducens, respectively.

Can EEG Predict Response to Atomoxetine in attention deficit hyperactivity disorder at 1 Week?

Objective: The objective of the study is to predict the early changes in electroencephalography (EEG) at 1 week and its correlation to clinical response at 6 weeks after treatment with atomoxetine in children with ADHD. Method: In 50 children (6-14 years) with ADHD (Diagnostic and Statistical Manual of Mental Disorders [5th ed.; DSM-5]), Vanderbilt ADHD Parent Rating Scale (VADPRS) and Vanderbilt ADHD Teachers Rating Scale (VADTRS) were applied at baseline, 1, 4, and 6 weeks. EEG was recorded using International 10-20 System of electrode placement at baseline and at 1 week after atomoxetine treatment. EEG changes at 1 week after atomoxetine therapy was correlated to clinical response at 6 weeks. Results: Patients were classified as responders or nonresponders based on the VADPRS/VADTRS findings. After 1 week of treatment, responders' theta cordance values were decreased, whereas nonresponders' values didn't decrease significantly. Conclusion: Patients with decreased theta cordance values, especially in the left temporoparietal region, at 1 week were likely to respond to atomoxetine while those without any such change were likely to be nonresponders.

Phase Space Reconstruction of EEG Signals for Classification of ADHD and Control Adults.

Attention deficit hyperactivity disorder (ADHD) is a childhood behavioral disorder that can persist into adulthood. Electroencephalography (EEG) plays a significant role in assessing the neurophysiology of ADHD because of its ability to reveal complex brain activity. The present study proposes an EEG-based diagnosis system using the phase space reconstruction technique to classify ADHD and control adults. Electric activity is recorded for 47 ADHD and 50 control adults during the eyes-open, eyes-closed, and Continuous Performance Test (CPT) condition. Various statistical features are extracted from Euclidean distances based on phase space reconstruction of signals. The proposed system is evaluated with 2 feature selection methods (correlation-based feature selection and particle swarm optimization) and 5 machine learning methods (neural dynamic classifier, support vector machine, enhanced probabilistic neural network, k-nearest neighbor, and naive-Bayes classifier). Experimental results showed the highest testing accuracy of 93.3% under the eyes-open, 90% under the eyes-closed, and 100% under the CPT condition. This study focused on the utility of phase space reconstruction of brain signals to discriminate between ADHD and control adults.

Event-Related Potential Analysis of ADHD and Control Adults During a Sustained Attention Task.

Background. Event-related potentials (ERPs) of attention deficit hyperactivity disorder (ADHD) population have been extensively studied using the time-domain representation of signals but time-frequency domain techniques are less explored. Although, adult ADHD is a proven disorder, most of the electrophysiological studies have focused only on children with ADHD. Methods. ERP data of 35 university students with ADHD and 35 control adults were recorded during visual continuous performance task (CPT). Gray level co-occurrence matrix-based texture features were extracted from time-frequency (t-f) images of event-related EEG epochs. Different ERP components measures, that is, amplitudes and latencies corresponding to N1, N2, and P3 components were also computed relative to standard and target stimuli. Results. Texture analysis has shown that the mean value of contrast, dissimilarity, and difference entropy is significantly reduced in adults with ADHD than in control adults. The mean correlation and homogeneity in adults with ADHD were significantly increased as compared with control adults. ERP components analysis has reported that adults with ADHD have reduced N1 amplitude to target stimuli, reduced N2 and P3 amplitude to both standard and target stimuli than controls. Conclusions. The differences in texture features obtained from t-f images of ERPs point toward altered information processing in adults with ADHD during a cognitive task. Findings of reduction in N1, N2, and P3 components highlight deficits of early sensory processing, stimulus categorization, and attentional resources, respectively, in adults with ADHD.

Pulpal perio relations: Interdisciplinary diagnostic approach - I.

Lesions of pulpal and periodontal origin may perpetuate from either the infections of dental pulp or periodontium or alveolar bone. This review focuses on interdisciplinary diagnostic approach towards lesions of periodontal or endodontic origin.