309 ADDITION OF BRAIN-DERIVED NEUROTROPHIC FACTOR IN MATURATION MEDIUM IMPROVED IN VITRO MEIOTIC COMPETENCE OF OVINE OOCYTES AND SUBSEQUENT PARTHENOGENETIC DEVELOPMENT

2015 ◽  
Vol 27 (1) ◽  
pp. 243
Author(s):  
A. H. Abazari-kia ◽  
A. Mohammadi-Sangcheshmeh ◽  
M. Salehi ◽  
M. Zhandi
Keyword(s):  
Embryonic Development ◽  
Neurotrophic Factor ◽  
Brain Derived Neurotrophic Factor ◽  
Culture Conditions ◽  
Control Group ◽  
P Value ◽  
Oocyte Competence ◽  
Maturation Medium ◽  
And Control

Overall efficiency of in vitro embryo production has remained low despite extensive effort to understand the effects of culture conditions, media composition, and supplementation. Brain-derived neurotrophic factor (BDNF), which is a physiologically important neurotrophin, has been used to enhance oocyte maturation in some previous studies (Lee et al. 2007; Zhang et al. 2010). However, the efficacy of BDNF to improve oocyte competence has not been fully established especially in ovine. Therefore, the present study aimed to evaluate the effect of BDNF during in vitro maturation (IVM) on maturation rate, intracellular glutathione (GSH) content, and embryonic development in sheep oocytes. Cumulus-oocyte complexes (COC) were obtained from ovaries of ewes. The COC were placed in maturation medium supplemented with either 10 (IVM-B10) or 100 (IVM-B100) ng mL–1 of BDNF (PeproTech, London, UK). Oocytes in control group were incubated in the same maturation medium without BDNF. The IVM was performed in a humidified atmosphere containing 5% CO2, 5% O2, and 90% N2 at 38.5°C for 24 h. After IVM, several oocytes from the IVM-B10 (n = 110), IVM-B100 (n = 124), and control (n = 110) groups were stained with Hoechst and were evaluated in relation to their metaphase-II rate. To measure GSH content, several oocytes from the IVM-B10 (n = 28), IVM-B100 (n = 33), and control (n = 37) groups were incubated in tyrodes medium containing 10 µM Cell Tracker blue for 30 min and transferred under fluorescence microscope, with digital images analysed by image J software. To evaluate the embryonic development, several oocytes from IVM-B10 (n = 145), IVM-B100 (n = 137), and control (n = 143) groups were subjected to parthenogenetic activation by applying 1 min of exposure to 2.5 µM ionomycin followed by 2 mM 6-DMAP treatment for 3 h. After stimulation, oocytes were cultured in CR1aa medium for 7 days under the conditions stated previously. Four replications were performed. The metaphase-II rate, cleavage, and blastocyst rates were compared by x2 analysis. The GSH content was analysed by one-way ANOVA. A P-value of less than 0.05 was considered significant. The results showed that metaphase-II rate was higher in the IVM-B100 group (88.7%), as compared with the control group (77.3%), but not significant as compared with that in the IVM-B10 group (84.5%). No difference was also found between the IVM-B10 group and control group in terms of the metaphase-II rate. Oocytes in the IVM-B10 group revealed a higher (96.8%) GSH content than both of the IVM-B100 (86.9%) and control (86.3%) groups. There was, however, no difference in the GSH content between the IVM-B100 group and control group. The proportion of cleaved embryos was not different between the groups; however, the blastocyst rate was higher in both the IVM-B10 (37.9%) and IVM-B100 (39.3%) groups compared with the control group (22.4%). Collectively, the results of this study showed that supplementation of IVM media with BDNF promoted nuclear maturation, increased GSH content, and stimulated in vitro embryonic development in ovine.

291 NOVEL OOCYTE SHIPPING AND MATURATION MEDIUM WITHOUT CO2 GAS PHASE IMPROVES IN VITRO MATURATION AND PREGNANCY OUTCOME IN CATTLE

2015 ◽  
Vol 27 (1) ◽  
pp. 234
Author(s):  
M. Barcelo-Fimbres ◽  
L. F. Campos-Chillon ◽  
N. R. Mtango ◽  
L. Bonilla ◽  
J. Verstegen
Keyword(s):  
Embryonic Development ◽  
Pregnancy Rates ◽  
Control Group ◽  
System Control ◽  
Co2 Gas ◽  
Oocyte Competence ◽  
N2 Atmosphere ◽  
Maturation Medium ◽  
Medium System

The aim of the present work was to evaluate embryonic development after shipping and maturation of bovine cumulus oocyte complexes (COC) collected by ovum pick up (OPU) in medium (SMM) that does not require CO2 gas for transport and maturation. Two experiments were conducted, experiment 1 stimulated nonlactating Holstein (n = 4), Jersey (n = 2), Angus (n = 4), and Wagyu (n = 2) donors with 6 pFSH injections (Pluset, MOFA Global LLC, Verona, WI, USA) were used. From each donor, some OPU sessions were delivered the same day (~3 h after collection) for IVM in conventional gas bicarbonate-equilibrated medium system (control), while COC from the other sessions were placed in a portable incubator at 38.5°C, delivered the next day allowing 24 h of maturation in SMM (BoviPro, Mofa Global, WI, USA). The COC were fertilized using commercial semen for each breed, and embryos were cultured in BBH7 medium (BoviPro, Mofa Global, WI, USA) at 38.5°C in 5% O2, 5% CO2, and 90% N2 atmosphere. Embryonic development was evaluated in this experiment. For experiment 2, Day 7 fresh Holstein and Jersey embryos (n = 610) from SMM (n = 550) and controls (n = 60) were transferred in synchronized virgin heifers and pregnancies were diagnosed by ultrasonography at d 35. Data were analysed by ANOVA using GLM, percentages were transformed using arcsin square root, and pregnancy rates were analysed by GenMod using SAS statistical software (Cary, NC, USA). Similar COC numbers were recovered for maturation treatments (P > 0.1; Table 1). The COC matured in SMM had higher cleavage and blastocyst rates than the control group (P < 0.01; Table 1), and this resulted in more transferable embryos per OPU session (P < 0.05; Table 1). We did not find breeds effects or interactions for any variable (P > 0.1; Table 1). After ET, SMM had similar pregnancy rates than control (53.8 v. 58.3%; P > 0.1); however, as more blastocysts were produced per OPU session in the SMM condition, more pregnancies were obtained per session (4.3 v. 2.1; P < 0.01). We conclude that COC matured in SMM had greater oocyte competence than control in commercial settings. The SMM resulted in greater embryonic development, similar pregnancy rates, but more transferable embryos and pregnancies per OPU session than the conventional maturation system. Table 1.Least squares means (± SE) of embryonic development of COC matured in SMM or control

scholarly journals Interleukin-10 and brain-derived neurotrophic factor responses to the Mat Pilates training in women with multiple sclerosis

2018 ◽  
Vol 28 (4) ◽  
pp. 31668
Author(s):  
Elham Eftekhari ◽  
Masoud Etemadifar
Keyword(s):  
Multiple Sclerosis ◽  
Neurotrophic Factor ◽  
Training Group ◽  
Serum Levels ◽  
Brain Derived Neurotrophic Factor ◽  
Interleukin 10 ◽  
Analysis Of Covariance ◽  
Control Group ◽  
P Value ◽  
Before And After

AIMS: To determine the effect of Mat Pilates on serum levels of interleukin-10 and brain-derived neurotrophic factor in women with multiple sclerosis.METHODS: Thirty women with multiple sclerosis with mild to moderate disability were recruited and randomly divided into equal Pilates training and Control groups. Patients in the training group accomplished a Pilates program three times a week for eight weeks. The Control group maintained their routine lifestyle. The serum level of interleukin-10 and brain-derived neurotrophic factor were measured before and after the protocol. The differences between groups were assessed by using analysis of covariance test to compare post-tests by considering covariate pre-tests (assuming a p-value <0.05 as significant).RESULTS: There were no significant changes in interleukin-10 (13.09±5.36 ng/ml in the Pilates training group compared to 13.21±4.76 ng/ml in the Control group, p= 0.81), whereas an increase in brain-derived neurotrophic factor was observed after eight-week Pilates training (11550.14±2619.60 ng/ml in the Pilates training group compared to 9664.35±3161.66 ng/ml in the Control group, p= 0.03).CONCLUSIONS: The results suggest that the intensity and duration of this protocol was not related to significant changes in interleukin-10, but was followed by an increase in brain-derived neurotrophic factor in these patients. Based on this finding, physical activity according to the individual’s ability is recommended for patients with multiple sclerosis, in parallel with drug therapy.

Erratum to: 264 EXOGENOUS LINOLENIC ACID IN OOCYTE MATURATION MEDIA PROMOTES NUCLEAR MATURATION AND PARTHENOGENETIC PREIMPLANTATION EMBRYONIC DEVELOPMENT IN THE GOAT

2013 ◽  
Vol 25 (1) ◽  
pp. 280
Author(s):  
A. Veshkini ◽  
A.-A. Khadem ◽  
M. Soleimani ◽  
R. Jahanbin ◽  
M. Salehi ◽  
...  
Keyword(s):  
Embryonic Development ◽  
Linolenic Acid ◽  
Oocyte Maturation ◽  
Mass Spectrometry Analysis ◽  
Control Group ◽  
Parthenogenetic Activation ◽  
Nuclear Maturation ◽  
Maturation Medium ◽  
Wide Range ◽  
And Control

Dietary intakes of polyunsaturated fatty acids are thought to mediate a wide range of actions in reproductive tissues. This includes the effects on ovarian follicle and corpus luteum functions via improved energy efficiency as well as providing precursors for the synthesis of signalling molecules such as steroids and prostaglandins. An appropriate level of α-linolenic acid (ALA) in the oocyte maturation medium has been shown to induce molecular changes associated with oocyte maturation and embryo developmental competence. In that light, we hypothesised that supplementation of exogenous ALA to maturation media could enhance nuclear maturation and embryonic development in the goat. A preliminary experiment was executed to measure the level of ALA in antral follicles by gas chromatography/mass spectrometry analysis. Our results revealed that the concentration of ALA in follicular fluids ranged from 0.006 to 0.02 mg mL–1 (21.5 to 71.8 µM, with a mean of ~50 µM). To test the effect of ALA on the competence of goat oocytes to complete meiotic maturation to metaphase II and sustain embryonic development, ovaries were obtained from a local abattoir. Cumulus–oocyte complexes were recovered by the slicing method followed by selection of oocytes with a homogenous cytoplasm and at least three layers of compact cumulus cells. The cumulus–oocyte complexes were placed in maturation media supplemented with 50 µM ALA. Oocytes in the control group were incubated in the same maturation medium without ALA. In vitro maturation (IVM) was performed in a humidified atmosphere containing 5% CO2, 5% O2, and 90% N2 at 38.5°C for 24 h. After IVM, several oocytes from the treatment (n = 170) and control (n = 166) groups were stained with Hoechst and were evaluated in relation to their metaphase-II rate. Other groups of oocytes from both the treatment (n = 70) and control (n = 61) groups were subjected to parthenogenetic activation by applying 1 min of exposure to 2.5 µM ionomycin followed by 2 mM 6-DMAP treatment for 3 h. After activation, oocytes were cultured in CR1aa medium for 7 days under the conditions stated above. Four replications were performed. Differences in developmental rates were analysed for significance by one-way ANOVA using SAS version 8.0 (SAS Institute Inc., Cary, NC, USA), considering P < 0.05 to be significant. As a result, supplementation of the maturation media with ALA did not appear to affect cumulus expansion. In contrast, IVM of goat oocytes in the presence of ALA resulted in a significantly higher maturation rate compared with maturation without ALA supplementation (66.4% v. 57.9%). Likewise, addition of ALA to the IVM medium significantly increased the rate of cleavage (60.1% v. 52.4%) and blastocyst formation (22.6% v. 14.9%), calculated from the activated oocytes. Collectively, the results of our study show that supplementation of IVM media with 50 µM ALA promotes nuclear maturation, increases cleavage rate, and results in higher blastocyst rate in goat oocytes after parthenogenetic activation. Thus, providing appropriate levels of ALA in maturation media could have beneficial effects on embryo development and reproductive efficiency in the goat.

286 EFFECTS OF PORCINE FOLLICULAR FLUID DURING IN VITRO MATURATION OF OOCYTES ON IN VITRO FERTILIZATION AND EMBRYONIC DEVELOPMENT

2007 ◽  
Vol 19 (1) ◽  
pp. 258
Author(s):  
B. Agung ◽  
T. Otoi ◽  
D. Fuchimoto ◽  
S. Senbon ◽  
A. Onishi ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Embryonic Development ◽  
Follicular Fluid ◽  
Culture System ◽  
In Vitro Maturation ◽  
Control Group ◽  
Rotating System ◽  
Vitro Fertilization ◽  
Maturation Medium

When used as a solo maturation medium for oocytes, porcine follicular fluid (pFF) promoted male pronucleus formation (MPF) of oocytes after in vitro maturation (IVM), using a static system, and in vitro fertilization (IVF) in pigs (Naito et al. 1988 Gamete Res. 21, 289–295). However, the developmental competence of oocytes matured in pFF after IVM/IVF has not been reported. This study was conducted to assess the ability of pFF as a maturation medium to support IVM/IVF of porcine oocytes and their subsequent in vitro development. pFF, including cumulus–oocyte complexes (COCs), was aspirated from follicles (2–5 mm in diameter) of prepubertal crossbred gilt ovaries, and large clusters of follicular cells (FC) were removed from pFF by filtration through 212 �m of mesh. All of the COCs in filtered pFF were collected, and COCs with compact cumulus cells were selected for IVM. Also, small clusters of FC were collected by centrifugation of the filtered pFF, and pFF without any cells was prepared by centrifugation and used as a maturation medium (MpFF) after supplementation with FSH and antibiotics. COCs were transferred to 3.5 mL (in a 15-mL test tube) of MpFF with FC (5.2 � 106 cells mL-1) and cultured for 44–48 h at 38.5�C in 5% O2 and 5% CO2 using the rotating culture system. As a control group, COCs were cultured in 2 mL of MpFF without FC in a 35-mm Petri dish by the standard static culture system. After maturation, culture oocytes were co-incubated (IVF) for 5 h with frozen–thawed sperm in vitro, as reported previously (Kikuchi et al. 2002 Biol. Reprod. 66, 1033–1041), and then some of them were fixed 10 h after IVF to assess the fertilization status; the rest of them were cultured in PZM (Yoshioka et al. 2002 Biol. Reprod. 60, 112–119) for 7 days to assess their early embryonic development. All of the data were analyzed by ANOVA. Oocytes cultured with FC in the rotating system (R group) showed significantly higher sperm penetration (71.0%), MPF formation (70.5%), and normal fertilization (monospermic fertilization with female and male pronuclei; 31.5%) rates than those in the control group (56.0%, 56.9%, and 17.6%, respectively; P &lt; 0.05). Also, the R group showed significantly higher rates of 8-cell embryos at 2 days after IVF and blastocyst formation at 7 days after IVF than those of the control group (17.2% vs. 8.3% and 10.9% vs. 4.5%, respectively; P &lt; 0.05). These results indicate that porcine oocytes matured in pFF supplemented with FC using the rotating system have the ability to be penetrated by sperm and form MPF, and to develop to the blastocyst stage at higher rates, than oocytes cultured in the standard static maturation culture system. In conclusion, the pFF can be a sole and simple maturation culture medium useful for the in vitro production of blastocysts in pigs.

182 MATURATION OF BOVINE CUMULUS-OOCYTE COMPLEXES WITH FOLLICLE FLUID VARYING IN ESTRADIOL CONTENT AFFECTS CUMULUS CELL EXPANSION WITHOUT AFFECTING SUBSEQUENT EMBRYO DEVELOPMENT IN VITRO

2017 ◽  
Vol 29 (1) ◽  
pp. 199
Author(s):  
A. W. Harl ◽  
E. L. Larimore ◽  
A. Al Naib ◽  
L. K. Wooldridge ◽  
A. D. Ealy ◽  
...  
Keyword(s):  
Cell Expansion ◽  
Cumulus Cell ◽  
Prostaglandin F2α ◽  
Blastocyst Stage ◽  
Control Group ◽  
Blastocyst Development ◽  
Cumulus Expansion ◽  
Oocyte Competence ◽  
Maturation Medium

The objective of this work was to determine how characteristics of bovine follicle fluid (FF; especially oestradiol content) affect cumulus cell expansion and oocyte competence. In the first study, FF was collected from abattoir-derived ovaries and pooled separately for large follicles (≥10 mm) and small follicles (≤3 mm). A portion of the FF from each category was charcoal stripped. These 4 types of FF were then used as the primary ingredient (75% vol/vol) in oocyte maturation media. A separate control group lacking FF but containing BSA was included to monitor potential impacts of protein on outcomes (control; without FF). Some of the cumulus-oocyte complexes (COC; n = 250) were matured in individual drops for analysis of cumulus expansion (photographed and measured at 0 and 21 h of maturation). Other COC (n = 770) were matured in groups of 12 to 25 in the previously described media, and then subjected to IVF procedures. Cleavage rates were recorded on Day 3, and blastocyst rates were recorded on Day 8 post-fertilization. Cumulus cell expansion was greatest when COC were matured in medium containing FF from large follicles, wherein it even exceeded the controls (P < 0.02). Maturation in FF from small follicles resulted in cumulus expansion that was intermediate between large and control. Maturation in charcoal-stripped FF severely restricted cumulus cell expansion (P < 0.05) compared with those matured in untreated FF. Despite the observed improvement in cumulus cell expansion, COC that had been matured in media containing FF were less likely to cleave (P < 0.05) and also less likely to develop to the blastocyst stage (P < 0.01) than those matured in control medium. Cleavage and blastocyst rates did not differ among any of the maturation media containing FF. In the second study, oestrous cycles of 9 crossbred cows were synchronized and FF samples were collected 36 to 42 h after prostaglandin F2α injection. Samples from individual cows were categorized as having high oestradiol (>800,000 pg mL−1; H) or low oestradiol concentrations (<800,000 pg mL−1; L). The FF was retained for use in in vitro experiments, where it was added to maturation media (20% vol/vol). Cumulus-oocyte complexes (n = 1,775) were randomly distributed into treatments across 12 in vitro maturation/fertilization replicates (H and L, balanced within replicate; 4 replicates/cow). Each replicate included the following 3 control groups: maturation medium containing BSA without FF, maturation medium without BSA with abattoir-derived FF, and maturation medium without BSA and without FF. The COC were matured in their assigned medium for 21 h, and then all COC were subjected to IVF procedures. Cleavage rates were recorded on Day 3, and blastocyst rates were recorded on Day 7 and 8 post-fertilization. Oestradiol content of the FF (H v. L) did not affect oocyte cleavage nor blastocyst rates on Day 7 or 8. The results of these studies indicate that although FF improves cumulus cell expansion during maturation in vitro, it does not result in higher rates of cleavage or blastocyst development regardless of oestradiol content.

Regulation of embryonic development and apoptotic-related gene expression by brain-derived neurotrophic factor in two different culture conditions in ovine

2015 ◽  
Vol 84 (1) ◽  
pp. 62-69 ◽  
Author(s):  
Amir Hossein Abazari-Kia ◽  
Maryam Dehghani-Mohammadabadi ◽  
Abdollah Mohammadi-Sangcheshmeh ◽  
Mahdi Zhandi ◽  
Mohammad Salehi
Keyword(s):  
Gene Expression ◽  
Embryonic Development ◽  
Neurotrophic Factor ◽  
Brain Derived Neurotrophic Factor ◽  
Culture Conditions ◽  
Related Gene

scholarly journals The supplementation of ovine oocyte maturation medium with triiodothyronine affects the embryo development and apoptotic gene expression

2021 ◽  
Vol 72 (3) ◽  
pp. 3195
Author(s):  
R ASADPOUR ◽  
F AHMADNEJAD ◽  
L ROSHANGAR ◽  
A SABERIVAND ◽  
A HAJIBEMANI
Keyword(s):  
Embryonic Development ◽  
Oocyte Maturation ◽  
Mrna Level ◽  
B Cell Lymphoma ◽  
Expression Level ◽  
Control Group ◽  
High Concentration ◽  
Chi Square ◽  
Maturation Medium

Triiodothyronine (T3) plays an essential role in different animal species’ embryonic development. The present research was designed to identify the effect of triiodothyronine on the in vitro ovine embryonic development and the expression of apoptotic genes.A total of 436 immature cumulus-oocyte complexes (COCs) were cultured for 24 h in the oocyte maturation medium supplemented with two concentrations of T3 (T-10 and T-100 ng/mL) or without T3(T-0: control group). Oocyte maturation, cleavage, and blastocyst rates were assessed under an inverted microscope as crucial indicators of embryo development.The relative mRNA abundance of BCL-2-associated X protein (BAX) and anti-apoptotic B-cell lymphoma-2 (BCL2) were determined at blastocysts (day 8 after IVF on day 0)by quantitative reverse transcription PCR.The data were analyzed by logistic regression using the GLIMMIX procedure followed by Chi-Square, and one-way ANOVA tests. The higher concentration of T3(100 ng/mL) significantly decreased cumulus expansion and blastocyst rate compared to controls (P<0.001). Additionally, a significantly higher expression level of BAX(P<0.001) and a dramatically lower expression level of BCL2 (P<0.01) were detected in the T-100ng/mL group compared to the controls.However, the relative mRNA level of BCL-2 was significantly higher in the T-10 ng/mL group compared to the control group (P<0.01).It appears that the supplementation of ovine oocyte maturation medium with T3 at high concentration (100 ng/mL) suppresses the ratio of blastocyst formation.

scholarly journals Erratum to: 264 EXOGENOUS LINOLENIC ACID IN OOCYTE MATURATION MEDIA PROMOTES NUCLEAR MATURATION AND PARTHENOGENETIC PREIMPLANTATION EMBRYONIC DEVELOPMENT IN THE GOAT

2013 ◽  
Vol 25 (3) ◽  
pp. 587
Author(s):  
A. Veshkini ◽  
A.-A. Khadem ◽  
M. Soleimani ◽  
R. Jahanbin ◽  
M. Salehi ◽  
...  
Keyword(s):  
Embryonic Development ◽  
Linolenic Acid ◽  
Oocyte Maturation ◽  
Mass Spectrometry Analysis ◽  
Control Group ◽  
Parthenogenetic Activation ◽  
Nuclear Maturation ◽  
Maturation Medium ◽  
Wide Range ◽  
And Control

Dietary intakes of polyunsaturated fatty acids are thought to mediate a wide range of actions in reproductive tissues. This includes the effects on ovarian follicle and corpus luteum functions via improved energy efficiency as well as providing precursors for the synthesis of signalling molecules such as steroids and prostaglandins. An appropriate level of α-linolenic acid (ALA) in the oocyte maturation medium has been shown to induce molecular changes associated with oocyte maturation and embryo developmental competence. In that light, we hypothesised that supplementation of exogenous ALA to maturation media could enhance nuclear maturation and embryonic development in the goat. A preliminary experiment was executed to measure the level of ALA in antral follicles by gas chromatography/mass spectrometry analysis. Our results revealed that the concentration of ALA in follicular fluids ranged from 0.006 to 0.02mgmL–1 (21.5 to 71.8µM, with a mean of ~50µM). To test the effect of ALA on the competence of goat oocytes to complete meiotic maturation to metaphase II and sustain embryonic development, ovaries were obtained from a local abattoir. Cumulus–oocyte complexes were recovered by the slicing method followed by selection of oocytes with a homogenous cytoplasm and at least three layers of compact cumulus cells. The cumulus–oocyte complexes were placed in maturation media supplemented with 50µM ALA. Oocytes in the control group were incubated in the same maturation medium without ALA. In vitro maturation (IVM) was performed in a humidified atmosphere containing 5% CO2, 5% O2, and 90% N2 at 38.5°C for 24h. After IVM, several oocytes from the treatment (n=170) and control (n=166) groups were stained with Hoechst and were evaluated in relation to their metaphase-II rate. Other groups of oocytes from both the treatment (n=70) and control (n=61) groups were subjected to parthenogenetic activation by applying 1min of exposure to 2.5µM ionomycin followed by 2mM 6-DMAP treatment for 3h. After activation, oocytes were cultured in CR1aa medium for 7 days under the conditions stated above. Four replications were performed. Differences in developmental rates were analysed for significance by one-way ANOVA using SAS version 8.0 (SAS Institute Inc., Cary, NC, USA), considering P&lt;0.05 to be significant. As a result, supplementation of the maturation media with ALA did not appear to affect cumulus expansion. In contrast, IVM of goat oocytes in the presence of ALA resulted in a significantly higher maturation rate compared with maturation without ALA supplementation (66.4% v. 57.9%). Likewise, addition of ALA to the IVM medium significantly increased the rate of cleavage (60.1% v. 52.4%) and blastocyst formation (22.6% v. 14.9%), calculated from the activated oocytes. Collectively, the results of our study show that supplementation of IVM media with 50µM ALA promotes nuclear maturation, increases cleavage rate, and results in higher blastocyst rate in goat oocytes after parthenogenetic activation. Thus, providing appropriate levels of ALA in maturation media could have beneficial effects on embryo development and reproductive efficiency in the goat.

134 TRANSCRIPTOME PROFILE OF BOVINE BLASTOCYSTS DERIVED FROM ALTERNATIVE IN VIVO AND IN VITRO CULTURE CONDITIONS AT SPECIFIC PHASES OF EARLY EMBRYONIC DEVELOPMENT

2012 ◽  
Vol 24 (1) ◽  
pp. 179 ◽  
Author(s):  
A. Gad ◽  
U. Besenfelder ◽  
V. Havlicek ◽  
M. Hölker ◽  
M. U. Cinar ◽  
...  
Keyword(s):  
Embryonic Development ◽  
In Vitro Culture ◽  
In Vitro Maturation ◽  
Expression Patterns ◽  
Culture Conditions ◽  
Control Group ◽  
Transcriptome Profile ◽  
Vitro Fertilization

An understanding of gene expression patterns due to altered environmental conditions during different time points of the pre-implantation period would improve our knowledge on regulation of embryonic development and improve success of embryo culture. The aim of this study was to examine the effect of alternative in vivo and in vitro culture conditions at specific phases of early embryonic development on transcriptome profile of bovine blastocysts. Using nonsurgical endoscopic oviducal transfer technology, 5 different blastocyst groups were produced. The first 2 groups were matured in vitro and then either transferred after maturation or after in vitro fertilization to synchronized recipients. The other 3 groups were matured, fertilized and cultured in vitro until 4-cell, 16-cell and morula stage before transfer. Blastocysts from each group were collected by uterine flushing at Day 7 and pooled in groups of 10. Complete in vitro (IVP) and in vivo blastocysts were produced and used as controls. A unique custom microarray (Agilent) containing 42 242 oligo probes (60-mers) was used over 6 replicates of each group vs the in vivo control group to examine the transcriptome profile of blastocysts. Compared with the in vivo control group, clear dramatic shifts were found in the number of differentially expressed genes (DEG, fold change ≥2) at 2 different time points. The first shift occurred for blastocyst groups that were transferred after in vitro fertilization and before embryonic genome activation (EGA). The second shift occurred for blastocyst groups that were transferred after EGA, as well as for the IVP group. Ontological classification of DEG showed that the more time spent under in vitro conditions, the higher the percentage of DEG involved in cell death and apoptotic processes. Moreover, lipid metabolism was the most significant process affected in the blastocysts transferred after in vitro maturation and blastocysts transferred at 16-cell stage. Most DEG involved in this process were down-regulated. Pathway analysis revealed that signalling pathways were the dominant pathways in all groups except the group that was transferred after in vitro maturation. That group showed significant down-regulation for genes involved in retinoic acid receptors activation pathways. These results showed that fertilization and EGA were the most critical developmental stages influenced by in vitro culture conditions and subsequently affect blastocyst quality, as measured in terms of gene expression patterns. Moreover, we identified molecular mechanisms and pathways that were influenced by altered culture conditions. These findings will enable the examination of strategies for modifying in vitro culture conditions at critical stages that will allow more efficient production of developmentally competent blastocysts.

Comparison of the Effect of 0.2% Chlorhexidine Mouthwash and Persica Mouthwash on the Color of Resin Composite

2021 ◽  
Author(s):  
Fatemeh Ebrahimzadeh ◽  
Hooman Fakhar ◽  
Hosein Akbari ◽  
Ramin Mosharraf ◽  
Azin Farzad
Keyword(s):  
Resin Composite ◽  
Control Group ◽  
P Value ◽  
Patient Dissatisfaction ◽  
Composite Restorations ◽  
School Of Dentistry ◽  
Initial Color ◽  
Group B ◽  
And Control

Introduction: Discoloration of resin composite restorations can lead to patient dissatisfaction. 0.2% Chlorhexidine and Persica mouthwashes are among the agents that cause discoloration. The aim of this study was to investigate the degree of discoloration caused by the 0.2% Chlorhexidine and Persica mouthwashes on resin composite samples. Materials and Methods: This in-vitro experimental study was conducted in Kashan and Isfahan School of Dentistry in 2020-2021. Number of 30 disc-shaped samples were fabricated from Charisma Diamond resin composite. The initial color of samples was measured by CIE Lab system in spectrophotometer. Then samples were divided into 3 groups (A, B, and C) (n = 10).The control group (A) was placed in distilled water, group B was immersed in the 0.2% Chlorhexidine mouthwash and group C was immersed in the Persica mouthwash. The color of the samples was measured again afterwards. Data were analyzed with One-way ANOVA and t-Test (α = 0.05). Results: The amount of l, a, b and ΔE after using 0.2% Chlorhexidine and Persica mouthwashes increased. The mean of Δl, Δa, Δb and ΔE showed significant differences between groups (p value < 0.05). Conclusion: The discoloration of Persica mouthwash was more than 0.2% Chlorhexidine mouthwash and control group. Therefore, for patients with resin composite restorations, 0.2% Chlorhexidine mouthwash is better.

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