scholarly journals Synergistic Effect of Insulin on in vitro Development of Immature Bovine Oocytes

2010 ◽  
Vol 5 (4) ◽  
pp. 258-265 ◽  
Author(s):  
Dashtizad
Keyword(s):  
Synergistic Effect ◽  
Bovine Oocytes

256 SYNERGISTIC EFFECT ON EMBRYO DEVELOPMENT AND EMBRYO QUALITY WHEN BOVINE DENUDED OOCYTES ARE CO-CULTURED WITH CUMULUS - OOCYTE COMPLEXES DURING IN VITRO MATURATION

2011 ◽  
Vol 23 (1) ◽  
pp. 226
Author(s):  
S. R. Dey ◽  
G. K. Deb ◽  
J. I. Bang ◽  
S. J. Cho ◽  
B. H. Choi ◽  
...  
Keyword(s):  
Synergistic Effect ◽  
In Vitro Culture ◽  
In Vitro Maturation ◽  
Cumulus Cell ◽  
Tunel Staining ◽  
Oocyte Growth ◽  
Respective Control ◽  
Bovine Oocytes

The oocyte and its surrounding somatic cells are metabolically coupled to each other through gap junctions. This phenomenon allows intercellular communication and transfer of different low-molecular-weight substrates between the cells necessary for oocyte growth. The oocyte itself regulates the cumulus cell microenvironment through oocyte-secreted factors. The development competence of the bovine oocytes is increased when denuded oocytes (DO) are co-cultured with cumulus–oocyte complexes (COC) during in vitro maturation (IVM). However, the fate of the DO, which are usually discarded after IVM, has not been determined. The present study aimed to investigate whether there is a synergistic effect of co-culturing COC and DO during IVM. We performed 3 IVM schemes: 1) COC and DO co-culture, with 12 COC and 60 DO; 2) COC control, with 12 COC; and 3) DO control, with 60 DO in 120-μL drop of TCM-199 for 22 to 24 h. Following IVM, IVF and in vitro culture were separately performed for the COC (COC co-culture) and DO (DO co-culture) from the IVM co-culture group. In vitro fertilization and in vitro culture (modified CR1aa) were done in 60-μL drops. Embryos were cultured at 38.5°C and 5% CO2 in air. Cleavage and blastocyst rates were checked at Day 3 and 8 from IVF on total COC/DO placed in IVM drop. Day 8 blastocysts were used for TUNEL staining using In Situ Cell Death Detection Kit (Roche, Budapest, Hungary). Data were analysed by one-way ANOVA, and significant differences among groups were tested by DMRT. Compared with the respective control treatments, co-culture has no effect on cleavage rates of COC and DO (see Table 1). However, blastocyst rates and total cell numbers of blastocysts were increased in COC co-culture and DO co-culture group compared with their respective control groups (see Table 1). Co-culture had no effect on apoptosis of blastocysts. These data show that co-culture of COC and DO improved developmental competence and quality of embryos from the COC co-culture and DO co-culture group. Table 1.Development competence and blastocyst quality of intact and denuded bovine oocytes This work was partly supported by the BK21 program, the KRF (KRF-2008-211-F00011), the IPET (108068-03-1-SB010), and the KOSEF (10525010001-05N2501-00110).

Benzisothiazolone Derivatives Exhibit Cytotoxicity in Hodgkin’s Lymphoma Cells through NF-κB Inhibition and are Synergistic with Doxorubicin and Etoposide

2020 ◽  
Vol 20 (6) ◽  
pp. 715-723
Author(s):  
Natarajan Nandakumar ◽  
Pushparathinam Gopinath ◽  
Jacob Gopas ◽  
Kannoth M. Muraleedharan
Keyword(s):  
Synergistic Effect ◽  
Hodgkin’S Lymphoma ◽  
A549 Cells ◽  
Hodgkin's Lymphoma ◽  
Luciferase Reporter ◽  
Luciferase Reporter Gene ◽  
Lymphoma Cells ◽  
Nuclear Extracts ◽  
Gene Assay

Background: The authors investigated the NF-κB inhibitory role of three Benzisothiazolone (BIT) derivatives (1, 2 and 3) in Hodgkin’s Lymphoma cells (L428) which constitutively express activated NF-κB. All three compounds showed dose-dependent NF-κB inhibition (78.3, 70.7 and 34.6%) in the luciferase reporter gene assay and were found cytotoxic at IC50 values of 3.3μg/ml, 4.35μg/ml and 13.8μg/ml, respectively by the XTT assay. BIT 1and BIT 2 (but not BIT 3) suppressed both NF-κB subunits p50 and p65 in cytoplasmic and nuclear extracts in a concentration-dependent manner. Furthermore, BIT 1 showed a moderate synergistic effect with the standard chemotherapy drugs etoposide and doxorubicin, whereas BIT 2 and 3 showed a moderate additive effect to antagonistic effect. Cisplatin exhibited an antagonist effect on all the compounds tested under various concentrations, except in the case of 1.56μg/ml of BIT 3 with 0.156μg/ml of cisplatin. The compounds also inhibited the migration of adherent human lung adenocarcinoma cells (A549) in vitro. We conclude that especially BIT 1 and BIT 2 have in vitro anti-inflammatory and anti-cancer activities, which can be further investigated for future potential therapeutic use. Methods: Inspired by the electrophilic sulfur in Nuphar alkaloids, monomeric and dimeric benzisothiazolones were synthesized from dithiodibenzoic acid and their NF-κB inhibitory role was explored. NF-κB inhibition and cytotoxicity of the synthesized derivatives were studied using luciferase reporter gene assay and XTTassay. Immunocytochemistry studies were performed using L428 cells. Cell migration assay was conducted using the A549 cell line. L428 cells were used to conduct combination studies and the results were plotted using CompuSyn software. Results: Benzisothiazolone derivatives exhibited cytotoxicity in Hodgkin’s Lymphoma cells through NF-κB inhibition. Potent compounds showed suppression of both NF-κB subunits p50 and p65 in a concentrationdependent manner, both in cytoplasmic and nuclear extracts. Combination studies suggest that benzisothiazolone derivatives possess a synergistic effect with etoposide and doxorubicin. Furthermore, the compounds also inhibited the migration of A549 cells. Conclusion: Benzisothiazolones bearing one or two electrophilic sulfur atoms as part of the heterocyclic framework exhibited cytotoxicity in Hodgkin’s Lymphoma cells through NF-κB inhibition. In addition, these derivatives also exhibited a synergistic effect with etoposide and doxorubicin along with the ability to inhibit the migration of A549 cells. Our study suggests that BIT-based new chemical entities could lead to potential anticancer agents.

Activation of bovine oocytes by protein synthesis inhibitors: new findings on the role of MPF/MAPKs†

2021 ◽  
Author(s):  
Cecilia Valencia ◽  
Felipe Alonso Pérez ◽  
Carola Matus ◽  
Ricardo Felmer ◽  
María Elena Arias
Keyword(s):  
Protein Synthesis ◽  
Kinase Activity ◽  
Cyclin B1 ◽  
Protein Synthesis Inhibitors ◽  
Vitro Fertilization ◽  
New Findings ◽  
Bovine Oocytes ◽  
Dependent Pathway

Abstract The present study evaluated the mechanism by which protein synthesis inhibitors activate bovine oocytes. The aim was to analyze the dynamics of MPF and MAPKs. MII oocytes were activated with ionomycin (Io), ionomycin+anisomycin (ANY) and ionomycin+cycloheximide (CHX) and by in vitro fertilization (IVF). The expression of cyclin B1, p-CDK1, p-ERK1/2, p-JNK, and p-P38 were evaluated by immunodetection and the kinase activity of ERK1/2 was measured by enzyme assay. Evaluations at 1, 4, and 15 hours postactivation (hpa) showed that the expression of cyclin B1 was not modified by the treatments. ANY inactivated MPF by p-CDK1Thr14-Tyr15 at 4 hpa (P < 0.05), CHX increased pre-MPF (p-CDK1Thr161 and p-CDK1Thr14-Tyr15) at 1 hpa and IVF increased p-CDK1Thr14-Tyr15 at 17 hours postfertilization (hpf) (P < 0.05). ANY and CHX reduced the levels of p-ERK1/2 at 4 hpa (P < 0.05) and its activity at 4 and 1 hpa, respectively (P < 0.05). Meanwhile, IVF increased p-ERK1/2 at 6 hpf (P < 0.05); however, its kinase activity decreased at 6 hpf (P < 0.05). p-JNK in ANY, CHX, and IVF oocytes decreased at 4 hpa (P < 0.05). p-P38 was only observed at 1 hpa, with no differences between treatments. In conclusion, activation of bovine oocytes by ANY, CHX, and IVF inactivates MPF by CDK1-dependent specific phosphorylation without cyclin B1 degradation. ANY or CHX promoted this inactivation, which seemed to be more delayed in the physiological activation (IVF). Both inhibitors modulated MPF activity via an ERK1/2-independent pathway, whereas IVF activated the bovine oocytes via an ERK1/2-dependent pathway. Finally, ANY does not activate the JNK and P38 kinase pathways.

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