THE RELEASE OF PLASMINOGEN ACTIVATORS FROM SPERMATOZOA DURING ACROSOME REACTION

PLASMINOGEN ACTIVATORS ARE ENZYMES THAT CONVERT THE PROENZYME PLASMINOGEN TO THE TRYPSIN-LIKE PROTEINASE PLASMIN. PLASMINOGEN ACTIVATORS AND PLASMIN ARE REGULATED BY SEVERAL PLASMINOGEN ACTIVATOR INHIBITORS AND PLASMIN INHIBITORS, RESPECTIVELY. IN ADDITION TO SEVERAL ENZYMES LOCATED IN THE HEAD OF SPERMATOZOON, THE PRESENCE OF PLASMINOGEN ACTIVATORS HAS RECENTLY BEEN SHOWN. PLASMINOGEN ACTIVATORS AND PLASMINOGEN ACTIVATORS INHIBITORS ARE LOCALIZED IN THE PLASMA MEMBRANE AND OUTER ACROSOMAL MEMBRANE OF SPERMATOZOA. THE PRESENT STUDY WAS UNDERTAKEN TO INVESTIGATE THE POSSIBLE RELEASE OF PLASMINOGEN ACTIVATORS,PLASMINOGEN ACTIVATOR INHIBITORS AND PLASMIN INHIBITORS FROM BULL, BOAR AND RAM SPERMATOZOA DURING THE ACROSOME REACTION INDUCED IN VITRO BY VARIOUS CHEMICAL AGENTS (HYAMINE 2389, TRITON X-100, CALCIUM IONOPHORE A23187). BOTH ACROSOMAL STATUS AND VIABILITY OF SPERMATOZOA WERE CHECKED BY USING THE STAINING PRECEDURES EOSIN-NIGROSIN FOR BOAR AND RAM SPERMATOZOA AND DUAL STAIN TECHNIQUE FOR BULL SPERMATOZOA. THE PLASMINOGEN ACTIVATOR ACTIVITY (PAA), PLASMINOGEN ACTIVATOR INHIBITION (PAI) AND PLASMIN INHIBITION (PI) WERE DETERMINED BY SPECTROPHOTOMETRIC METHODS. THE IDENTIFICATION OF PLASMINOGEN ACTIVATORS (T-PA, U-PA) WAS ATTAINED BY A COMBINATION OF IMMUNOLOGICAL AND SPECTROPHOTOMETRIC METHODS. (SHORTENED)ΣΤΗΝ ΠΕΙΡΑΜΑΤΙΚΗ ΑΥΤΗ ΕΡΓΑΣΙΑ ΔΙΕΡΕΥΝΗΘΗΚΕ Η ΠΙΘΑΝΟΤΗΤΑ ΑΠΕΛΕΥΘΕΡΩΣΗΣ ΤΩΝ ΕΝΕΡΓΟΠΟΙΩΝ ΤΟΥ ΠΛΑΣΜΙΝΟΓΟΝΟΥ, ΤΩΝ ΑΔΡΑΝΟΠΟΙΩΝ ΤΩΝ ΕΝΕΡΓΟΠΟΙΩΝ ΤΟΥ ΠΛΑΣΜΙΝΟΓΟΝΟΥ ΚΑΙ ΤΩΝ ΑΔΡΑΝΟΠΟΙΩΝ ΤΗΣ ΠΛΑΣΜΙΝΗΣ ΚΑΤΑ ΤΗΝ ΑΝΤΙΔΡΑΣΗ ΤΟΥ ΑΚΡΟΣΩΜΑΤΟΣ ΤΟΥ ΣΠΕΡΜΑΤΟΖΩΑΡΙΟΥ. ΓΙΑ ΤΗ ΣΥΓΚΕΚΡΙΜΕΝΗ ΕΡΕΥΝΑ ΧΡΗΣΙΜΟΠΟΙΗΘΗΚΑΝ ΔΕΙΓΜΑΤΑ ΣΠΕΡΜΑΤΟΣ ΑΠΟ 4 ΚΑΠΡΟΥΣ ΦΥΛΗΣ LARGE WHITE, 10 ΚΡΙΟΥΣ ΦΥΛΗΣ ΧΙΟΥ ΚΑΙ 8 ΤΑΥΡΟΥΣ ΦΥΛΗΣ BROWN SWISS (129 ΔΕΙΓΜΑΤΑ). ΠΡΙΝ ΑΚΟΜΗ ΤΑ ΣΠΕΡΜΑΤΟΖΩΑΡΙΑ ΥΠΟΣΤΟΥΝ ΤΗΝ ΕΠΙΔΡΑΣΗ ΤΩΝ ΟΥΣΙΩΝ ΠΟΥ ΠΡΟΚΑΛΟΥΝ ΤΗΝ ΑΝΤΙΔΡΑΣΗ ΤΟΥ ΑΚΡΟΣΩΜΑΤΟΣ, ΑΠΑΛΛΑΣΟΝΤΑΝ ΑΠΟ ΤΟ ΠΛΑΣΜΑ ΤΟΥ ΣΠΕΡΜΑΤΟΣ ΜΕ ΦΥΓΟΚΕΝΤΡΗΣΗ, ΠΛΕΝΟΝΤΑΝ ΚΑΙ ΑΡΑΙΩΝΟΝΤΑΝ ΜΕ ΕΙΔΙΚΟ ΡΥΘΜΙΣΤΙΚΟ-ΘΡΕΠΤΙΚΟ ΥΠΟΣΤΡΩΜΑ ΔΙΑΦΟΡΕΤΙΚΟ ΓΙΑ ΚΑΘΕ ΖΩΙΚΟ ΕΙΔΟΣ. Η ΑΝΤΙΔΡΑΣΗ ΤΟΥΑΚΡΟΣΩΜΑΤΟΣ ΠΡΟΚΛΗΘΗΚΕ ΣΤΑ ΣΠΕΡΜΑΤΟΖΩΑΡΙΑ ΤΟΥ ΚΑΠΡΟΥ, ΤΟΥ ΚΡΙΟΥ ΚΑΙ ΤΟΥ ΤΑΥΡΟΥ ΜΕΤΑ ΑΠΟ ΕΠΙΔΡΑΣΗ ΥΑΜΙΝΗΣ 2389, TRITON X-100 ΚΑΙ CALCIUM IONOPHORE A23187. ΟΙ ΣΥΓΚΕΝΤΡΩΣΕΙΣ ΤΩΝ ΟΥΣΙΩΝ ΠΟΥ ΧΡΗΣΙΜΟΠΟΙΗΘΗΚΑΝ ΓΙΑ ΤΗΝ ΠΡΟΚΛΗΣΗ ΤΗΣ ΑΝΤΙΔΡΑΣΗΣ ΤΟΥ ΑΚΡΟΣΩΜΑΤΟΣ, ΚΑΘΩΣ ΚΑΙ Ο ΧΡΟΝΟΣ ΕΚΘΕΣΗΣ ΤΩΝ ΣΠΕΡΜΑΤΟΖΩΑΡΙΩΝ ΣΤΙΣ ΟΥΣΙΕΣ ΑΥΤΕΣ, ΚΑΘΟΡΙΣΘΗΚΑΝ ΕΤΣΙ, ΜΕΤΑ ΑΠΟ ΠΟΛΛΟΥΣ ΠΕΙΡΑΜΑΤΙΣΜΟΥΣ, ΩΣΤΕ ΝΑ ΕΠΙΤΕΥΧΘΕΙ ΤΟ ΜΕΓΙΣΤΟ ΠΟΣΟΣΤΟ ΤΗΣ ΑΝΤΙΔΡΑΣΗΣ ΤΟΥ ΑΚΡΟΣΩΜΑΤΟΣ ΧΩΡΙΣ ΣΗΜΑΝΤΙΚΗ ΑΥΞΗΣΗ ΤΟΥ ΠΟΣΟΣΤΟΥ ΤΩΝ ΝΕΚΡΩΝ ΣΠΕΡΜΑΤΟΖΩΑΡΙΩΝ. (ΠΕΡΙΚΟΠΗ

Impact of Exercise and Aging on Rat Urine and Blood Metabolome. An LC-MS Based Metabolomics Longitudinal Study

Aging is an inevitable condition leading to health deterioration and death. Regular physical exercise can moderate the metabolic phenotype changes of aging. However, only a small number of metabolomics-based studies provide data on the effect of exercise along with aging. Here, urine and whole blood samples from Wistar rats were analyzed in a longitudinal study to explore metabolic alterations due to exercise and aging. The study comprised three different programs of exercises, including a life-long protocol which started at the age of 5 months and ended at the age of 21 months. An acute exercise session was also evaluated. Urine and whole blood samples were collected at different time points and were analyzed by LC-MS/MS (Liquid Chromatography–tandem Mass Spectrometry). Based on their metabolic profiles, samples from trained and sedentary rats were differentiated. The impact on the metabolome was found to depend on the length of exercise period with acute exercise also showing significant changes. Metabolic alterations due to aging were equally pronounced in sedentary and trained rats in both urine and blood analyzed samples

The addition of crocin in the freezing medium extender improves post-thaw semen quality

Semen cryopreservation is arguably the most important method or technique contributing to the advancement of modern animal production. However, the quality of sperm after thawing is still highly variable. The addition of antioxidant compounds to the freezing medium has been used customarily to counteract the harmful effects of Reactive Oxygen Species (ROS) that are produced during the freeze/thaw process. Crocin, a potent antioxidant, improves the fertilizing capacity of spermatozoa. In this study, we evaluated the potential of crocin (0, 0.5 and 1 mM) as an extender additive to diminish the damaging effects of cryopreservation on bovine spermatozoa. Post-thaw semen quality was assessed by means of motility, viability and lipid peroxidation (LPO). We further investigated the effect of crocin supplementation upon freezing on sperm quality parameters during their incubation at 37°C for up to 2 hr. Overall, the data assessment indicates that crocin facilitated a general improvement of the quality of freeze/thawed spermatozoa, under the present experimental conditions. Crocin (1 mM) maintained a higher percentage of alive spermatozoa with intact acrosome with rapid and progressive motility, compared to the control extender. Moreover, the spermatozoa cryopreserved in the presence of crocin exhibited higher values in CASA kinematic parameters (VCL, VSL, VAP, ALH) immediately after thawing. Furthermore, the positive effect of crocin on motility parameters was also sustained over a period of 2 hr incubation at 37°C. This effect of crocin may be attributed to the observed inhibition of LPO during the incubation period. Thus, the results indicate that the addition of crocin (especially at a final concentration of 1 mM) in the freezing extender medium may benefit the preservation of the quality parameters of spermatozoa that are compromised by the freeze/thaw heat shock and the stress during handling for IVF or artificial insemination

Omega 3 fatty acids supplementation has an ameliorative effect in experimental ulcerative colitis despite increased colonic neutrophil infiltration Los suplementos de ácidos grasos omega 3 tienen efectos beneficiosos en colitis ulcerosa a pesar del aumento de la infiltracción por neutrófilos del colon

Purpose: omega 3 polyunsaturated fatty acids have anti-inflammatory properties and can be beneficial in the treatment of inflammatory diseases, such as ulcerative colitis. Dextran sodium sulphate (DSS) colitis in rats appears to mimic nearly all of the morphological characteristics and lesion distributions of ulcerative colitis. The purpose of the current study was to investigate the efficacy of omega 3 fatty acids in the treatment of experimental ulcerative colitis. Methods: thirty-six Wistar rats were randomly assigned to group A or group B receiving 5% dextran sulfate sodium (DSS) in their drinking water for eight days. For the next eight days post-DSS, group A animals received tap-water, and group B animals were fed a nutritional solution containing high levels of omega 3 polyunsaturated fatty acids (ProSure®, Abbott Laboratories, Zwolle, Netherlands) once per day, administrated with a orogastric feeding tube. Results: animals fed an omega 3 rich diet exhibited a statistically significant increase in hematocrit and hemoglobin levels, compared to animals drinking tap water, and a trend towards histopathological and clinical improvement, with the administration of omega 3 fatty acids ameliorating epithelial erosion by day 8 post-DSS, but no statistically significant difference was observed between group A and group B animals at 4 or 8 days post-DSS. Also, a statistically significant increase in neutrophil infiltration was observed, as depicted by myelohyperoxidase activity. Conclusion: our findings support a positive role of omega 3 polyunsaturated fatty acids supplementation in an experimental model of ulcerative colitis despite the increased colonic neutrophil infiltration. Further studies are needed in order to investigate the role of increased neutrophils in colonic mucosa

The Strong Antioxidant Sheep/Goat Whey Protein Protects Against mTOR Overactivation in Rats: A Mode of Action Mimicking Fasting

Whey protein, a by-product of the cheese industry, can be putatively used as a functional food due to its beneficial health properties. The main objective of the present study was to assess in vivo the effect of a sheep/goat whey protein on the plasma amino acid profile and mammalian target of rapamycin (mTOR), a regulator of skeletal myogenesis. A control group was fed with a standard commercial diet while the experimental group received a standard commercial diet plus sheep/goat whey protein for 28 days. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was conducted to determine plasma amino acid levels while the expression of p70-S6 Kinase 1 (p70-S6K1) in liver and quadriceps muscles was quantified and used as a biomarker of mTOR activity. The results obtained showed a decrease in the levels of essential and branched-chain amino acids (BCAAs) in the experimental group. Furthermore, p70-S6K1 expression was decreased in the liver of rats consumed whey protein. In conclusion, the reduction of amino acid levels and the concomitant inactivation of mTOR imply that whey could potentially act protectively against disorders induced by mTOR overactivation. Intriguingly, this mode of action mimics fasting, an approach with established advantageous health effects

Effects of Aging, Long-Term and Lifelong Exercise on the Urinary Metabolic Footprint of Rats

Life expectancy has risen in the past decades, resulting in an increase in the number of aged individuals. Exercise remains one of the most cost-effective treatments against disease and the physical consequences of aging. The purpose of this research was to investigate the effects of aging, long-term and lifelong exercise on the rat urinary metabolome. Thirty-six male Wistar rats were divided into four equal groups: exercise from 3 to 12 months of age (A), lifelong exercise from 3 to 21 months of age (B), no exercise (C), and exercise from 12 to 21 months of age (D). Exercise consisted in swimming for 20 min/day, 5 days/week. Urine samples collection was performed at 3, 12 and 21 months of life and their analysis was conducted by liquid chromatography-mass spectrometry. Multivariate analysis of the metabolite data did not show any discrimination between groups at any of the three aforementioned ages. However, multivariate analysis discriminated the three ages clearly when the groups were treated as one. Univariate analysis showed that training increased the levels of urinary amino acids and possibly protected against sarcopenia, as evidenced by the higher levels of creatine in the exercising groups. Aging was accompanied by decreased levels of urinary amino acids and signs of increased glycolysis. Concluding, both aging and, to a lesser degree, exercise affected the rat urinary metabolome, including metabolites related to energy metabolism, with exercise showing a potential to mitigate the consequences of aging

Redox Biomarkers Assessment after Oral Administration of Wine Extract and Grape Stem Extract in Rats and Mice

Wine and by-products of the winemaking process, such as grape stems, are rich in bioactive polyphenolic compounds that might be beneficial for animal and human health. In recent years, the administration of dietary polyphenols with strong antioxidant and cytoprotective properties has constituted an emerging line of research interest toward disease prevention. However, in scientific literature, only a limited number of studies have investigated the safety and the toxicological risks of polyphenolic compounds in vivo. Based on the above, the purpose of the present study was two-fold: first, to examine the effects of oral administration of a grape stem extract, derived from the Greek red wine Mavrodaphne, on mice redox biomarkers; and second, to investigate the biological effects of oral administration of a wine extract, derived from the emblematic Greek red wine Xinomavro, on rats. Toward this purpose, body weight, growth rate, hematological, biochemical, and histopathological parameters, as well as a panel of redox biomarkers, were examined. According to our results, the administration of Mavrodaphne grape stem extract in mice induced alterations in redox homeostasis, preventing mice from the adverse effects of lipid peroxidation. Contrariwise, the administration of Xinomavro wine extract induced both beneficial and harmful outcomes on rat redox status determined by the examined tissue. Collectively, our study reports that the Mavrodaphne grape stem extract, a serious pollutant when disposed in environmental matrices, is an important source of bioactive polyphenolic compounds that could protect from oxidative damage and improve animal and human health. Finally, the Xinomavro wine extract exerts tissue-specific changes in redox balance, which are indicative of the complexity that characterizes the biological systems

Redox Biomarkers Assessment after Oral Administration of Wine Extract and Grape Stem Extract in Rats and Mice

Wine and by-products of the winemaking process, such as grape stems, are rich in bioactive polyphenolic compounds that might be beneficial for animal and human health. In recent years, the administration of dietary polyphenols with strong antioxidant and cytoprotective properties has constituted an emerging line of research interest toward disease prevention. However, in scientific literature, only a limited number of studies have investigated the safety and the toxicological risks of polyphenolic compounds in vivo. Based on the above, the purpose of the present study was two-fold: first, to examine the effects of oral administration of a grape stem extract, derived from the Greek red wine Mavrodaphne, on mice redox biomarkers; and second, to investigate the biological effects of oral administration of a wine extract, derived from the emblematic Greek red wine Xinomavro, on rats. Toward this purpose, body weight, growth rate, hematological, biochemical, and histopathological parameters, as well as a panel of redox biomarkers, were examined. According to our results, the administration of Mavrodaphne grape stem extract in mice induced alterations in redox homeostasis, preventing mice from the adverse effects of lipid peroxidation. Contrariwise, the administration of Xinomavro wine extract induced both beneficial and harmful outcomes on rat redox status determined by the examined tissue. Collectively, our study reports that the Mavrodaphne grape stem extract, a serious pollutant when disposed in environmental matrices, is an important source of bioactive polyphenolic compounds that could protect from oxidative damage and improve animal and human health. Finally, the Xinomavro wine extract exerts tissue-specific changes in redox balance, which are indicative of the complexity that characterizes the biological systems

Additional file 1: of The NAD+ precursor nicotinamide riboside decreases exercise performance in rats

Raw data of time to exhaustion in the incremental swimming test for the control and the nicotinamide riboside group. (XLSX 9 kb