scholarly journals IN VITRO EFFECTS OF ISOQUERCITRIN ON SELECTED VITALITY MARKERS OF BOVINE SPERMATOZOA

2019 ◽  
Vol 1 (2) ◽  
pp. 18-23
Author(s):  
Filip Benko ◽  
Hana Greifová ◽  
Eva Tvrdá

The aim of this study was to evaluate the dose- and time-dependent in vitroeffects of isoquercitrin (ISO), a natural flavonoid with numerous biological properties on bovine spermatozoa during three different time periods (0 h, 2 h, 24 h). Bovine semen samples were diluted and cultivated in physiological saline solution containing 0.5% DMSO together with 200, 100, 50, 10, 5 and 1 μmol/L ISO. Spermatozoa motility was measured using the HTM IVOS CASA (Computer Assisted Semen Analyzer) system. The viability of spermatozoa was assessed by the metabolic (MTT) assay, production of superoxide radicals was quantified using the nitroblue tetrazolium (NBT) test, and chemiluminescence was used to evaluate the generation of reactive oxygen species (ROS). The results of the movement activity showed a significant increase in the motility during long term cultivation in case of concentrationsranging between 5 and 50 μmol/L ISO (P<0.05; 24 h). At the same time, supplementation of several concentrations of ISO led to a significant preservation of the cell viability (P<0.05 in the case of 50 μmol/L, P<0.01 with respect to 1 and 5 μmol/L, and P<0.001 in relation to 10 μmol/L; 24 h). ISO addition at 10 and 50 μmol/L also provided a significantly higher protection against superoxide (P<0.05) and ROS (P<0.001) overgeneration after a 24 h cultivation. We may suggest that supplementation of ISO to bovine spermatozoa, particularly at concentrations ranging between 10 and 50 μmol/L, may offer protection to the motility, viability and oxidative status of the sperm cells, particularly notable at 24 h.

PLoS ONE ◽  
2021 ◽  
Vol 16 (9) ◽  
pp. e0257766
Author(s):  
Zuzana Knazicka ◽  
Hana Duranova ◽  
Veronika Fialkova ◽  
Michal Miskeje ◽  
Tomas Jambor ◽  
...  

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.


1994 ◽  
Vol 74 (1) ◽  
pp. 53-58 ◽  
Author(s):  
J. L. Bailey ◽  
M. M. Buhr ◽  
L. Robertson

Correlations among computer-assisted spermatozoa motility analyses, Ca2+ fluxes and in vivo fertility of bovine spermatozoa based on a total of 4482 inseminations were investigated in each of four ejaculates from six bulls. The Ca2+ parameters assessed the rate of change in intra- and extracellular Ca + in fresh and cryopreserved spermatozoa from the same ejaculates and were described in another study. Of the seven motility parameters of cryopreserved semen investigated, all differed significantly among bulls but none were related to the in vivo fertility of cryopreserved semen. The amplitude of lateral head displacement, a motility parameter associated with hyperactivation, was positively correlated with the intracellular Ca2+ levels and the rate of Ca2+ accumulation of cyropreserved spermatozoa. The highest fertility was observed when initial extracellular Ca2+ for cryopreserved spermatozoa was high and when Ca2+ efflux rates of cryopreserved cells approached the higher efflux rates of fresh spermatozoa. Fertility was reduced when cryopreserved spermatozoa had initial internal Ca2+ levels greater than those of fresh spermatozoa or when cryopreserved spermatozoa internalized Ca2+ rapidly. Calcium flux, but not motility, may predict fertilizing capacity of cryopreserved bovine spermatozoa. Key words: Calcium, computer-analysed motility, fertility, spermatozoa, bull


Author(s):  
Ann Chidester Van Orden ◽  
John L. Chidester ◽  
Anna C. Fraker ◽  
Pei Sung

The influence of small variations in the composition on the corrosion behavior of Co-Cr-Mo alloys has been studied using scanning electron microscopy (SEM), energy dispersive x-ray analysis (EDX), and electrochemical measurements. SEM and EDX data were correlated with data from in vitro corrosion measurements involving repassivation and also potentiostatic anodic polarization measurements. Specimens studied included the four alloys shown in Table 1. Corrosion tests were conducted in Hanks' physiological saline solution which has a pH of 7.4 and was held at a temperature of 37°C. Specimens were mechanically polished to a surface finish with 0.05 µm A1203, then exposed to the solution and anodically polarized at a rate of 0.006 v/min. All voltages were measured vs. the saturated calomel electrode (s.c.e.).. Specimens had breakdown potentials near 0.47V vs. s.c.e.


2018 ◽  
Vol 63 (No. 4) ◽  
pp. 127-135 ◽  
Author(s):  
T. Slanina ◽  
M. Miškeje ◽  
F. Tirpák ◽  
M. Błaszczyk ◽  
R. Stawarz ◽  
...  

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.


2009 ◽  
Vol 2009 ◽  
pp. 1-7 ◽  
Author(s):  
Razika Zeghir-Bouteldja ◽  
Manel Amri ◽  
Saliha Aitaissa ◽  
Samia Bouaziz ◽  
Dalila Mezioug ◽  
...  

Hydatidosis is characterized by the long-term coexistence of larvaEchinococcus granulosusand its host without effective rejection. Previous studies demonstrated nitric oxide (NO) production (in vivo and in vitro) during hydatidosis. In this study, we investigated the direct in vitro effects of NO species: nitrite (NO2−), nitrate (NO3−) and peroxynitrite (ONOO−) on protoscolices (PSCs) viability and hydatid cyst layers integrity for 24 hours and 48 hours. Our results showed protoscolicidal activity ofNO2−andONOO−24 hours and 3 hours after treatment with 320 μM and 80 μM respectively. Degenerative effects were observed on germinal and laminated layers. The comparison of the in vitro effects of NO species on the PSCs viability indicated thatONOO−is more cytotoxic thanNO2−. In contrast,NO3−has no effect. These results suggest possible involvement ofNO2−andONOO−in antihydatic action and point the efficacy of these metabolites as scolicidal agents.


Author(s):  
Büşra Yazıcılar ◽  
Fatma Böke ◽  
Azize Alaylı ◽  
Hayrunisa Nadaroglu ◽  
Semin Gedikli ◽  
...  

1989 ◽  
Vol 257 (2) ◽  
pp. F177-F181 ◽  
Author(s):  
C. Khadouri ◽  
S. Marsy ◽  
C. Barlet-Bas ◽  
A. Doucet

Because previous studies indicated that in the collecting tubule, N-ethylmaleimide (NEM)-sensitive ATPase, the biochemical equivalent of the proton pump, is controlled by mineralocorticoids in the long term, the present study was designed to investigate whether such control also exists in the short term. Therefore we investigated the in vivo and in vitro effects of aldosterone on the enzyme activity in cortical and outer medullary collecting tubules (CCT and MCT, respectively) from adrenalectomized rats. Administration of aldosterone (10 micrograms/kg body wt) markedly stimulated NEM-sensitive ATPase activity in the CCT and MCT within 3 h. Similarly, incubating CCT or MCT for 3 h in the presence of 10(-8) M aldosterone enhanced NEM-sensitive ATPase activity up to values similar to those previously measured in the corresponding nephron segments of normal rats. In vitro stimulation of NEM-sensitive ATPase was dose dependent in regard to aldosterone (apparent affinity constant approximately 10(-9) M), appeared after a 30-min lag period, and reached its maximum after 2-2.5 h. Finally, actinomycin D and cycloheximide totally abolished the in vitro action of aldosterone, demonstrating the involvement of protein synthesis in this process.


2019 ◽  
Vol 54 (2) ◽  
pp. 139-146 ◽  
Author(s):  
MA Kilani ◽  
AZ Hassan ◽  
ST Fadason ◽  
AM Obalowu ◽  
A Aliyu ◽  
...  

Phytochemical constituents of Croton lobatus L. (C. lobatus) water extracts and quantitative analysis were carried out following standard procedures. The antibacterial activity against Staphylococcus aureus (ATCC 33591); Streptococcus Spp; Pseudomonas aeruginosa (ATCC 9028); Proteus vulgaris; Escherichia coli (ATCC 43895); and Salmonella Spp (ATCC 4932) was carried out at the concentration of 0.5g/mL, 0.05 g/mL and 0.00 5g/mL of water. In vivo antimicrobial assay was carried out by creating four wounds of 0.5 by 0.5 cm on dorsal surface of a male albino rat under anesthesia. The wounds were left for 48 hrs, after which they were accessed and samples were collected for culture, identification and colony forming unit counts (CFU). Respective treatment using dried C. lobatus, C. lobatus (water extract), Physiological saline solution and Cicatrin powder was carried out and samples were collected at day one, three, five and seven after initiation of treatments for CFU counts on nutrient and MacConkey agar. The phytochemical studies revealed that C. lobatus contains carbohydrates, glycoside, saponins, steroids, triterpenes, flavonoids, alkaloids and tannins. Croton lobatus L. showed a dose dependent activity against micro organisms with C. lobatus 0.5 inhibited the growth of most bacteria at the zone of inhibition ≤ 21mm. This was also supported by in vivo antimicrobial assay. Secondary metabolite tannins, triterpenoids, flavonoids, crotonic acids and saponin were responsible for its antimicrobial activity against the tested microorganisms thereby supporting its usage by the traditional medicine practitioner in Nigeria to treat chronic wounds. Bangladesh J. Sci. Ind. Res.54(2), 139-146, 2019


1985 ◽  
Vol 63 (1) ◽  
pp. 63-66 ◽  
Author(s):  
J. E. Steele ◽  
T. Paul

Cockroach fat body incubated in a simple physiological saline solution releases trehalose to the surrounding medium. The output of trehalose occurs in the absence of ambient Ca2+ and decreases slowly with time. In two separate experiments, 0.1 mM CaCl2 added to the saline increased the output of trehalose on average by 70% but higher concentrations of Ca2+ did not further increase the efflux of trehalose. Stimulation of trehalose efflux by corpus cardiacum extract is absolutely dependent on extracellular Ca2+, no increase occurring beyond the basal level in the absence of the ion. The activity of corpus cardiacum extract increases as the concentration of CaCl2 is increased to 0.5 mM. This concentration of Ca2+ in the saline permits the extract to increase trehalose efflux by as much as 60% above the basal level. Corpus cardiacum extract, as well as the hypertrehalocaemic agents cyclic AMP and theophylline, increase significantly the influx of Ca2+ into fat body in vitro. The basal efflux of trehalose from fat body and that stimulated by corpus cardiacum extract is not dependent on extracellular Mg2+.


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