Purification and Characterization of Acetylcholinesterase from Clarias Batrachus and Oreochromis Mossambica Brain Tissues

This study reports on the purification and characterization of a soluble AChE (EC 3.1.1.7) from Clarias batrachus and Oreochromis mossambica brain tissues. The purification protocol involved homogenization, centrifugation, ultrafiltration, application of customsynthesized affinity chromatography gel (Edrophonium– Sephacryl S400) and the use of high performance liquid chromatography system (HPLC). The affinity matrix was synthesized by coupling an AChEspecific inhibitor, edrophonium chloride to epoxyactivated Sephacryl S400 matrix. Soluble AChE from C. batrachus and O. mossambica were purified 26.4 and 27.9 fold with a specific activity of 59.7 × 10 3 and 73.1 × 10 3 U/mg proteins, respectively. The molecular weight of AChE for C. batrachus estimated on Superose TM gel filtration column under nondenaturing conditions is 311 kDa. Native polyacrylamide gel electrophoresis (NativePAGE) under nondenaturing conditions showed only one major molecular form of protein for C. batrachus with a molecular weight of about 309 kDa, while AChE from O. mossambica could not be purified. Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate and betamercaptoethanol (SDSPAGE) gave only one band for C. batrachus with an estimated molecular weight of 74 kDa. Based on the molecular weights obtained for C. batrachus from both SDSPAGE and NativePAGE, the purified AChE can be postulated as being a tetramer form linked with disulfide bonds. Acetylcholinesterases purified from brain tissues samples of C. batrachus and partially purified from O. mossambica have been analyzed further on substrate and sensitivity to inhibitors to distinguish from butrylcholinesterase (BuChE). The AChE from C. batrachus and O. mossambica were most active against acetylthiocholine (ATC) and shows less activity against propionylthiocholine (PTC) and butyrylthiocholine (BTC). From a kinetic point of view, the purified AChE from C. batrachus exhibit the Michaelis constants Km, for ATC, PTC and BTC in the range of 97, 138 and 238 μM and the maximum velocities Vmax were 347, 64 and 25 μmol/min/mg protein, respectively. Meanwhile, partially purified AChE from O. mossambica exhibit Km(app) for ATC, PTC and BTC in the range of 125, 260 and 600 μM and Vmax(app) were 276, 59 and 36 μmol/min/mg protein, respectively. The turnover number (kcat) for purified AChE from C. batrachus with ATC as a substrate was 0.19 × 10 5 min 1 . The inhibition constant (ki) values of eserine, propidium and carbofuran were 0.34, 81 and 0.51 μM 1 min 1 for C. batrachus and 0.24, 65 and 0.41 μM 1 min 1 for O. mossambica, respectively. This enzyme is apparently an AChE since it hydrolyzes ATC at a higher rate than other substrates, such as BTC and PTC, at pH 7.0 and 25ºC, and is inhibited by eserine but not by isoOMP

GREEN TEA CATECHIN LOADED NANODELIVERY SYSTEMS FOR THE TREATMENT OF PANDEMIC DISEASES

Tea (Camellia sinensis, Family: Theaceae) is one of the extremely consumed beverages around the world, behind to water. The brew tea is the merely food product contains abundant quantity of the catechins. Green tea is the least processed and thus contains rich antioxidant, polyphenols, especially catechin called epigallocatechin-3-gallate (EGCG), which is whispered to be responsible for a wide range of the health benefits. The key to the amazing health benefits that are derived from green tea is that the leaves are steamed which preserves the EGCG compound from being oxidized. However, the other varieties of teas are under go fermentation process, which breaks down the potential EGCG and destroy from its healing properties. In reality, green tea has very extensive history dating back thousands and thousands of years ago. However, the pharmacological efficacy and stability of green tea catchiness are primarily depended on the formulation and way to drink to alleviate the deadly diseases with scientific evidence. Nanotechnology is a vibrantly emerging field especially in the pharmaceutical industry to explore a lot of application. The promising nano-delivery system used to enhance the therapeutic efficacy with a minimal dose, minimize the dose-related toxicity, target delivery, site-specific delivery, and controlled/sustain the delivery application. In recent decades, the application of nanotechnology has been utilized for phytopharmaceutical industry including green tea catechins to maximize the health benefits. In this review, we tried our level best retrieve the value of information on nanodelivery application of green tea catchiness for various devastating diseases

Biogas Production From a Mixture of Cow Manure with Chicken Manure

Biogas technology with zero waste concept is expected to be the alternative energy and to reduce environmental problems. The purpose of this study is to know the biogas yield per kilogram of each chicken and cow manure comparison. The study was conducted in six treatments with the addition of chicken manure of 0, 100, 300, 500, 700 and 1000 grams. The fermentation process is done using a batch system and biogas measurement was taken daily. The parameters to be observed were organic matter, the degree of acidity (pH), temperature, volume of biogas, biogas productivity, and C / N ratio of each treatment. The results showed that the overall pH at the beginning and end of the study tend to be close to neutral. The highest biogas yield was resulted from a mixture of chicken manure and cow manure at the composition of 1:1 or 50%:50% with biogas total amount of 35.690 ml and biogas productivity of 0,33 liters/g (volatile solid)

Dissecting the biochemical and hormonal changes of thidiazuron on defoliation of cotton CO17 (Gossypium hirsutum) to enhance mechanical harvest efficiency

Using chemical defoliants to remove cotton leaves represents a groundbreaking shift in cotton cultivation. The mechanization of cotton harvest is increasing, but a substantial amount of foliage that remains on the plant even at maturity is the major barrier for mechanical harvest. Properly completing mechanical and manual harvests requires artificial leaf detachment through defoliants. Still there is no proper defoliant concentration, application times and mechanism of action available. Therefore, the present study aimed to find an effective defoliant and application time to enhance mechanical harvest efficiency, along with a clear description of the mechanism of actions in cotton CO17 (Gossypium hirsutum). The field experiment was conducted during the year 2019-20 and used five concentrations of Thidiazuron defoliant (100, 150, 200, 250 and 300ppm) and [email protected]% (T2) in cotton variety CO17 to study the physiological, biochemical and hormonal responses at 120, 127 and 134 days after sowing. As a result, the concentrations of plant growth hormones, indole-3 acetic acid (4.9 fold), zeatin (32.7%) and gibberellic acid (7 fold) reduced. In contrast, abscisic acid (48.6%), jasmonic acid (34.9%), salicylic acid (2.15 fold) increased in the T7- Thidiazuron + Diuron (300 ppm) treatment followed by T5-Thidiazuron + Diuron (200 ppm). Additionally, the antioxidant enzymes ascorbate peroxidase, peroxidase, catalase, superoxide dismutase, cellulase in leaves, petiole and bolls were decreased due to defoliant T5- Thidiazuron + Diuron (200 ppm) followed by T7-Thidiazuron + Diuron (300 ppm), indicating that the hormone concentration, antioxidative and hydrolytic enzymes are ruled out and forces the defoliation process.

Salt stress and its impact on rice physiology with special reference to India- A review

With the increasing population, by 2030, the population of India will have seen an unprecedented rise of 1.43 billion and require food grains of around 311 million tones. Of the total area, nearly 5% of the area in India is affected by soil salinity. It is said that about 10% of soil is salinized every year. At this rate, 50% of the land area will be salinized by 2050.These repercussions challenge us to expand the area under cultivation or to increase the yield per unit area to maintain food security and sustainability. In order to meet the growing demands of the increased population, two major approaches can be met. Firstly, the available area under cultivation must be increased, which can be done by the reclamation of various problematic soils and making them suitable for cultivation. The second and holistic approach is to employ various biotechnological and breeding aspects in the development of resistant varieties surviving the harsh and unfavourable environment and showing no subsequent reduction in the yield parameters. For this, one must understand the various physiological aspects of tolerance for screening the elite varieties suited for a particular ecosystem or environment. Thus, the present study vividly explains the various physiological aspects of salt stress on rice. Employing these techniques, one can screen superior genotypes resistant to various stresses, thus keeping the Malthus predictions at bay

Application of defoliants alters leaf growth and gas exchange parameters for cotton defoliation

The goal of the current study was to determine how different chemical defoliants and application timing affected defoliation in cotton variety CO 17. The studies were conducted using a split-plot design with three applications at three different times as the main plot and seven defoliants as the subplot. Thidiazuron + Diuron (0.03%) defoliant reduced the gas exchange parameters, photosynthetic rate, transpiration rate and stomatal conductance by reducing plant growth parameters, leaf area, leaf area index, specific leaf weight which significantly increased the defoliation percentage. The negative correlation of cotton growth and gas exchange parameters with defoliation percentage was observed in correlation studies. In conclusion, Thidiazuron + Diuron (0.03%) defoliant was found to be superior in action for improving leaf defoliation and its associated parameters. And also it may be a cost-effective cotton defoliant for aiding the mechanical picking of cotton bolls

Assay for heavy metals using an inhibitive assay based on the acetylcholinesterase from Clarias batrachus

Acetylcholinesterase (AChE) is usually used as an inhibitive assay for insecticides. A lesser-known property of AChE is its inhibition by heavy metals. In this work, we evaluate an AChE from brains of Clarias batrachus (catfish) exposed to wastes from aquaculture industry as an inhibitive assay for heavy metals. We discovered that the AChE was inhibited completely by Hg2+, Ag2+, Pb2+, Cu2+, Cd2+, Cr6+ and Zn2+ during initial screening. When tested at various concentrations, the heavy metals exhibited exponential decay type inhibition curves. The calculated IC50 (mg/L) for the heavy metals Ag2+, Cu2+, Hg2+, Cr6+ and Cd2+ were 0.088, 0.078, 0.071, 0.87 and 0.913, respectively. The IC50 for these heavy metals are comparable, and some are lower than the IC50 values from the cholinesterases from previously studied fish. The assay can be carried out in less than 30 minutes at ambient temperature

Microbial succession during the transition from active to inactive stages of deep-sea hydrothermal vent sulfide chimneys

© The Author(s), 2020. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Hou, J., Sievert, S. M., Wang, Y., Seewald, J. S., Natarajan, V. P., Wang, F., & Xiao, X. Microbial succession during the transition from active to inactive stages of deep-sea hydrothermal vent sulfide chimneys. Microbiome, 8(1), (2020): 102, doi:10.1186/s40168-020-00851-8.Background Deep-sea hydrothermal vents are highly productive biodiversity hotspots in the deep ocean supported by chemosynthetic microorganisms. Prominent features of these systems are sulfide chimneys emanating high-temperature hydrothermal fluids. While several studies have investigated the microbial diversity in both active and inactive sulfide chimneys that have been extinct for up to thousands of years, little is known about chimneys that have ceased activity more recently, as well as the microbial succession occurring during the transition from active to inactive chimneys. Results Genome-resolved metagenomics was applied to an active and a recently extinct (~ 7 years) sulfide chimney from the 9–10° N hydrothermal vent field on the East Pacific Rise. Full-length 16S rRNA gene and a total of 173 high-quality metagenome assembled genomes (MAGs) were retrieved for comparative analysis. In the active chimney (L-vent), sulfide- and/or hydrogen-oxidizing Campylobacteria and Aquificae with the potential for denitrification were identified as the dominant community members and primary producers, fixing carbon through the reductive tricarboxylic acid (rTCA) cycle. In contrast, the microbiome of the recently extinct chimney (M-vent) was largely composed of heterotrophs from various bacterial phyla, including Delta-/Beta-/Alphaproteobacteria and Bacteroidetes. Gammaproteobacteria were identified as the main primary producers, using the oxidation of metal sulfides and/or iron oxidation coupled to nitrate reduction to fix carbon through the Calvin-Benson-Bassham (CBB) cycle. Further analysis revealed a phylogenetically distinct Nitrospirae cluster that has the potential to oxidize sulfide minerals coupled to oxygen and/or nitrite reduction, as well as for sulfate reduction, and that might serve as an indicator for the early stages of chimneys after venting has ceased. Conclusions This study sheds light on the composition, metabolic functions, and succession of microbial communities inhabiting deep-sea hydrothermal vent sulfide chimneys. Collectively, microbial succession during the life span of a chimney could be described to proceed from a “fluid-shaped” microbial community in newly formed and actively venting chimneys supported by the oxidation of reductants in the hydrothermal fluid to a “mineral-shaped” community supported by the oxidation of minerals after hydrothermal activity has ceased. Remarkably, the transition appears to occur within the first few years, after which the communities stay stable for thousands of years.This work was supported by the China Ocean Mineral Resources R&D Association (grant No. DY135-B2-12), the Natural Science Foundation of China (grant No. 41530967, 41921006, 91751205), the Senior User Project of RV KEXUE (KEXUE2019GZ06), and by the US National Science Foundation grant OCE-1136727 and the WHOI Investment in Science Fund to S.M.S

Autoantibody Biomarker Discovery in Primary Open Angle Glaucoma Using Serological Proteome Analysis (SERPA)

Glaucoma is an optic neurological disorder and the leading cause of irreversible blindness worldwide, with primary open angle glaucoma (POAG) as its most prevalent form. An early diagnosis of the disease is crucial to prevent loss of vision. Mechanisms behind glaucoma pathogenesis are not completely understood, but disease related alterations in the serological autoantibody profile indicate an immunologic component. These changes in immunoreactivity may serve as potential biomarkers for glaucoma diagnostics. We aimed to identify novel disease related autoantibodies targeting antigens in the trabecular meshwork as biomarkers to support early detection of POAG. We used serological proteome analysis (SERPA) for initial autoantibody profiling in a discovery sample set. The identified autoantibodies were validated by protein microarray analysis in a larger cohort with 60 POAG patients and 45 control subjects. In this study, we discovered CALD1, PGAM1, and VDAC2 as new biomarker candidates. With the use of artificial neural networks, the panel of these candidates and the already known markers HSPD1 and VIM was able to classify subjects into POAG patients and non-glaucomatous controls with a sensitivity of 81% and a specificity of 93%. These results suggest the benefit of these potential autoantibody biomarkers for utilization in glaucoma diagnostics

Assay for heavy metals using an inhibitive assay based on the acetylcholinesterase from Channa striatus

Acetylcholinesterase (AChE) is usually used as an inhibitive assay for insecticides. A lesser known property of AChE is its inhibition by heavy metals. In this work we evaluate an AChE from brains of striped snakehead (Channa striatus) wastes from aquaculture industry as an inhibitive assay for heavy metals. We discovered that the AChE was inhibited almost completely by Hg2+, Ag2+ and Cu2+ during an initial screening. When tested at various concentrations, the heavy metals exhibited exponential decay type inhibition curves. The calculated IC50 for the heavy metals Hg2+, Ag2+, Pb2+, Cu2+ and Cr6+ were 0.08432, 0.1008, 0.1255, 0.0871, and 0.1771, respectively. The IC50 for these heavy metals are comparable and some are lower than the IC50 values from the cholinesterases from previously studied fish. The assay can be carried out in less than 30 minutes at ambient temperature