Effect of supplementation with inorganic and organic selenium on sperm quality and quantity in north-east Bulgarian merino rams

Aim. To determine whether 5-aminolevulinic acid-based photodynamic therapy (ALA-PDT) is effective in combating ultraviolet A- (UVA-) induced oxidative photodamage of hairless mice skin in vivo and human epidermal keratinocytes in vitro. Methods. In in vitro experiments, the human keratinocyte cell line (HaCaT cells) was divided into two groups: the experimental group was treated with ALA-PDT and the control group was left untreated. Then, the experimental group and the control group of cells were exposed to 10 J/m2 of UVA radiation. ROS, O2− species, and MMP were determined by fluorescence microscopy; p53, OGG1, and XPC were determined by Western blot analysis; apoptosis was determined by flow cytometry; and 8-oxo-dG was determined by immunofluorescence. Moreover, HaCaT cells were also treated with ALA-PDT. Then, SOD1 and SOD2 were examined by Western blot analysis. In in vivo experiments, the dorsal skin of hairless mice was treated with ALA-PDT or saline-PDT, and then, they were exposed to 20 J/m2 UVA light. The compound 8-oxo-dG was detected by immunofluorescence. Conclusion. In human epidermal keratinocytes and hairless mice skin, UVA-induced oxidative damage can be prevented effectively with ALA-PDT pretreatment.

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Piglets produced by transfer of embryos obtained by in vitro fertilization of oocytes matured in vitro with thymosin: A case report

Medycyna Weterynaryjna ◽

10.21521/mw.6356 ◽

2020 ◽

Vol 76 (03) ◽

pp. 6356-2020 ◽

Cited By ~ 2

Author(s):

KATARZYNA PONIEDZIAŁEK-KEMPNY ◽

BARBARA GAJDA ◽

IWONA RAJSKA ◽

LECHOSŁAW GAJDA ◽

ZDZISŁAW SMORĄG

Keyword(s):

Case Report ◽

In Vitro Fertilization ◽

Control Group ◽

First Birth ◽

Vitro Fertilization ◽

Research Material ◽

Preliminary Study ◽

Experimental Group

The aim of the study was to examine the in vivo viability of in vitro-produced (IVP) porcine embryos obtained from oocytes matured with thymosin. The research material for this study consisted of immature pig oocytes obtained from ovaries after slaughter and ejaculated semen obtained from one boar. The immature oocytes were cultured in vitro until the metaphase II stage in a medium supplemented with thymosin (TMS). The presumptive zygotes obtained were cultured in vitro for 4-40 hours. The presumptive zygotes and 2-4-cell embryos were evaluated in vivo after transferring them to synchronized recipients. After the transfer of embryos from the experimental group into 2 recipients (50 embryos into each gilt) and the transfer of 50 embryos from the control group into 1 recipient, both gilts that had received embryos obtained by in vitro fertilization of oocytes matured with TMS became pregnant and delivered a total of 16 live piglets. After the transfer of embryos from the control group, no pregnancy was achieved. In conclusion, the results of our preliminary study suggest that the maturation of pig oocytes with thymosin supports the in vivo survival of in vitro produced embryos. It is important to note, that this was the first birth of piglets obtained after transfer of IVP embryos in Poland.

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hPMSCs protects against D-galactose-induced oxidative damage of CD4+ T cells through activating Akt-mediated Nrf2 antioxidant signaling

10.21203/rs.3.rs-46702/v3 ◽

2020 ◽

Author(s):

Yanlian Xiong ◽

Yueming Wang ◽

Jiashen Zhang ◽

Nannan Zhao ◽

Hengchao Zhang ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Nuclear Translocation ◽

Cell Senescence ◽

Antioxidant Defenses ◽

Control Group ◽

Akt Inhibitor ◽

Gsk 3Β ◽

T Cell Senescence

Abstract Background: Mesenchymal stem cells (MSCs) was considered as regenerative therapeutic approach in both acute and chronic diseases. However, whether MSCs regulate the antioxidant metabolism of CD4+ T cells and weaken immunosenescence remains unclear. Here, we reported the protective effects of hPMSCs in aging-related CD4+ T cell senescence and identified the underlying mechanisms using a D-gal induced mouse aging model.Methods: In vivo study, 40 male C57BL/6 mice (8 weeks) were randomly divided into four groups: control group, D-gal group, hPMSC group and PBS group. In in vitro experiment, human naive CD4+ T (CD4CD45RA) cells were prepared using a naive CD4+ T cell isolation kit II and pretreated with the Akt inhibitor LY294002 and Nrf2 inhibitor ML385. Then, isolated naive CD4+ T cell were cocultured with hPMSCs for 72 h in the absence or presence of anti-CD3/CD28 Dynabeads and IL-2 as a mitogenic stimulus. Intracellular ROS changes were detected by flow cytometry. The activities of the antioxidant enzymes superoxide dismutase, glutathione peroxidase and catalase were measured by colorimetric analysis. The senescent T cells were detected SA-β-gal stain. The expression of aging related proteins were detected by Western blotting, RT-PCR and confocal microscopy.Results: We found that hPMSC treatment markedly decreased the ROS level, SA-β-gal positive cells number, senescence-associated secretory phenotype (IL-6 and OPN) expression and aging-related protein (P16 and P21) expression in senescent CD4+ T cells. Furthermore, hPMSC treatment effectively upregulated Nrf2 nuclear translocation and the expression of downstream target genes (HO-1, CAT, GCLC and NQO1) in senescent CD4+ T cells. Moreover, in vitro studies revealed that hPMSCs attenuated CD4+ T cell senescence by upregulating the Akt/GSK-3β/Fyn pathway to activate Nrf2 functions. Conversely, the antioxidant effects of hPMSCs were blocked by the Akt inhibitor LY294002 and Nrf2 inhibitor ML385 in senescent CD4+ T cells.Conclusions: Our results indicate that hPMSCs attenuate D-gal induced CD4+ T cell senescence by activating Nrf2-mediated antioxidant defenses and that upregulation of Nrf2 by hPMSCs is regulated via the Akt/GSK-3β/Fyn pathway.

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81 EFFECT OF SUCROSE CONCENTRATION IN WARMING MEDIUM ON THE DEVELOPMENT POTENTIAL OF VITRIFIED BOVINE OOCYTES

Reproduction Fertility and Development ◽

10.1071/rdv17n2ab81 ◽

2005 ◽

Vol 17 (2) ◽

pp. 190

Author(s):

W.C. Chang ◽

J. Xu ◽

S. Jiang ◽

X.C. Tian ◽

X. Yang ◽

...

Keyword(s):

Sucrose Concentration ◽

Control Group ◽

Bovine Oocyte ◽

Cell Stage ◽

Significant Difference ◽

Humidified Air ◽

Bovine Oocytes ◽

Experimental Group ◽

One Way Anova

The aim of this experiment was to determine the effect of the sucrose concentration (0 to 0.33 M) in the dilution medium on the viability, fertilizability, and development of vitrified bovine oocytes. Bovine oocyte-cumulus complexes were collected from slaughterhouse ovaries and in vitro-matured as reported previously. After 24-h maturation in TCM199-based medium under 5% CO2 humidified air at 39°C, these were exposed to hyaluronidase and carefully pipetted to remove all except the 3–5 innermost layers of cumulus. Oocytes were put into the pre-equilibration medium for 3 min and then into vitrification solution containing HEPES-buffered TCM199 supplemented with 20% FBS, ethylene glycol, and dimethylsulphoxide for 25–30 s; they were then vitrified by modified solid surface vitrification (Dinnyes et al. 2000 Biol. Reprod. 63, 513–518).The oocytes were warmed at 39°C by placing them in holding medium with 0, 0.08, 0.17, 0.25, or 0.33 M sucrose. Non-vitrified oocytes were used as controls. Oocytes were inseminated 30 min after warming, and the presumptive zygotes were cultured in CR1-aa medium supplemented with 6 mg/mL BSA at 39°C in a humidified atmosphere of 5% CO2, 5% O2, and 90% N2 for eight days. Data were analyzed by one-way ANOVA. As shown in Table 1, there was no significant difference in survival rate (P > 0.05) of the vitrified oocytes that were placed in dilution solution containing 0.17, 0.25, or 0.33 M sucrose and the non-treated controls. On Day 2 (fertilized on Day 0), cleavage to the 8-cell stage was similar for the 0.17, 0.25, and 0.33 M dilution groups, but the rates for all three were significantly lower (P < 0.05) than for the control group. The blastocyst rate on Day 8 was significantly higher for the 0.25 M group than for any other experimental group but still significantly lower than for the control. In conclusion, this study suggests that with this vitrification/warming procedure the optimum concentration of sucrose in the dilution solution is 0.25 M. Table 1. Oocyte survival after vitrification/warming and subsequent embryo development The authors would like to thank Ms Colleen Shaffer for the preparation of bovine oocytes.

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Experimental Application of Sage in Rabbit Husbandry

Acta Veterinaria Brno ◽

10.2754/avb200877040581 ◽

2008 ◽

Vol 77 (4) ◽

pp. 581-588 ◽

Cited By ~ 10

Author(s):

R. Szabóová ◽

A. Lauková ◽

Ľ. Chrastinová ◽

M. Simonová ◽

V. Strompfová ◽

...

Keyword(s):

Inhibitory Effect ◽

Negative Influence ◽

Feed Consumption ◽

Control Group ◽

Coagulase Negative Staphylococci ◽

E Coli ◽

Commercial Feed ◽

Experimental Group

Salvia spp. belongs to the Labiatae family and is characterized by antimicrobial and antiinflammatory effect. The aim of this study was to test its in vitro and in vivo inhibitory effect against bacteria as well as to find an alternative possibility to use sage in the rabbit ecosystem examining biochemical, zootechnical and inmunological indicators, compared to the commercial feed mixture Xtract. Using the sage extract in in vitro tests, its inhibitory effect was noted. Under in vivo conditions, in the experimental group with sage (EG1), reduction of Pseudomonas-like sp. (p < 0.01) and E. coli (p < 0.01) was noted after 7 days of sage application compared to the control group CG2 (with Robenidin) as well as after 21 days of sage extract application, when the reduction of coagulase-negative staphylococci (p < 0.01) was detected (in comparison with the experimental group-EG2, Xtract group). In the caecum of rabbits from EG1, higher values of lactic, acetic and butyric acids were noted. The values of propionic acid were not influenced. Biochemical indicators were not influenced; however, the values of GSH Px were lower in EG1 compared to EG2. Higher phagocytic activity (18%) was noted in EG1 than in EG2 (13%) after 21 days of additives application. The reduction of Eimeria sp. oocysts was demonstrated in EG1 (sage group) after 7 days of sage application comparing to CG2 (217 OPG to 566 OPG). The animals in both experimental groups achieved higher feed consumption and weight gain, lower mortality compared to both controls. Neither of the additives had a negative influence on the health status and growth performance of rabbits.

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hPMSCs protects against aging-induced oxidative damage of CD4+ T cells through activating Akt-mediated Nrf2 antioxidant signaling

10.21203/rs.3.rs-46702/v1 ◽

2020 ◽

Author(s):

Yanlian Xiong ◽

Yueming Wang ◽

Jiashen Zhang ◽

Nannan Zhao ◽

Aiping Zhang ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Nuclear Translocation ◽

Cell Senescence ◽

Antioxidant Defenses ◽

Control Group ◽

Akt Inhibitor ◽

Gsk 3Β ◽

T Cell Senescence

Abstract Background: Mesenchymal stem cells (MSCs) was considered as regenerative therapeutic approach in both acute and chronic diseases. However, whether MSCs regulate the antioxidant metabolism of CD4+ T cells and weaken immunosenescence remains unclear. Here, we reported the protective effects of hPMSCs in aging-related CD4+ T cell senescence and identified the underlying mechanisms using a D-gal induced mouse aging model.Methods: In vivo study, 40 male C57BL/6 mice (8 weeks) were randomly divided into four groups: control group, D-gal group, hPMSC group and PBS group. In in vitro experiment, human naive CD4+ T (CD4CD45RA) cells were prepared using a naive CD4+ T cell isolation kit II and pretreated with the Akt inhibitor LY294002 and Nrf2 inhibitor ML385. Then, isolated naive CD4+ T cell were cocultured with hPMSCs for 72 h in the absence or presence of anti-CD3/CD28 Dynabeads and IL-2 as a mitogenic stimulus. Intracellular ROS changes were detected by flow cytometry. The activities of the antioxidant enzymes superoxide dismutase, glutathione peroxidase and catalase were measured by colorimetric analysis. The senescent T cells were detected SA-β-gal stain. The expression of aging related proteins were detected by Western blotting, RT-PCR and confocal microscopy.Results: We found that hPMSC treatment markedly decreased the ROS level, SA-β-gal positive cells number, senescence-associated secretory phenotype (IL-6 and OPN) expression and aging-related protein (P16 and P21) expression in senescent CD4+ T cells. Furthermore, hPMSC treatment effectively upregulated Nrf2 nuclear translocation and the expression of downstream target genes (HO-1, CAT, GCLC and NQO1) in senescent CD4+ T cells. Moreover, in vitro studies revealed that hPMSCs attenuated CD4+ T cell senescence by upregulating the Akt/GSK-3β/Fyn pathway to activate Nrf2 functions. Conversely, the antioxidant effects of hPMSCs were blocked by the Akt inhibitor LY294002 and Nrf2 inhibitor ML385 in senescent CD4+ T cells.Conclusions: Our results indicate that hPMSCs attenuate D-gal induced CD4+ T cell senescence by activating Nrf2-mediated antioxidant defenses and that upregulation of Nrf2 by hPMSCs is regulated via the Akt/GSK-3β/Fyn pathway.

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Cortical Piezo-Puncture as a Minimally Invasive Method for Reducing MiniScrew Implant Insertion Torque: A Preliminary in vitro Study

APOS Trends in Orthodontics ◽

10.4103/apos.apos_53_18 ◽

2018 ◽

Vol 8 ◽

pp. 124-130

Author(s):

Juan Pablo Gomez ◽

Lisbetd Giraldo ◽

Daniela Montoya ◽

Miguel Urrea

Keyword(s):

In Vitro Study ◽

Control Group ◽

Insertion Torque ◽

Miniscrew Implants ◽

Mini Implants ◽

Intact Bone ◽

Invasive Method ◽

Miniscrew Implant ◽

Experimental Group

Objective The objective of this study was to determine the effect of cortical piezo-puncture (CPP) on maximum insertion torque (MIT), maximum removal torque (MRT), and maximum axial load (MAL) during the insertion of self-drilling miniscrew implants (MSI), in an experimental model with proximal epiphysis of bovine tibia. Materials and Methods A comparative study was conducted using two groups of 20 self-drilling MSI inserted in intact bone (control group) and in bone with previous CPP (experimental group). MIT, MRT, and MAL of the 20 mini implants of each group were measured. Using SPSS software, Student’s t-test was applied to compare MIT and MRT and the U-test Mann–Whitney test was applied to compare MAL in both groups as well as Pearson and Spearman correlation. Results In the experimental group, average values of 12.85 (±4,32) Newton x centimeters (Ncm), 13.7 (±4,54) Ncm, and 22,474 (±895,95) gF for MIT, MRT, and MAL were found, respectively. In the control group, average values found for MIT, MRT, and MAL were 20.2 (±4,7) Ncm, 22.3 (±5,17) Ncm, and 4688,7 (±320,18) gF, respectively. Statistically significant differences were observed in MIT, MRT, and MAL between control and experimental groups (P < 0,001). Conclusions CPP before insertion of orthodontic MSI in bovine tibia significantly reduces MIT, MRT, and MAL.

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Evaluation of silver nanoparticles for the prevention of SARS-CoV-2 infection in health workers: in vitro and in vivo

10.1101/2021.05.20.21256197 ◽

2021 ◽

Author(s):

Horacio Almanza-Reyes ◽

Sandra Moreno ◽

Ismael Plascencia-Lopez ◽

Martha Alvarado-Vera ◽

Leslie Patron-Romero ◽

...

Keyword(s):

Silver Nanoparticles ◽

High Risk ◽

Cultured Cells ◽

Health Workers ◽

Randomized Study ◽

Healthcare Personnel ◽

Control Group ◽

Experimental Group

SARS-CoV-2 infection in hospital areas is of a particular concern, since the close interaction between health care personnel and patients diagnosed with COVID-19, which allows virus to be easily spread between them and subsequently to their families and communities. Preventing SARS-CoV-2 infection among healthcare personnel is essential to reduce the frequency of infections and outbreaks during the pandemic considering that they work in high-risk areas. In this research, silver nanoparticles (AgNPs) were tested in vitro and shown to have an inhibitory effect on SARS-CoV-2 infection in cultured cells. Subsequently, we assess the effects of mouthwash and nose rinse with ARGOVIT silver nanoparticles (AgNPs), in the prevention of SARS-CoV-2 contagion in health workers consider as high-risk group of acquiring the infection in the General Tijuana Hospital, Mexico, a hospital for the exclusive recruitment of patients diagnosed with COVID-19. We present a prospective randomized study of 231 participants that was carried out for 9 weeks (during the declaration of a pandemic). The "experimental" group was instructed to do mouthwash and nose rinse with the AgNPs solution; the "control" group was instructed to do mouthwashes and nose rinse in a conventional way. The incidence of SARS-CoV-2 infection was significantly lower in the "experimental" group (two participants of 114, 1.8%) compared to the "control" group (thirty-three participants of 117, 28.2%), with a 84.8% efficiency. We conclude that the mouth and nasal rinse with AgNPs helps in the prevention of SARS-CoV-2 infection in health personnel who are exposed to patients diagnosed with COVID-19.

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DYNAMICS OF HAEMATOLOGICAL AND FUNCTIONAL PARAMETERS OF PEKIN DUCK BLOOD DURING ADMINISTRATION OF ORGANIC SELENIUM PREPARATION

Veterinary Science Today ◽

10.29326/2304-196x-2018-2-25-64-68 ◽

2018 ◽

pp. 64-68

Author(s):

Ye. O. Anisimova ◽

V. V. Pronin ◽

L. V. Kletikova ◽

N. N. Yakimenko

Keyword(s):

Total Protein ◽

Color Index ◽

Mineral Metabolism ◽

Pekin Duck ◽

Control Group ◽

Axillary Vein ◽

Functional Parameters ◽

Organic Selenium ◽

Age Related ◽

Experimental Group

Age-related dynamics of hematological and functional parameters of Pekin duck blood during administration of organic selenium preparation DAFS-25k (at a 1.3 mg/kg dose) was presented. In order to study morphological parameters from the age of one day old to 120 days old blood was collected from axillary vein of birds prior to feeding each 15 days in the morning. To assess hematological parameters RBC and WBC count was performed, hemoglobin contents and hematocrit value were determined and color index of blood was calculated. The experiment demonstrated gradual increase in these parameters in the experimental and control groups. Birds from the experimental group demonstrated higher parameters than those of the control group. It was determined that hemoglobin contents was 9.86% higher in the experimental group in comparison with the control group due to selenium administration. The blood color index in this group was also higher (р ≤ 0.05). Biochemical blood analysis included the following parameters: total protein, albumin, uric acid, glucose, calcium, phosphorus, ALT and AST transaminase. The test results showed that during all age periods total protein and albumin contents in ducks of the experimental group exceeded those of the control group. The selenium-containing preparation had a positive influence on the carbohydrate metabolism ensuring high level of metabolic processes in birds, contributed to optimal calcium to phosphorus ratio which positively influenced mineral metabolism and development of the locomotor system. Obtained AST and ALT values in both groups are not beyond reference values which indirectly confirms absence of toxic effect on ducks’ organism if selenium is administered at the specified doses. It was determined that DAFS-25k has not affected hematological and functional parameters of Pekin duck blood and it contributed to mineral metabolism correction, reduction of stress factor effect within critical periods of development and improvement of the excretory system function.

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The mitochondrial calcium uniporter induces apoptosis in cardiomyocytes cultured with high-glucose medium by affecting mitochondrial function

10.21203/rs.3.rs-27647/v1 ◽

2020 ◽

Author(s):

Xia Chen ◽

Wenyun Guo ◽

Zhe Jing ◽

Tao Zhang ◽

Zhaoqi Wu ◽

...

Keyword(s):

Oxidative Stress ◽

High Glucose ◽

Caspase 3 ◽

Normal Group ◽

Control Group ◽

Caspase 9 ◽

Mitochondrial Ros ◽

H9c2 Cardiomyocytes ◽

Experimental Group

Abstract Background As the number of diabetics worldwide continues to increase, diabetic cardiomyopathy has become one of the main causes of cardiovascular disease risk in diabetic patients. Currently, the pathophysiological mechanism of DCM has not been fully elucidated. In the present study, relevant pathological changes of cardiomyocytes in the high glucose environment were simulated by in vitro culture of rat H9C2 cardiomyocytes, to explore the mechanism by which MCU induces apoptosis in cardiomyocytes. Method: Cultured rat myocardium H9C2 cells in vitro and divided into high glucose group (glucose concentration 33 mmol/L), normal group (glucose concentration 5.5 mmol/L), experimental group (5.5 mmol/L glucose and transfected with MCU siRNA) and control group (5.5 mmol/L glucose and transfected negative control siRNA). Comparative analysis of MCU expression, Ca2+ uptake, mitochondrial function, oxidative stress and apoptosis of two groups of cells. Results (1) Compared with normal group, in the high glucose group the MCU expression of myocardial cells in H9C2 rats decreased, The Ca2+ levels, membrane potential and mitochondrial ATP levels decreased, mitochondrial ROS levels increased, NADH+/NADPH ratio in cardiomyocytes increased, GSH/GSSG ratio decreased, the expression levels of cleaved caspase-3 and cleaved caspase-9 increased, bcl-2 expression decreased, the number of cardiomyocytes apoptotic cells increases. (2) Compared with the normal group and the control group, the experimental group MCU expression of myocardial cells in H9C2 rats decreased, The Ca2+ levels, membrane potential and mitochondrial ATP levels decreased, mitochondrial ROS levels increased, NADH+/NADPH ratio in cardiomyocytes increased, GSH/GSSG ratio decreased, the expression levels of cleaved caspase-3 and cleaved caspase-9 increased, bcl-2 expression decreased, the number of cardiomyocytes apoptotic cells increases. Discussion This study suggested that MCU expression in rat H9C2 cardiomyocytes was decreased in the high glucose environment, causing abnormal mitochondrial calcium uptake and imbalanced calcium homeostasis, which may further contribute to mitochondrial dysfunction and enhanced oxidative stress in cardiomyocytes. Mitochondrial dysfunction and enhanced oxidative stress ultimately led to apoptosis in cardiomyocytes.

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