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Nov 2024 DOI 10.14302/issn.2690-4829.jen-24-5291
The perpendicular orientation of the proximal alpha helix to the heme plane in chloroperoxidase (CPO) maximizes the influence of its intrinsic helix dipole that has been shown to reduce the “push-effect”, thereby increasing the heme redox potential and fine-tuning the catalytic capabilities of CPO. We investigated the effects of a disruption of the hydrogen bonding network between R26-N37 and A27-N33, formed by the proximal alpha helix, on the CPO structural stability and catalytic profile using site-directed mutagenesis and spectroscopy. The mutant CPO (R26A, N33A, and R26A/N33A) exhibited significant tertiary structural changes and distinct heme coordination, likely, due to destabilization of the proximal helix as a result of the disruption of the proximal hydrogen bonding network. In line with these observations, biochemical characterizations showed that all mutants displayed dramatically different activity profiles relative to that of the WT CPO. Mutant epoxidation and peroxidation activities were markedly enhanced, especially in the R26A/N33A CPO mutant. Moreover, all mutant CPO enzymes exhibited broader pH profiles in both epoxidation and peroxidation activities, including a shift in the optimal peroxidation activity towards pH 3.5 as opposed to pH 2.75. Conversely, the dismutation activity (pH 3.0-5.5) was almost completely lost while chlorination activity (pH 2.75-5.0) was virtually non-existent in all CPO mutants. Our results demonstrate the important role the R26-N37 and A27-N33 hydrogen bond pairs play in the heme coordination and tertiary structure of CPO defining its catalytic capabilities, and also suggest the importance of the proximal helix stability and orientation.
Apr 2023 DOI 10.14302/issn.2377-2549.jndc-23-4494
A planned experimental study on the production of green hydrogen by alkaline electrolysis is carried out by the Plan Composite Centered (PCC) method. The parameters studied are the concentration of the electrolyte, the distance between electrodes, the height of the electrodes, the total supply voltage of the electrolyser, temperature, and the electrolyte type. The results show that the effect of concentration, height, voltage and temperature are positive. However, the effect of the distance between the electrodes is negative. Electrolysis with potassium hydroxide (KOH) is more efficient than with sodium hydroxide (NaOH). The second-order interactions are weak, except for the voltage-temperature interaction which is significant. The results of the experimental study conducted in this work are in agreement with previous studies. Two a polynomial modeling (with KOH and with NaOH) suitable for predicting the flow of hydrogen produced are presented. Three optimizations of ascending constraints on the operating parameters to have a maximum hydrogen production and with a minimum of electrical energy and a minimum of concentration consumed are carried out.
Dec 2019 DOI 10.14302/issn.2641-7669.ject-19-3125
Emission of Hydrogen Sulphide (H2S) from dumpsites has become a global threat due to its impact on global climate change. This study assessed the spatial and seasonal levels of H2S emissions from 6 dumpsites (LA - LF), with portable air quality meter (AEROQUAL-Series 300). Results showed that the spatial level of H2S ranged from 1.40 ppm - 14.34 ppm. Based on seasonal variation level of H2S ranged from 1.88 ppm – 3.86 ppm (p<0.05), with higher values in wet season. Meanwhile H2S were not detected in the control station (LX). Based on model for Air Quality Index (AQI), H2S emission was predominantly rated as safe and moderate, except for the two stations in the central dumpsite (LE and LF). These results confirmed the emission of H2S from the dumpsite due to anthropogenic activities. We therefore recommend policies aimed at sequestration of H2S, by the reduce, reuse and recycle policy of waste stream.
Oct 2018 DOI 10.14302/issn.2471-2140.jaa-18-2400
Antioxidants can reduce oxidative stress in cells is used for the treatment of several disorders such as cancer, cardiovascular, and inflammatory diseases. The present study was evaluated the antioxidant potential of the Consciousness Energy Healing (The Trivedi Effect®) Treated human hepatoma cell line (HepG2) and Dulbecco's Modified Eagle Medium (DMEM) for the assessment of cell viability under hydrogen peroxide-induced oxidative stress. The Biofield Energy Treated HepG2 cells group was maintained for 23 days under standard conditions. On the next day, the cells were challenged with 1 mM of H2O2 for the generation of oxidative stress. The ability of the Biofield Energy Healing Treatment to protect from the oxidative stress was determined by MTT cell viability assay and compared with the negative control group. The percentage of cell viability was significantly (p≤0.001) increased by 13.6% in the Biofield Energy Treated DMEM group; while altered by 3.2% in the Biofield Energy Treated HepG2 cells group compared to the negative control groussp. Overall, the Biofield Energy Treated DMEM showed a better antioxidative protection against oxidative stress than HepG2 cells group, which was induced by H2O2. Therefore, the results envisaged that The Trivedi Effect®- Biofield Energy Healing Treatment has an impact on the protection of various vital organs from oxidative stress; which might be helpful in the development of powerful/energized growth medium for the accelerated study with a cost-effective manner.
Sep 2018 DOI 10.14302/issn.2377-2549.jndc-18-2224
This paper describes the theoretical foundations for the electric power and hydrogen generator that functions with zero energy input without violating the laws of thermodynamics. This generation system is a combined energy cycle consisting of the H2O=H2+1/2O2 reduction reaction performed by the water electrolytic cell and the H2+1/2O2=H2O oxidation reaction performed by the fuel cell. This electrolytic method differs from the conventional electrolytic scheme in that if a quasi-static process is assumed, so that the theoretical power requirement is only 17% of the total energy required. This method performs electrostatic-to-chemical energy conversion by electrostatic-induction potential-superposed electrolytic scheme. If this electrolytic cell that delivers the pure stoichiometric H2-O2 mixture is combined with a fuel cell to form an energy cycle, then this may lead to the concepts of a hydrogen redox electric power generator and a hydrogen redox hydrogen generator that use alkaline water electrolyte or solid polymer electrolyte membrane (PEM) for both electrolytic cell and fuel cell. In the power generator, part of power delivered by the fuel cell is returned to the electrolytic cell, and the remainder represents the net power output. According to calculations based on data from the operational conditions for commercially available electrolytic cell and fuel cell, more than 70% of the power delivered from the fuel cell can be extracted outside the cycle as net power output without the use of any external source of energy.
Jul 2017 DOI 10.14302/issn.2372-6601.jhor-17-1473
Objective: This study aimed to evaluate the relation of Lactate dehydrogenase (LDH) levels with stage of the disease and it role in monitoring tumor response to therapy in lymphoma patients. Methods. LDH levels were evaluated on 65 diagnosed Algerian children and compared to healthy control. Results: Our results revealed that LDH levels were significantly higher in untreated children with both hodgkin’s and non hodgkin’s lymphomas compared to control. Moreover, it was observed that the higher is the stage of disease, the more serum LDH level will be. However, there was a significant fall in serum LDH activity by completion of the chemotherapeutic courses. Conclusion: LDH plays an important role in tumor initiation and maintenance. The elevated serum LDH may reflect, release of the enzyme from malignant cells and suggest that they may reflect tumor burden and therefore correlate with disease progression.
Oct 2016 DOI 10.14302/issn.2377-2549.jndc-16-1212
Application of nickel in different industries has been developed and so contamination of natural water is a great concern due to its potentially toxic effects on living beings. Therefore, fast monitoring of Ni2+ in aqueous samples is important. In this work, we fabricated a sensitive optical sensor for determination of nickel in mineral water samples and hydrogen peroxide solutions. The optode was prepared by incorporation of 1-(2-pyridylazo)-2-naphthol and sodium tetraphenylborate in a plasticized poly (vinyl chloride) membranes containing dioctyladipate as a plasticizer. The influence of several parameters such as pH, base matrix, solvent mediator and ligand concentration were optimized. Comparison the obtained results with previously reported sensors revealed that the proposed method, in addition to fast and simplicity, provided good linear range (1.70–85.20 µmol L-1) and low detection limit (0.17 µmol L-1). The precision (relative standard deviation) was better than 1.55% for 7 replicate determinations of 17.10 µmol L-1 of Ni in various membranes.
Aug 2025 DOI 10.14302/issn.3070-1937.ijbt-25-5408
Background and Objectives Platelets are small, anucleate blood cells produced in the bone marrow, primarily involved in blood coagulation. Platelet concentrate is a vital blood product with extensive applications. However, its short lifespan and limited donor availability pose global challenges. This study aimed to follow the trend of platelets 5 during days of storage. Material and Methods We studied on 40 platelet bags and analyzed glucose levels, lactate dehydrogenase (LDH), bacterial culture, and apoptosis using flow cytometry with Annexin V-PI over three consecutive days (first, third, and fifth) post-blood collection. Data were analyzed using SPSS. Results No significant correlations were found between age, blood group, or gender and the variables studied. No bacterial growth was detected. Glucose levels decreased significantly from day 1 (382 mg/dl) to day 5 (298 mg/dl). The average platelet apoptosis increased significantly from 3.65% on day 1 to 9.06% on day 5. Significant correlations were observed between glucose levels and apoptosis on days 3 (p<0.05) and 5 (p<0.01). No correlation found between LDH and apoptosis or necrosis, although a significant relationship between necrosis and apoptosis was noted on day 5 (p=0.003). Conclusion These findings suggest that while demographic factors do not influence the studied variables, the significant decrease in glucose levels correlates with increased platelet apoptosis over time, highlighting potential metabolic interactions that warrant further investigation. Highlights 1. The study revealed subtle variations in metabolic markers related to donor demographics, particularly gender and age. Understanding these differences can inform targeted donor selection strategies to optimize platelet quality. 2. A significant negative correlation was found between glucose levels and apoptosis rates, indicating that as glucose decreases, platelet viability declines. This relationship highlights the need for careful monitoring of glucose levels during storage to maintain platelet function. 3. Fluctuations in lactate dehydrogenase (LDH) levels were correlated with increasing rates of apoptosis, suggesting that LDH could serve as a valuable biomarker for assessing platelet quality throughout the storage period. This finding could lead to improved storage protocols and enhanced transfusion safety.
Aug 2025 DOI 10.14302/issn.2690-4829.jen-25-5617
Redox enzymes are a type of enzyme that catalyzes redox reactions, that is, electron transfer reactions between two chemical species. Redox enzymes are essential for many biological processes, including cellular respiration, photosynthesis, energy production, and the elimination of free radicals. They are divided into two main types: oxidoreductases and electron transferases. Oxidoreductases catalyze the direct transfer of electrons between two chemical species, while electron transferases catalyze electron transfer by cofactors. Examples of redox enzymes include cytochrome c oxidase, NADH dehydrogenase, succinate dehydrogenase, and catalase. Each of these enzymes play an important role in cellular metabolism and organism homeostasis.
Sep 2024
A new approach to protecting the side chain of lysine during solid-phase peptide synthesis in an aqueous environment is explored. The objectives of this study are twofold: 1) to reduce the consumption of chemical solvents in peptide synthesis, and 2) to lower the costs associated with lysine side chain protection. In an effort to minimize the usage of organic solvents in amino acid side chain protection for peptide synthesis,18-crown-6 molecule (18C6) was introduced as protective agent for Lysine (Lys) side chain protection. 18C6 is utilized to protect ε-amino group of on side chain of Lys in peptide synthesis. 18C6 features cavity at its center. The 18C6 molecules exhibit a unique and distinctive host-guest affinity towards ε-amino group on side chain of Lys. 18C6 bind to Lys side chain through hydrogen bonds with ε-amino group of Lys, eliminating the need for chemical reactions in organic solvents to attach protective molecules to the side chain groups. This simplifies the side chain protection procedure significantly for Lys, reducing the consumption of organic solvents. Consequently, this procedure offers advantages for contributing to a greener planet by minimizing chemical reactions and organic solvent usage. Additionally, the cost of side chain protection of Lys is dramatically reduced.
Apr 2021 DOI 10.14302/issn.2576-6694.jbbs-21-3773
L-tryptophan is an essential α-amino acid, necessary for the normal growth in newborns, nitrogen balance in adults, protein synthesis, precursor of serotonin, melatonin, niacin, and albeit inefficiently in human, also the precursor of indole alkaloids and auxins in plants. This current study was designed to investigate the impact of the Trivedi Effect®-Biofield Energy Healing Treatment (Blessing) on the structural properties and the isotopic abundance ratio of L-tryptophan using LC-MS analytical technique. L-tryptophan sample was divided into two parts, one part of L-tryptophan was considered as the control sample (no Biofield Energy Treatment was provided), while the second part was treated with the Trivedi Effect®-Consciousness Energy Healing Treatment/Blessing remotely by a renowned Biofield Energy Healer, Dahryn Trivedi and termed as the treated sample. The mass spectra of both the control and treated samples with respect to the chromatographic peak at retention time (Rt) 2.1 minutes exhibited the mass of the molecular ion peak adduct with hydrogen ion at m/z 205.08 (calcd for C11H13N2O2+, 205.1), along with low molecular fragmented mass peaks at m/z 188, 159, and 102 for C11H12N2O2+, C10H11N2+, and C8H6+, respectively were also observed. The isotopic abundance ratio of PM+1/PM (2H/1H or 13C/12C or 15N/14Nor17O/16O) in the treated L-tryptophan was significantly increased by 35.93% compared with the control sample. Hence,the 13C, 2H, 15N, and 17O contributions from C11H13N2O2+ to m/z 206.08 in the treated L-tryptophan was significantly increased compared to the control sample. It could be hypothesized that the changes in the isotopic abundance and mass peak intensities due to the modification in nuclei possibly through the interference of neutrino particles using the Trivedi Effect®-Consciousness Energy Healing Treatment. The Biofield Energy Treated/Blessed L-tryptophan with increased stable isotopic abundance ratio might have changed the physicochemical properties with higher force constant in the molecule. The new form of treated L-tryptophan would be a better and more stable in the supplements, nutraceutical, and pharmaceutical formulations, which would be advantageous for the prevention and treatment of pellagra, depression, kynurenine. It could also maintain the normal label of tryptophan and avoid increase of its metabolite, lower the neurotoxin and a metabotoxin behavior, glutaric aciduria type I (glutaric acidemia type I) disorder, eosinophilia-myalgia syndrome (EMS), incurable and sometimes fatal flu-like neurological condition, etc. As tryptophan is the precursor for the plant hormones like indole alkaloids and auxins, hence, this treated L-tryptophan would be advantageous for the improvement of yield, productivity, and quality of crops and other plants.
Mar 2020 DOI 10.14302/issn.2766-869X.jfd-20-3193
Fusarium solani NVS671 identified from infected sugarcane stem of Co 671 as a new pathogen was subjected to various cultural conditions to understand its physiological profile. In continuation with our previous work, cultural analysis was carried out under in vitro condition by supplementing various carbon and nitrogen sources in Czapek-Dox agar (CDA). Under different hydrogen ion concentrations, it was found that the growth of Fusarium solaniNVS671 was less at pH 4 and pH 10 and could reach up to 5 cm after 7 days of incubation on Potato Dextrose Agar (PDA). It is observed that the pH around 7 to 8 was optimum for the growth of F. solani. Ten different nitrogenous (N) sources and nine different carbon sources were tested on CDA medium to know their effect on the mycelial growth rate and characteristics of the fungus. Among the N sources evaluated, ammonium chloride (7.96±0.11 cm) was found to be most efficient for mycelial growth promotion followed by ammonium nitrate (7.7±0.1 cm) and ammonium sulphate (7.3±0.1 cm). The most preferred carbon source recorded to promote best radial mycelial growth was starch (7.96±0.05 cm) and sucrose (7.93±0.05 cm). Capabilities of using different carbon and nitrogen sources and ability to grow at different pH levels may allow species to adapt to specific soil conditions.This study is important to understand the physiology and metabolite preference of F. solani.
Jan 2020 DOI 10.14302/issn.2379-7835.ijn-20-3159
Pyruvate holds superior biomedical properties in increase of hypoxia tolerance, correction of severe acidosis, exertion of anti-oxidative stress and protection of mitochondria against apoptosis, so that it improves multi-organ function in various pathogenic insults. Particularly, pyruvate preserves key enzyme: pyruvate dehydrogenase (PDH) activity through direct inhibition of pyruvate dehydrogenase kinas (PDK), as a PDH activator, in hypoxia. Therefore, pyruvate is robustly beneficial for cell/organ function over citrate, acetate, lactate, bicarbonate and chloride as anions in current medical fluids. Pyruvate-enriched oral rehydration salt/solution (Pyr-ORS) and pyruvate-based intravenous (IV) fluids would be more beneficial than WHO-ORS and current IV fluids in both crystalloids and colloids, respectively. Pyruvate-containing fluids as the new generation would be not only a volume expander, but also a therapeutic agent simultaneously in fluid resuscitation in critical care patients. Pyruvate may be also beneficial in prevent and treatment of diabetes, aging and even cancer. Pyruvate clinical applications indicates a new revolutionary medical advance, following the WHO-ORS prevalence, this century.
Jun 2019 DOI 10.14302/issn.2377-2549.jndc-19-2765
Tetrodotoxin (TTX) is a potent neurotoxin. Its name derives from Tetraodontiformes, an order that includes pufferfish, porcupinefish, ocean sunfish, and triggerfish; several of these species carry the toxin. Although tetrodotoxin was discovered in these fish and found in several other aquatic animals (e.g., in blue–ringed octopuses, rough–skinned newts, and moon snails), it is actually produced by certain infecting or symbiotic bacteria like Pseudoalteromonas, Pseudomonas, and Vibrio as well as other species found in animals. Parameters such as FT–IR and Raman vibrational wavelengths and intensities for single crystal Tetrodotoxin (TTX)are calculated using density functional theory and were compared with empirical results. The investigation about vibrational spectrum of cycle dimers in crystal with carboxyl groups from each molecule of acid was shown that it leads to create Hydrogen bounds for adjacent molecules. The current study aimed to investigate the possibility of simulating the empirical values. Analysis of vibrational spectrum of Tetrodotoxin (TTX) is performed based on theoretical simulation and FT–IR empirical spectrum and Raman empirical spectrum using density functional theory in levels of F/6–31G*, HF/6–31++G**, MP2/6–31G, MP2/6–31++G**, BLYP/6–31G, BLYP/6–31++G**, B3LYP/6–31G and B3LYP6–31–HEG**. Vibration modes of methylene, carboxyl acid and phenyl cycle are separately investigated. The obtained values confirm high accuracy and validity of results obtained from calculations. Molecular structure of Tetrodotoxin (TTX) 1–42.
Feb 2019 DOI 10.14302/issn.2329-9487.jhc-19-2582
Introduction Heart disorders are the major concern of population health worldwide. According to WHO estimates 2018, 17.9 million peoples were died due to cardiovascular disorders. Aim The aim of this study was to investigate the cardioprotective activity of Biofield Energy Treated test item, Dulbecco's Modified Eagle Medium (DMEM) using rat cardiomyocytes (H9c2). Methods The test item (DMEM) was divided into three parts, first part received one-time Biofield Energy Treatment by a renowned Biofield Energy Healer, Mahendra Kumar Trivedi and was labeled as the one-time Biofield Energy Treated (BT-I) DMEM, while second part received the two-times Biofield Energy Treatment and is denoted as BT-II DMEM. The third part did not receive any treatment and defined as the untreated DMEM group. Results Cell viability of the test samples by 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay showed 89.03% and 98.49% in the BT-I and BT-II groups, respectively suggested a nontoxic and safe in nature of the tested test item. The BT-I group showed 16.01% restoration of cell viability. The level of lactate dehydrogenase (LDH) was significantly inhibited by 50.37% and 49.35% in the BT-I and BT-II groups, respectively compared to the untreated DMEM group. Moreover, percent protection of creatine kinase-myocardial band (CK-MB) by 49.48% and 59.79% in the BT-I and BT-II groups, respectively, compared to the untreated DMEM group. Reactive oxygen species (ROS) level in terms of mean fluorescence unit (FU) was reduced by 6.64% in the BT-I group than untreated DMEM. Besides, BT-I and BT-II groups significantly increased the level of % apoptotic cells by 63.16% and 97.37% (p≤0.05), respectively than untreated DMEM. Conclusion Allover, results envisaged that Biofield Treatment significantly improved different cardiac parameters. Thus, Biofield Energy Treatment (The Trivedi Effect®) could be utilized as a cardio-protectant against several cardiac disorders such as coronary artery disease, heart attack, arrhythmias, heart failure, congenital heart disease, cardiomyopathy, etc.
Aug 2018 DOI 10.14302/issn.2377-2549.jndc-18-2220
Diphenhydramine HClis a weakly fluorescent drug having tertiary amine group forming ion pair complex with eosin Y in dichloromethane at pH 5 in disodium hydrogen phosphate-citric acid buffer solution. The complex formation was the basis for the development of new analytical method for determination of active diphenhydramine in pharmaceutical formulations. The stoichiometric ratio between diphenhydramine and eosin Y was studied by mole ratio method and found to be 2:1. The ion-pair complex showed maximum fluorescence emission intensity at 554 nm with excitation at 259 nm. The linear dynamic range was obtained in the concentration range of 2-22 µg mL-1 with a linear equation of FI = 0.361 + 13.675 C. The apparent Gibb’s free energy (ΔGº) was calculated and found to be -80.783 KJ mol-1, confirmed the feasibility of the reaction. The proposed method was successfully applied to the determination of diphenhydramine HCl in pharmaceutical formulations and in good agreement with the reference method.
Mar 2018 DOI 10.14302/issn.2575-1212.jvhc-18-2012
Bisphenol A (BPA) is an endocrine disruptor with a weak estrogenic effect used in industry as a component of food cans. We aimed to study the toxic effects of BPA on mRNA expression of steroidogenic genes and testicular structure in mature male rats. Animals were divided into 3 groups: vehicle control rats as first group, while second group received 10 µg/kg BW and third group received BPA 15 µg/kg BW orally every alternate day for a period of 105 successive days. Serum testosterone level, mRNA expression of genes related to steroid synthesis, histopathological examination, spermatogenesis index and number of Leydig cells were evaluated in this study. Lower serum hormone levels were observed in both BPA-treated groups as compared to the control group. The gene expression patterns of steroidogenic acute regulatory protein (StAR), cytochrome P450 17a(CYP17a) and 3β-Hydroxysteroid dehydrogenase (3β-HSD) were significantly down-regulated in BPA-treated rats compared to control group. Meanwhile, the expression of aromatase (CYP19) and lutinizing hormone receptor (LHR) was significantly up-regulated. Histopathological lesions were observed in the testes and epididymis of BPA-treated rats. Spermatogenesis index and the number of Leydig cells were significantly decreased in BPA-treated groups compared with the control group. This study highlights negative effect of BPA on steroidogenic genes and testicular structure in male rats.
Aug 2017 DOI 10.14302/issn.2638-4469.japb-17-1534
Algae are a class of photosynthetic organisms found in both marine and freshwaters habitats. As these organisms have a short doubling time, they are considered among fastest growing creatures. They have different pathways to fix atmospheric carbon dioxide and to efficiently utilize the nutrients to convert it into biomass. In few years, a focus has been shifted towards these organisms due to their food and fuel production capability. In fuel industry algae biofuels have been emerged as a clean, nature friendly, cost effective solution to other fuels. Algae fuels are categorized into bio-ethanol, biogas, bio-hydrogen, biodiesel and bio-oil. Algae as a food have been explored for different applications as in production of single cell proteins, pigments, bioactive substances, pharmaceuticals and cosmetics. The present review has been prepared to throw a light on enormous applications of algae as food and fuel and also to provide some information about different commercially available algae products.
Dec 2016 DOI 10.14302/issn.2471-2140.jaa-16-1378
Methionine (Met) is a nutritionally essential amino acid and has been widely demonstrated to improve cellular oxidative balance and mediate oxidative stress. Met targets reactive oxygen species (ROS) directly by being oxidized to Met sulfoxide (MetO) 1. Met can be metabolized to cysteine (Cys) through transsulfuration pathway, which is further metabolized to glutathione (GSH), taurine, and hydrogen sulfide (H2S). All these metabolites exhibit antioxidant functions in various models (reviewed at 2). More recently, Met also has been demonstrated to enhance cellular oxidative tolerance via pentose phosphate pathway (PPP) 3, which contributes to the balance of cellular reducing power and accelerates the reduction reaction of MetO and GSH oxidative product GSSH back to Met and GSH.
Aug 2016 DOI 10.14302/issn.2474-9273.jbtm-16-1151
Oxidative stress mediated neural cell death is thought to be involved in the progression of secondary cell injury following brain trauma. Agents that can block oxidative stress-related injury could be potential therapies for TBI. Resveratrol, a polyphenol found in plants and red wine, is cytoprotective due to its potent antioxidant activities. To further understand how resveratrol could affect oxidative stress-induced injury, we hypothesized that the cytoprotective activities of resveratrol could be dose-dependent. In this study, resveratrol-induced cytoprotection was evaluated in cultured astrocytes. Primary rat astrocytes were cultured in T-75 flasks to a confluence of 80% before being plated onto 96-well plates. After 24 hours of acclimation, astrocytes were treated with various doses of hydrogen peroxide (H2O2) (0.1, 0.25, 0.5 and 1 µM) and resveratrol (25, 50, 75, 100 µM), respectively. Cell viability was determined 24 hours later using Alamar Blue Assay. Treatment of astrocytes with 0.5 mM H2O2, left 65% of astrocytes non-viable whereas treatment of astrocytes with 0.1 mM H2O2 had no effect on astrocytes viability; whereas 1 mM, H2O2 caused total loss of astrocyte viability. Resveratrol treatment at 75 µM and 100 µM has reduced 0.5 mM H2O2-induced cytotoxicity in astrocytes by 50%. Immunostaining with GFAP also confirmed these findings about the cytoprotective effects of resveratrol in astrocytes exposed to H2O2. These results suggest that resveratrol could be a potential neuroprotective agent in TBI due to its antioxidant properties. Further studies are needed to evaluate the long- term effects of resveratrol in animal models of TBI.
Jun 2014 DOI 10.14302/issn.2328-0182.japst-13-288
ALS is the neurodegenerative disease which is caused due to breakdown in interaction between UBL and rpn1. In this study, we explore the interaction of UBL and rpn1 which is involved in protein degradation. Protein recycling system plays a crucial role in degradation of deformed or damaged proteins. Task of degradation of damaged ubiquitinated proteins is completed by proteasome with the help of ubiquilin2 protein which links 19s proteasome and poly-Ub chain attached to damaged protein. More specifically, N-terminal UBL domain interacts with rpn1 subunit of base complex of 19s proteasome and C-terminal UBA domain interacts with tetra poly-Ub chain attached to damaged protein. In present study, UBL domains are docked against homology modeled rpn1 with the help of Patch dock server. Further the docked structures are refined using fire dock server and best docked structure is chosen having global energy -16.71. Best docked structures are analyzed using swiss-pdb viewer software to show hydrogen bonds between interacting proteins. Here we explore a mutation E6A and P11A in UBL structure with the help of YASARA which is significantly increasing the interaction between interacting proteins in terms of hydrogen bonds.