Protein Oxidation Levels Research Articles

The investigation of the efficacy of the prodrug DDI-10 against Newcastle disease virus infection in young chicken

Introduction. Newcastle disease virus (NDV) is the deadliest virus in the poultry industry. Many RNA viruses induce oxidative stress on the host during its pathogenesis, NDV being one among them. The present study aims to evaluate the protective property of novel phosphorylated DDI-10 in vivo in experimentally infected chicken. Material and methods. NDV induced oxidative damage in the liver and lung were measured by determining antioxidant enzyme levels, protein oxidation and nitration using ELISA, Western blot and immuno co-localization assay. Results. Glutathione dependent enzymes GPx, GST, and GR were significantly decreased in the NDV infected group due to pathogenesis; DDI-10 treatment was shown to significantly increase this reduced enzyme activity. In comparison to the healthy control group, protein oxidation and nitration levels were significantly increased in the NDV infected group. In the DDI-10 treated group, the oxidation and nitration levels were decreased compared to the NDV infected group. Further estimation of protein nitration and oxidation in western blot and immune co-localization assays correlated with the ELISA results; an intensified band was demonstrated in the NDV infected tissue group, in addition to a high number of co-localized cells being present in immunofluorescence-stained sections compared to control chicken tissues. These alterations were noticeably reduced in novel phosphorylated DDI-10 treatment group. Conclusions. These results suggest that DDI-10 mitigates NDV induced oxidative stress, subsequently exerting an ameliorative effect against NDV pathogenesis.

Effect of chitosan/starch aldehyde-catechin conjugate composite coating on the quality and shelf life of fresh pork loins.

Fresh pork is susceptible to oxidation and spoilage. Edible coating containing antioxidant and antimicrobial agents can create moisture and oxygen barriers around pork and inhibit oxidation and microbial growth in the pork. In this study, chitosan in combination with starch aldehyde-catechin conjugate (SACC) was used as a novel edible coating material for preserving fresh pork loins at chilled storage (4 ± 1°C) for 14 days. Effect of chitosan/SACC composite coating on the quality of pork loins including weight loss, colour, pH value, microbial spoilage, lipid oxidation, protein oxidation, texture and sensory attributes during chilled storage was determined. Chitosan and SACC had synergistic antioxidant and antimicrobial actions. As compared with uncoated and chitosan coated pork loins, chitosan/SACC coated pork loins showed lower weight loss (7.16%), pH value (5.99), total viable count (7.11 log CFU g-1 ), total volatile base nitrogen content (130.2mg kg-1 ), lipid oxidation level (0.47 mg malondialdehyde kg-1 ), protein oxidation level (0.047 mmol free thiol group g-1 ) and shear force (27.40 N) on day 14. Meanwhile, chitosan/SACC composite coating effectively maintained the colour, micro-structure and sensory attributes of pork loins throughout chilled storage period. The shelf life of pork loins was extended from 8 days (uncoated samples) to 14 days by chitosan/SACC composite coating. Chitosan/SACC composite coating effectively retarded the oxidation and spoilage of pork loins during chilled storage. © 2022 Society of Chemical Industry.

Pathomorphological and biochemical study of the golden grey mullet Chelon auratus (Risso, 1810) in the waters of the southwestern Crimea (the Black Sea)

The golden grey mullet Chelon auratus (Risso, 1810) (Mugilidae) is a valuable commercial and recreational species ranking first in terms of catch volume of the Black Sea indigenous mullets. The importance of this species in the regional fishery among demersal fish requires the development of a system for assessing its health status. Such research is based on an integrated approach involving biochemical and pathomorphological methods: these allow to investigate the alterations in fish prior the occurrence of visible manifestations, disruption of the processes of growth and reproduction, reduction of commercial size, and decrease in abundance. The aim of our work was to study both pathomorphological alterations and several biochemical parameters of golden grey mullet tissues for assessing its health status. Fish visual examination and pathological autopsy were carried out. For histological analysis, samples of the gills, liver, kidneys, gastrointestinal tract, spleen, and pancreas were fixed in Davidson’s solution and processed by standard methods. Based on the histological studies, the fish health status was investigated by a modified semi-quantitative analysis of alterations according to the Bernet et al. protocol and by assessing the distribution of lesion in organs using a scoring system. We determined the importance factors of alterations for C. auratus, the values of organ alteration indices, and the total index of fish pathology. The biochemical studies permitted to reveal the level of protein oxidation, lipid and urea peroxidation, and the activity of aminotransferases and alkaline phosphatase in the liver; moreover, we quantified albumin and glucose concentration in the blood serum. In the organs of the golden grey mullet, the histopathological alterations referring to four types of the reaction patterns were detected (circulatory disorders, regressive and progressive alterations, and inflammatory processes). Furthermore, parasites representing several species of different systematic groups (Protozoa, Monogenea, Trematoda, and Nematoda) were identified. It was established that the most severe histopathological alterations were caused by a parasitic protozoan, presumably Ichthyophonus sp. When carrying out a semi-quantitative analysis of alterations, the mullets were conventionally divided into conditionally healthy individuals and infected ones. Pathomorphological data were obtained, and the set of biochemical parameters was compared in these two groups. Significant differences were revealed in the values of organ alteration indices in C. auratus in the kidneys, liver, gastrointestinal tract, and pancreas. The values of the total index of fish pathology also differed significantly. The biochemical studies revealed a significant increase in urea content in the liver of fish from the group 2, that may indicate the kidney and gill excretory dysfunction (it was confirmed histologically). No significant differences were found in the level of lipid peroxidation, protein oxidation, and activity of aminotransferases in the liver of conditionally healthy and infected fish. The results of our investigation confirm high informativeness of the studied parameters for assessing the health status of the golden grey mullet.

Ketorolac tromethamine alleviates IL-1β-induced chondrocyte injury by inhibiting COX-2 expression.

Osteoarthritis (OA) is one of the most frequently diagnosed chronic diseases, and its prevalence is rising as life expectancy increases. The present study was designed to investigate the role of ketorolac tromethamine (KT) in OA by establishing an in vitro model in ATDC5 cells. The OA model was established through induction using 10 ng/ml IL-1β. KT was then used to treat the ATDC5 cells. An MTT assay was adopted to detect the viability of ATDC5 cells with or without IL-1β induction, and cyclo-oxygenase-2 (COX-2) expression in IL-1β-induced ATDC5 cells was measured via reverse transcription-quantitative (RT-q)PCR and western blotting. To explore the effects of KT on proliferation and apoptosis in IL-1β-induced ATDC5 cells, COX-2 was overexpressed and RT-qPCR was employed to detect the mRNA expression of COX-2. The viability of IL-1β-induced ATDC5 cells was detected by using a Cell Counting Kit-8 assay. In addition, levels of apoptosis and apoptosis-related proteins were determined using TUNEL staining and western blotting, respectively. Additionally, the effects of KT on oxidative stress in IL-1β-induced ATDC5 cells were also investigated. The expression levels of nitric oxide (NO) and inducible NO synthase (iNOS) were detected via NO kit assay and western blotting, respectively. In addition, the expression levels of oxidative stress-related proteins, including reactive oxygen species (ROS), superoxide dismutase (SOD) and prostaglandin E2 (PGE2), were determined using ELISA. To investigate the effects of KT on the inflammatory response and extracellular matrix (ECM) degradation, ELISA and western blotting were adopted to detect inflammatory-related proteins and ECM degradation-related proteins. Results from MTT assay indicated that KT decreased ATDC5 cell viability in a concentration-dependent manner. The expression of COX-2 was found to be downregulated in IL-1β-induced ATDC5 cells after treatment with KT, according to RT-qPCR and western blotting results. KT inhibited apoptosis and the expression levels of NO, iNOS and inflammatory-related proteins in IL-1β-induced ATDC5 cells, while COX-2 overexpression reversed these inhibitory effects. However, the increased proliferation of IL-1β-induced ATDC5 cells after the stimulation of KT was decreased by COX-2 overexpression. Additionally, KT upregulated Bcl-2, SOD, type II collagen and aggrecan expression levels in IL-1β-induced ATDC5 cells, whereas Bax, ROS, matrix metallopeptidase (MMP)1 and MMP13 expression levels were downregulated. KT promoted the proliferation of IL-1β-induced ATDC5 cells, whereas COX-2 overexpression reversed the promotive effects of KT, revealing that KT could alleviate IL-1β-induced chondrocyte injury by suppressing COX-2 expression.

Chinese Tea Alleviates CCl4-Induced Liver Injury through the NF-κBorNrf2Signaling Pathway in C57BL-6J Mice.

Liver injury is a life-threatening condition that is usually caused by excessive alcohol consumption, improperdiet, and stressful lifestyle and can even progress to liver cancer. Tea is a popular beverage with proven health benefits and is known to exert a protective effect on the liver, intestines, and stomach. In this study, we analyzed the therapeutic effects of six kinds of tea on carbon tetrachloride (CCl4)-induced liver injury in a mouse model. The mice were injected with 10 mL/kg 5% CCl4 to induce liver injury and then given oral gavage of green tea, yellow tea, oolong tea, white tea, black tea, and dark tea, respectively. The serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured, and the expression levels of inflammation and oxidative stress-related proteins in the liver tissues were quantified. All six kinds of tea partly reduced the liver index, restored the size of the enlarged liver in the CCl4 model, and decreased the serum levels of ALT and AST. Furthermore, the highly fermented dark tea significantly reduced the expression levels of NF-κB and the downstream inflammatory factors, whereas the unfermented green tea inhibited oxidative stress by activating the antioxidant Nrf2 pathway. Taken together, tea can protect against liver inflammation, and unfermented tea can improve antioxidant levels. Further studies are needed on the bioactive components of tea to develop drugs against liver injury.

Biochemical Parameters and Protein Oxidation Relationship for Hepatitis C Patients and Healthy Ones

Globally hepatitis C virus is recognized as one of the basic health issues of liver and result in chronic liver diseases whose diagnosis is difficult and possible only with symptoms until it may leads to liver cancer. DNA damage along with lipids and protein oxidation is caused as result of oxidative stress in hepatitis patients. This work was carried out to search out the oxidation of protein and lipids per-oxidation with relative to DNA damage for hepatitis C patients. Sampling was done in 50 suspected patients among them 20 were positive for HCV, while 30 were negative. With the help of 2-4, dinitrophenyl hydrazine assay quantification of protein carbonyls was done and results showed much increased values than normal. Biochemical parameters of HCV such as complete blood count, triglycerides and cholesterol level were recorded. The results were significant in positive cases than the control group. The study elaborated a new window in the research of chronic HCV patients for severe biochemical parameters alterations, over oxidation of proteins and liver dysfunction. It was therefore concluded that oxidative stress was responsible for increased level of protein oxidation and altered biochemical parameters that leads to damage of liver

Effect of alpha-mangostin on olanzapine-induced metabolic disorders in rats.

Objective(s):As olanzapine has side effects such as weight gain and metabolic disorders, and alpha-mangostin has been shown to control metabolic disorders, the effects of alpha-mangostin on metabolic disorders induced by olanzapine were investigated in this study.Materials and Methods:Obesity was induced in female Wistar rats by daily administration of olanzapine (5 mg/kg/day, IP, 14 days). Rats were divided into 6 groups:1) vehicle (control); 2) olanzapine (5 mg/kg/day); 3,4,5) olanzapine+ alpha-mangostin (10, 20, 40 mg/kg/day, IP); 6) alpha-mangostin (40 mg/kg/day). Weight changes were measured every 3 days and food intake was assessed every day. Systolic blood pressure, plasma levels of blood sugar, triglycerides, total cholesterol, HDL, LDL, leptin, oxidative stress markers (MDA, GSH), AMPK, and P-AMPK protein levels in liver tissue were assessed on the last day of the study. Results:Administration of olanzapine significantly increased weight gain, food intake, blood pressure, triglycerides, LDL, blood sugar, leptin, and MDA in rat liver tissue and also decreased GSH, AMPK, and P-AMPK in liver tissue compared with the control group. Different doses of alpha-mangostin significantly reduced weight gain, food intake, systolic blood pressure, triglycerides, LDL, blood sugar, leptin, and MDA. Also, they significantly increased GSH, AMPK, and P-AMPK in liver tissue compared with the olanzapine group.Conclusion:Olanzapine increases leptin levels, food intake, and weight, induces oxidative stress, decreases the levels of AMPK and P-AMPK proteins in liver tissue, and causes metabolic disorders. But, alpha-mangostin reduces the negative effects of olanzapine by activation of AMPK.

Dietary intake of diosgenin delays aging of male fish Nothobranchius guentheri through modulation of multiple pathways that play prominent roles in ROS production.

Oxidative stress including DNA damage, increased lipid and protein oxidation, is an important feature of aging. Diosgenin (DG) has been shown to have diverse biological effects, including amelioration of aging-related cognition deficits, but the anti-aging activity of DG has not been tested before in animal models. In the present study, we clearly demonstrated that dietary intake of DG extended both mean and maximum lifespans of the male fish Nothobranchius guentheri by approximately 3.23 and 3.67weeks, respectively, reduced the accumulation of lipofuscin (LF) in the gills and senescence-associated-β-galactosidase (SA-β-Gal) in the caudal fins, and lowered the levels of protein oxidation, lipid peroxidation and reactive oxygen species (ROS) in the muscles, indicating that DG possesses rejuvenation and anti-aging property. We also showed that DG enhanced the activity of antioxidant enzymes, including catalase, superoxide dismutase and glutathione peroxidase, promoted the proteolytic activity of the ubiquitin-proteasome pathway, and suppressed the phosphatidylinositol 3-kinase/protein kinase/molecular target of rapamycin (PI3K/AKT/mTOR) signaling pathway. Altogether, this study highlights for the first time the rejuvenation and anti-aging property of the naturally occurring steroidal sapogenin DG. It also suggests that DG exerts its rejuvenation and anti-aging activity through modulation of multiple signaling pathways that play prominent roles in ROS production.

The crucial role of oxidative stress in non-alcoholic fatty liver disease-induced male reproductive toxicity: the ameliorative effects of Iranian indigenous probiotics.

Several studies have focused on the high potential effects of probiotics on the reproductive system. However, there is a paucity of information regarding the ameliorative intracellular roles of indigenous Iranian yogurt-extracted/cultured probiotics on animals' reproductive health suffering from obesity and/or fatty liver disease, such as non-alcoholic fatty liver disease (NAFLD). For this purpose, simultaneously with the consumption of D-fructose (200g/1000mL water, induction of NAFLD model), all pubertal animals were also gavaged every day for 63 consecutive days with extracted probiotics, including 1 × 109CFU/mL of Lactobacillus acidophilus (LA), Bifidobacterium spp. (BIF), Bacillus coagulans (BC), Lactobacillus rhamnosus (LR), and a mixture form (LA + BIF + BC + LR). At the end of the ninth week, the indices of epididymal sperm, and oxidative stress, as well as histopathological changes, were assessed. The results show that NAFLD could induce robust oxidative stress, highlighted as considerable increments in ROS level, TBARS content, total oxidized protein levels, along with severe decrements in reduced glutathione reservoirs, total antioxidant capacity in the hepatic and testicular tissues, as well as testicular and hepatic histopathological alterations. Moreover, a significant decrease in the percentage of sperm progressive motility, sperm count, and membrane integrity along with an increment in the percentage of sperm abnormality was detected in NAFLD animals. The observed adverse effects were significantly reversed upon probiotics treatment, especially in the group challenged with a mixture of all probiotics. Taken together, these findings indicate that the indigenous yogurt-isolated/cultured probiotics had a high potential antioxidant activity and the ameliorative effect against reprotoxicity and blood biochemical alterations induced by the NAFLD model. Highlights: 1. Reproductive indices could be reversely affected by xenobiotics and diseases. 2. NAFLD and cholestasis considerably affect the reproductive system in both genders. 3. NAFLD induced hepatic and testicular oxidative stress (OS). 4. NAFLD induced histopathological alterations and spermatotoxicity through OS. 5. The adverse effects were significantly reversed upon exposure to probiotics.

Subchronic exposure to Epoxiconazole induced-heart damage in male Wistar rats

Epoxiconazole is a worldwide fungicide used to control fungal diseases. Although to its hazardous effects in non-target species, little information is available in the literature to show the cardiotoxic effects of EPX in male rats. Thus, our investigation aimed to assess the outcomes of EPX exposure on some biochemical parameters, the generation of oxidative stress, DNA fragmentation and histopathological alterations in the heart tissue. EPX was administered orally at doses of 8, 24, 40 and 56 mg/kg body weight, representing, respectively NOEL (No observed effect level), NOEL× 3, NOEL× 5 and NOEL× 7 for 28 consecutive days in male Wistar rats. Our results show that EPX induced a significant decrease of cardiac acetylcholinesterase, an increase of biochemical markers, such as creatinine phosphokinase (CPK) and a perturbation of the lipid profile. Furthermore, EPX caused diverse histological modifications in the myocardium, including congestion of cardiac blood vessels, cytoplasmic vacuolization, leucocytic infiltration and hemorrhage. Indeed, we have shown that EPX induces increase of lipid peroxidation, protein oxidation levels and DNA damage. On the other hand, we have found an increase of the antioxidant enzymes activity such as catalase (CAT) and superoxide dismutase (SOD) activities. The glutathione peroxidase and glutathione S tranferase initially enhanced at the doses of 8, 24, and 40 mg/kg b.w. and then decreased at the dose of 56 mg/kg b.w. In conclusion, our work has shown that EPX causes cardiotoxic effects by altering redox status and damaging heart tissue.

Blood biochemistry and antioxidant status altered by anthropogenic impact in Adélie penguins (Pygoscelis adeliae).

Human activities are increased in Antarctica during decades, primarily due to the logistic and tourism operations, which consequent negative impact on penguin populations, altering their physiological responses. Therefore, we aimed to assess the blood biochemistry and oxidant/antioxidant balance of Adélie penguins (Pygoscelis adeliae) inhabiting two selected colonies: Potter Peninsula, considered as a low impacted colony, and Esperanza/Hope Bay as a high impacted colony. The levels of calcium, uric acid, and fructosamine, showed significant high values (p<0.05) for the Potter Peninsula´s penguins. Besides, this population showed high levels of plasma protein oxidation and erythrocyte lipid peroxidation (p<0.005) while the Esperanza/Hope Bay population presented high levels of erythrocyte protein oxidation and plasma lipid peroxidation (p<0.005). The oxidative damage values were similar in the Potter Peninsula population and slightly lower in the Esperanza/Hope Bay penguins if the results were compared to previous reports. The enzymes superoxide dismutase and glutathione peroxidase had significantly (p<0.005) high activity in the Esperanza/Hope Bay population, which also showed high reduced glutathione levels. The glutathione S-transferase activity was significantly high (p<0.005) in the Potter Peninsula population. The obtained results might take into account for making decisions about management and protection plans for the different penguin nesting areas in Antarctica.

Evaluation of the toxic effects of thimerosal and/or aluminum hydroxide in SH-SY5Y cell line.

In this study, we aimed to evaluate possible toxic effects of thimerosal, aluminum and combination of thimerosal and aluminum in SH-SY5Y cells. Inhibitory concentrations were determined by MTT assay; reactive oxygen species (ROS) were determined by a fluorometric kit and antioxidant/oxidant parameters were measured by spectrophotometric kits. Nuclear factor erythroid 2-associated factor 2 (Nrf2), norepinephrine (NE), dopamine transporter (DAT) and dopamine beta β-hydroxylase (DBH) levels were measured by sandwich ELISA kits while 8-hydroxy deoxyguanosine (8-OHdG) and dopamine levels were determined by competitive ELISA kits. Thimerosal (1.15μM) and aluminum (362μM) were applied to cells at inhibitory concentrations 20 (IC20s) for 24 h. ROS increased significantly in cells aluminum- and aluminum+thimerosal-treated cells. Glutathione levels decreased in aluminum group while total antioxidant capacity and protein oxidation levels increased significantly in aluminum and aluminum+thimerosal groups. Lipid peroxidation increased significantly in groups treated with aluminum and aluminum+thimerosal. Nrf2 levels and DNA damage were significantly higher in all groups while dopamine levels significantly increased in cells treated with thimerosal and aluminum+thimerosal, DAT levels were found to be higher in all experimental groups compared to the control. These findings showed that both thimerosal and aluminum can change oxidant/antioxidant status, cause DNA damage, alter dopamine and DAT levels. Changes seen in cells treated with combined exposure to aluminum and thimerosal are more pronounced. Special care should be taken while vaccinating sensitive populations and safer alternatives for aluminum and thimerosal should used.

Ellagic Acid Alleviates Rheumatoid Arthritis in Rats through Inhibiting MTA1/HDAC1-Mediated Nur77 Deacetylation.

Ellagic acid (EA) was reported to play protective roles in rheumatoid arthritis (RA). It was found that the level of metastasis-associated gene 1 (MTA1)/histone deacetylase 1 (HDAC1) protein complex was downregulated by polyphenols in several human disorders. Notably, inhibition of MTA1 or HDAC1 has anti-inflammatory effects on RA. Therefore, our study is aimed at investigating whether EA prevents RA progression through regulating the MTA1/HDAC1 complex. Herein, the human fibroblast-like synoviocyte (FLS) cell line MH7A was treated with TNF-α to induce an inflammation model in vitro and then incubated with different concentrations of EA. Western blot analysis showed that EA reduced MTA1 expression in a dose-dependent manner in MH7A cells. Then, TNF-α-treated MH7A cells were incubated with EA alone or together with MTA1 overexpression plasmid (pcDNA-MTA1), and we found that EA inhibited proliferation, inflammation cytokine levels, and oxidative stress marker protein levels and promoted apoptosis in MH7A cells, while MTA1 overexpression abolished these effects. Moreover, coimmunoprecipitation assay verified the interaction between MTA1 and HDAC1. EA downregulated the MTA1/HDAC1 complex in MH7A cells. MTA1 knockdown inhibited proliferation, inflammation, and oxidative stress and promoted apoptosis in MH7A cells, while HDAC1 overexpression reversed these effects. Moreover, chromatin immunoprecipitation assay indicated that EA inhibited HDAC1-mediated Nur77 deacetylation. Rescue experiments demonstrated that Nur77 knockdown reversed the effects of EA on MH7A cell biological behaviors. Additionally, EA treatment attenuated arthritis index, paw swelling, synovial hyperplasia, and inflammation in collagen-induced arthritis (CIA) rats. In conclusion, EA inhibited proliferation, inflammation, and oxidative stress and promoted apoptosis in MH7A cells and alleviated the severity of RA in CIA rats though downregulating MTA1/HDAC1 complex and promoting HDAC1 deacetylation-mediated Nur77 expression.

Acute RyR1 Ca2+ leak enhances NADH-linked mitochondrial respiratory capacity

Sustained ryanodine receptor (RyR) Ca2+ leak is associated with pathological conditions such as heart failure or skeletal muscle weakness. We report that a single session of sprint interval training (SIT), but not of moderate intensity continuous training (MICT), triggers RyR1 protein oxidation and nitrosylation leading to calstabin1 dissociation in healthy human muscle and in in vitro SIT models (simulated SIT or S-SIT). This is accompanied by decreased sarcoplasmic reticulum Ca2+ content, increased levels of mitochondrial oxidative phosphorylation proteins, supercomplex formation and enhanced NADH-linked mitochondrial respiratory capacity. Mechanistically, (S-)SIT increases mitochondrial Ca2+ uptake in mouse myotubes and muscle fibres, and decreases pyruvate dehydrogenase phosphorylation in human muscle and mouse myotubes. Countering Ca2+ leak or preventing mitochondrial Ca2+ uptake blunts S-SIT-induced adaptations, a result supported by proteomic analyses. Here we show that triggering acute transient Ca2+ leak through RyR1 in healthy muscle may contribute to the multiple health promoting benefits of exercise.

BENEFICIALL EFFECT OF THE EUROPEAN RASPBERRY (RUBUS IADEUS) EXTRACT ON THE ACTIVITY AND OXIDATIVE PROFILE OF BOVINE SPERMATOZOA

Berries belong to the best dietary sources of bioactive compounds. These fruits contains bioactive compounds such as antioxidants (phenolic compounds and fruit colorants), vitamins (ascorbic acids) and minerals. This study aimed to estimate the effect of three concentrations (75 µg/mL, 150 µg/mL and 300 µg/mL) of the European raspberry (Rubus iadeus) on the selected quality parameters of bovine spermatozoa after 0h, 2h, and 24h of in vitro culture. Sperm motility was evaluated using the Computer-assisted sperm analysis (CASA) system. The mitochondrial activity was detected by the metabolic (MTT) assay. The determination of ROS (Reactive Oxygen Species) quantity was observed through the luminol method. The protein and lipid oxidation were evaluated using spectrophotometric assays. The experiment showed that the highest applied concentration of European raspberry extract at 2h (p&lt;0.05) and all concentration after 24h (p&lt;0.05; p&lt;0.001) exhibited motility-promoting effects. We observed the highest increase of mitochondrial activity using 300 µg/mL by 2h (p&lt;0.05) and using 150 µg/mL (p&lt;0.05) and 300 µg/mL (p&lt;0.01) after 24h. Exposure to this extract led to decreasing levels of ROS generation at 2h of incubation (p&lt;0.05) with the highest dose as well as after 24h using 150 µg/mL (p&lt;0.01) and 300 µg/mL (p&lt;0.001) applied doses. The protein oxidation levels showed decreasing rates at 2h (p&lt;0.05; 300 µg/mL) and at time of 24h (p&lt;0.01; 150 µg/mL and p&lt;0.001; 300 µg/mL). All selected raspberry concentration prevented oxidative degeneration of lipids during 2h (p&lt;0.05) and after 24h (p&lt;0.05; p&lt;0.001). Our results indicate, that European raspberry extract has beneficial effect on the measured parameters of the bovine spermatozoa during in vitro incubation but depending on time and applied dose.

Chemical and Biological Characterization of the Anticancer Potency of Salvia fruticosa in a Model of Human Malignant Melanoma.

Malignant melanoma is one of the most aggressive types of skin cancer with an increasing incidence worldwide. Thus, the development of innovative therapeutic approaches is of great importance. Salvia fruticosa (SF) is known for its anticancer properties and in this context, we aimed to investigate its potential anti-melanoma activity in an in vitro model of human malignant melanoma. Cytotoxicity was assessed through a colorimetric-based sulforhodamine-B (SRB) assay in primary malignant melanoma (A375), non-malignant melanoma epidermoid carcinoma (A431) and non-tumorigenic melanocyte neighbouring keratinocyte (HaCaT) cells. Among eight (8) different fractions of S. fruticosa extracts (SF1-SF8) tested, SF3 was found to possess significant cytotoxic activity against A375 cells, while A431 and HaCaT cells remained relatively resistant or exerted no cytotoxicity, respectively. In addition, the total phenolic (Folin–Ciocalteu assay) and total flavonoid content of SF extracts was estimated, whereas the antioxidant capacity was measured via the inhibition of tert-butyl hydroperoxide-induced lipid peroxidation and protein oxidation levels. Finally, apoptotic cell death was assessed by utilizing a commercially available kit for the activation of caspases - 3, - 8 and - 9. In conclusion, the anti-melanoma properties of SF3 involve the induction of both extrinsic and intrinsic apoptotic pathway(s), as evidenced by the increased activity levels of caspases - 8, and - 9, respectively.

Gonadal Rejuvenation of Mice by Growth Differentiation Factor 11.

Growth differentiation factor 11 (GDF11), also known as bone morphogenetic protein 11, has been shown to have rejuvenation and antiaging properties, but little information is available regarding the role of GDF11 in reproductive system to date. In this study, we first confirmed the bioavailability of recombinant GDF11 (rGDF11) by oral delivery in mice. We also showed that dietary intake of rGDF11 had little influence on body and gonadal (ovary/testis) weights of recipient mice, indicating their general condition and physiology were not affected. Based on these findings, we started to test the function of rGDF11 in ovary and testis of mice and to explore the underlying mechanisms. It was found that to some extent, rGDF11 could attenuate the senescence of ovarian and testicular cells, and contribute to the recovery of ovarian and testicular endocrine functions. Moreover, rGDF11 could rescue the diminished ovarian reserve in female mice and enhance the activities of marker enzymes of testicular function (sorbitol dehydrogenase and glucose-6-phosphate dehydrogenase) in male mice, suggesting a potential improvement of fertility. Notably, rGDF11 markedly promoted the activities of antioxidant enzymes in the ovary and testis, and remarkably reduced the levels of lipid peroxidation, protein oxidation, and reactive oxygen species (ROS) in the ovary and testis. Collectively, these results suggest that GDF11 can protect ovarian and testicular functions of aged mice via slowing down the generation of ROS through enhancing activities of antioxidant enzymes.

Anti-inflammatory Effects of a Novel Herbal Extract in the Muscle and Spinal Cord of an Amyotrophic Lateral Sclerosis Animal Model.

Amyotrophic lateral sclerosis (ALS) is a complex disease characterized by motor neuron loss and muscle atrophy. There is no prominent treatment for ALS as the pathogenic process in the skeletal muscle and spinal cord is complex and multifactorial. Therefore, we investigated the effects of a herbal formula on the multi-target effects in the skeletal muscle and spinal cord in hSOD1G93A transgenic mice. We prepared a herbal extract (HE) from Glycyrrhiza uralensis, Atractylodes macrocephala Koidzumi, Panax ginseng, and Astragalus membranaceus. Control and HE-treated mice underwent rotarod and footprint tests. We also performed immunohistochemical and Western blotting analyses to assess expression of inflammation-related and oxidative stress-related proteins in the muscle and spinal cord tissues. We found that the HE increased motor activity and reduced motor neuron loss in hSOD1G93A mice. In addition, the HE significantly reduced the levels of inflammatory proteins and oxidative stress-related proteins in the skeletal muscles and spinal cord of hSOD1G93A mice. Furthermore, we demonstrated that the HE regulated autophagy function and augmented neuromuscular junction in the muscle of hSOD1G93A mice. Based on these results, we propose that the HE formula may be a potential therapeutic strategy for multi-target treatment in complex and multifactorial pathological diseases.

Genotoxicity and Cytotoxicity Induced in Zygophyllum fabago by Low Pb Doses Depends on the Population’s Redox Plasticity

Lead (Pb) soil contamination remains a major ecological challenge. Zygophyllum fabago is a candidate for the Pb phytostabilisation of mining tailings; nevertheless, the cytogenotoxic effects of low doses of Pb on this species are still unknown. Therefore, Z. fabago seeds collected from non-mining (NM) and mining (M) areas were exposed to 0, 5 and 20 µM Pb for four weeks, after which seedling growth, Pb cytogenotoxic effects and redox status were analyzed. The data revealed that Pb did not affect seedling growth in M populations, in contrast to the NM population. Cell cycle progression delay/arrest was detected in both NM and M seedlings, mostly in the roots. DNA damage (DNAd) was induced by Pb, particularly in NM seedlings. In contrast, M populations, which showed a higher Pb content, exhibited lower levels of DNAd and protein oxidation, together with higher levels of antioxidants. Upon Pb exposure, reduced glutathione (GSH) and non-protein thiols were upregulated in shoots and were unaffected/decreased in roots from the NM population, whereas M populations maintained higher levels of flavanols and hydroxycinnamic acids in shoots and triggered GSH in roots and shoots. These differential organ-specific mechanisms seem to be a competitive strategy that allows M populations to overcome Pb toxicity, contrarily to NM, thus stressing the importance of seed provenance in phytostabilisation programs.

Antibacterial Mechanisms of Reduced Iron-Containing Smectite-Illite Clay Minerals.

Reduced nontronite has been demonstrated to be antibacterial through the production of hydroxyl radical (•OH) from the oxidation of structural Fe(II). Herein, we investigated the antibacterial activity of more common smectite-illite (S-I) clays toward Escherichia coli cells, including montmorillonite SWy-3, illite IMt-2, 50-50 S-I rectorite RAr-1, 30-70 S-I ISCz-1, and nontronite NAu-2. Under an oxic condition, reduced clays (with a prefix r before mineral names) produced reactive oxygen species (ROS), and the antibacterial activity followed the order of rRAr-1 > rSWy-3 ≥ rNAu-2 ≫ rIMt-2 ≥ rISCz-1. The strongest antibacterial activity of rRAr-1 was contributed by a combination of •OH and Fe(IV) generated from structural Fe(II)/adsorbed Fe2+ and soluble Fe2+, respectively. Higher levels of lipid and protein oxidation, intracellular ROS accumulation, and membrane disruption were consistent with this antibacterial mechanism of rRAr-1. The antibacterial activity of other S-I clays depended on layer expandability, which determined the reactivity of structural Fe(II) and the production of •OH, with the expandable smectite being the most antibacterial and nonexpandable illite the least. Our results provide new insights into the antibacterial mechanisms of clay minerals.