Purification and characterization of L-arginine deiminase from Penicillium chrysogenum.

L-arginine deiminase (ADI, EC 3.5.3.6) hydrolyzes arginine to ammonia and citrulline which is a natural supplement in health care. ADI was purified from Penicillium chrysogenum using 85% ammonium sulfate, DEAE-cellulose and Sephadex G200. ADI was purified 17.2-fold and 4.6% yield with a specific activity of 50 Umg- 1 protein. The molecular weight was 49 kDa. ADI expressed maximum activity at 40oC and an optimum pH of 6.0. ADI thermostability was investigated and the values of both t0.5 and D were determined. Kd increased by temperature and the Z value was 38oC. ATP, ADP and AMP activated ADI up to 0.6 mM. Cysteine and dithiothreitol activated ADI up to 60 µmol whereas the activation by thioglycolate and reduced glutathione (GSH) prolonged to 80 µmol. EDTA, α,α-dipyridyl, and o-phenanthroline inactivated ADI indicating that ADI is a metalloenzyme. N-ethylmaleimide (NEM), N-bromosuccinimide (NBS), butanedione (BD), dansyl chloride (DC), diethylpyrocarbonate (DEPC) and N-acetyl-imidazole (NAI) inhibited ADI activity indicating the necessity of sulfhydryl, tryptophanyl, arginyl, lysyl, histidyl and tyrosyl groups, respectively for ADI catalysis. The obtained results show that ADI from P. chrysogenum could be a potential candidate for industrial and biotechnological applications.

Application of a novel biological-nanoparticle pretreatment to Oscillatoria acuminata biomass and coculture dark fermentation for improving hydrogen production.

BACKGROUND: Energy is the basis and assurance for a world's stable development; however, as traditional non-renewable energy sources deplete, the development and study of renewable clean energy have emerged. Using microalgae as a carbon source for anaerobic bacteria to generate biohydrogen is a clean energy generation system that both local and global peers see as promising. RESULTS: Klebsiella pneumonia, Enterobacter cloacae, and their coculture were used to synthesize biohydrogen using Oscillatoria acuminata biomass via dark fermentation. The total carbohydrate content in O. acuminata was 237.39 mg/L. To enhance the content of fermentable reducing sugars, thermochemical, biological, and biological with magnesium zinc ferrite nanoparticles (Mg-Zn Fe2O4-NPs) pretreatments were applied. Crude hydrolytic enzymes extracted from Trichoderma harzianum of biological pretreatment were enhanced by Mg-Zn Fe2O4-NPs and significantly increased reducing sugars (230.48 mg/g) four times than thermochemical pretreatment (45.34 mg/g). K. pneumonia demonstrated a greater accumulated hydrogen level (1022 mLH2/L) than E. cloacae (813 mLH2/L), while their coculture showed superior results (1520 mLH2/L) and shortened the production time to 48 h instead of 72 h in single culture pretreatments. Biological pretreatment + Mg-Zn Fe2O4 NPs using coculture significantly stimulated hydrogen yield (3254 mLH2/L), hydrogen efficiency)216.9 mL H2/g reducing sugar( and hydrogen production rate (67.7 mL/L/h) to the maximum among all pretreatments. CONCLUSION: These results confirm the effectiveness of biological treatments + Mg-Zn Fe2O4-NPs and coculture dark fermentation in upregulating biohydrogen production.

Heavy Metals Assessment and Health Risk to Consumers of Two Commercial Fish Species from Polyculture Fishponds in El-Sharkia and Kafr El-Sheikh, Egypt: Physiological and Biochemical Study.

Metal pollution is a major environmental concern worldwide, especially in Egypt. The aquaculture industry uses widespread artificial feeds to stimulate fish production, leading to metal accumulation in the aquatic environment. Heavy metal concentrations (HMCs) in sediments, water, and tissues were studied to study the effect of pollution levels on heamatological, and biochemical, immunological aspects of farmed fish as well as on human health. Results declared that the HMC levels in the water and sediment were significantly different between El-Sharkia and Kafr El-Sheikh fishponds (T-test, p < 0.05). This was supported by the metal pollution index in the water and sediment, indicating that El-Sharkia fishponds (ES fishponds) were more contaminated than Kafr El-Sheikh fishponds (KES fishponds). Also, HMCs in fish tissues were significantly increased in fish cultivated in ES fishponds than in KES fishponds. Haematological, immunological, and biochemical alterations of Bolti (Oreochromis niloticus) and Topara (Chelon ramada) fish were significantly different within the different fish species as well as the different fishponds. From the human health perspective, the THQ-HMC and HI-HMC associated with the consumption of muscle suggest a safe non-carcinogenic risk to human health. In contrast, cadmium poses a cancer risk to children who consume the muscular tissue of Bolti fish from ES fishponds, which should be regarded as a warning sign based on data indices and a human health perspective. In order to minimise HMC pollution in the aquaculture sector, it is advisable to take possible assessments and carry out continuous monitoring considering international WHO/FAO assessments.

Plastic wastes biodegradation: Mechanisms, challenges and future prospects.

The growing accumulation of plastic wastes is one of the main environmental challenges currently faced by modern societies. These wastes are considered a serious global problem because of their effects on all forms of life. There is thus an urgent need to demonstrate effective eco-environmental techniques to overcome the hazardous environmental impacts of traditional disposal paths. However, our current knowledge on the prevailing mechanisms and the efficacy of synthetic plastics' biodegradation still appears limited. Under this scope, our review aims to comprehensively highlight the role of microbes, with special emphasis on algae, on the entire plastic biodegradation process focusing on the depolarization of various synthetic plastic types. Moreover, our review emphasizes on the ability of insects' gut microbial consortium to degrade synthetic plastic wastes. In this view, we discuss the schematic pathway of the biodegradation process of six types of synthetic plastics. These findings may contribute to establishing bio-upcycling processes of plastic wastes towards biosynthesis of valuable metabolic products. Finally, we discuss the challenges and opportunities for microbial valorization of degraded plastic wastes.

Clozapine-Induced Cardiotoxicity: Role of Oxidative Stress, Tumour Necrosis Factor Alpha and NF-κß.

Safety concerns have been raised about clozapine-induced cardiotoxicity particularly in young patients. The exact mechanism of clozapine cardiotoxicity has not yet been thoroughly studied. This study aimed to investigate the possible mechanisms of clozapine-induced cardiotoxicity in a rat model. Young male Wistar rats were treated with clozapine (10, 15 and 25 mg/kg/day, i.p.) for 21 days. Haemodynamic and echocardiographic studies were performed for assessment of cardiac functions. Heart sections were studied histopathologically and immunohistochemically. Serum and cardiac markers of cardiotoxicity, oxidative stress, inflammation and apoptosis were evaluated. Heart sections of clozapine-treated animals showed increased cardiac inflammation that correlated with the clozapine dose. Serum levels of CK-MB and LDH levels increased, as did cardiac levels of TNF-α, MDA, NO, myeloperoxidase, 8-OHdG, caspase-3 and NF-κB p65. In contrast, GSH levels and GSH-Px activity decreased. Furthermore, immunohistochemical examination of the heart sections showed positive immunostaining for both 3-nitrotyrosine and caspase-3 in all clozapine-treated groups. Clozapine, particularly in relatively high doses, has a clear cardiotoxic effect. This cardiotoxicity is accompanied by increased myocardial oxidative stress, inflammatory cytokines, DNA damage and apoptosis with attenuation in antioxidant defences, thus explaining the previously reported myocarditis and pericarditis during clozapine therapy in clinical studies.

Clozapine-induced cardiotoxicity in rats: Involvement of tumour necrosis factor alpha, NF-κß and caspase-3.

Clozapine, an ideal antipsychotic drug for the treatment of resistant schizophrenia, is considered the most underutilised treatment for schizophrenia. However, safety concerns have been raised about clozapine-induced cardiotoxicity, which may lead to sudden death, particularly in young patients. The exact mechanism of clozapine cardiotoxicity has not yet been thoroughly studied. This study aimed to investigate the possible mechanisms of clozapine-induced cardiotoxicity in a rat model. Young male Wistar rats were treated with clozapine (10, 15 and 25 mg/kg/day, i.p.) for 21 days. Haemodynamic and echocardiographic studies were performed for assessment of cardiac functions. Heart sections were studied histopathologically and immunohistochemically. Serum and cardiac markers of cardiotoxicity, oxidative stress, inflammation and apoptosis were evaluated. Heart sections of CLZ-treated animals showed increased cardiac inflammation that correlated with the clozapine dose. Serum levels of CK-MB and LDH levels increased, as did cardiac levels of TNF-α, MDA, NO, myeloperoxidase (MPO), 8-OHdG, caspase-3 and NF-κB p65. In contrast, GSH levels and GSH-Px activity decreased. Furthermore, immunohistochemical examination of the heart sections showed positive immunostaining for both 3-nitrotyrosine and caspase-3 in all clozapine-treated groups. Clozapine, particularly in relatively high doses, has a clear cardiotoxic effect. This cardiotoxicity is accompanied by increased myocardial oxidative stress, inflammatory cytokines, DNA damage and apoptosis with attenuation in antioxidant defences, thus explaining the previously reported myocarditis and pericarditis during clozapine therapy in clinical studies.

Protective effect of captopril against clozapine-induced myocarditis in rats: role of oxidative stress, proinflammatory cytokines and DNA damage.

Clozapine (CLZ) is the most effective therapeutic alternative in the treatment of resistant schizophrenia. However, the cardiotoxicity of CLZ, particularly in young patients, has raised concerns about its safety. Captopril is a well-known angiotensin-converting enzyme inhibitor with antioxidant properties effective in treating hypertension and heart failure. The aim of this study was to investigate the protective effect of captopril against clozapine-induced myocarditis in rats and the possible mechanisms behind this effect. The effect of captopril treatment [5 or 10mg/kg/d, injected intraperitoneally (i.p.) for 21days] on the cardiotoxic effect of coadministered CLZ (25mg/kg/d, i.p.) was assessed. Myocarditis was assessed histopathologically, immunohistochemically and biochemically. Frozen heart specimens were used to determine the amount of lipid peroxides product (MDA), nitric oxide (NO), reduced glutathione (GSH), glutathione peroxidase (GSH-Px) activity, proinflammatory cytokines (TNF-α and IL-10) and DNA degradation product(8-OHdG). Coadministration of captopril with the tested doses of CLZ decreased the histological hallmarks and biochemical markers (CK-MP and LDH) of myocarditis. In addition, captopril attenuated the effects of CLZ on oxidative stress parameters, NO and serum and cardiac 8-OHdG levels. Captopril significantly attenuated the effect of CLZ on all measured parameters in a dose-dependent manner. These results suggested that captopril exerts a protective action against CLZ-induced myocarditis. Multiple mechanisms contribute to this effect, including a decrease in cardiac oxidative stress and proinflammatory cytokines production, modulation of antioxidant status and protection from oxidative DNA damage. Hence, captopril may be effective in reducing the incidence and severity of CLZ-induced myocarditis in humans.