Proteome analysis of pseudomonas putida KT2440 using 2D gel electrophoresis and LC/ESI-Q-TOF mass spectrometry

The proteome can be defined as the complete set of global protein expression by an organism at any given time. It is this gene expression rather than genome itself that is responsible for most of the reactions taking place within a cell. Tools such as two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and mass spectroscopy (MS) are often utilized for the analysis and identification of the proteins that constitute the proteome. This thesis provides a general overview of proteomics and describes the underlying processes, mechanisms and technologies associated with modern proteomic analysis with specific emphasis on the use of electrospray ionization time of flight mass spectrometry. The use of MS/MS analysis of ESI-QTOF data for the identification of proteins is also explained in detail. Other chromatographic approaches such as high performance liquid chromatography (HPLC), where separation is based on the polarity of the mobile phase, and gel filtration chromatography systems, where separation is based on the molecular size, are also described in detail. The thesis work focuses on identifying the proteomic signature in the soil bacterium Pseudomonas putida using these analytical tools, particularly LC/ESI-Q-TOF MS (Electrospray Ionization Time of Flight Mass Spectrometer). Protein digest analysis was done with lactoperoxidase (LPO), lysozyme, and ribonuclease on HPLC. Whole protein and tryptic peptides were analyzed on the system. Peaks corresponding to the whole proteins as well as the peptides generated from the tryptic digestion proteins were observed. The goal of the entire project is to isolate proteins from the bacteria Pseudomonas Putida strain KT2440 using biochemical techniques to first separate them using 2-D PAGE, subsequently performing in-gel tryptic digest, and finally identifying individual protein spots on the gel using LC / ESI-Q-TOF MS and protein databases

PMMA-CNT matrices for vacuum electronic, biosensing and energy applications

Carbon nanotubes (CNTs) are interesting materials with extraordinary properties for various applications. Here, vertically-aligned multiwalled CNTs (VA-MWCNTs) are grown by our dual radio frequency plasma enhanced chemical vapor deposition (PECVD). After optimizing the synthesis processes, these VA-MWCNTs were fabricated in to a series of devices for applications in vacuum electronics, glucose biosensors, glucose biofuel cells, and supercapacitors In particular, we have created the so-called PMMA-CNT matrices (opened-tip CNTs embedded in poly-methyl methacrylate) that are promising components in a novel energy sensing, generation and storage (SGS) system that integrate glucose biosensors, biofuel cells, and supercapacitors. The content of this thesis work is described as follows: 1. We have first optimized the synthesis of VA-MWCNTs by our PECVD technique. The effects of CH4 flow rate and growth duration on the lengths of these CNTs were studied. 2. We have characterized these VA-MWCNTs for electron field emission. We noticed that as grown CNTs suffers from high emission threshold, poor emission density and poor long-term stability. We attempted a series of experiments to understand ways to overcome these problems. First, we decrease the screening effects on VA-MWCNTs by creating arrays of self-assembled CNT bundles that are catalyst-free and opened tips. These bundles are found to enhance the field emission stability and emission density. Subsequently, we have created PMMA-CNT matrices that are excellent electron field emitters with an emission threshold field of more than two-fold lower than that of the as-grown sample. Furthermore, no significant emission degradation was observed after a continuous emission test of 40 hours (versus much shorter tests in reported literatures). Based on the new understanding we learnt from the PMMA-CNT matrices, we further created PMMA-STO-CNT matrices by embedding opened-tip VA-MWCNTs that are coated with strontium titanate (SrTiO3) with PMMA. We found that the PMMA-STO-CNT matrices have all the desired properties of the PMMA-CNT matrices. Furthermore, PMMA-STO-CNT matrices offer much lower emission threshold field, about five-fold lower than that of as grown VA-MWCNTs. The new understandings we obtained are important for practical application of VA-MWCNTs in field emission devices. 3. Subsequently, we have functionalized PMMA-CNT matrices for glucose biosensing. Our biosensor was developed by immobilized glucose oxidase (GOχ) on the opened-tip CNTs exposed on the matrices. The durability, stability and sensitivity of the biosensor were studied. In order to understand the performance of miniaturized glucose biosensors, we have then investigated the effect of working electrode area on the sensitivity and current level of our biosensors. 4. Next, functionalized PMMA-CNT matrices were utilized for energy generation and storage. We found that PMMA-CNT matrices are promising component in glucose/O2 biofuel cells (BFCs) for energy generation. The construction of these BFCs and the effect of the electrode area on the power density of these BFCs were investigated. Then, we have attempted to use PMMA-CNT matrices as supercapacitors for energy storage devices. The performance of these supercapacitors and ways to enhance their performance are discussed. 5. Finally, we further evaluated the concept of energy SGS system that integrated glucose biosensors, biofuel cells, and supercapacitors. This SGS system may be implantable to monitor and control the blood glucose level in our body

Role of antioxidative enzymes in toxic bloom forming Cyanobacteria

The antioxidative enzymes catalase and ascorbate peroxidase in toxic bloom forming cyanobacteria play a pioneer role in scavenging free radicals, which are generated as an outcome of photosynthesis and respiration. Ten Bloom forming toxic cyanobacterial strains Synechococcus elongates, Synechocystis aquatalis, Merismopedia glauca, Microcystis aeruginosa, M. aeruginosa (O), Arthrospira (=Spirulina) platensis, Nostoc paludosum, Anabaena iyengarii, A. variabilis and Cylindrospermum muscicola were isolated from natural blooms and purified. The results depict the significant activity of antioxidative enzymes. In A. platensis the activity of catalase (28.7 ± 2.3 mM/min/ìg protein), and Ascorbate peroxidase (1.91±0.12 mM/min/ìg protein) enzyme was maximum. It was also observed that the activity of ascorbate perooxidase was very less compared to activity of catalase. The growth analysis and pigment profile were also studied. Growth measurements revealed that cells attain maximum growth nearly after 15-20 days of inoculation except S. aquatalis, A. platensis and N. paludosum maximum, where optimum growth was achieved after 25th day. Chlorophyll-a content of A.platensis showed maximum concentration (14.47 ± 1.17 mg/L) and minimum concentration in S. elongates (2.30 ± 0.138 mg/L). The concentration of phycobiliproteins was found maximum in A. platensis (2.09 mg/ml). The detection of antioxidative potential of cyanobacterial strains will help to broaden the knowledge about their survival and can pave path for their utilization in biotechnology

A Resilient and Optimal Microgrid Scheduling Portfolio in Linear Programming Platform

In recent year, alarming rate of natural disasters around the world have demanded the need for operative solution in field of power generation, to control polluted energy sources which are major cause of global warming. Microgrid facilitates penetration of renewable energy sources into the existing distribution systems to reduce the overall carbon footprint of the globe by reducing the dependency on the main grid. Efficient but linear microgrid resource scheduling algorithms are gaining interest in present time due to its simplicity and fast computation. This research paper aims to serve the purposes by designing a mixed integer linear programming based microgrid scheduling problem while various types of scenario, minimize the electricity cost for the utilities also maintain the generation and load balance. The strategy is implemented on a small microgrid  to prove its efficacy .In this paper mainly optimal scheduling of microgrid has been done in various scenario, and obtained there global minimum electricity cost to the help of mixed integer linear programming (MILP) algorithm.  Microgrid is a small scale type of power grid, which provides the energy locally, its offers integration between distributed energy resources and the locally connected loads. Microgrid able to be operated with the main grid and in standalone mode also ability to transitions between these two modes, the mode of operation of microgrid is depends on the system operating condition. The reliability of power grid is improving more when it’s integrated with the Microgrid and works together. Power exchange with the upper stream grid is done through the point of common coupling (PCC). Microgrid having renewable energy resources i.e. PV, Wind and non-renewable energy resources, DG, FC, and MC connected with the Battery storage type system and locally connected loads. So it is a very reliable scenario, main grid with the microgrid, it is more beneficial, economy and stable types of system. Keywords: Optimal Microgrid Scheduling, Linear Programming DOI: 10.7176/APTA/76-05 Publication date:March 31st 2019

A CLINICAL STUDY TO EVALUATE THE EFFICACY OF SAINDHAVADI TAILA MATRA VASTI IN THE MANAGEMENT OF AMAVATA W.S.R. RHEUMATOID ARTHRITIS

  Objective: The objective of the study is to find the efficacy of Saindhavadi Taila Matra Vasti in the management of Amavata (rheumatoid arthritis [RA]).Methods: For the present clinical study 15 patients of Amavata (RA) were registered from the Outpatient Department, PG Department of Kayachikitsa, Rishikul Campus, Haridwar. Saindhavadi Taila Matra Vasti was given 60 ml once daily for 8 days, followed by an interval of 7 days. Again Vasti was given once daily for 8 days followed by gap of 7 days. Same cycle was repeated next month. Assessment of the patients was done on the basis of subjective, objective, and functional parameters at the interval of 15 days.Results: Statistically significant result was found in subjective parameters such as pain intensity, Sandhishotha, Gaurav, Apaka (p<0.01 in each), Jwara, Aruchi, and Utsahahani (p<0.05 in each). Statistically non-significant result was found in all the functional parameters (p>0.05), that is, in grip strength, foot pressure, and goniometry. In biochemical parameters, statistically significant result was found in erythrocyte sedimentation rate only (p<0.05). Although non-significant result was found in other biochemical parameters such as hemoglobin, RA factor, and C-reactive protein (CRP) concentration, the mean scores of RA factor and CRP were reduced from 48.7 IU/ml and 10.4 mg/L before treatment to 25.8 IU/ml, 8.2 mg/L after treatment, respectively.Conclusion: In the clinical study, patients got symptomatic relief in many of the complaints but no significant result was found in functional parameters and most of the biochemical parameters. Thus, it can be concluded that Saindhavadi Taila Matra Vasti alone is effective in mild-to-moderate cases of Amavata (RA) and in severe cases it can be used along with oral Ayurvedic formulations for better results

A New Spectrophotometric Method for the Determination of Itraconazole Based on the Oxidation Reaction

The solubility of Itraconazole (Itcz) in various solvents such as distilled water, hydrochloric acid and sulphuric acid in presence of cationic and anionic surfactant i.e. cetyl trimethyl ammonium bromide (CTAB) and sodium lauryl sulphate (SLS) was examined. It has been found that SLS greatly enhanced the solubility of Itraconazole by 13 fold. A simple spectrophotometric method has been proposed for the determination of Itraconazole in pure and pharmaceutical formulation by ammonium metavanadate in presence of SLS. The method is based on the formation of wine red colored species on treating Itcz with ammonium metavanadate at CMC of SLS i.e. 0.0008M. The wine red species (due to reduced V(v)) exhibiting maximum absorbance at 385 nm. Keywords: Itraconazole, sodium lauryl sulphate, solubilit

Evaluation of Antimicrobial Activity of Ruta graveolens Stem Extracts by Disc Diffusion Method

In the present study, we evaluated the antimicrobial activity of ethanolic, methanolic, chloroform and water extracts of Ruta graveolens stem against Escherichia coli (MTCC 40), Pseudomonas aeruginosa (MTCC 424), Klebsiella pneumoniae (MTCC432), Staphyloccous aureus (MTCC 96), Bacillus subtilis (MTCC 619), Salmonella typhimurium (MTCC 98) and Aeromonas culicicola (MTCC 3249) as well as fungi such as Aspergillus niger, Aspergillus flavus, Penicilium crysogenum, Rhizopus stolonifer and Fusarium oxosporium by agar-disc diffusion  and (MIC) method. The ethanolic stem extract showed pronounced inhibition of growth than other extracts. The ethanolic extract showed most susceptible activity against S. aureous and B. subtilis (zone of inhibition was 22.0±0.04mm and19.4±0.24mm) where as P. aeruginosa (Zone of inhibition was 12.5±0.17mm) was the most resistant bacterial strain at a concentration of 100µg/ml. All the extract showed moderate antifungal activity except Fusarium oxosporium. Phytochemical analysis showed mainly the presence of alkaloids and steroids, which may the active compounds

Long-term treatment and poor management of psychiatric manifestations in mesial temporal sclerosis leading to suicidality in a young male

AbstractPsychiatric disorders including anxiety, psychosis, and aggressive behaviors are frequently diagnosed in patients with epilepsy. In this communication, we report a patients with mesial temporal lobe sclerosis with interictal affective-somatoform (dysphoric) disorders who was never treated for psychiatric manifestations, and who deliberately took a massive dose of phenytoin and phenobarbitone with a motive of suicide, resulting in severe combined toxicity. Such unfortunate incidences may be prevented, and quality of life can be improved with early diagnosis, through the selection of the right antiepileptic drugs, reasonable psychiatric consultation, and appropriate biological and psychological treatments