scholarly journals The effect of resorcinol on bovine spermatozoa parameters in vitro

2020 ◽  
pp. 675-686
Author(s):  
M Massányi ◽  
M Halo ◽  
L Strapáková ◽  
T Slanina ◽  
P Ivanič ◽  
...  
Keyword(s):  
Cultivation Temperature ◽  
Negative Effects ◽  
Mtt Test ◽  
Progressive Motility ◽  
Time Period ◽  
Entire Duration ◽  
Bovine Spermatozoa ◽  
Time Periods ◽  
Motility Parameters

The goal of this study was to observe the effect of resorcinol on motility, viability and morphology of bovine spermatozoa. The semen was used from six randomly chosen breeding bulls. Ejaculate was diluted by different solutions of resorcinol in 1:40 ratio. Samples were divided into 7 groups with different concentrations of resorcinol (Control, RES1 – 4 mg/ml, RES2 – 2 mg/ml, RES3 – 1 mg/ml, RES4 – 0.5 mg/ml, RES5 – 0.25 mg/ml and RES6 – 0.125 mg/ml). Motility of spermatozoa was detected using CASA method at temperature of 37 °C in time periods 0, 1, 2, 3, 4 hours from the start of the experiment. Significant motility differences between all groups except control and RES6 with difference of 5.58 %, as well as between RES1 and RES2 groups with difference of 2.17 % were found. Progressive motility had the same significant differences. Spermatozoa viability (MTT test) decreased compared to control in all experimental groups during the entire duration of experiment. Observing morphologically changed spermatozoa, no significant changes were observed and a higher percentage of spermatozoa with separated flagellum in all experimental resorcinol groups compared to control were detected. Also, increased number of spermatozoa with broken flagellum, acrosomal changes and other morphological forms in the group with the highest concentration of resorcinol (RES1) were found. Results of our study clearly show negative effects on motility parameters of spermatozoa which depend on concentration, cultivation temperature and time period.

scholarly journals Effect of taurine on turkey (Meleagris gallopavo) spermatozoa viability and motility

2018 ◽  
Vol 63 (No. 4) ◽  
pp. 127-135 ◽  
Author(s):  
T. Slanina ◽  
M. Miškeje ◽  
F. Tirpák ◽  
M. Błaszczyk ◽  
R. Stawarz ◽  
...  
Keyword(s):  
Meleagris Gallopavo ◽  
Lower Percentage ◽  
Computer Assisted ◽  
Viability Assessment ◽  
Spermatozoa Motility ◽  
In Vitro Incubation ◽  
Time Periods ◽  
Casa System ◽  
Motility Parameters

The effect of taurine on the turkey spermatozoa motility and viability during the in vitro incubation was assessed. Experimental samples were prepared by diluting the raw semen in nine different concentrations of taurine – from 10 mg/ml to 0.078125 mg/ml. The motility parameters were evaluated by the CASA system (Computer Assisted Semen Analyser) using the program Sperm Vision<sup>®</sup> and for spermatozoa viability assessment the eosin-nigrosin staining was performed. Selected parameters were evaluated at six time periods: 0, 1, 2, 3, 4, and 5 h at 5°C and 41°C. At 5°C, a significantly lower percentage of motility and progressive motility was detected only in the samples with the highest concentration of taurine (10 mg/ml) at time 0 and 1. After 2 h of incubation a significant preventive effect of taurine on spermatozoa parameters was observed. The tendency of the taurine effect on motility parameters was different during the in vitro incubation at 41°C. Significantly lower values of motility parameters were detected in all experimental samples in comparison to the control after 5 h. The analysed concentrations of taurine did not significantly affect viability of turkey spermatozoa during all time periods. A higher percentage of dead spermatozoa were observed at 41°C (4.87–9.90%) if compared to 5°C (2.12–4.88%). The results indicated that the addition of taurine (from 2.5 to 7.5 mg/ml) to turkey spermatozoa positively affected the monitored spermatozoa parameters incubated at 5°C.

scholarly journals Antibacterial Effects of Levofloxacin, Erythromycin, and Rifampin in a Human Monocyte System againstLegionella pneumophila

1998 ◽  
Vol 42 (12) ◽  
pp. 3153-3156 ◽  
Author(s):  
Aldona L. Baltch ◽  
Raymond P. Smith ◽  
Mary A. Franke ◽  
Phyllis B. Michelsen
Keyword(s):  
Antibacterial Activity ◽  
Legionella Pneumophila ◽  
Human Monocyte ◽  
Antibacterial Activities ◽  
Human Monocytes ◽  
Antibacterial Effects ◽  
In Vitro System ◽  
Time Period ◽  
Time Periods

ABSTRACT The antibacterial activities of levofloxacin, erythromycin, and rifampin against intracellular Legionella pneumophilaL-1033, serogroup 1, were studied. In an in vitro system utilizing adherent human monocytes, L. pneumophila L-1033, a phagocytosis time period of 1 h, and antibiotic (levofloxacin, erythromycin, and/or rifampin) at 1 to 10 times the MIC, the CFU/ml values for the monocyte lysate were determined during 0- to 4-day time periods. The decrease in CFU/ml with levofloxacin at pH 7.4 was rapid, occurring within 24 h, and was drug concentration dependent (P < 0.01). The decrease in CFU with rifampin was first observed at 48 h (P < 0.01), while only a minimal decrease in CFU/ml was observed with erythromycin. Combination of levofloxacin and rifampin and of levofloxacin and erythromycin at ten times their MICs significantly decreased the CFU/ml value (P < 0.01), to the value attained by levofloxacin alone, while combination of rifampin and erythromycin did not. Removal of levofloxacin after 24 h of incubation resulted in regrowth ofL. pneumophila L-1033, while a continued slow decrease in CFU/ml was seen following rifampin removal; CFU/ml values were unaffected by the removal of erythromycin. At 4 days, and even in assays performed following antibiotic removal, the CFU/ml value continued to be lower in the levofloxacin and rifampin assays than in the assays with erythromycin. Levofloxacin had a significantly higher bactericidal activity against L. pneumophila L-1033 than erythromycin or rifampin. In these assays, the addition of erythromycin or rifampin did not affect the antibacterial activity of levofloxacin.

Platelet Factor 4 (PF4) Release In Vitro From Unstimulated Human Platelets

1981 ◽  
Author(s):  
D S Cohen ◽  
J M Strohschein ◽  
R N Saunders ◽  
D I Cargill
Keyword(s):  
Platelet Rich Plasma ◽  
Venous Blood ◽  
Human Platelets ◽  
Platelet Factor 4 ◽  
Platelet Factor ◽  
Time Period ◽  
Time Periods ◽  
Endogenous Release ◽  
Two Populations

Human venous blood was collected (9.5:0.5) in an anticoagulant (provided in the PF4 RIA kit, Abbott Labs.) consisting of 2.5% EDTA, 0.025% adenosine and 7.0% procaine HCl. Platelet rich plasma (PRP) was stirred (900 RPM) at 37°C in a Payton aggregometer for intervals of 3, 6, 9 and 12 minutes. Unincubated aliquots of PRP were taken as the 0 time controls.A significant (p < 0.02) elevation in PF4 was observed with increasing intervals. Furthermore, from the data obtained, two populations of PF4 releasers were delineated: A high releasing (HR) group and a low releasing (LR) group. HR (n=10) displayed a baseline release of 10.3 ± 0.8 ng/ml and values of 36.1 ± 9.0 (p < 0.05 compared to baseline), 46.8 ± 8.2 (p < 0.01), 54.6 ± 7.9 (p < 0.01) and 87 ± 13.1 (p < 0.01) for the corresponding time periods. LR (n=7) exhibited significant but diminished release at each time period with a basal level of 10.1 ± 1.0 and 14.7 ± 1.4 (p < 0.05 compared to baseline), 15.3 ± 1.5 (p < 0.05), 19.7 ± 2.4 (p < 0.05) and 23.6 ± 2.0 (p < 0.01) respectively. Comparison of both populations revealed significant (p < 0.01) differences at the 6, 9 and 12 minute intervals. Total PF4 content from Triton lysed PRP displayed similar levels between groups: 360.0 ±22.6 (HR) and 380.2 ± 15.2 (LR).We conclude that an endogenous release of PF4 occurs in vitro in unstimulated human PRP even in a milieu designed to deter release. This should be a concern when PF4 values are to be assessed.

scholarly journals Is there a role for endocannabinoids in sperm–oviduct interaction?

Reproduction ◽  
2010 ◽  
Vol 140 (2) ◽  
pp. 247-257 ◽  
Author(s):  
R Talevi ◽  
V Barbato ◽  
S De Iorio ◽  
V Mollo ◽  
T Capriglione ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Phospholipase D ◽  
Zona Pellucida ◽  
Endocannabinoid System ◽  
Sperm Storage ◽  
Sperm Viability ◽  
Progressive Motility ◽  
Bovine Spermatozoa ◽  
Sperm Reservoir

The endocannabinoid system (ECS) has been found in reproductive cells and tissues in several mammals. Spermatozoa are able to respond to anandamide, and the oviduct is able to synthesize and modulate the concentration of this endocannabinoid along the isthmic and ampullary regions. The main aim of this study was to understand whether the ECS has a role during sperm storage and release within the oviduct in cattle. Data showed that 1) the endocannabinoid receptors 1 and 2 (CB1 and CB2) are present in bovine spermatozoa both in the initial ejaculate and in spermatozoa bound to the oviduct in vitro; 2) CB1 receptor is still detectable in spermatozoa released from the oviduct through penicillamine but not in those released through heparin; 3) arachidonylethanolamide (AEA) does not affect sperm viability, whereas it depresses sperm progressive motility and kinetic values; 4) sperm–oviduct binding and release in vitro are not influenced by AEA; 5) AEA depresses sperm–zona pellucida (ZP) binding; 6) binding of heparin-capacitated spermatozoa to the ZP is not affected by AEA; 7) N-acylphosphatidylethanolamine-selective phospholipase D, the main enzyme involved in anandamide synthesis, is expressed in oviductal epithelial cells. In conclusion, secretion of AEA from epithelial cells might contribute to the oviduct sperm-reservoir function, prolonging the sperm fertile life through the depression of motility and capacitation. Capacitation signals, such as heparin, that promote sperm release, might remodel the sperm surface and cause a loss of the sperm sensitivity to AEA.

scholarly journals RELATIONSHIP BETWEEN COPPER IN DIFFERENT CULTURE MEDIA AND BOVINE SPERMATOZOA MOTILITY PARAMETERS IN VITRO

2017 ◽  
Vol 7 (3) ◽  
pp. 226-234 ◽  
Author(s):  
Zuzana Kňažická ◽  
Eva Tušimová ◽  
Jana Bezáková ◽  
Michal Miškeje ◽  
Tatiana Bojňanská ◽  
...  
Keyword(s):  
Culture Media ◽  
Spermatozoa Motility ◽  
Bovine Spermatozoa ◽  
Motility Parameters

scholarly journals In vitro effect of ferrous sulphate on bovine spermatozoa motility parameters, viability and Annexin V-labeled membrane changes

PLoS ONE ◽  
2021 ◽  
Vol 16 (9) ◽  
pp. e0257766
Author(s):  
Zuzana Knazicka ◽  
Hana Duranova ◽  
Veronika Fialkova ◽  
Michal Miskeje ◽  
Tomas Jambor ◽  
...  
Keyword(s):  
Cell Viability ◽  
Ferrous Sulphate ◽  
Annexin V ◽  
Computer Assisted ◽  
Spermatozoa Motility ◽  
Bovine Spermatozoa ◽  
High Concentrations ◽  
Vitro Effect ◽  
Motility Parameters

The aim of this study was to assess the dose- and time-dependent in vitro effects of ferrous sulphate (FeSO4.7H2O) on the motility parameters, viability, structural and functional activity of bovine spermatozoa. Spermatozoa motility parameters were determined after exposure to concentrations (3.90, 7.80, 15.60, 31.20, 62.50, 125, 250, 500 and 1000 μM) of FeSO4.7H2O using the SpermVisionTM CASA (Computer Assisted Semen Analyzer) system in different time periods. Cell viability was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) assay, and the Annexin V-Fluos was applied to detect the membrane integrity of spermatozoa. The initial spermatozoa motility showed increased average values at all experimental concentrations compared to the control group (culture medium without FeSO4.7H2O). After 2 h, FeSO4.7H2O stimulated the overall percentage of spermatozoa motility at the concentrations of ≤ 125 μM. However, experimental administration of 250 μM of FeSO4.7H2O significantly (P < 0.001) decreased the spermatozoa motility but had no negative effect on the cell viability (P < 0.05) (Time 2 h). The lowest viability was noted after the addition of ≥ 500 μM of FeSO4.7H2O (P < 0.001). The concentrations of ≤ 62.50 μM of FeSO4.7H2O markedly stimulated (P < 0.001) spermatozoa activity after 24 h of exposure, while at high concentrations of ≥ 500 μM of FeSO4.7H2O the overall percentage of spermatozoa motility was significantly inhibited (P < 0.001) and it elicited cytotoxic action. Fluorescence analysis confirmed that spermatozoa incubated with higher concentrations (≥ 500 μM) of FeSO4.7H2O displayed apoptotic changes, as detected in head membrane (acrosomal part) and mitochondrial portion of spermatozoa. Moreover, the highest concentration and the longest time of exposure (1000 μM of FeSO4.7H2O; Time 6 h) induced even necrotic alterations to spermatozoa. These results suggest that high concentrations of FeSO4.7H2O are able to induce toxic effects on the structure and function of spermatozoa, while low concentrations may have the positive effect on the fertilization potential of spermatozoa.

Negative effects of oxidative stress in bovine spermatozoa on in vitro development and DNA integrity of embryos

2018 ◽  
Vol 30 (10) ◽  
pp. 1359 ◽  
Author(s):  
L. Bittner ◽  
S. Wyck ◽  
C. Herrera ◽  
M. Siuda ◽  
C. Wrenzycki ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Dna Damage ◽  
Sperm Chromatin ◽  
Control Group ◽  
Dna Integrity ◽  
Negative Effects ◽  
Developmental Abnormalities ◽  
Developmental Rates ◽  
Bovine Spermatozoa

Oxidative stress in spermatozoa has effects on subsequent embryo development. The aim of the present study was to elucidate whether sperm oxidative stress results in increased DNA damage in the embryo. To this end, bovine spermatozoa were incubated for 1 h at 37°C without or with 100 µM H2O2, resulting in non-oxidised (NOX-S) and oxidised (OX-S) spermatozoa respectively. Non-incubated spermatozoa served as the control group (CON-S). After IVF, developmental rates 30, 46 and 60 h and 7 days after IVF were assessed. DNA damage was analysed in embryos using the comet assay and a DNA damage marker (γH2AX immunostaining); the apoptotic index was determined in blastocysts. Exposure of spermatozoa to H2O2 induced a significant amount of sperm chromatin damage. The use of OX-S in IVF resulted in significantly reduced cleavage and blastocyst rates compared with the use of CON-S and NOX-S. Furthermore, in embryos resulting from the use of OX-S, a developmental delay was evident 30 and 46 h after IVF. γH2AX immunostaining was lower in blastocysts than in early embryos. In blastocysts, the comet and apoptotic indices were significantly higher in embryos resulting from the use of OX-S than CON-S and NOX-S. In conclusion, oxidative stress in spermatozoa induces developmental abnormalities and is a source of DNA damage in the resulting embryos.

scholarly journals Caffeine strongly improves motility parameters of turkey spermatozoa with no effect on cell viability

2018 ◽  
Vol 66 (1) ◽  
pp. 137-150 ◽  
Author(s):  
Tomáš Slanina ◽  
Michal Miškeje ◽  
Filip Tirpák ◽  
Martyna Błaszczyk ◽  
Grzegorz Formicki ◽  
...  
Keyword(s):  
Incubation Temperature ◽  
Control Sample ◽  
Computer Assisted ◽  
Time Period ◽  
The Individual ◽  
The Impact ◽  
Casa System ◽  
Positive Effect ◽  
Motility Parameters

The purpose of this study was to evaluate the impact of caffeine on turkey spermatozoa during in vitro incubation. Experimental samples were prepared by diluting the raw semen with nine different concentrations of caffeine – from 0.078125 mg/mL to 10 mg/mL. The individual motility parameters were evaluated by the Computer Assisted Semen Analyser (CASA) system, and the viability of spermatozoa was evaluated using eosin-nigrosin staining. Selected parameters were recorded at six time periods: 0, 1, 2, 3, 4 and 5 h at 5 °C and 41 °C. A significantly higher motility and progressive motility of spermatozoa (P < 0.01 and P < 0.001, respectively) was detected in the samples containing caffeine ranging from 0.15625 to 7.5 mg/mL as compared to the control sample at 5 °C. At an incubation temperature of 41 °C the positive effect of caffeine on motility parameters was observed only at the beginning of incubation (at times 0 and 1). The tested caffeine concentrations showed no significant effect on the viability of turkey spermatozoa at any time period of incubation. A higher percentage of dead spermatozoa was observed for incubation at 41 °C (from 5.96% to 11.1%) in comparison to 5 °C (from 1.62% to 5.79%). The results suggest that caffeine can be used as a suitable component of turkey semen extenders and has the potential to improve fertility.

Properties and Biotechnological Application of mutant Derivatives of Mini-intein PRP8 from Penicillium chrysogenum with Improved Control of C-terminal Processing

2019 ◽  
Vol 35 (6) ◽  
pp. 91-101
Author(s):  
F.A. Klebanov ◽  
S.E. Cheperegin ◽  
D.G. Kozlov
Keyword(s):  
Penicillium Chrysogenum ◽  
Protein Denaturation ◽  
Biologically Active ◽  
Cultivation Temperature ◽  
Target Proteins ◽  
E Coli ◽  
Complete Inactivation ◽  
Target Molecules ◽  
Proteins And Peptides

Mutant variants of mini-intein PRP8 from Penicillium chrysogenum (Int4b) with improved control of C-terminal processing were characterized. The presented variants can serve as a basis for self-removed polypeptide tags capable of carrying an affine label and allowing to optimize the process of obtaining target proteins and peptides in E. coli cells. They allow to synthesize target molecules in the composition of soluble and insoluble hybrid proteins (fusions), provide their afnne purification, autocatalytic processing and obtaining mature target products. The presented variants have a number of features in comparison with the known prototypes. In particular the mutant mini-intein Int4bPRO, containing the L93P mutation, has temperature-dependent properties. At cultivation temperature below 30 °C it allows the production of target molecules as part of soluble fusions, but after increasing of cultivation temperature to 37 °C it directs the most of synthesized fusions into insoluble intracellular aggregates. The transition of Int4bPRO into insoluble form is accompanied by complete inactivation of C-terminal processing. Further application of standard protein denaturation-renaturation procedures enable efficiently reactivate Int4bPRO and to carry out processing of its fusions in vitro. Two other variants, Int4b56 and Int4b36, containing a point mutation T62N or combination of mutations D144N and L146T respectively, have a reduced rate of C-terminal processing. Their use in E. coli cells allows to optimize the biosynthesis of biologically active target proteins and peptides in the composition of soluble fusions, suitable for afnne purification and subsequent intein-dependent processing without the use of protein denaturation-renaturation procedures. intein, fusion, processing, processing rate, gelonin The work was supported within the framework of the State Assignment no. 595-00003-19 PR.

Diabetes and Heart Failure: Is it Hyperglycemia or Hyperinsulinemia?

2020 ◽  
Vol 18 (2) ◽  
pp. 148-157 ◽  
Author(s):  
Triantafyllos Didangelos ◽  
Konstantinos Kantartzis
Keyword(s):  
Heart Failure ◽  
Insulin Treatment ◽  
Close Association ◽  
Adverse Outcomes ◽  
Negative Effects ◽  
Beneficial Effects ◽  
Appropriate Treatment ◽  
Increased Risk ◽  
Treatment Of Diabetes

The cardiac effects of exogenously administered insulin for the treatment of diabetes (DM) have recently attracted much attention. In particular, it has been questioned whether insulin is the appropriate treatment for patients with type 2 diabetes mellitus and heart failure. While several old and some new studies suggested that insulin treatment has beneficial effects on the heart, recent observational studies indicate associations of insulin treatment with an increased risk of developing or worsening of pre-existing heart failure and higher mortality rates. However, there is actually little evidence that the associations of insulin administration with any adverse outcomes are causal. On the other hand, insulin clearly causes weight gain and may also cause serious episodes of hypoglycemia. Moreover, excess of insulin (hyperinsulinemia), as often seen with the use of injected insulin, seems to predispose to inflammation, hypertension, dyslipidemia, atherosclerosis, heart failure, and arrhythmias. Nevertheless, it should be stressed that most of the data concerning the effects of insulin on cardiac function derive from in vitro studies with isolated animal hearts. Therefore, the relevance of the findings of such studies for humans should be considered with caution. In the present review, we summarize the existing data about the potential positive and negative effects of insulin on the heart and attempt to answer the question whether any adverse effects of insulin or the consequences of hyperglycemia are more important and may provide a better explanation of the close association of DM with heart failure.

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