Cryopreservation of ram semen: baicalein efficiency on oxidative stress, chromatin integrity, viability and motility post thaw.

Baicalein (B) has potential antioxidant properties, but it has not been tested as a ram semen extender. This study aimed to assess the impact of B on various sperm parameters and determine its potential influence on semen quality after the freeze-thawing process. During the breeding season, ejaculates were obtained from four rams with the aid of an artificial vagina. The collected mixed semen samples were divided into four groups: control (C; 0), B0.5 (0.5 mM), B1 (1 mM), and B2 (2 mM). After semen extension, the samples were loaded into 0.25 mL straws and stored for 2 h at 4°C prior to freezing in liquid nitrogen vapor and thawed in a water bath at 37°C. Among the groups, B0.5 demonstrated the highest progressive motility results, while B1 and B2 exhibited reduced motility (p < 0.05). In terms of high mitochondrial membrane potential, plasma membrane and acrosome integrity, and viability, B0.5 showed significantly superior outcomes to the other B groups (p < 0.05), although it was not significantly better than C. B1 displayed the highest plasma membrane integrity levels (p < 0.05). Notably, B2 displayed the lowest total antioxidant status levels among the groups (p < 0.05). The findings of this study suggested that the in vitro spermatological characteristics of ram spermatozoa such as progressive motility and chromatin integrity can be protected from the freeze-thawing process by using the 0.5 mM dose of baicalein as a semen extender. The treatment of sperm freezing might benefit from further in-depth research on the role of B in the improvement of cryoinjury and its underlying processes.

Investigation of the effects of syringic acid supplemented to Tris semen diluent on ram semen freezability.

To the best of our knowledge, no research has been conducted to test the effects of syringic acid (SA) on ram semen freezing within the scope of natural antioxidants added to semen extenders. Therefore, this study had two main objectives. First, to test whether adding SA to ram semen freezing extender has a protective effect and contributes positively to sperm kinetic, plasma and acrosome integrity, mitochondrial membrane potential, lipid peroxidation, oxidant and antioxidant and DNA damage parameters after thawing. Second, it was to determine at what concentration the SA supplemented to the extender could be applied by in vitro studies by preserving the fertilization ability of frozen semen at the highest level. In the study, six individuals of Sönmez rams were used. The semen was collected from the rams using an artificial vagina and pooled. The pooled semen was divided into five different groups and extended with 0, 0.5, 1, 2 and 4 mM SA (control C, SA0.5, SA1, SA2 and SA4, respectively). After dilution, the semen samples were kept at 4°C for 3 h, then loaded into 0.25 mL straws and frozen in liquid nitrogen vapour. The SA1 and SA2 groups were higher plasma membrane and acrosome integrity (PMAI), high mitochondrial membrane potential (HMMP), plasma membrane integrity and motility compared to other groups (p < .05). It was observed that SA supplemented to the Tris extender significantly reduced DNA damage, and the lowest values were obtained especially in the SA1 and SA2 treatments (p < .05). Also, lowest MDA level was determined at the SA1 and this was statistically significant compared to SA4 and C (p < .05). In conclusion, it was revealed that SA added to Tris semen extender at 1 and 2 mM treatment doses increased progressive and total motility and preserved PMAI, plasma membrane integrity, HMMP and DNA integrity.

A New Substrate and Nitrogen Source for Traditional Kombucha Beverage: Stevia rebaudiana Leaves.

Recently, the use of different herbal products as carbon sources instead of black and green tea in the preparation of traditional kombucha has been investigated. In this study, functional kombucha was prepared by adding Stevia rebaudiana Bertoni leaves, which have special organoleptic properties, to kombucha medium, and some properties of the beverage were analyzed. Tea blends were determined as 100% green tea (control = C), 75% green tea (GT) + 25% Stevia (ST), 50% GT + 50% ST, and 100% ST. On the 15th day of fermentation, gluconic acid (43.12 ± 0.01 g/L) was detected as dominant organic acid in GT75 + ST25 samples compared to group C (p < 0.05). According to physicochemical parameters that determine the drinkability properties of prepared teas, the best results were in GT25 + ST75 compared to group C (p < 0.05). It proved that the highest activity was in GT25 + ST75 on the 10th day in the groups that applied different antioxidant tests (DPPH, MCA, and CUPRAC). The antimicrobial activities of kombucha at 25, 50, 75, and 100% concentrations of GT and ST reached the highest levels in the GT25 + ST75 group in samples after 10 days of fermentation for all selected microorganisms. The results prove that GT25 + ST75 kombucha is a functional product with high drinkability on the 10th day of fermentation and also more beneficial for health due to the phenolic compounds from both green tea and Stevia. Stevia rebaudiana leaves can be suggested that be used as a new substrate and nitrogen source for kombucha production.

Investigation of Changes in Spermatozoon Characteristics, Chromatin Structure, and Antioxidant/Oxidant Parameters after Freeze-Thawing of Hesperidin (Vitamin P) Doses Added to Ram Semen.

We conducted this study to determine the potential cryopreservative effects of different hesperidin (vitamin P; H) doses on ram semen after freeze-thawing. Semen samples were obtained from Sönmez rams using an artificial vagina. The samples were divided into six groups: control, 10, 50, 100, 250, and 500 µg/mL H (C, H10, H50, H100, H250, and H500, respectively). At the end of the study, sperm motility and kinetic parameters, acrosome integrity (AI), mitochondrial membrane potential (MMP), viability, lipid peroxidation levels (LPL), chromatin damage, oxidant parameters, and antioxidant parameters were assayed. None of the doses of H added to the semen extender showed any enhancing effects on progressive motility compared to C (p > 0.05). In fact, H500 had negative effects (p < 0.05). Moreover, AI was the highest at the H10 dose, while LPL values were the lowest at the same dose (p < 0.05). The doses of H10 and H50 added to the Tris extender medium showed positive effects on sperm cell chromatin damage. Consequently, we can say that H doses used in this study are not effective on semen progressive motility, but the H10 dose is effective on AI and chromatin damage by reducing LPL.

Thymoquinone improves motility, plasma membrane integrity and DNA integrity of frozen-thawed ram semen.

The aim of this study was to determine the effects of thymoquinone (TQ), which is the most essential active compound of Nigella sativa, on the spermatological parameters of ram semen during cryopreservation. Ejaculates were collected from five Sonmez rams using an artificial vagina and extended with Tris-based extender not containing TQ (control, 0 µg/ml TQ) and containing 10, 25, 50 and 100 µg/ml TQ. The extended semen samples were equilibrated in a + 4°C cold cabinet for 2 h. After 2 h, the samples were loaded into 0.25 ml French straws. The straws were frozen by liquid nitrogen vapour and stored in a liquid nitrogen container (-196°C). The frozen straws were thawed in a water bath (37°C for 30 s) and evaluated in terms of motility characteristics, plasma membrane and acrosome integrity, mitochondrial reactive oxygen species levels, lipid peroxidation levels, DNA damage and biochemical alterations (oxidative stress index, malondialdehyde and glutathione). TQ100 had higher total motility (53.59 ± 3.01) and progressive motility (19.84 ± 1.44; not significantly different from TQ25 and TQ50) compared to the control and TQ10 (p Ë‚ 0.05). According to the results of the analyses on motility characteristics, there were significant differences between the groups in terms of curvilinear velocity (VCL), amplitude of lateral head displacement (ALH) and linearity (LIN; p Ë‚ 0.05). The highest DNA damage was detected in the control group (p Ë‚ 0.05). TQ50 had higher plasma membrane and acrosome integrity (59.56 ± 5.92) compared to the control and TQ25 (p < 0.05) but not significantly different from TQ10 and TQ100. The lowest mitochondrial reactive oxygen species levels were detected in TQ50 and TQ100 (p Ë‚ 0.05). There were no significant differences among the groups in terms of their oxidative stress index, lipid peroxidation, malondialdehyde and glutathione levels (p > 0.05). According to the results, it could be concluded that supplementing 50 or 100 µg/ml TQ to Tris extenders that were used for ram semen cryopreservation showed a positive effect on motility, plasma membrane integrity and acrosome integrity, and it reduced DNA damage and mitochondrial reactive oxygen species levels.

The investigation of antioxidant and anti-inflammatory potentials of apitherapeutic agents on heart tissues in nitric oxide synthase inhibited rats via Nω-nitro-L-arginine methyl ester.

BACKGROUND: High blood pressure effects heart and vessels. Development of pathogenesis is the result of oxidative stress. We aimed to investigate the antioxidant effects of propolis, caffeic acid phenethyl ester (CAPE), and pollen on the hearts of rats which chronic nitric oxide synthase (NOS) inhibited through Nω-nitro-L-arginine methyl ester (L-NAME). Paraoxonase 1 (PON1), total antioxidant status (TAS), total oxidant status (TOS), oxidative stress index (OSI), asymmetric dimethylarginine (ADMA), and nuclear factor-κB (NF-κB) were analyzed on the heart. MATERIAL AND METHODS: Sprague-Dawley rats were divided five groups of seven rats in every group; Group I: Control, Group II: L-NAME, Group III: L-NAME+propolis, Group IV: L-NAME+CAPE and Group V: L-NAME+pollen. L-NAME become dissolved in regular saline (0.9% NaCl w/v). The ethanolic extract of propolis (200 mg/kg/days, gavage), pollen (100 mg/kg/days, by gavage), CAPE (50 µM/kg/days, intraperitoneally), and the NOS inhibitor L-NAME (40 mg/kg, intraperitoneally) had been administered. RESULTS: Blood pressure (BP) of rats treated with propolis, CAP,E and pollen statistically significant decreased. Decreasing in BP of the rats of pollen group was more than CAPE and propolis groups (P < .05). PON1 and TAS levels decreased in L-NAME-treated groups (P < .05), but ranges have been better in propolis, CAPE and pollen groups. TOS, ADMA and NF-κB levels increased (P < .05) in L-NAME group; however, these parameters were lower (P < .05) in propolis and CAPE groups (P < .05). CONCLUSIONS: Vasorelaxant properties and free radical scavenging actions of propolis, CAPE, and pollen may reduce the oxidative stress and blood pressure in the rats chronic NOS inhibited through L-NAME.

Therapeutic potentials of propolis and pollen on biochemical changes in reproductive function of L-NAME induced hypertensive male rats.

BACKGROUND: Hypertension is described as increased blood pressure based on changed hemodynamics and associated with an increased oxidative damage to reproductive function. This work is to determine therapeutic and protective effects of apitherapy products (propolis and pollen) on reproductive functions of L-NAME-induced hypertensive male rats. METHODS: Experimental animals were indiscriminately separated into four groups of seven rats in each group: (I) Control, (II) L-NAME, (III) L-NAME+ propolis and (IV) L-NAME+ pollen. At the end of the experimental applications, the rats were decapitated by anesthesia and biochemical analyzes were performed on the removed testicular tissues. RESULTS: The levels of TOS, NF-κB and MDA in the L-NAME group compared to control have increased (p < 0.05). The levels of these parameters in L-NAME+ propolis and L-NAME+ pollen groups compared to the L-NAME group have decreased (p < 0.05). TAS levels, PON1 and CAT activities were significantly decreased in testis tissue samples in the L-NAME-induced group (p < 0.05). However, these parameters were significantly lower in the L-NAME plus propolis and pollen groups (p < 0.05) compared with rats administered L-NAME alone (p < 0.05). NO level significantly reduced (p < 0.05) in L-NAME group compared with control group. There was no statistically significant changes in the NO level of the L-NAME+ propolis group compared with the L-NAME-treated group (p > 0.05). CONCLUSION: It has been determined that ethanolic extracts of propolis and pollen, which are natural bee products in the regulation of rising blood pressure. Propolis or pollen is thought to help regulate reproductive function by inhibiting the functioning of inflammatory pathways leading to hypertension.

The effects of pollen, propolis, and caffeic acid phenethyl ester on tyrosine hydroxylase activity and total RNA levels in hypertensive rats caused by nitric oxide synthase inhibition: experimental, docking and molecular dynamic studies.

The objective of the present study was to evaluate the effects of propolis, pollen, and caffeic acid phenethyl ester (CAPE) on tyrosine hydroxylase (TH) activity and total RNA levels of Nω-nitro-L-arginine methyl ester (L-NAME) inhibition of nitric oxide synthase in the heart, adrenal medulla, and hypothalamus of hypertensive male Sprague dawley rats. The TH activity in the adrenal medulla, heart, and hypothalamus of the rats was significantly increased in the L-NAME group vs. control (p < 0.05). Treatment with L-NAME led to a significant increase in blood pressure (BP) in the L-NAME group compared to control (p < 0.05). These data suggest that propolis, pollen, and CAPE may mediate diminished TH activity in the heart, adrenal medulla, and hypothalamus in hypertensive rats. The decreased TH activity may be due to the modulation and synthesis of catecholamines and BP effects. In addition, the binding mechanism of CAPE within the catalytic domain of TH was investigated by means of molecular modeling approaches. These data suggest that the amino acid residues, Glu429 and Ser354 of TH may play a pivotal role in the stabilization of CAPE within the active site as evaluated by molecular dynamics (MD) simulations. Gibbs binding free energy (ΔGbinding) of CAPE in complex with TH was also determined by post-processing MD analysis approaches (i.e. Poisson-Boltzmann Surface Area (MM-PBSA) method).

Effects of propolis, caffeic acid phenethyl ester, and pollen on renal injury in hypertensive rat: An experimental and theoretical approach.

The objective of this study was to evaluate the antioxidant effects of propolis, caffeic acid phenethyl ester (CAPE; active compound in propolis), and pollen on biochemical oxidative stress biomarkers in rat kidney tissue inhibited by Nω -nitro-L-arginine methyl ester (L-NAME). The biomarkers evaluated were paraoxonase (PON1), oxidative stress index (OSI), total antioxidant status (TAS), total oxidant status (TOS), asymmetric dimethylarginine (ADMA), and nuclear factor kappa B (NF-κB). TAS levels and PON1 activity were significantly decreased in kidney tissue samples in the L-NAME-treated group (P < 0.05). The levels of TAS and PONI were higher in the L-NAME plus propolis, CAPE, and pollen groups compared with the L-NAME-treated group. TOS, ADMA, and NF-κB levels were significantly increased in the kidney tissue samples of the L-NAME-treated group (P < 0.05). However, these parameters were significantly lower in the L-NAME plus propolis, CAPE, and pollen groups (P < 0.05) compared with rats administered L-NAME alone (P < 0.05). Furthermore, the binding energy of CAPE within catalytic domain of glutathione reductase (GR) enzyme as well as its inhibitory mechanism was determined using molecular modeling approaches. In conclusion, experimental and theoretical data suggested that oxidative alterations occurring in the kidney tissue of chronic hypertensive rats may be prevented via active compound of propolis, CAPE administration.

In-vitro Antioxidant Activities of the Ethanolic Extracts of Some Contained-Allantoin Plants.

It has been investigated the in-vitro antioxidant properties of ethanol extracts of the contained-allantoin plants in this study. Contained-allantoined plant samples Plantago lanceolata, Plantago major, Robinia pseudoacacia, Platanus orientalis and Aesculushippocastanum were tested at different concentrations. The antioxidant activities of plant samples were analysed by 1,1- diphenyl-2-picrylhydrazyl (DPPH) radical scavenging method, cupric reducing antioxidant capacity (CUPRAC), reducing power assay and ß-carotene bleaching method. Plantago major plant showed the highest antioxidant capacity compared to other plant extracts in results of the in-vitro assays including 1,1- diphenyl-2-picrylhydrazyl (DPPH) radical scavenging method with 90.25 %, cupric reducing antioxidant capacity (CUPRAC) with 1.789 %, reducing power assay (FRAP) with 1.321 % and ß-carotene bleaching method with 78.01 % in 1 mg/mL. The lowest antioxidant activity was determined in Robinia pseudoacacia plant. In conclusion, allantoin shows antioxidant properties and it has the positive effect on total antioxidant capacity.

Melatonin and Respiratory Diseases: A Review.

Melatonin is an indoleamine with potent multifunctional biological and pharmacological effects, both receptor dependent and receptor-independent effects, including antioxidant, anticancer, antitumor, anti-inflammatory, anti-aging, anti-diabetic, antiviral, neuroprotective activities. Melatonin mitigates tissue injury via modification of abnormalities in redox status and other biochemical markers. At the molecular level, the biological and pharmacological activities of melatonin are attributed to the inhibition of nuclear factor-κappa beta (NF-κß), c-Fos over expression and down-regulation of matrix metalloproteinases-3 (MMP-3), which are regulators of pro-inflammatory and pro-fibrotic cytokines. There are numerous scientific reports on the therapeutic potential of melatonin in treatment of asthma, respiratory diseases for infections, chronic obstructive pulmonary disease, lung cancer, pleural cavity diseases, as well as vascular pulmonary disease. In the present communication, we systematically review the therapeutic potential of melatonin in the treatment of respiratory diseases along with its molecular mechanism of actions.

Antioxidant effects of propolis on carp Cyprinus carpio exposed to arsenic: biochemical and histopathologic findings.

Propolis, a resinous material produced by worker bees from the leaf buds and exudates of plants, is reported to possess various therapeutic properties. The aim of this study was to investigate the effects of propolis on biochemical parameters and histopathologic findings in carp Cyprinus carpio L. exposed to arsenic. A sublethal concentration of arsenic (0.01 mg l-1) and/or 10 mg l-1 propolis were administered to fish for 1 wk. Catalase (CAT) activities and malondialdehyde (MDA) levels were determined in liver, gill and muscle tissues in control, arsenic only, propolis only and arsenic+propolis treatment groups. Results showed that CAT activity decreased in the arsenic group compared to the control and propolis groups. CAT activity in the arsenic+propolis group was significantly higher compared to the arsenic group. MDA levels in fish exposed to 0.01 mg l-1 arsenic significantly increased compared to the control group. However, MDA levels in the arsenic+propolis group were significantly lower compared to the arsenic group. Histopathological changes in the liver, gill and muscle tissues of carp were examined by light microscopy: various changes were observed in all tissues of fish in the arsenic group. Propolis showed important antioxidant effects against arsenic toxicity in all fish tissues.

Role of propolis on biochemical parameters in kidney and heart tissues against L-NAME induced oxidative injury in rats.

Nitric oxide (NO), produced by endothelial NO synthase, is recognised as a central antiinflammatory and antiatherogenic principle in the vasculature. Epidemiological and clinical studies have demonstrated that a growing list of natural products, as components of the daily diet or phytomedical preparations, may improve vascular function by enhancing NO bioavailability. In this article, we investigated antioxidant effects of propolis on biochemical parameters in kidney and heart tissues of acute NO synthase inhibited rats by Nω-nitro-l-arginine methyl ester (l-NAME). There was increase (p < 0.001) in the activities of catalase and malondialdehyde levels in the l-NAME treatment groups when compared with control rats, but NO levels were decreased in both kidney and heart tissues. There were statistically significant changes (p < 0.001) in these parameters of l-NAME + propolis treated rats as compared with l-NAME-treated group. In summary, propolis may influence endothelial NO production.

Effects of various propolis concentrations on biochemical and hematological parameters of rainbow trout (Oncorhynchus mykiss).

Biochemical and hematological parameters in blood of rainbow trout treated to various concentrations of propolis for 96 h were determined. Total leukocyte count and granulocytes values increased (p<0.05) in 0.02 and 0.03 g/L propolis groups. There was a decrease in agranulocytes (p<0.05) erythrocytes, hemoglobin and hematocrit values for fish exposed to 0.02 and 0.03 g/L propolis. MCV and MCH values (p<0.05) were significantly increased; 0.02 and 0.03 g/L propolis caused an increase (p<0.05) in the levels of glucose, blood urea nitrogen, triglyceride, total cholesterol, lactate dehydrogenase, amylase and gamma glutamyltransferase. There was a decrease in the levels of aspartate aminotransferase and alkaline phosphatase. Hematological and biochemical protective effects of 0.01 g/L propolis were investigated. Dose-dependent effects of propolis on blood of fish can be favorable, opening new perspectives of investigation on their biological properties and utilization.