Effect of Different Cryoprotectants (Glycerol, Methanol and Dimethyl Sulfoxide) on Post-thaw Quality, Viability, Fertilization Ability and DNA Damage of Cryopreserved Nile Tilapia (Oreochromis niloticus) Spermatozoa.

BACKGROUND: Cryopreservation of sperm from different fish species requires different protocols. Therefore, it is necessary to perform studies to establish reliable procedures for each species. OBJECTIVE: Experiments were designed to analyse the effect of different types of cryoprotectants on post-thaw motility, viability and fertility as well as cryoresistance of cryopreserved Nile tilapia (Oreochromis niloticus) sperm. MATERIALS AND METHODS: Sperm samples were diluted with an ionic extender containing glycerol (Gly), methanol (MeOH) and dimethyl sulfoxide (DMSO) at ratios of 5, 10 and 15 % respectively. Diluted samples were aspirated into 0.25 ml French straws and frozen 3 cm above the surface of liquid nitrogen (LN) in a styrofoam box and stored in a LN tank. DNA damage was evaluated with the comet assay technique following cryopreservation. RESULTS: Supplementation of extender with 10% glycerol gave the highest motility rate compared with the other cryoprotectant groups (P<0.05). Differences in terms of post-thaw motility duration, cell viability and fertilization rates were not significant among treatments (P>0.05). Although Gly gave the best score (5.0 ± 0.1%, P>0.05) at the concentration of 10%, 5% Me2SO caused significant DNA damage (15.0 ± 1.0%, P<0.05) with the comet test. CONCLUSION: Gly or MeOH are more suitable cryoprotectants than DMSO for the cryopreservation of Nile tilapia sperm.

Effect of Extender Supplemented with Boron on Post-Thaw Motility, Viability, DNA Damage and Fertilization Ability of Cryopreserved Brown Trout (Salmo trutta macrostigma) Spermatozoa.

BACKGROUND: Boron has been considered as an essential nutrient for decreasing lipid peroxidation and improving antioxidant mechanism in different animal species. On the other hand, its effect on quality or DNA damage following cryopreservation process in fish sperm is still unclear. OBJECTIVE: Experiments were designed to analyse the effect of an ionic based extender supplemented with boron on post-thawed motility, viability, fertility and DNA integrity of cryopreserved brown trout (Salmo trutta macrostigma) sperm. MATERIALS AND METHODS: Sperm samples were cryopreserved with the ionic extender containing different boron concentrations (0.1, 0.2, 0.3 and 0.4 mM) using a controlled freezer at two different freezing rates (FR-I: 10°C min-1 from +4°C to -40°C and FR-II: 15°C min-1 from +4°C to -40°C). Sperm motility, viability, fertilization, eyeing and DNA fragmentations were determined in post-thawed samples. RESULTS: Freezing rate-I provided significantly higher fertilization and eyeing rates compared to freezing rate-II (p<0.05). Higher post-thaw motility (62.8±1.4%) and fertilization (75.2±0.9%) rates were obtained with the 0.4 mM boron concentration at freezing rate-I. CONCLUSION: Supplementation of the extender with boron increased fertilization and eyeing rates and also decreased DNA damages at both freezing rates.

Supplementation of quercetin for advanced DNA integrity in bull semen cryopreservation.

The aim of this study was to identify the effects of adding quercetin (Q) to Tris extender in order to identify levels of oxidative stress in bull sperm after freeze thawing. Ejaculates were collected via artificial vagina from Holstein bulls. Semen was divided into five tools and diluted to a final concentration of 15 × 106 spermatozoa/ml with the Tris extender containing Q (25, 50, 100 and 200 µg/ml) and no additive (control; C). All examples were equilibrated at 4°C during 4 hr then were loaded into 0.25-ml straws and frozen using a controlled rate. Sperm motility and motility characteristics were determined using the computer-assisted semen analyser. Sperm membrane integrity was assessed using the hypoosmotic swelling test. Sperm chromatin integrity was investigated using the single cell gel electrophoresis. Total antioxidant capacities were performed colorimetrically. Q supplementation used as an antioxidant did not produce better results in the proportion of sperm progressive and total motility, plasma membrane integrity and sperm abnormalities. Q supplementation exhibited the favourable tail length, tail DNA and tail moment. In conclusion, when whole parameters are considered, Q25 can be added to the Tris extender due to its positive effect on sperm DNA integrity and no adverse effect on the progressive and total motilities of sperm.

Supplementation of Rosmarinic Acid Has Reduced Oxidative Stress on Bull Spermatozoa Following the Freeze Thawing Process.

BACKGROUND: Cryopreservation has a side effect on the motility, chromatin integrity and viability of sperm cells. OBJECTIVE: The present study investigated the effects of supplementation with rosmarinic acid (RA) Tris extender on sperm quality parameters, plasma and acrosome membrane damage, antioxidant enzyme activity and chromatin integrity following the freeze thawing process on bull spermatozoa. MATERIALS AND METHODS: Ejaculates were split into five aliquots and diluted to a final concentration of 15x106 spermatozoa per ml with the Tris extender containing RA (25, 50, 100 and 200 microgram per ml) and (control) and then frozen at a controlled rate. RESULT: Treatments did not give better results on the percentages of sperm progressive, total motility and sperm motion characters (P >0.05); however, RA25 and RA50 exhibited favourable chromatin integrity. In conclusion, RA25 and RA50 increased total antioxidant activity. As a consequence, the amount of MDA and chromatin damage were reduced in sperm cells.

Adsorption of Hydrogen Molecules on Carbon Nanotubes Using Quantum Chemistry and Molecular Dynamics.

Physisorption and storage of molecular hydrogen on single-walled carbon nanotube (SWCNT) of various diameters and chiralities are studied by means of classical molecular dynamics (MD) simulations and a force field validated using DFT-D2 and CCSD(T) calculations. A nonrigid carbon nanotube model is implemented with stretching (C-C) and valence angle potentials (C-C-C) formulated as Morse and Harmonic cosine potentials, respectively. Our results evidence that the standard Lennard-Jones potential fails to describe the H2-H2 binding energies. Therefore, our simulations make use of a potential that contains two-body term with parameters obtained from fitting CCSD(T)/CBS binding energies. From our MD simulations, we have analyzed the interaction energies, radial distribution functions, gravimetric densities (% wt), and the distances of the adsorbed H2 layers to the three zigzag type of nanotubes (5,0), (10,0), and (15,0) at 100 and 300 K.

The effects of proanthocyanidin on testicular toxicity in rats exposed to a glyphosate-based herbicide.

The purpose of this study is to determine the effects of proanthocyanidin (PA) on spermatological parameters and testicular toxicity in male rats exposed to glyphosate (GLP). In our study, four groups were formed out of 24 male rats, each group would include 6 rats. The rats in the PA group were given a dose of 400 mg/kg/day dissolved in DMSO via gastric gavage. The rats in the GLP+PA groups were first given GLP at the LD50/10 dose of 787.85 mg/kg/day, followed by administering PA at a dose of 400 mg/kg/day dissolved in DMSO via gastric gavage. The rats in the GLP group were given GLP at the LD50/10 dose of 787.85 mg/kg/day dissolved in DMSO via gastric gavage. It was determined that in terms of motility, in comparison to the control group, the decreases in the GLP group and the increases in the PA and GLP+PA groups were statistically significant (p<0.001). The administration of GLP increased DNA damage compared to the control group, but the GLP+PA and PA applications reduced DNA damage (p<0.001). The analysis of testosterone levels indicated a statistically significant reduction in the GLP group compared to the other groups. Consequently, it was determined that PA effectively prevented the decreases in the spermatological parameters lowered as a result of GLP exposure and the oxidative stress and toxicity in testicular tissue.

Effect of α-lipoic acid on sperm quality, reproductive tract measures in thinner exposed rats.

The aim of this study was to investigate the effect of alpha lipoic acid (ALA, an universal antioxidant) on thinner-induced testicular toxicity regarding spermatological features, body and reproductive tract measures in rats. Adult male Sprague-Dawley rats were divided into five treatment groups, eight rats in each. Control group was treated with placebo. Group O was given only olive oil. The group L received only α-lipoic acid. Thinner + Lipoic Acid group received thinner + α-lipoic acid and group T received only thinner. Thinner alone administration caused significant decreases in body and some reproductive organ weights, sperm count, motility and sperm membrane integrity, and significant increases in seminal vesicle weight and abnormal sperm rates compared with the values in the control group. However, concomitant administration of thinner with α-lipoic acid provided significant improvements in sperm parameters compared with values in alone group T. In conclusion, the results of this study suggest that α-lipoic acid has a protective effect against thinner-induced reproductive dysfunction in male rats.

The cryoprotective potential of propolis supplemented in frozen-thawed bull semen; biochemical and physiological findings.

In this study, the cryoprotective effect of different doses of propolis (P) on bull semen, which has solid pharmacological properties thanks to its rich phenolic components, was investigated biochemically and physiologically. Semen samples were collected from Simmental breed bulls via the artificial vagina and pooled. After dividing into five groups, control (C: no additive) and four different P (200, 100, 50, and 25 µg/mL) groups, the final concentration was diluted to 16×106 per straw. Semen samples were equilibrated at 4°C for approximately 4 hours, then placed in French straws and frozen. After thawing, sperm motility and kinetic parameters, DNA integrity by single-cell gel electrophoresis, sperm abnormalities by liquid fixation, and lipid peroxidation levels by the colorimetric method was analyzed by Computer-Assisted Semen Analyzer. P added to the diluent showed no effect on motility and kinetic parameters at P25 and P50 (p>0.05), while P100 and P200 had a negative effect (p⟨0.001). The addition of P (25 and 50) showed a treatment effect on tail abnormality compared to C (p⟨0.05). Especially P50 had a positive effect on tail length, tail DNA, and tail movement, while P100 and P200 caused DNA damage (p⟨0.001). MDA levels increased in all P dose groups compared to C (p⟨0.001). This study has clearly demonstrated that P25 and P50 supplements could be used therapeutically to treat sperm tail abnormalities and prevent DNA damage in post-thawed bull sperm.

Influence of sperm concentration on the motility, morphology, membrane and DNA integrity along with oxidative stress parameters of ram sperm during liquid storage.

The aim of this study was to determine the influences of two different concentrations in terms of motility, morphology, membrane integrity (viability and HOST response: HE-test; modified hypoosmotic swelling test (HOST) associated with supravital eosin staining test), DNA integrity (COMET assay) and oxidative stress parameters (MDA, malondialdehyde; AOA, total antioxidant activity; GSH, reduced glutathione; NOx, nitric oxide) of liquid stored ram sperm for 5 days. Two different concentrations suitable for laparoscopic and cervical inseminations were employed. A total of 5 Pirlak rams (Daglic × Kivircik, local breed) with satisfactory breeding potential were selected. Semen samples were collected by artificial vagina. Ejaculates were extended to 25 and 100 million sperm per ml with Tris-based egg-yolk (T) extender at room temperature and stored at 4°C. The concentration significantly had effects on motility, morphology, membrane and DNA integrity along with oxidative stress parameters (P<0.05). Overall changes in the motility, morphology, membrane and DNA integrity along with oxidative stress parameters of ram sperm diluted with T in the 100 × 10(6)ml(-1) concentration and preserved at 4°C for a short term was found to be better preservation than that of diluted with T in the 25 × 10(6)ml(-1) concentration. Significant concentration × days of storage interactions were found for all parameters except the NOx. Further studies are required to elucidate the oxidative stress changes on sperm quality parameters in different concentrations during the liquid storage of ram semen.

Calidad percibida de la atención de enfermería por usuarios hospitalizados, Tunja, 2011 / Perceived quality of nursing care hospital by users, Tunja 2011

Objetivo: evaluar la calidad percibida de los usuarios en servicios de hospitalización respecto a la atención de enfermería recibida. Metodología: estudio cuantitativo, descriptivo de corte transversal. Análisis univariado y bivariado. La muestra n=183 fue recolectada en un Hospital de Tunja en los servicios de Medicina Interna y Quirúrgicas, a través del instrumento servqhos-e. Resultados: la medición de la calidad objetiva, subjetiva y el nivel de satisfacción de los usuarios de los servicios del hospital, reportó puntuaciones con medias de 4,08 para amabilidad y 3,98 para preparación del personal de enfermería y un nivel de satisfacción global del 92,9%. Discusión: la medición de la calidad percibida por los usuarios mediante el instrumento servqhos-e, permite evaluar desde la percepción de los usuarios, la calidad de la atención brindada por profesionales de enfermería en aspectos objetivos y subjetivos, los cuales son predictores de la satisfacción; los aspectos mejor evaluados corresponden a variables subjetivas, sin embargo, las expectativas de los usuarios no se superan...

Objective: to evaluate the usersÆ perception regarding the quality of the nursing care received. Methodology: a quantitative, cross-sectional descriptive study with univariate and bivariate analyses. The sample (n = 183) was collected from the Internal Medicine and Surgery units of the Tunja Hospital using the servqhos-e instrument. Results: the measurement of the objective and subjective quality, as well as the measurement of the usersÆ satisfaction level regarding hospital services had scores with a mean of 4.08 for kindness, and 3.98 for nursing staff training. In addition, the overall satisfaction level was 92.9%. Discussion: using the instrument servqhos-e to measure the quality perceived by the users made it possible to assess the quality of the care provided by nursing professionals. This measurement was conducted from the userÆs own point of view and focused on the objective and subjective aspects, which are predictors of satisfaction. Additionally, the highest scores belonged to subjective variables, but the usersÆ expectations were not met...

Relationship between some reproductive parameters and biochemical properties of blood serum in rams.

The objective of the present study was to describe the relationship between some reproductive parameters, hormonal levels, and some biochemical properties of blood serum in rams under the Afyon province conditions. The total protein and globulin were positively (P < 0.01) correlated with sperm motility and sperm concentration in all of rams. Total lipid was negatively (P < 0.05) correlated with sperm motility and concentration in Daglic, and it was positively (P < 0.01) correlated with percentage of abnormal spermatozoa in Chios rams. Total lipid and cholesterol was positively (P < 0.01) correlated with triiodothyronine in all of rams and was negatively (P < 0.05) correlated with testosterone in Daglic rams. Percentage of abnormal spermatozoa were negatively (P < 0.01) correlated with alanine amino transaminase (ALT) level, but were positively (P < 0.01) associated with aspartate amino transaminase (AST) level and AST/ALT ratio in all of rams.