84 PTEROSTILBENE CAN REDUCE THE PERCENTAGE OF LIPIDS AND REACTIVE OXYGEN SPECIES IN IN VITRO-PRODUCED BOVINE EMBRYOS

2017 ◽  
Vol 29 (1) ◽  
pp. 149
Author(s):  
F. Sosa ◽  
J. Fernando de la Torre ◽  
H. Álvarez ◽  
S. Pérez ◽  
M. E. Kjelland ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Culture Medium ◽  
Statistical Significance ◽  
Cell Mass ◽  
Reactive Oxygen ◽  
Oxygen Species ◽  
Embryo Production ◽  
Cell Counts ◽  
Significant Difference ◽  
And Control

The actual challenge for the majority of research centers involves the embryo culture medium, since it is known that the culture medium plays a large role in determining embryo quality. Pterostilbene (PT) is a natural analogue of Resveratrol, an antioxidant that can reduce lipids in embryos, but no reports exist of PT being used with IVF-produced embryos or gametes. The objective of the present research was to evaluate the effect of PT in culture media CDM1 and CDM2 on embryo production, cell count, lipid accumulation, and reactive oxygen species (ROS). A total of 4 concentrations of PT and a control were evaluated, i.e. 3, 1, 0.33, 0.11, and 0 μM, in 2 separate experiments. The first experiment was performed using 6 replicates (n = 204) to evaluate blastocyst production (n = 201) and determine the percentage of lipids using the stain Sudan-Black, (n = 100). Hoechst 33258 and propidium iodide were used for determining cell counts. The second experiment was performed using 7 replicates, the effect of using PT (0.33 μM) was compared with a control with 2 O2 concentrations (5 and 20%) for evaluating ROS production (n = 124). Blastocysts without zona pellucida were incubated 48 h at 38.5°C in PBS (without polyvinyl alcohol) with 60 μL of ro-green fluorescent protein. After incubation, 25 μL of 4′,6-diamidino-2-phenylindole (1 mg mL−1) was added and incubated for 5 min. A fluorescence microscope was used and positive ROS particles digitized using Photoshop CS6 and quantified using the program ImageJ®. The data were transformed to arcsin values for subsequent analysis. In the first experiment, ANOVA and least significant difference tests were used to determine statistical significance (Statgraphics). No significant differences were found among treatments (P > 0.05) for the internal cell mass: 3, 1, 0.33, 0.11 μM PT (29.2 ± 5.2; 28.9 ± 3.8; 22.2 ± 3.2; 29.0 ± 1.7, respectively) and the control (27.0 ± 1.7). The cells of the trophectoderm did not differ (P > 0.05) between treatments (31.7 ± 4.8; 31.3 ± 3.9; 38.6 ± 3.5; 30.8 ± 1.8) and control (33.4 ± 2.1). Total cells did not differ (P > 0.05) between treatments (72.0 ± 9.8; 82.6 ± 4.3; 94.8 ± 12.8; 73.2 ± 9.2) and control (83.8 ± 7.7). Embryo production (Day 8) was greater for control (33.5 ± 3.0) versus treatments (14.1 ± 1.7; 19.4 ± 1.9; 21.1 ± 2.6; 20.8 ± 2.1) (P < 0.05); however, PT reduced the percentage of lipids (11.0 ± 0.8; 10.7 ± 0.9; 11.6 ± 1.3; 11.3 ± 1.1) within the cytoplasm of the embryos (P < 0.05) versus control (17.01 ± 1.20). In the second experiment, a factorial 2 × 2 matrix demonstrated that the O2 concentration did not have an effect on ROS (P > 0.05); however, the PT had a significant effect on the reduction of ROS (P < 0.05), i.e. a negative correlation, r = −0.835. In summary, we determined that PT did not improve the production of blastocysts but resulted in a significant reduction of ROS and lipids.

scholarly journals Correlation between Serum Interleukin-17 level and Serum Reactive Oxygen Species levels among Children experiencing Otitis Media with Effusion

2021 ◽  
Author(s):  
Marwa Abdelhafeez ◽  
Noura Mostafa Mohamed
Keyword(s):  
Reactive Oxygen Species ◽  
Otitis Media ◽  
Otitis Media With Effusion ◽  
Reactive Oxygen ◽  
Serum Levels ◽  
Control Group ◽  
Interleukin 17 ◽  
Oxygen Species ◽  
Significant Difference ◽  
And Control

Abstract Introduction The detection of inflammatory mediators in the serum of children with have otitis media with effusion (OME) and their correspondence with clinical considerations may enable the use of a modern nonsurgical curative treatment for OME. Objective To determine the relation between interleukin-17 (IL-17) serum level and reactive oxygen species (ROS) serum levels in children suffering from OME and to disclose if any variation occurs in the level of IL-17 Will affect the ROS and antioxidant equilibrium in the serum, which indicates the entire body's reaction to OME. Methods The present study was a case-control study. A total of 24 children experienced OME, and 24 healthy controls were recruited.All participants in the study were subjected to a systematic clinical investigation including otoscopic, audiometric, and tympanometric examination. Also, venous blood samples were collected from all children to determine the levels of IL-17 and ROS. Results The mean ± standard deviation (SD) age ranges of the patients and the control group were 6.8 ± 2.7 and 6.2 ± 3.4 years, respectively. A stylistically significant difference in the levels of serum nitric oxide (NO), catalase (CT), myeloperoxidase (MPO), and malondialdehyde (MDA) (p < 0.05) was detected between OME and control patients. No significant difference was found in serum levels of superoxide dismutase (SOD) and glutathione peroxidase (GPX) between OME and control patients. The serum levels of MDA, NO, and MPO positively correlated with the serum levels of IL-17 in OME patients. Conclusion In the present study, there is a reasonable role of the IL-17 pathway in OME pathogenesis through an increase in ROS levels.

scholarly journals Reactive oxygen species formation during tetanic contractions in single isolated Xenopus myofibers

2011 ◽  
Vol 111 (3) ◽  
pp. 898-904 ◽  
Author(s):  
Li Zuo ◽  
Leonardo Nogueira ◽  
Michael C. Hogan
Keyword(s):  
Reactive Oxygen Species ◽  
Skeletal Muscle ◽  
Real Time ◽  
Contractile Activity ◽  
Reactive Oxygen ◽  
Ros Generation ◽  
Oxygen Species ◽  
Fatigue Development ◽  
Species Formation ◽  
Significant Difference

Contracting skeletal muscle produces reactive oxygen species (ROS) that have been shown to affect muscle function and adaptation. However, real-time measurement of ROS in contracting myofibers has proven to be difficult. We used amphibian ( Xenopus laevis) muscle to test the hypothesis that ROS are formed during contractile activity in isolated single skeletal muscle fibers and that this contraction-induced ROS formation affects fatigue development. Single myofibers were loaded with 5 μM dihydrofluorescein-DA (Hfluor-DA), a fluorescent probe that reacts with ROS and results in the formation of fluorescein (Fluor) to precisely monitor ROS generation within single myofibers in real time using confocal miscroscopy. Three identical periods of maximal tetanic contractions (1 contraction/3 s for 2 min, separated by 60 min of rest) were conducted by each myofiber ( n = 6) at 20°C. Ebselen (an antioxidant) was present in the perfusate (10 μM) during the second contractile period. Force was reduced by ∼30% during each of the three contraction periods, with no significant difference in fatigue development among the three periods. The Fluor signal, indicative of ROS generation, increased significantly above baseline in both the first (42 ± 14%) and third periods (39 ± 10%), with no significant difference in the increase in fluorescence between the first and third periods. There was no increase of Fluor in the presence of ebselen during the second contractile period. These results demonstrated that, in isolated intact Xenopus myofibers, 1) ROS can be measured in real time during tetanic contractions, 2) contractile activity induced a significant increase above resting levels of ROS production, and 3) ebselen treatment reduced ROS generation to baseline levels but had no effect on myofiber contractility and fatigue development.

scholarly journals aflN Is Involved in the Biosynthesis of Aflatoxin and Conidiation in Aspergillus flavus

Toxins ◽  
2021 ◽  
Vol 13 (11) ◽  
pp. 831
Author(s):  
Kunzhi Jia ◽  
Lijuan Yan ◽  
Yipu Jia ◽  
Shuting Xu ◽  
Zhaoqi Yan ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Aspergillus Flavus ◽  
Prevention And Control ◽  
Reactive Oxygen Species Generation ◽  
Reactive Oxygen ◽  
Hyphal Growth ◽  
Aflatoxin Production ◽  
Oxygen Species ◽  
Aflatoxin Biosynthesis ◽  
And Control

Aspergillus flavus poses a threat to society economy and public health due to aflatoxin production. aflN is a gene located in the aflatoxin gene cluster, but the function of AflN is undefined in Aspergillus flavus. In this study, aflN is knocked out and overexpressed to study the function of AflN. The results indicated that the loss of AflN leads to the defect of aflatoxin biosynthesis. AflN is also found to play a role in conidiation but not hyphal growth and sclerotia development. Moreover, AlfN is related to the response to environmental oxidative stress and intracellular levels of reactive oxygen species. At last, AflN is involved in the pathogenicity of Aspergillus flavus to host. These results suggested that AflN played important roles in aflatoxin biosynthesis, conidiation and reactive oxygen species generation in Aspergillus flavus, which will be helpful for the understanding of aflN function, and will be beneficial to the prevention and control of Aspergillus flavus and aflatoxins contamination.

scholarly journals Elevated Serum Reactive Oxygen Species Level Predicts Early Abortion

2021 ◽  
Vol 5 (1) ◽  
pp. 37
Author(s):  
Joserizal Serudji ◽  
Nuzulia Irawati ◽  
Johanes Cornelius Mose ◽  
Hirowati Ali ◽  
Yusrawati Yusrawati
Keyword(s):  
Reactive Oxygen Species ◽  
Gestational Age ◽  
Elevated Serum ◽  
Reactive Oxygen ◽  
Normal Pregnancy ◽  
Trophoblast Invasion ◽  
Enzyme Linked Immunosorbent Assay ◽  
Oxygen Species ◽  
Significant Difference ◽  
Early Abortion

Background: Impaired trophoblast invasion is associated with early abortion. The calorie needed for the trophoblast cell (TC) invasion is mainly met by adenosine triphosphate (ATP) produced in the mitochondria. Reactive oxygen species (ROS), byproduct of ATP synthesis, plays an important role in cellular physiology, but a high level of ROS may result in deoxyribonucleic acid (DNA) damage or cell dysfunction, thereby impaired TC invasion leading to early abortion. The study aims to determine elevated serum ROS level to predicts early abortion.Materials and method: This was an observational study with a cross-sectional design. Fifty subjects with gestational age less than 12 weeks, consist of 25 early abortions and 25 normal pregnancies subjects, were included in this study. Clinical examination and diagnosis are carried out in 2 Hospitals and 5 Public Health Centers in Padang. Examination of ROS levels was carried out by enzyme-linked immunosorbent assay (ELISA) in the Biomedical Laboratory, Faculty of Medicine, Universitas Andalas. The Mann-Whitney test was used to analyze the difference of serum ROS levels, with a significance level of 0.05.Results: The subjects of the two study groups were equivalent in terms of age, gestational age, and gravidity (p=0.051, p=0.453, and p=1.000). The median ROS levels were found to be 1.36 (1.02-26.30) ng/mL in the early abortion and 1.20 (0.43-2.75) ng/mL in the normal pregnancy (p=0.003).Conclusion: There is a significant difference between ROS levels in early abortion and normal pregnancy.Keywords: ROS, early abortion, normal pregnancy

136 GameteGuard in maturation medium improves bovine embryo quality and quantity

2021 ◽  
Vol 33 (2) ◽  
pp. 176
Author(s):  
A. Scharf ◽  
J. Barfield ◽  
M. Shepherd ◽  
L. Herickhoff
Keyword(s):  
Reactive Oxygen Species ◽  
Reactive Oxygen ◽  
Bovine Embryo ◽  
Apoptotic Cells ◽  
Oxygen Species ◽  
Bovine Embryos ◽  
High Quality ◽  
Embryo Production ◽  
Maturation Medium ◽  
The Impact

The number of invitro-produced (IVP) bovine embryos generated in the United States increased 11% from 2017 to 2018, demonstrating a demand for new technology to rapidly and efficiently improve herd genetics. Unfortunately, embryo production on a per oocyte basis remains low, limiting the number of high-quality commercial embryos available for transfer or freezing. Reactive oxygen species (ROS) are a major contributor to low embryo production rates of invitro gametes and are especially problematic in maturation media. This is due to high numbers of mitochondria in the bovine oocyte producing ROS exacerbated by the invitro environment. GameteGuard® is an organic plant-based additive that provides protection against reactive oxygen species. The aims of this research were to evaluate the effects of GameteGuard addition to invitro bovine embryo production media to (1) mitigate the impact of ROS on oocyte maturation and competence, and (2) increase the production of high-quality embryos in our IVP system. Four rounds of invitro embryo production (Colorado State University protocol and media) with or without maturation medium supplemented with GameteGuard were performed (n=1175 total oocytes). Oocytes were aspirated from abattoir ovaries; those with homogeneous ooplasm and multiple layers of even cumulus cells were matured using control (n=585) or GameteGuard-supplemented (n=590) maturation medium. Oocytes were fertilized with frozen/thawed semen from proven IVF bulls (n=4) and then cultured in a two-step media system to Day 7. Blastocysts were evaluated on Day 7 according to IETS embryo grading standards. A subset of 80 blastocysts were fixed and stained after grading using an In Situ Cell Death Detection Kit (Fluorescein; Millipore Sigma) and Hoechst 33342 (Invitrogen) as apoptotic and counter-stains, respectively. Stained blastocysts were assessed by immunofluorescence microscopy to obtain the percentage of apoptotic cells and total cell count. Because multiple bulls were used, a linear mixed model was required for data analysis. Treatment was considered the fixed effect and bulls as a random effect to determine the impact of the treatment on stage and quality of blastocysts. Cleavage rates were not different (P&gt;0.5) but GameteGuard supplementation of maturation medium resulted in a 22% increase in grade-1 embryos per total blastocysts produced (P&lt;0.01; n=79/106 control vs. n=120/137 GameteGuard grade 1 blastocysts). Additionally, the percent of grade 2 and 3 embryos per total blastocysts decreased, (P&lt;0.01; 27%, n=27/106 control and 11%, n=14/137 GameteGuard grade 2 and 3 blastocysts), suggesting that supplementation with GameteGuard promoted development of better-quality embryos overall. The percentage of apoptotic cells was also significantly higher in control blastocysts compared with treated blastocysts (P&lt;0.02; 11.4%, n=38 control and 12.8%, n=42 GameteGuard). GameteGuard supplementation of the maturation medium significantly increased the production of high-quality IVP bovine embryos, thereby providing potential marketable gain for embryo producers.

80 REACTIVE OXYGEN SPECIES LEVEL IN PIG EMBRYOS CULTURED IN PRESENCE OF HYALURONAN

2016 ◽  
Vol 28 (2) ◽  
pp. 169
Author(s):  
B. Gajda ◽  
M. Kucia ◽  
Z. Smorag ◽  
M. Romek
Keyword(s):  
Reactive Oxygen Species ◽  
Embryo Culture ◽  
Culture Medium ◽  
Reactive Oxygen ◽  
Blastocyst Stage ◽  
Molecular Probes ◽  
Control Group ◽  
Oxygen Species ◽  
Science Center ◽  
Experimental Group

It has been reported that during in vitro embryo culture reactive oxygen species (ROS) are generated and are detrimental to embryo development. A recent study (Smorag et al., Proc. 9th ICPR, 2013, 110) demonstrated that an addition of 1 mg mL–1 of hyaluronan (HA) to porcine embryo culture medium improves the development of zygote to blastocyst stage and the quality of produced embryos. Moreover, the embryos cultured with HA showed lower inner mitochondrial membrane potential (Romek et al., 2015 Proc. Symp. Progress in Cell Biology: Mitochondria and Chloroplast, Krakow, 31). Based on the beneficial effect provided by supplementation of HA during embryo culture, we investigated the ROS level in porcine embryos cultured with HA. Porcine zygotes were obtained surgically after flushing the oviducts of superovulated and inseminated gilts. In the experimental group, zygotes were cultured up to the blastocyst stage in NCSU-23 medium supplemented with 1 mg mL–1 of HA (CROMA, Pharma GmbH, Leobendorf, Austria), in an atmosphere containing 5% CO2 in air, at 39°C. In the control group, HA supplementation was omitted. To measure ROS level, embryos at the stages 2–4 and 8–16 cell, morula, and blastocyst (experimental group) and zygote, 2–4 and 8–16 cell, morula, and blastocyst (control group) were labelled with 5 μM CM-H2DCFDA dye (Molecular Probes Inc., OR, USA) for 30 min at 39°C. Labelled embryos were then examined under a Nikon Eclipse microscope with a CCD camera. The total amount of fluorescence emitted from each individual embryos and proportional to the ROS level was measured in arbitrary units. The data were analysed using one-way ANOVA and post-hoc Tukey test. ROS level (mean ± standard error of the mean) in the experimental group was 8.21 ± 2.65 (n = 25), 10.31 ± 3.13 (n = 18), 9.08 ± 2.89 (n = 21), and 20.45 ± 2.38 (n = 31) for 2–4 cell, 8–16 cell, morula, and blastocyst, respectively, whereas in the control group was 9.15 ± 3.43 (n = 15), 7.11 ± 3.13 (n = 18), 8.67 ± 3.04 (n = 19), 11.47 ± 2.46 (n = 29), and 54.74 ± 2.89 (n = 21) for zygote, 2–4 cell, 8–16 cell, morula, and blastocyst, respectively. For experimental and control groups, ROS levels remained unchanged up to morula. On the contrary, at the blastocyst stage from the experimental group ROS level decreased significantly (P ≤ 0.05) in comparison with blastocysts from the control group. In conclusion, porcine blastocysts derived from zygotes cultured with supplementation of 1 mg of HA possess a significantly lower ROS level than blastocysts cultured without HA. This suggests that HA supplementation in culture medium can reduce the ROS level in porcine cultured blastocysts. The project was funded by the National Science Center based on decision number DEC-2012/07/B/NZ9/01326.

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