117 EFFECT OF RESVERATROL ON THE CULTURE OF PORCINE EMBRYOS

2009 ◽  
Vol 21 (1) ◽  
pp. 158
Author(s):  
K. Lee ◽  
C. Wang ◽  
D. Miller ◽  
Z. Machaty
Keyword(s):  
Caspase 3 ◽  
Total Cell ◽  
Control Group ◽  
Blastocyst Formation ◽  
Dependent Effect ◽  
Expression Levels ◽  
Cell Numbers ◽  
Apoptotic Activity ◽  
Dose Dependent ◽  
Dose Dependent Effect

Resveratrol (3,4′,5-trihydroxystilbene) is a phytoalexin present in a variety of dietary sources including grapes, plums and peanuts. It was shown to exert a wide variety of pharmacological activities ranging from anti-inflammatory effects to immunomodulation and chemoprevention. The aim of this study was to investigate the effect of resveratrol on porcine embryo development. In vitro-matured pig oocytes were activated by two direct current electric pulses (60 μ seconds each) followed by an incubation with 10 μg mL–1 cycloheximide and 7.5 μg mL–1 cytochalasin B, for 5 h. Activated oocytes (in groups of 10) were then placed in 20 μL drops of Porcine Zygote Medium 3 (PZM-3) supplemented with various concentrations of resveratrol (0 μm, 0.5 μm, 25 μm) and cultured for 7 days. The frequency of blastocyst formation in each group was recorded and compared by Chi-square test; the total cell numbers of the blastocysts from the two best groups (control and 0.5 μm) were counted and compared by Student’s t-test. In addition, expression levels of three genes related to apoptosis (Bax, Bcl-2, and Caspase-3) were quantified in the blastocysts by quantitative real-time RT-PCR. The analysis was repeated three times, and differences in gene expression were compared by ANOVA. Differences at P < 0.05 were considered significant. Incubation of parthenotes with 25 μm resveratrol decreased blastocyst formation from 28.1% (n = 89; control group) to 4.5% (n = 66; P < 0.05). However, when parthenogenetic embryos were cultured in the presence of 0.5 μm resveratrol, blastocyst formation increased significantly: whereas only 33.0% of control embryos (n = 312) reached the blastocyst stage, this percentage in the resveratrol-treated group was 43.5% (n = 301; P < 0.05). The average total cell numbers in control and 0.5 μm resveratrol-treated blastocysts were 42.4 ± 2.1 and 44.0 ± 1.9, respectively; the difference was not statistically significant. Finally, lower expression of the Bcl-2 and Caspase-3 genes was observed in the embryos cultured with 0.5 μm resveratrol. Evidences indicated that resveratrol has a dose-dependent effect on cells. At high concentrations resveratrol exerted an antiproliferation effect, whereas at low concentration it promoted cell division. In this study a similar dose-dependent effect of resveratrol was found. The presence of 25 μm resveratrol had a negative effect on embryonic development. However, 0.5 μm resveratrol in the culture medium induced higher blastocyst formation compared with the control group. The presence of resveratrol also affected expression levels of genes related to apoptosis. Decreased levels of Bcl-2 suggest that resveratrol may suppress the mitochondria-related anti-apoptotic activity; whereas down-regulated Caspase-3 indicates a decreased apoptotic activity in the embryos treated with resveratrol. Our results suggest that the treatment of pig embryos with 0.5 μm of resveratrol has a beneficial effect on preimplantation development leading to improved blastocyst formation. The impact of resveratrol on apoptosis needs further investigation.

41 EFFICIENT STRATEGY FOR INTERSPECIFIC CLONING IN FELIDS

2014 ◽  
Vol 26 (1) ◽  
pp. 134
Author(s):  
L. N. Moro ◽  
M. I. Hiriart ◽  
J. Jarazo ◽  
C. Buemo ◽  
A. Sestelo ◽  
...  
Keyword(s):  
Species Conservation ◽  
Total Cell ◽  
Domestic Cat ◽  
Control Group ◽  
Felis Silvestris ◽  
Blastocyst Formation ◽  
Acinonyx Jubatus ◽  
Cell Numbers ◽  
Number Of Cells

Most of the 36 species of wild felids are at a level of threat, and interspecific SCNT (iSCNT) comes as a strategy to contribute to these species conservation. The aim of this study was to evaluate the effect of embryo aggregation in cheetah (Ch, Acinonyx jubatus), bengal (Ben, a hybrid between Felis silvestris and Prionailurus bengalensis), and domestic cat (DC, Felis silvestris) embryos generated by cloning. DC oocytes were in vitro matured and zona-free SCNT (with DC fibroblasts) or iSCNT (with Ch or Ben fibroblasts) was performed. The reconstructed embryos were activated with 5 μM ionomycin and 1.9 mM 6-DMAP, and cultured in SOF using microwells. Cloned embryos were cultured individually or as 2-embryo aggregates. The experimental groups were Ch1X, Ch2X, Ben1X, Ben2X, and the control groups were DC1X and DC2X. Embryo development was compared by Fisher's exact test (P ≤ 0.05). Embryo aggregation improved cleavage (Day 2) and blastocyst (Day 7) rates per well in all the groups (87.2% v. 96.7%, 83.8% v. 93.3% and 87.6% v. 98.2% for cleavage; and 13.7% v. 28.6%, 33.3% v. 43.8% and 27.4% v. 47.7% for blastocyst, for Ch1X (n = 102), Ch2X (n = 91), Ben1X (n = 154), Ben2X (n = 105), DC1X (n = 113), and DC2X (n = 109), respectively. Moreover, the Ch2X blastocyst rate was statistically similar as the control group DC1X. The mean total cell numbers of the blastocysts obtained were 264 ± 211 and 400.8 ± 97 for Ch1X and Ch2X, 278 ± 62 and 517 ± 104 for Ben1X and Ben2X, 385 ± 127 and 625 ± 183 for DC1X and DC2X, respectively. Although no statistical differences were obtained between the 1X and 2X groups, the 2X groups nearly doubled the average number of cells compared with the 1X groups. Blastocysts were also classified as grade 1 (expanded blastocysts with a well-defined ICM), grade 2 (expanded blastocysts without a well-defined ICM), and grade 3 (not expanded blastocysts). This classification showed an increase in grade 1 DC2X blastocyst compared with DC1X blastocysts (36.7% v. 16.1%), but no differences were observed in the other species. Expression of OCT-4 was assessed by inmunocytochemistry. The cheetah blastocysts markedly over-expressed this protein: the percentage of cells that expressed OCT-4 in Ch1X, Ch2X, Ben1X, Ben2X, DC1X, and DC2X was 88.2, 80.2, 46.3, 45.4, 51, and 47.4%, respectively, with statistical differences among all the groups except Ben1X and Ben2X. The proportion of OCT-4 expressing cells over total cell numbers was analysed by the difference of proportions test (P ≤ 0.05). In conclusion, iSCNT resulted in high rates of blastocyst formation, especially when embryo aggregation was applied. This strategy has not been previously evaluated in felids or iSCNT procedures, and has been demonstrated to improve blastocyst formation, the number of cells in the 3 groups, and the blastocyst quality in the DC. Other pluripotent genes besides OCT-4 should be studied to determine whether the overexpression of this gene in cheetah embryos is the consequence of an inefficient nuclear reprogramming that prevents a correct regulation. Finally, the iSCNT and embryo aggregation could contribute to species conservation in felids.

133 EFFECT OF PROGESTERONE SUPPLEMENTATION OF MATURATION MEDIUM ON THE DEVELOPMENT OF IN VITRO-MATURED-IN VITRO-FERTILIZED-IN VITRO-CULTURED BOVINE EMBRYOS

2014 ◽  
Vol 26 (1) ◽  
pp. 180 ◽  
Author(s):  
K. Syoji ◽  
K. Imai ◽  
H. Koyama ◽  
O. Dochi
Keyword(s):  
Cell Mass ◽  
Calf Serum ◽  
Total Cell ◽  
Differential Staining ◽  
Control Group ◽  
Blastocyst Formation ◽  
Cell Numbers ◽  
Maturation Medium ◽  
Significant Difference

The objective of this study was to investigate whether progesterone (P4) supplementation to in vitro maturation (IVM) medium could affect the competence of bovine oocyte to develop into blastocysts in vitro. Cumulus–oocyte complexes (COC) were collected by aspiration of ovarian follicles (2 to 6 mm in diameter) obtained from a local abattoir. The COC were matured for 20 h in TCM-199 supplemented with 5% calf serum and 0.02 AU mL–1 FSH at 38.5° in an atmosphere of 5% CO2 in air. After 18 h of gamete co-culture (5 × 106 sperms mL–1), presumptive zygotes were cultured in CRlaa containing 5% calf serum at 38.5°C in an atmosphere of 5% CO2, 5% O2, and 90% N2 for 9 days (fertilization = Day 0). Progesterone was added to the IVM medium 10 h after the start of the culture (1 μg group = 1 μg mL–1 of P4; 5 μg group = 5 μg mL–1 of P4; control group = no P4). The maturation (MII) rates were investigated after 20 h of starting the IVM culture. After maturation, the COC were denuded mechanically, and a part of the oocytes were mounted on slides, fixed with aceto-alcohol (1 : 3) solution for 48 h, stained with aceto-orcein, and observed under a phase-contrast microscope to determine their nuclear status (1 μg group: n = 32; 5 μg group: n = 28; control group: n = 31). The remaining COC were used for IVF. The cleavage rates were investigated on day 2, and the blastocyst formation rates were investigated on Days 7 to 9, respectively (1 μg group: n = 264; 5 μg group: n = 274; control group: n = 277). The blastocysts from Day 7 were used for differential staining of the inner cell mass (ICM) and trophectoderm cells (TE). The total cell numbers, ICM, and TE in the blastocysts were counted (1 μg group: n = 28; 5 μg group: n = 24; control group: n = 24). The rates of MII, cleavage, and blastocyst formation were expressed and analysed by the chi-squared test. Each set of cell numbers (mean ± standard error) was analysed by the unpaired t-test. The MII rate in the control group (76.7%) was significantly lower (P < 0.05) than that in the 1 μg group (93.8%). The cleavage rate in the 1 μg group (85.6%) was significantly higher (P < 0.05) than those in the control group (74.7%) and 5 μg group (77.4%). Further, the blastocyst formation rate in the 1 μg group (47.7%) and 5 μg group (43.4%) were significantly higher (P < 0.05) than that in the control group (35.0%). The ICM numbers (mean ± s.e.) were 39.5 ± 13.8 to 36.2 ± 8.9, the TE numbers were 74.4 ± 22.4 to 66.2 ± 12.9, the total cell numbers of blastocysts were 110.6 ± 28.2 to 103.0 ± 13.8. There was no significant difference in cell numbers among the groups. These results indicate that the cleavage and blastocyst formation rates can be improved by the addition of 1 μg mL–1 of P4 to the maturation medium.

Dose-dependent effect of testosterone on the induction of eclipse coloration in castrated wild mallard drakes (Anas platyrhynchos L.)

1992 ◽  
Vol 70 (3) ◽  
pp. 428-431 ◽  
Author(s):  
Eberhard Haase ◽  
Reiner Schmedemann
Keyword(s):  
Breeding Season ◽  
Control Group ◽  
Dependent Effect ◽  
Daily Dose ◽  
Natural Breeding ◽  
Dose Dependent ◽  
Dose Dependent Effect ◽  
Wild Mallard ◽  
Intermediate Pattern

Four groups of adult wild mallard drakes that had been previously castrated were injected daily with 5, 1, or 0.2 mg testosterone dissolved in propanediol or with the vehicle alone. The injections started on February 29 and continued for 64 days during the natural breeding season of the species. The birds were kept in outdoor aviaries in a seminatural environment in Kiel (54°N). By the end of the postnuptial molt in mid-July the groups differed markedly in coloration. The vehicle-injected control group had regained an almost typical nuptial plumage, whereas the castrates that had received the highest daily dose of testosterone exhibited an almost complete eclipse plumage. The two other groups showed an intermediate pattern, with about 40% eclipse coloration in the group injected with 1 mg testosterone/day and about 10% eclipse coloration in the group that had received 0.2 mg testosterone/day. The possible role of the aromatization of testosterone to estrogens in the induction of the eclipse coloration is discussed.

Increasing Concentration of Sinovial: Effect on Cartilage Protection in a Rabbit ACLT Model

Cartilage ◽  
2020 ◽  
pp. 194760352090536
Author(s):  
Pomme Boissier ◽  
Pierre Mainil-Varlet ◽  
Giuseppe R. Mautone
Keyword(s):  
Cruciate Ligament ◽  
Intraoperative Complications ◽  
Cartilage Degradation ◽  
Research Society ◽  
Control Group ◽  
Dependent Effect ◽  
India Ink ◽  
Anterior Cruciate ◽  
Dose Dependent ◽  
Dose Dependent Effect

Objective This study aimed to test the hypothesis that administration of increasing doses of Sinovial (hyaluronic acid [HA]), would exhibit a dose-dependent effect on the prevention of cartilage degradation, without local and systemic toxicity. Methods Twenty-seven adult rabbits were subjected to anterior cruciate ligament transection (ACLT). Two Sinovial products containing HA concentrations of 1.6% and 2.4% were used as active treatment, and 0.9% saline was used as control and injected intra-articularly 7 days post ACLT. Radiographs were taken prior to surgery, at injection and sacrifice times. After euthanasia, 8 weeks postsurgery, knee joints were observed macroscopically using India ink staining with OARSI (Osteoarthritis Research Society International) scoring and histologically using modified Mankin scoring. The synovial membranes were analyzed using Cake classification. Results No intraoperative complications were observed. One week postinjection, 4 animals in the HA 2.4% group developed subcutaneous nodules that disappeared spontaneously. No inflammation of the synovial membrane was ever observed. The control group exhibited the maximum uptake of India ink 2.22 ± 0.14. HA groups exhibited a dose-dependent ( P = 0.02) reduction in India ink uptake: 1.75 ± 0.17 for HA 1.6% and 1.58 ± 0.14 for HA 2.4%. The most marked dose-dependent effect of this study was a reduction of modified Mankin score for HA groups, with the 2.4% treatment achieving a statistically significant improvement as compared with the control group (7.19 ± 0.85 for saline, 4.65 ± 0.66 for HA 1.6%, and 3.53 ± 0.59 for HA 2.4%; P = 0.005). Conclusions A dose-dependent protective effect on cartilage was observed after injection of both HA solutions.

scholarly journals Silver‐doped sol‐gel borate glasses: Dose‐dependent effect on Pseudomonas aeruginosa biofilms and keratinocyte function

2021 ◽  
Author(s):  
Shiva Naseri ◽  
Gabriele Griffanti ◽  
William C. Lepry ◽  
Vimal B. Maisuria ◽  
Nathalie Tufenkji ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Sol Gel ◽  
Borate Glasses ◽  
Dependent Effect ◽  
Dose Dependent ◽  
Dose Dependent Effect

Dose-Dependent Effect of Statins on the Incidence of Acute Kidney Injury After Cardiac Surgery

2011 ◽  
Vol 91 (2) ◽  
pp. 520-525 ◽  
Author(s):  
Salima Mithani ◽  
Michael Kuskowski ◽  
Yelena Slinin ◽  
Areef Ishani ◽  
Edward McFalls ◽  
...  
Keyword(s):  
Acute Kidney Injury ◽  
Cardiac Surgery ◽  
Kidney Injury ◽  
Dependent Effect ◽  
Dose Dependent ◽  
Dose Dependent Effect

Enlargement of cerebrospinal fluid spaces in long-term benzodiazepine abusers

1987 ◽  
Vol 17 (4) ◽  
pp. 869-873 ◽  
Author(s):  
C. Schmauss ◽  
J.-C. Krieg
Keyword(s):  
Low Dose ◽  
Matched Control ◽  
Control Group ◽  
High Dose ◽  
Withdrawal Therapy ◽  
The Mean ◽  
Dose Dependent ◽  
Dose Dependent Effect ◽  
Age And Sex

SynopsisIn 17 benzodiazepine (BDZ) dependent in-patients a CT scan was performed before initiation of withdrawal therapy. The evaluation of the ventricular to brain ratio (VBR) by standardized and computerized measurements revealed significantly higher mean VBRs for both high-and low-dose BDZ-dependent patients compared to the mean VBR of an age- and sex-matched control group. In addition, the mean VBR of high-dose BDZ-dependent patients (N = 8) was significantly higher than the mean VBR of low-dose BDZ-dependent patients (N = 9). This difference could not be accounted for by the age of the patients or duration of BDZ-dependency and, therefore, suggests a dose-dependent effect of BDZs on the enlargement of internal CSF-spaces. On the other hand, higher values for the width of external CSF-spaces were found to be related to increasing age of the patients and duration of BDZ-dependency.

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