Protective effects of melatonin on bovine sperm characteristics and subsequent in vitro embryo development

In assisted reproduction by ICSI, PVP has been successfully used to replicate the viscosity of sperm solution, thus facilitating the handling and immobilization of spermatozoa. Sperm is suspended in medium containing polyvinylpyrrolidone (PVP), then injected into the oocytes together with a small amount of the medium in ICSI. However the effects of PVP on sperm function and embryo development have not been investigated in detail. In the present study, we investigated the effects of PVP solution on sperm function and embryonic development. Frozen–thawed spermatozoa from a Japanese Black bull and immature COCs from slaughterhouse bovine ovaries were used for all experiments. In experiment 1, bovine sperm was cultured in SOF or SOF containing 10% PVP. For detection of sperm acrosomal and chromatin integrity, sperm cultured in each medium were stained by the triple staining method and acridine orange after 0, 15, 30, and 60 min of culture. In experiment 2, zygotes were injected with PVP solution and cultured in vitro; subsequent cleavage and development to blastocysts were examined. In experiment 3, zygote injected with PVP solution was fixed by 4% paraformaldehyde after 1–3 h of PVP injection. The location of PVP solution in zygote was observed. In experiment 4, two-cell embryos were microinjected with a solution of dextran conjugated with fluorescein (FITC-dextran) and cultured in vitro. The location of FITC-dextran in the embryo was examined. In experiment 1, acrosome reactions of the sperm were enhanced after 15 min of incubation in PVP solution (P < 0.05), but chromatin integrity of the sperm was not influenced (P > 0.05). In experiment 2, PVP suppressed the development of the zygote to 2-cell, morula and blastocyst (75.0%, 35.1%, and 26.3% vs. 61.3%, 20.2%, and 12.9% for control and PVP group, respectively, P < 0.05). In experiment 3, the locations of PVP solution in the zygote were observed 1–3 h after injection. In experiment 4, FITC-dextran was observed in ICM at the blastocyst stage. These findings suggest that PVP affects the acrosome but not the chromatin of sperm in ICSI. PVP solution exists locally in embryos injected and affects the developmental capacity of the embryos.

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7 THE EFFECT OF A NOVEL RECOMBINANT PROTEASE TREATMENT ON BOVINE SPERM FERTILIZING CAPACITY AND EMBRYO DEVELOPMENT IN VITRO

Reproduction Fertility and Development ◽

10.1071/rdv21n1ab7 ◽

2009 ◽

Vol 21 (1) ◽

pp. 104

Author(s):

J. T. Aaltonen ◽

K. J. Mattson ◽

N. M. Loskutoff

Keyword(s):

Embryo Development ◽

Disease Transmission ◽

Bos Taurus ◽

Gradient Centrifugation ◽

Control Group ◽

Bovine Embryo ◽

Human Sequence ◽

Bovine Sperm

As described in the IETS Manual (Stringfellow and Seidel, 1995), and endorsed by the OIE, trypsin can be used (for specific pathogens and livestock) to effectively remove certain infectious agents from in vivo-derived embryos for international transport. Because of the multimillion-dollar AI industry for livestock, the OIE has encouraged more research in developing similar decontamination techniques for semen as an added safeguard to animal quarantine for the prevention of disease transmission. Most or all of the earlier studies on embryos used a porcine pancreatic-derived trypsin. Because of more stringent guidelines from international regulatory agencies on the use of animal products, several serine protease recombinants are now available. Previous experiments comparing the porcine pancreatic extract with a recombinant bovine sequence trypsin developed in corn resulted in no statistical difference in cleavage or morula/blastocyst rates. (Mattson et al. 2008 Theriogenology 69, 724–727). An additional in vivo study treating bovine sperm with a yeast-derived human-sequence trypsin resulted in significantly more transferable-quality embryos after the AI of superovulated cows as compared with sperm not treated with trypsin (Blevins et al. 2008 Reprod. Fertil. Dev. 20, 84). The goal of this experiment was to examine the in vitro development of bovine embryos produced from sperm treated with a recombinant trypsin found in a commercially available density gradient centrifugation (DGC) product (Bovipure, Nidacon, Sweden) compared with DGC without trypsin. Oocyte aspiration, maturation, fertilization, and embryo culture were performed using standard methods in 5 replications (n = 2220 oocytes). Semen was collected and pooled from 2 Bos taurus bulls and frozen in an egg-yolk cryodiluent (Biladyl, Minitube). The semen was processed using Bovipure DGC composed of 2 mL of 40% colloid of silane-coated silica particles containing either a yeast-derived human sequence recombinant trypsin containing no animal by-products (n = 1126 oocytes) or the same colloid without trypsin as the control group (n = 1094 oocytes). Both 40% concentrations were layered over 2 mL of an 80% concentration of the same colloid without any additives. The density gradients were centrifuged at 300g for 20 min, after which time the pellets were washed in 5 mL of prewarmed TL Hepes solution (Cambrex) and centrifuged at 500g for 10 min. The resulting sperm pellets were then resuspended in a volume calculated to provide 1 × 106 sperm mL–1, to be used for in vitro inseminations. Results were compared using a 2-tailed unpaired t-test. Cleavage rates for the trypsin-treated sperm (n = 969, 35.8%) and the control (n = 950, 44.3%) groups were not statistically different (P = 0.20). Although more embryos reached the morula to blastocyst stages in the control group (n = 421, 61.0%) than in the trypsinized group (n = 347, 54.7%), these differences also were not statistically significant (P = 0.85). In conclusion, trypsinized Bovipure DGC of sperm before insemination showed no detrimental effects on IVF-derived bovine embryo development.

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Impacts of oxidative stress on bovine sperm function and subsequent in vitro embryo development

Animal Reproduction ◽

10.21451/1984-3143-ar2018-0041 ◽

2018 ◽

Vol 15 (Suppl. 1) ◽

pp. 703-710 ◽

Cited By ~ 1

Author(s):

Heinrich Bollwein ◽

Lilli Bittner

Keyword(s):

Oxidative Stress ◽

Embryo Development ◽

Sperm Function ◽

Bovine Sperm

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Influence of post‐thawing thermal environment on bovine sperm characteristics and in vitro fertility

Andrologia ◽

10.1111/and.13266 ◽

2019 ◽

Vol 51 (6) ◽

pp. e13266 ◽

Cited By ~ 2

Author(s):

Daniela Botta ◽

Rubens Paes Arruda ◽

Yeda Fumie Watanabe ◽

Júlio Cesar Balieiro ◽

Narian Romanello ◽

...

Keyword(s):

Thermal Environment ◽

Bovine Sperm ◽

Sperm Characteristics

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Effect of royal jelly on bovine sperm characteristics during post–thaw incubation in vitro

Revista Veterinaria ◽

10.30972/vet.2121747 ◽

2010 ◽

Vol 21 (2) ◽

pp. 81 ◽

Cited By ~ 3

Author(s):

Saber Mohammed Abd–Allah

Keyword(s):

Royal Jelly ◽

Bovine Sperm ◽

Sperm Characteristics ◽

Para Determinar

<p>El semen bovino criopreservado generalmente es considerado de menor poder fertilizante comparado con el semen fresco. Tal reducción abarca tanto una más baja viabilidad post–descongelado como una posible disfunción subletal de la población de espermatozoides sobrevivientes. El presente estudio fue realizado para determinar el efecto de la jalea real (JR) sobre la motilidad, viabilidad e integridad acrosomal de espermatozoides durante la etapa de incubación post–descongelado. Las muestras de semen congelado–descongelado se lavaron e incubaron a 37°C en buffer Tris conteniendo JR en proporción de 0,1; 0,2; 0,3; 0,4; 0,5% o ninguna (control). La motilidad y la viabilidad espermáticas, así como la integridad acrosomal, fueron evaluadas a las 0; 0,5; 1; 1,5 y 2 h. Comparados con los controles, los porcentajes de motilidad, viabilidad e integridad acrosomal fueron más altos en los espermatozoides incubados en Tris conteniendo 0,4% de JR (p < 0,05). Después de 2 h de incubación, los porcentajes de motilidad, viabilidad e integridad acrosomal de los espermatozoides fueron respectivamente de 52,3; 52,5 y 19,8% en el buffer que contenía JR al 0,4%. Los resultados indican que la adición de 0,4% de JR en los medios de incubación fue capaz de mantener la buena calidad y longevidad de los espermatozoides. La jalea real puede ser usada como aditivo seminal para mejorar la viabilidad y fertilidad de los espermatozoides.</p>

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In vitro exposure to CPF affects bovine sperm epigenetic gene methylation pattern and the ability of sperm to support fertilization and embryo development

Environmental and Molecular Mutagenesis ◽

10.1002/em.22242 ◽

2018 ◽

Vol 60 (1) ◽

pp. 85-95 ◽

Cited By ~ 9

Author(s):

Maria Michela Pallotta ◽

Vincenza Barbato ◽

Alain Pinton ◽

Hervè Acloque ◽

Roberto Gualtieri ◽

...

Keyword(s):

Embryo Development ◽

Methylation Pattern ◽

Gene Methylation ◽

Bovine Sperm ◽

In Vitro Exposure

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Protective effects of Dioscorea membranacea rhizome ethanolic extract against doxorubicin-induced genotoxicity in human lymphocytes in vitro

Planta Medica ◽

10.1055/s-0035-1565576 ◽

2015 ◽

Vol 81 (16) ◽

Author(s):

T Ratanavalachai ◽

S Thitiorul ◽

A Itharat ◽

N Runraksa ◽

S Ruangnoo

Keyword(s):

Human Lymphocytes ◽

Ethanolic Extract ◽

Protective Effects

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Antioxidant Activity, Phenolic Content and Hematoprotective Effects of Cleome arabica Polyphenolic Leaf Extract

Phytothérapie ◽

10.3166/phyto-2019-0206 ◽

2019 ◽

Author(s):

C. Tigrine ◽

A. Kameli

Keyword(s):

Antioxidant Activity ◽

Biological Effects ◽

Reducing Power ◽

Hematological Toxicity ◽

Protective Effects ◽

Ferrous Ions ◽

Polyphenolic Extract ◽

Cleome Arabica

In this study a polyphenolic extract from Cleome arabica leaves (CALE) was investigated for its antioxidant activity in vitro using DPPH•, metal chelating and reducing power methods and for its protective effects against AraC-induced hematological toxicity in vivo using Balb C mice. Results indicated that CALE exhibited a strong and dose-dependent scavenging activity against the DPPH• free radical (IC50 = 4.88 μg/ml) and a high reducing power activity (EC50 = 4.85 μg/ml). Furthermore, it showed a good chelating effects against ferrous ions (IC50 = 377.75 μg/ml). The analysis of blood showed that subcutaneous injection of AraC (50 mg/kg) to mice during three consecutive days caused a significant myelosupression (P < 0.05). The combination of CALE and AraC protected blood cells from a veritable toxicity. Where, the number of the red cells, the amount of hemoglobin and the percentage of the hematocrite were significantly high. On the other hand, AraC cause an elevation of body temperature (39 °C) in mice. However, the temperature of the group treated with CALE and AraC remained normal and did not exceed 37.5 °C. The observed biological effects of CALE, in vitro as well as in vivo, could be due to the high polyphenol and flavonoid contents. In addition, the antioxidant activity of CALE suggested to be responsible for its hematoprotective effect.

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Healing effects of curcumin nanoparticles in deep tissue injury mouse model.

Current Drug Delivery ◽

10.2174/1567201818666201214125237 ◽

2020 ◽

Vol 18 ◽

Author(s):

Zirui Zhang ◽

Shangcong Han ◽

Panpan Liu ◽

Xu Yang ◽

Jing Han ◽

...

Keyword(s):

Wound Healing ◽

Mrna Expression ◽

Tissue Injury ◽

Inflammatory Cells ◽

Pcr Analysis ◽

Protective Effects ◽

Deep Tissue ◽

Deep Tissue Injury

Background: Chronic inflammation and lack of angiogenesis are the important pathological mechanisms in deep tissue injury (DTI). Curcumin is a well-known anti-inflammatory and antioxidant agent. However, curcumin is unstable under acidic and alkaline conditions, and can be rapidly metabolized and excreted in the bile, which shortens its bioactivity and efficacy. Objective: This study aimed to prepare curcumin-loaded poly (lactic-co-glycolic acid) nanoparticles (CPNPs) and to elucidate the protective effects and underlying mechanisms of wound healing in DTI models. Methods: CPNPs were evaluated for particle size, biocompatibility, in vitro drug release and their effect on in vivo wound healing. Results : The results of in vivo wound closure analysis revealed that CPNP treatments significantly improved wound contraction rates (p<0.01) at a faster rate than other three treatment groups. H&E staining revealed that CPNP treatments resulted in complete epithelialization and thick granulation tissue formation, whereas control groups resulted in a lack of compact epithelialization and persistence of inflammatory cells within the wound sites. Quantitative real-time PCR analysis showed that treatment with CPNPs suppressed IL-6 and TNF-α mRNA expression, and up-regulated TGF-β, VEGF-A and IL-10 mRNA expression. Western blot analysis showed up-regulated protein expression of TGF-β, VEGF-A and phosphorylatedSTAT3. Conclusion: Our results showed that CPNPs enhanced wound healing in DTI models, through modulation of the JAK2/STAT3 signalling pathway and subsequent upregulation of pro-healing factors.

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Newly Developed Recombinant Antithrombin Protects the Endothelial Glycocalyx in an Endotoxin-Induced Rat Model of Sepsis

International Journal of Molecular Sciences ◽

10.3390/ijms22010176 ◽

2020 ◽

Vol 22 (1) ◽

pp. 176

Author(s):

Toshiaki Iba ◽

Jerrold H. Levy ◽

Koichiro Aihara ◽

Katsuhiko Kadota ◽

Hiroshi Tanaka ◽

...

Keyword(s):

Dose Group ◽

Low Dose ◽

Leukocyte Adhesion ◽

Endothelial Glycocalyx ◽

High Dose Group ◽

Control Group ◽

High Dose ◽

Protective Effects ◽

Circulating Levels

(1) Background: The endothelial glycocalyx is a primary target during the early phase of sepsis. We previously reported a newly developed recombinant non-fucosylated antithrombin has protective effects in vitro. We further evaluated the effects of this recombinant antithrombin on the glycocalyx damage in an animal model of sepsis. (2) Methods: Following endotoxin injection, in Wistar rats, circulating levels of hyaluronan, syndecan-1 and other biomarkers were evaluated in low-dose or high-dose recombinant antithrombin-treated animals and a control group (n = 7 per group). Leukocyte adhesion and blood flow were evaluated with intravital microscopy. The glycocalyx was also examined using side-stream dark-field imaging. (3) Results: The activation of coagulation was inhibited by recombinant antithrombin, leukocyte adhesion was significantly decreased, and flow was better maintained in the high-dose group (both p < 0.05). Circulating levels of syndecan-1 (p < 0.01, high-dose group) and hyaluronan (p < 0.05, low-dose group; p < 0.01, high-dose group) were significantly reduced by recombinant antithrombin treatment. Increases in lactate and decreases in albumin levels were significantly attenuated in the high-dose group (p < 0.05, respectively). The glycocalyx thickness was reduced over time in control animals, but the derangement was attenuated and microvascular perfusion was better maintained in the high-dose group recombinant antithrombin group (p < 0.05). (4) Conclusions: Recombinant antithrombin maintained vascular integrity and the microcirculation by preserving the glycocalyx in this sepsis model, effects that were more prominent with high-dose therapy.

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