180 EXPRESSION OF TUMOR NECROSIS FACTOR-STIMULATED GENE 6 PROTEIN IN BOVINE CUMULUS–OOCYTE COMPLEXES AT DIFFERENT MATURATION STATUS

2013 ◽  
Vol 25 (1) ◽  
pp. 239
Author(s):  
P. Trzeciak ◽  
R. Starzyński ◽  
Ł. Rąpała ◽  
S. Dąbrowski ◽  
E. Nałęcz-Nieniewska ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Protein Localization ◽  
Statistical Significance ◽  
Mrna Level ◽  
Fluorescent Microscopy ◽  
Cumulus Cells ◽  
Mean Values ◽  
Significant Difference ◽  
Tukey Honestly Significant Difference

The TSG-6 is a ~35-kDa protein belonging to hyaluronan binding superfamily proteins. The TSG-6 plays role in inflammation and in inflammation-like processes (i.e. ovulation). The tsg-6 expression is induced in cumulus–oocyte complex (COC) cells just before ovulation. It is involved in the migration of cumulus cells though the formation and stabilisation of the extracellular matrix. Disturbances in secretion of this protein lead to a reduction in the number of ovulated oocytes. Although studies of the prevalence and role of TSG-6 in COC were conducted in several animal models, little is known about TSG-6 in cattle. The aim of this study was to assess tsg-6 mRNA expression and protein localization in bovine cumulus cells from COC at different maturation status. Ovaries were collected from the slaughterhouse. Cumulus cells were isolated from COCs in different stages of maturity. In variant I, COC were isolated from small follicles of Φ 2 to 6 mm. In variant II, COC were also isolated from small follicles and matured in vitro in TCM199 HEPES with 10% FBS and 0.02 IU NIH-pFSH mL–1, 1 µg mL–1 β-oestradiol, 0.2 µM sodium pyruvate, 50 µg mL–1 gentamicin in 5% CO2, 38.5°C for 24 h. In variant III, COCs were isolated from large follicles of Φ >15 mm. Analysis of tsg-6 expression in cumulus cells was performed using real-time PCR. Expression of Tsg-6 was normalized to that of s18. Statistical analysis of tsg-6 mRNA level in all variants was carried out by 1-way ANOVA, and comparisons of mean values were made with the Tukey honestly significant difference test (Statgraphic 5.1 Centurion); P < 0.05 was considered to reflect the presence of statistical significance. For all variants, paraffin-embedded slices of COC were carried out to localise TSG-6 by indirect method of immunofluorescence. Immunostaining of the TSG-6 protein was performed using primary polyclonal antibody raised against bovine TSG-6. Antigen–antibody complexes were visualised after incubation with secondary IgG conjugated with fluorescein isothiocynanate. Immunolocalization of TSG-6 was performed using fluorescent microscopy. The relative expression of tsg-6 mRNA in variant II was more than 2 times higher (1.887 ± 0.797 a.u.) compared with variant III (0.760 ± 0.130 a.u.). The difference was statistically significant (P < 0.05). No tsg-6 expression in variant I was detected. The presence of TSG-6 protein in the extracellular matrix of cumulus cells from variant II as well as from variant III was detected. The present data suggest that tsg-6 gene expression and its protein presence are stimulated during COC maturation in cattle as previously demonstrated for other species. Supported by Warsaw University of Life Sciences, Faculty of Veterinary Medicine: 505-10-02330050.

56 COMPARISON OF THE MORPHOLOGY OF VARIOUS REGIONS OF THE CATTLE OVIDUCT IN FOUR PHASES OF THE OVARIAN CYCLE

2012 ◽  
Vol 24 (1) ◽  
pp. 140
Author(s):  
A. M. Duszewska ◽  
A. Compa ◽  
M. Zelechowska ◽  
A. Piliszek ◽  
A. Rynkowska ◽  
...  
Keyword(s):  
Cross Sections ◽  
Statistical Significance ◽  
Ovarian Cycle ◽  
Mean Values ◽  
Honestly Significant Difference ◽  
Significant Difference ◽  
Ovarian Morphology ◽  
Tukey Honestly Significant Difference ◽  
Hematoxylin And Eosin ◽  
Mucosal Folds

The oviduct provides the environment necessary for the gamete transport, completion of spermatozoa capacitation, oocyte fertilization and the early development of embryos. In cattle, all of these processes take place between Day 0 to 4 of the ovarian cycle (Day 0 is the day of ovulation). In previous studies, temporal changes in the bovine oviduct morphology were evaluated by dividing the ovarian cycle into luteal and follicular phases. In order to understand the relation between the bovine oviduct morphology and processes occurring there, the ovarian cycle has been further divided into four phases: I (Day 0–4), II (Day 5–10), III (Day 11–17) and IV (Day 18–20), with the day of ovulation considered Day 0 (1980 J. Dairy Sci. 63, 155–160). The aim of the study was to evaluate the oviduct morphology of the infundibulum, ampulla and isthmus in 4 phases of the ovarian cycle. Research material comprised cattle oviducts (classified into 1 of the 4 phases of the cycle based on ovarian morphology), dissected into infundibulum, ampulla and isthmus and subsequently sectioned and processed for histological preparations (hematoxylin and eosin, H&E, staining). Diameters of transverse cross-sections of oviduct and its lumen and thickness of tunicas: mucosa, muscularis and serosa were evaluated in relation to the region of oviduct and the phase of ovarian cycle. Values are given in μm. Statistical analysis was carried out by 1-way analysis of variance and comparisons of mean values were made with the Tukey honestly significant difference test (Statgraphics Plus 5); P < 0.05 was considered to reflect the presence of statistical significance. The comparison of the diameters of transverse cross-sections (A) of oviduct and its lumen (B) shows significant statistical differences between ampulla and isthmus within the phases: A-I (4507.26 vs 2524.47), II (4510.53 vs 2540.67), III (4503.28 vs 2534.07), IV (4500.73 vs 2533.90); B-I (4191.10 vs 1950.88), II (4173.63 vs 1986.33), III (4198.53 vs 1966.88) and IV (4192.50 vs 1959.33). There are no differences among 4 phases of the ovarian cycle. The thickness of tunicas muscularis and serosa of infundibulum (I: 26.81 vs 196.85; II: 27.03 vs 201.80; III: 26.22 vs 199.45; IV: 23.97 vs 198.01), ampulla (I: 91.51 vs 214.50; II: 90.72 vs 212.55; III: 88.61 vs 213.30; IV: 89.65 vs 206.28) and isthmus (I: 202.29 vs 216.52; II: 199.24 vs 207.74; III: 200.90 vs 212.38; IV: 200.38 vs 210.86) show only statistically significant differences within the phases, whereas the tunica mucosa shows only statistically significant differences between phases and the term of the height of epithelium at the base of mucosal folds (I: 26.49; II: 25.20; III: 24.14; IV: 29.96) and their apical parts (I: 28.09; II: 26.01; III: 25.45; IV: 30.96). In conclusion, differences in oviduct morphology are mainly region specific, whereas the epithelium morphologically infundibulum, ampulla and isthmus show variation in the 4 phases of the ovarian cycle. Supported by Grant MNiSW N311236137.

165 In vitro maturation of ovine and caprine oocytes during breeding and nonbreeding seasons

2019 ◽  
Vol 31 (1) ◽  
pp. 207
Author(s):  
M. Markle ◽  
C. K. Mak ◽  
V. Medina ◽  
C. R. F. Pinto
Keyword(s):  
Breeding Season ◽  
In Vitro Maturation ◽  
Statistical Significance ◽  
Polar Body ◽  
Cumulus Cells ◽  
Nonbreeding Season ◽  
Nuclear Maturation ◽  
Significant Difference ◽  
Maturation Rate

The current study investigated the in vitro meiotic competence of ovine and caprine oocytes that underwent nuclear maturation during the breeding and nonbreeding seasons. We hypothesised that maturation rates of ovine and caprine oocyte would be significantly lower during the nonbreeding season. Ovine (Katahdin crossbred) and caprine (mainly Spanish crossbred) ovaries were collected from a local abattoir in the southern United States. Age of the animals was not determined. Cumulus-oocyte complexes (COC) were harvested by slicing the ovaries and searching using a stereomicroscope. Oocytes with more than 3 layers of unexpanded cumulus cells and with evenly granulated cytoplasm were selected for in vitro maturation (IVM). A commercial bovine IVM media (IVF Bioscience, Falmouth, United Kingdom) was used throughout the study. After 24h of IVM, ovine and caprine oocytes were denuded and oocytes with an extruded polar body (meiotic metaphase II oocytes) were considered to have reached nuclear maturation. The seasons in this study were defined as follows: breeding season=September to April and nonbreeding season=May to July. The presence of corpus hemorrhagicum or corpus luteum in at least 70% of the ovaries indicated the breeding season for the animals. Proportions of oocytes undergoing nuclear maturation were analysed using a two-tailed Chi-squared test. Statistical significance was set at P ≤ 0.05. The ovine maturation rate was 59% (65/111) and 49% (254/519) and the caprine maturation rate was 70% (39/56) and 40% (64/162) during the breeding and nonbreeding seasons, respectively. These results show a significant difference in nuclear maturation for caprine oocytes (P&lt;0.001) during the breeding and nonbreeding seasons; however, there was no significant difference in nuclear maturation for ovine oocytes (P=0.06) during the breeding and nonbreeding seasons. High environmental temperatures during the nonbreeding season may have had detrimental effects on oocyte nuclear maturation in caprine but not in ovine oocytes. Why oocytes from these 2 species differ on how they are adversely affected by season remains to be elucidated.

scholarly journals Introducing intrauterine antibiotic infusion as a novel approach in effectively treating chronic endometritis and restoring reproductive dynamics: a randomized pilot study

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Konstantinos Pantos ◽  
Mara Simopoulou ◽  
Evangelos Maziotis ◽  
Anna Rapani ◽  
Sokratis Grigoriadis ◽  
...  
Keyword(s):  
Fertility Restoration ◽  
Statistical Significance ◽  
Antibiotic Administration ◽  
Oral Antibiotic ◽  
Treatment Rate ◽  
Chronic Endometritis ◽  
Vitro Fertilization ◽  
Significant Difference ◽  
Antibiotic Infusion

AbstractThe chronic nature of Chronic Endometritis (CE) along with the challenging management and infertility entailed, call for cutting-edge therapeutic approaches. This study introduces the novel treatment of intrauterine antibiotic infusion (IAI) combined with oral antibiotic administration (OAA), and it assesses respective performance against the gold standard treatment of OAA. Data sourced herein reports on treatment efficiency and fertility restoration for both patients aiming to conceive naturally or via In Vitro fertilization. Eighty CE patients, 40 presenting with recurrent implantation failure, and 40 with recurrent pregnancy loss, were enrolled in the IVF and the natural conception arm respectively. Treatment was subjected to randomization. Effectively treated patients proceeded with either a single IVF cycle or were invited to conceive naturally over a 6-month period. Combination of IAI and OAA provided a statistically significant enhanced effectiveness treatment rate (RR 1.40; 95%CI 1.07–1.82; p = 0.01). No statistically significant difference was observed regarding the side-effects rate (RR 1.33; 95%CI 0.80–2.22; p = 0.52). No statistically significant difference was observed for either arm regarding live-birth rate. Following an intention-to-treat analysis, employment of IAI corresponds to improved clinical pregnancy rate-albeit not reaching statistical significance. In conclusion, complimentary implementation of IAI could provide a statistically significant enhanced clinical treatment outcome.

scholarly journals Comparison of Shear Bond Strength and Microleakage of Various Bulk-fill Bioactive Dentin substitutes: An in vitro Study

2016 ◽  
Vol 17 (12) ◽  
pp. 997-1002 ◽  
Author(s):  
Fahad I Alkhudhairy ◽  
Zeeshan H Ahmad
Keyword(s):  
Bond Strength ◽  
Shear Bond Strength ◽  
Physical And Mechanical Properties ◽  
Pairwise Comparison ◽  
In Vitro Study ◽  
Composite Resins ◽  
Intergroup Comparison ◽  
Significant Difference ◽  
Tukey Honestly Significant Difference

ABSTRACT Introduction Various bulk-fill materials depending on their composition, viscosity, and flow ability have different physical and mechanical properties. The aim of this in vitro study was to determine and compare the shear bond strength and microleakage properties of activa restorative with other bulk-fill restorative materials surefil (SDR), Biodentine, ever X posterior. Materials and methods Forty permanent premolars were selected for shear bond strength, and 20 permanent premolars were selected with class II cavities on mesial and distal side for microleakage. Universal testing device was used to assess the shear bond strength. Microleakage was checked using dye penetration method under a stereomicroscope. Mean and standard deviation values were calculated from the recorded values. Intergroup comparison was done by one-way analysis of variance (ANOVA) followed by pairwise comparison using Tukey honestly significant difference (HSD) post hoc test. Results The mean shear bond strength was highest for SDR surefil followed by Ever X posterior, Bioactive restorative, and Biodentine respectively. In this study, SDR (surefil) showed better shear bond strength and better microleakage properties compared with the other test materials (F = 186.7157, p < 0.05). Conclusion The result of this study showed that flowable and fiber-reinforced composites have better shear bond strength and microleakage properties. Clinical significance Flowable bulk-fill composite resins can be used as dentin substitutes because of its superior properties. How to cite this article Alkhudhairy FI, Ahmad ZH. Comparison of Shear Bond Strength and Microleakage of Various Bulk-fill Bioactive Dentin substitutes: An in vitro study. J Contemp Dent Pract 2016;17(12):997-1002.

186 THE EFFECT OF MATRIGEL ON THE DEVELOPMENT OF IN VITRO-FERTILIZED PORCINE EMBRYOS

2007 ◽  
Vol 19 (1) ◽  
pp. 209
Author(s):  
S.-W. Kim ◽  
M.-J. Lee ◽  
B.-C. Yang ◽  
G.-S. Im ◽  
H.-H. Seong ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Statistical Significance ◽  
Control Group ◽  
Fertilization In Vitro ◽  
Mouse Tumor ◽  
Matrix Product ◽  
Vitro Fertilization ◽  
Culture Systems

The application of matrix proteins to culture systems for growth of embryos is a logical extension in the quest to better simulate the in vivo culture environment. Matrigel, a commercially available extracellular matrix product containing collagen IV, laminin, entactin, and proteoglycans isolated from mouse tumor cells, has been tested. Development of mouse pre-implantation embryos cultivated in conventional culture medium was contrasted to that of embryos grown in solubilized Matrigel medium. In the solubilized Matrigel medium, in vitro blastocyst formation and hatching were significantly enhanced over that observed in the medium alone control. Therefore, the aim of this study was to investigate the effect of solubilized Matrigel on the development of porcine embryos after in vitro fertilization. In vitro-matured oocytes were fertilized in mTBM medium with fresh spermatozoa for 6 h. Putative zygotes were cultured in PZM-3 medium supplemented with (matrigel group) or without (control group) 0.8% Matrigel for 6 days. The number of cells in blastocysts was determined by staining with Hoechst 33342. Assessment of apoptosis in blastocysts was examined by TUNEL. The statistical significance of the data was analyzed using chi-square test and Student&apos;s t-test. The addition of Matrigel appeared not to increase the proportion of blastocysts (control: 71/219, 21.8 � 2.2% vs. Matrigel: 69/220, 23.5 � 5.8%). However, the mean cell numbers were significantly increased by Matrigel (Matrigel: n = 31, 52.9 � 18.1 vs. control: n = 30, 42.3 � 14.4; P &lt; 0.01). The proportion of apoptotic cells was significantly lower in the Matrigel group (Matrigel: 4.5 � 4.2% vs. control: 6.6 � 5.5%; P &lt; 0.05). In this experiment, Matrigel appeared to increase blastocyst quality of porcine embryos. Results suggest that Matrigel, as an extracellular matrix component, may be another avenue for formulating more physiological culture systems.

326 EVALUATION OF CULTURE DURATION AND PARTIAL CUMULUS CELL REMOVAL DURING CANINE OOCYTE MATURATION

2006 ◽  
Vol 18 (2) ◽  
pp. 270
Author(s):  
C. Hanna ◽  
C. Long ◽  
M. Westhusin ◽  
D. Kraemer
Keyword(s):  
In Vitro Maturation ◽  
Calf Serum ◽  
Cumulus Cell ◽  
Germinal Vesicle ◽  
Cumulus Cells ◽  
Culture Conditions ◽  
Germinal Vesicle Breakdown ◽  
Significant Difference ◽  
Culture Duration

The objectives of this study were to determine whether the percentage of canine oocytes that resume meiosis during in vitro maturation could be increased by either increasing culture duration or by removing approximately one-half of the cumulus cells 24 h after oocytes were placed into culture. Canine female reproductive tracts were collected from a local clinic and ovaries were minced in warm TL-HEPES. Oocytes with a consistently dark ooplasm and at least two layers of cumulus cells were selected, cultured in a basic canine oocyte in vitro maturation medium consisting of TCM-199 with Earl's salts, 2.92 mM Ca-lactate, 20 mM pyruvic acid, 4.43 mM HEPES, 10% fetal calf serum, 1% Penicillin/Streptomycin (GibcoBRL, Grand Island, NY, USA), and 5 μg/mL porcine somatotropin, and incubated at 38.5°C in 5% CO2 in humidified air. Treatment groups were randomly assigned and oocytes were cultured for 60, 84, or 132 h (Basic). From each of these groups, one-half of the oocytes were pipetted through a fine bore pipette to partially remove the cumulus cells 24 h after the start of culture (Basic–1/2). At the end of culture, all oocytes were denuded and the nuclear status was observed with Hoechst 33342 under ultraviolet fluorescence. All data were analyzed by ANOVA with P < 0.05. Since the canine oocyte is ovulated at the germinal vesicle (GV) stage of meiosis and requires up to five days to mature in the oviduct, it was hypothesized that an increased culture time would allow for more oocytes to undergo nuclear maturation to metaphase II (MII). It was also hypothesized that partial removal of cumulus cells would decrease the cumulus cell component in the ooplasm that sustains meiotic arrest, allowing for more oocytes to resume meiosis (RM = germinal vesicle breakdown to MII). Results within each treatment group indicate that there is no significant difference between culture duration and the percent of oocytes that mature to MII. Additionally, there was no significance in the percent of oocytes that resumed meiosis after partial cumulus cell removal. Taken together, these data suggest that neither treatment is effective in canine in vitro maturation systems, given the current maturation culture conditions. Table 1. Nuclear status* of oocytes for three time periods with or without partial cumulus cell removal

229 EXPRESSION OF mRNA ENCODING GLYCOLYTIC ENZYMES IN BOVINE CUMULUS CELLS DURING IN VITRO MATURATION: EFFECTS OF TIME AND FSH

2010 ◽  
Vol 22 (1) ◽  
pp. 272
Author(s):  
E. S. Caixeta ◽  
P. Ripamonte ◽  
M. F. Machado ◽  
R. B. da Silva ◽  
C. Price ◽  
...  
Keyword(s):  
Gene Expression ◽  
Expression Pattern ◽  
In Vitro Maturation ◽  
Housekeeping Gene ◽  
Cumulus Cells ◽  
Glycolytic Enzymes ◽  
Mrna Abundance ◽  
Relative Expression ◽  
Significant Difference

Mammalian oocytes require pyruvate as an energy source for growth and resumption of meiosis. Because oocytes are not competent to carry out glycolysis, cumulus cells (CC) are responsible for metabolizing glucose into pyruvate and providing it to the oocyte through gap junctions. The understanding of the energetic metabolism of CC in culture conditions might provide basis for the improvement of COC in vitro maturation. The aim of this study was to determine the temporal patterns of mRNA expression of glycolytic enzymes [phosphofructokinase (PFKP), aldolase (ALDOA), triosephosphate isomerase (TPI), enolase (ENO1), pyruvate kinase (PKM2), and lactate dehydrogenase (LDHA)] in bovine CC during COC in vitro maturation with or without FSH. Immature COC (grades 1 and 2) were obtained from 2- to 8-mm follicles from abattoir ovaries (predominantly Bos indicus). Cumulus cells were separated from COC and frozen before (immature group) or after COC culture for 4, 8, 12, 16, and 20 hours with (10 ng/mL) or without FSH. Total RNA was extracted using RNeasy® (Qiagen, Valencia, CA, USA), and 100 ng of RNA was reverse transcribed using oligo dT primers and Omniscript® (Qiagen). Relative expression of target genes was assessed by real-time PCR using bovine-specific primers and Power SYBR green master mix in an ABI Prism® 7300. To select the most stable housekeeping gene for expression normalization, cyclophilin-A (CYC-A), GAPDH, and histone H2AFZ amplification profiles were compared using the geNorm applet for Microsoft Excel (Vandesompele J et al. 2002 Genome Biol. 3, 1-11); the most stable housekeeping gene was CYC-A. Relative expression values were calculated using the AACt method with efficiency correction (Pfaffl MW 2001 Nucleic Acids Res. 29, 2002-2007). Effects of time in culture and of FSH treatment were tested by ANOVA, and groups were compared by Tukey-Kramer Honestly Significant Difference test. Nonparametric analysis was used when data were not normally distributed. Abundance of mRNA of all glycolytic enzymes decreased during in vitro maturation with or without FSH. Expression of PFKP, ALDOA, TPI1, ENO1, and LDHA genes was decreased to around half of the initial value (time 0) by 4 to 8 h of culture (P < 0.05) and did not increase thereafter. A similar expression pattern was observed for PKM2, although mRNA abundance was reduced later in comparison with other enzymes; levels were decreased by 16 (without FSH) to 20 h (with FSH) of culture. The presence of FSH did not alter the overall temporal pattern of gene expression but decreased mRNA abundance for PFKP, ALDOA, and TPI1 at 20, 16 and 16 h of culture, respectively. In conclusion, gene expression of glycolytic enzymes decreased with time during COC in vitro maturation in cattle, and FSH did not have a major influence on this expression pattern. This study was supported by CAPES and FAPESP.

175 Nobiletin enhances the quality of in vitro-matured bovine oocytes and blastocysts by altering the transcription of key developmental genes

2019 ◽  
Vol 31 (1) ◽  
pp. 212
Author(s):  
Y. N. Cajas ◽  
K. Cañón-Beltrán ◽  
M. E. González ◽  
P. Ramos-Ibeas ◽  
A. Gutierrez-Adán ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Gene Expression ◽  
Expression Analysis ◽  
Gene Expression Analysis ◽  
Statistical Significance ◽  
Housekeeping Genes ◽  
Cumulus Cells ◽  
Cell Junction ◽  
In Vitro Production

One of the problems associated with in vitro production of embryos in bovine is the increase in reactive oxygen species (ROS), which leads to cell alterations and death. Nobiletin is a polymethoxyflavone isolated from citrus fruits with various beneficial effects on cell cycle regulation and inhibition of ROS production. In a preliminary study, we demonstrated that supplementation of 25 or 50 µM nobiletin to the in vitro maturation (IVM) medium reduces oxidative stress and improves oocyte nuclear and cytoplasmic maturation and embryo development. Thus, in this study, we aimed to evaluate the antioxidant activity of nobiletin during IVM on bovine matured oocytes, their cumulus cells (CC), and blastocysts by quantitative changes of gene expression. Immature cumulus oocytes complexes (COC) were aspirated from ovaries of slaughtered heifers. Selected COC underwent IVM in TCM-199+10% FCS and 10ng mL−1 epidermal growth factor (EGF; Control) supplemented with 25 µM (Nob25) or 50 µM (Nob50) nobiletin (MedChemExpress, Monmouth Junction, NJ, USA) or 0.001% dimethyl sulfoxide (DMSO control), a vehicle for nobiletin dilution, in 5% CO2 in air at 38.5°C. After 24h, 50 matured oocytes/group and their CC were snap-frozen in LN2 for gene expression analysis. The remaining oocytes were fertilized (Day 0) and cultured in vitro. Blastocysts (Day 7; n=50/group) were snap-frozen in LN2 for gene expression analysis (5 replicates). The mRNA abundance of candidate genes related with oxidative stress (SOD2, CYP51); apoptosis (BAX); quality (BMP15, BMP7, CLIC1, MAPK1, ABCB1); and cell junction (GJA1) was measured by quantitative PCR; H2AFZ and 18S rRNA were used as housekeeping genes. Statistical significance was assessed by one-way ANOVA. Supplementation of IVM medium with Nob25 or Nob50 produced changes in the expression levels of genes related to oxidative stress and apoptosis during IVM compared with controls. SOD2 and CYP51 were down-regulated in oocytes and CC (P&lt;0.05) but not in blastocysts, whereas BAX was down-regulated only in CC (P&lt;0.05). Nobiletin supplementation in IVM increased the expression of MAPK1 in oocytes and blastocysts (P&lt;0.05); however, no differences were observed in CC. BMP15 for oocytes and their CC and GJA1 for CC were up-regulated in Nob25 and Nob50 groups compared with controls (P&lt;0.05). The relative abundance of CLIC1 decreased in blastocysts from both nobiletin groups compared with controls (P&lt;0.05). No significant differences in the expression in ABCB1 and BMP7 were detected. In conclusion, our results suggest that supplementation of 25 or 50 µM nobiletin to the IVM medium reduces oxidative stress in oocytes and CC, decreases CC apoptosis, and provokes positive changes in the expression of genes related to oocyte and embryo quality. This research was supported by Spanish MINECO (AGL2015-70140-R and AGL2015-66145-R). Y. N. Cajas was supported by a grant from SENESCYT-Ecuador.

160 Exposure to Cadmium Affects Oocyte and Embryo Competence in Cattle

2018 ◽  
Vol 30 (1) ◽  
pp. 219
Author(s):  
C. De Canditiis ◽  
N. Pagano ◽  
V. Franco ◽  
I. Paradiso ◽  
É. C. Dos Santos ◽  
...  
Keyword(s):  
Embryo Development ◽  
Cumulus Cells ◽  
Early Embryo ◽  
Dapi Staining ◽  
Vitro Fertilization ◽  
Nuclear Maturation ◽  
Significant Difference ◽  
Vitro Model ◽  
Hoechst Staining

There is a growing worldwide concern regarding the increased release of the heavy metal cadmium (Cd) in the environment, due to several industrial processes, as it is known to affect health. Among other heavy metals, Cd is widely recognised to influence the reproductive system at different levels, interfering with both gametes and embryo functions in several species (Thompson and Bannigan, 2008 Reprod. Toxicol. 25, 304-315). The in vitro model can be used to mimic environmental conditions allowing us to evaluate their effect on oocyte maturation and early embryo development. Therefore, the aim of this study was to evaluate the influence of different Cd concentrations on nuclear maturation, apoptosis in cumulus cells, and cleavage and blastocyst yields in cattle. For this purpose, abattoir-derived bovine oocytes were in vitro matured, fertilized, and cultured according to standard procedures (Rubessa et al. 2011 Theriogenology 76, 1347-1355). In particular, oocytes were matured with 0 (control; n = 126), 0.1 μM (n = 139), 1 μM (n = 134), and 10 μM of Cd (n = 135), at 39°C under humidified air with 5% CO2, 7% O2, and 88% N2. For each replicate, after 22 h of maturation, a representative sample of oocytes (n = 10 per each group) was used to evaluate nuclear maturation by 4′,6-diamidino-2-phenylindole (DAPI) staining and another sample (n = 10 per each group) to assess cumulus-cells complex apoptosis by TUNEL/Hoechst staining (Pocar et al. 2005 Reproduction 130, 857-868). The remaining oocytes were in vitro fertilized and cultured with 0 (n = 106), 0.1 μM (n = 119), 1 μM (n = 114), and 10 μM (n = 115) Cd. The experiment was repeated 3 times. On Day 8 post-IVF, the blastocyst yields were recorded. Differences among groups were analysed by ANOVA, with the least significant difference method used as a post hoc test. Data are presented as means ± SE. Unexpectedly, the exposure of oocytes to Cd during IVM did not affect the percentage of oocytes undergoing nuclear maturation (on average 96.3 ± 2.3). In contrast, concentrations of 1 and 10 μM Cd increased the percentage of apoptotic cumulus-cells in cumulus–oocyte complexes (COC) compared with the control (3.4 ± 0.4, 10.6 ± 1.8, 15.0 ± 0.9, 16.7 ± 4.0, respectively, with 0, 0.1, 1, and 10 μM; P < 0.05). It is worth pointing out that with the highest concentration, cumulus expansion did not occur and cumulus cells appeared detached from the oocyte. Likewise, 1 and 10 μM Cd decreased cleavage rates compared with the control (68.7 ± 1.8, 54.3 ± 5.0, 58.5 ± 4.2 and 2.8 ± 2.6, respectively, with 0, 0.1, 1, and 10 μM Cd; P < 0.01). Finally, blastocyst yields decreased when oocytes were treated with 0.1 μM Cd and no development to blastocyst was observed at the 2 higher concentrations (35.1 ± 1.7, 26.2 ± 3.1, 0, 0, respectively, with 0, 0.1, 1, and 10 μM; P < 0.01). In conclusion, exposure to Cd during maturation negatively affects bovine COC, as indicated by the increased apoptotic index in cumulus cells, without influencing the nuclear maturation process. Furthermore, the presence of Cd during in vitro fertilization and culture severely impairs both the fertilization and post-fertilization embryo development.

scholarly journals Effect of Denture Base Reinforcement Using Light Cured E- Glass Fibers on the Level of Salivary Immunoglobulin A

2018 ◽  
Vol 6 (11) ◽  
pp. 2168-2172
Author(s):  
Shady M. El Naggar ◽  
Mohamed I. Seif El Nasr ◽  
Hassan M. Sakr ◽  
Sherihan M. Eissa ◽  
Asmaa N. Elboraey ◽  
...  
Keyword(s):  
Immunoglobulin A ◽  
Acrylic Resin ◽  
Enzyme Linked Immunosorbent Assay ◽  
Complete Dentures ◽  
Denture Base ◽  
Mean Values ◽  
Glass Fibres ◽  
Significant Difference ◽  
Salivary Immunoglobulin A

BACKGROUND: A gap still exists between in vitro and clinical studies concerning the biocompatibility of the material in the oral environment and their potential to cause immunological undesirable side effects. The uses of glass fibres to improve the mechanical properties of acrylic resin denture base polymers are well documented in vitro. AIM: The present study aimed to evaluate the effect of denture base reinforcement using light-cured E- glass fibres mesh on the level of salivary immunoglobulin A (S-IgA) in patients wearing complete dentures. MATERIAL AND METHODS: Fourteen completely edentulous patients, in need of complete dentures, participated in the study. The patients were divided into two groups (n = 7) according to the treatment protocol. In the first group, patients received conventional heat-cured acrylic resin dentures. In the second group, the mandibular dentures were reinforced using light cured resin impregnated E glass fibres mesh. In both groups, salivary samples were collected using passive drool technique. The level IgA was assessed by enzyme-linked immunosorbent assay (ELISA) technique at different time intervals. Statistical analysis was carried out using one-way ANOVA followed by Tukey`s post-hoc test and independent t-test. The significant level was set at P ≤ 0.05. RESULTS: Acrylic resin dentures and reinforced ones demonstrated an increase in the mean values of IgA level at the end of the follow-up intervals. And this increase was statistically significant (P ≤ 0.05). Although, the reinforced dentures revealed higher mean values, there was no statistically significant difference between the two groups (P > 0.05) CONCLUSIONS: Within the limitations of the present study, the following could be concluded: (1) the insertion of complete dentures induced changes in the level of IgA; and (2) denture base reinforcement using light cured resin impregnated E-glass fibres mesh had a similar effect to that of heat cured acrylic resin on the level of IgA.

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