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Jun 2023
S. Prakasha Gowda A.Corresponding author
Insulin is a frequent peptide hormone addition in serum-free mammalian cell culture media. It contributes in a variety of biological functions, including as promoting cell proliferation, cell cycle progression, and glucose uptake. However, it is unknown how stable insulin is under in vitro cell culture media treatment conditions. The instability of insulin in aqueous solutions has caused a number of issues, necessitating the development of new therapeutic strategies that can keep insulin stable and functioning. Such choices are required to accommodate updated insulin delivery guidelines as well as the storage and transportation of insulin. To preserve structural and functional integrity, protein medicines are frequently stabilized with antioxidants in aqueous solutions. In the present study, the effects of the antioxidants disodium ethylenediaminetetraacetic acid dihydrate (EDTA) and sodium selenite (Se) and their ability to scavenge free radicals on insulin stability in the medium Dulbecco's Modified Eagle Medium (DMEM) and Roswell Park Memorial Institute (RPMI) were examined. To investigate the stability of human recombinant insulin, in vitro serum-free DMEM and RPMI media were utilized for 5 days at 37˚C containing different EDTA and Se concentrations. Reversed phase high performance liquid chromatography (RP-HPLC) was used to detect and quantify insulin. Sodium dodecyl sulfate polyacrylamide gel (SDS-PAGE) electrophoresis was used to assess conformational stability. The results demonstrated that, when EDTA and Se were added separately to DMEM and RPMI media, insulin stability was improved compared to when neither compound was added.
Aug 2020 DOI 10.14302/issn.2694-2275.jzr-20-3435
Radwan EHCorresponding author
Damanhour University, Faculty of Science, Zoology department, Egypt
Background Nephrotoxicity is a complication due to the effect of some toxic chemicals on kidney. Current study planned to screen the effect of Trigonella foenumaqueous seeds extracts on EDTA induced nephrotoxicity. Trigonella foenum known for its various medicinal properties is also a natural antioxidant and a free radical scavenger with no documented evidence as a nephron-protective agent. Objective To investigate the protective effects of aqueous seed extracts of Trigonella foenum. Material and Methods The present study was used 40 male albino rats (Rattus albinus) with weight of (150 ± 10) g with divided into four groups: control gp; EDTA gp (95 mg/kg); Trigonella foenum gp (500 mg/kg) and EDTA + Trigonella f oenum gp by gastric tube daily for 4 weeks. Blood urea, creatinine, GFR, creatinine clearance, MDA and GPx analyses and microscopic examination of kidney were performed. Results In the present study, Blood samples were taken from all groups and concentration of serum urea, creatinine, GFR, Creatinine clearance, MDA and GPx were determined. Histopathological observations were observed in kidney tissue. Data were analyzed using analysis of variance (ANOVA). EDTA induced an increase in urea and creatinine as well as there was a decrease in the concentration of GFR and creatinine clearance. The level of MDA was increase while the concentration of GPx was decrease in the serum of EDTA group. The aqueous extracts of Trigonella seeds significantly prevented renal damage by normalizing increased levels of renal markers. The correction of oxidative stress biomarkers was consistent with amelioration of the histopathological changes induced by EDTA. Hence, it is suggested that ameliorative effect of aqueous extract of Trigonella foenumagainst EDTA induced nephrotoxicity. Conclusion The present data suggest that aqueous extract of Trigonella foenum exhibits reno-protective effect in EDTA induced renal damage.
Jan 2020 DOI 10.14302/issn.2379-7835.ijn-19-3139
Q. Almulaiky YaaserCorresponding author
Chemistry Department, Faculty of Sciences and Arts, University of Jeddah, Khulais, P.O. Box 355, Khulais, 21921, Saudi Arabia
In this study, a peroxidase from new source was purified using ion exchange and gel filtration techniques. The recovery for peroxidase activity was 19% with 11-fold purification and specific activity of 749 unit/mg protein. Purified peroxidase demonstrated a molecular mass of 39 kDa using gel filtration and was confirmed as a single band on SDS-PAGE. The purified peroxidase revealed a broad optimum pH activity at 6.0-6.5 and 50°C temperature. The kinetic parameters for purified peroxidase toward H2O2 and guaiacol as substrates were found to be Km = 3.355, 5.395 mM, Kcat = 99.52, 79.56 s-1 and Vmax =1.531, 1.242 µmole ml-1 min-1, respectively. The catalytic efficiency (kcat/Km) of the purified peroxidase was 14.75 and 29.66 s−1 mM−1 for guaiacol and H2O2, respectively. Peroxidase activity was observed to be enhanced by Cu2+, Co2+, Ni2+ and inhibited in the presence of Sn2+, Al3+, Hg2+, NaN3, EDTA and urea. Characterization showed that peroxidase purified from C. forskohlii has the ability to be used for food industrial applications.
Aug 2017 DOI 10.14302/issn.2690-4721.ijcm-17-1676
W. Taylor-Robinson AndrewCorresponding author
School of Health, Medical & Applied Sciences, Central Queensland University, Brisbane, QLD 4000, Australia
Malaria is a mosquito-transmitted infectious disease caused by intracellular protozoan parasites of the genus Plasmodium. In the absence of prompt and appropriate treatment contraction of primary infection by a human being often represents a medical emergency since it may progress rapidly to life-threatening complications. Exposure to parasites activates the immune system resulting in, among other effects, the release of reactive oxygen intermediates (ROI). This has the potential to induce oxidative damage, thereby causing cellular destruction, and hence to have a severe effect on vital organs of the body. Overexpression of ROI leads to immunosuppression and is a causal factor in the development of malaria-related disease symptoms. However, the body possesses various defence mechanisms, notably including the production of antioxidants, which are capable of reducing the cellular effects of ROI. Antioxidants are either sourced exogenously from the diet or synthesized through different intracellular mechanisms. Antioxidants that include glutathione peroxidase, catalase, EDTA and vitamin C suppress the initial production of ROI. Others such as uric acid, superoxide dismutase and vitamin E may also inhibit potentially damaging products of ROI metabolism. Current anti-malarial drugs often have damaging side-effects, as exemplified by memory impairment following treatment for cerebral malaria. Recent studies have explored the potential use of antioxidants alone or in combination with anti-malarials as a therapeutic means to negate Plasmodium-induced oxidative stress and its associated metabolic complications. It is indicated that when utilized in an adjuvant capacity antioxidants of natural and synthetic origin may improve anti-malarial therapy by causing less damage to the host during malaria infection.