• Journal of Embryo Transfer
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• v.26 no.3
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• pp.171-180
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• 2011

In this study I report that in vitro development rates of bovine nuclear transfer embryos activated either with boar sperm cytosolic factor (SCF) or with ionomycin followed by cycloheximide (CHX) and subsequent in vivo developmental rates after embryo transfer are related to blastocyst quality as evaluated by apoptosis analysis. SCF was extracted from porcine semen then purified for post-activation injection after nuclear transfer. The optimal timing for SCF injection was determined to be at least 22 h post-IVM for parthenogenetic activation of bovine oocyies. A total of 364 oocytes were successfully enucleated and 268 (73.6%) fused and were injected with SCF. The survival rate of fused and injected embryos was 109/113 (96.5%) after 2 h. The cleavage rates of nuclear transfer embryos after 3 d of culture in the ionomycin/CHX treated group were significantly higher than those of the SCF-activated group (93.3% vs 81.7%, p<0.01, respectively). However, at 7 d and 9 d there was no significant difference between the total developmental rates to blastocyst for either treatment group. Total blastocyst cell numbers were also not significantly different between the two activation treatments (ionomycin/CHX: 149.5${\pm}$7.7 vs. SCF: 139.3${\pm}$4.4 cells). In contrast, the apoptotic levels in the SCF blastocysts were higher than those produced after the chemical treatment (28.2${\pm}$5.1% vs. 8.8${\pm}$0.6%, respectively). A total of 18 expanded or hatching blastocysts was transferred to nine synchronized recipients in each activation group; 5/9 (55.5%) and 2/9 (22.2%) were pregnant at 40 d in the ionomycin/CHX treatment and SCF activated group, respectively. However, only one went to term in the ionomycin/CHX treatment while none of the pregnancies from the SCF group were maintained by 90 d. In conclusion, these results suggest that SCF derived from different species is a limited activator to be used for activation after bovine nuclear transfer in lieu of a chemical activation protocol.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1996.04a
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• pp.180-180
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• 1996

Previous studies have shown that kainic acid (KA) causes an elevation of hippocampal proenkephalin mRNA level. However, the role of proto-oncogene products, such as c-Fos, c-Jun and Fra proteins in the regulation of KA-induced proenkephalin mRNA increase in the hippocampus has not been well characterized. Thus, in the present study, the effect of cycloheximide (CHX) on KA-induced proenkephalin mRNA and immediate early gene products induction was examined. After pretreating with either vehicle or CHX (20 mg/kg, s.c.) for 30 min, KA (10 mg/kg) was administered s.c. The animals were sacrificed 1,2, or 8 hrs after KA administration. Total RNA and were isolated for Northern blot assay, and proteins were isolated for Western and electrophoretic gel-shift assays. First, we found that CHX inhibited KA-induced proenkephalin mRNA increase without altering intracellular proenkephalin protein level. Secondly, Western blot assays showed that KA increased c-Fos, c-Jun and Fra proteins at 1,2, and 8 hrs and CHX inhibited these immediate early gene products. Finally, electrophoretic gel shift assays revealed that KA increased both AP-1 and ENKCRE-2 DNA binding activities. Furthermore, CHX attenuated KA-induced AP-1 and ENKCRE-2 DNA binding activities. Both AP-1 and ENKCRE-2 DNA binding activities were abolished by cold AP-1 or ENKCRE-2 oligonucleotides, and further reduced by antibodies against c-Fos or c-Jun. Antibody against CREB reduced ENKCRE-2, but not AP-1, DNA binding activity. Our results suggest that on-going protein synthesis is required for elevation of hippocampal proenkephalin mRNA level induced by KA. All c-Fos, c-Jun, and Fra proteins appears to be involved in the regulation of hippocampal proenkephalin mRNA level induced by KA (This study was supported by a grant from KOSEF).

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.35 no.5
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• pp.312-315
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• 2009

Objective: It is important to sterilize oral cavity with antibacterial agent before surgery for preventing infection. The object of this study was to compare the effect on reduction of salivary bacterial counts according to applied time when povidone-iodine (PVI) and chlorhexidine gluconate (CHX), most broadly used materials in dentistry, were applied intraorally before the surgery. Methods: Sixty subjects were divided into 6 groups. PVI and CHX were applied in each group for 1, 2 and 3 minutes, respectively. Then salivary microbacteria taken before and after applying the materials were cultured using 5% sheep blood agar plate. Results: There was significant difference in reduction of microbacteria in both PVI and CHX and the effect did not show differences depending on time. When applied for a minute, PVI showed somewhat higher reduction rate than CHX, but in the other groups, there was no difference in reduction rate. Conclusion: We found that there was no significant difference in sterilization ability of PVI and CHX in all groups in this study. Therefore, both agents would get sufficient effect when applied for a minute.

• Restorative Dentistry and Endodontics
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• v.44 no.1
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• pp.8.1-8.7
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• 2019

Objectives: Irrigants are imperative in endodontic therapy for the elimination of pathogens from the infected root canal. The present study compared the antimicrobial efficacy of octenidine dihydrochloride (OCT) with chlorhexidine (CHX) and sodium hypochlorite (NaOCl) against Staphylococcus epidermidis (S. epidermidis) for root canal disinfection. Materials and Methods: The minimum inhibitory concentration (MIC) was obtained using serial dilution method. The agar diffusion method was then used to determine the zones of inhibition for each irrigant. Lastly, forty 6-mm dentin blocks were prepared from human mandibular premolars and inoculated with S. epidermidis. Samples were randomly divided into 4 groups of 10 blocks and irrigated for 3 minutes with saline (control), 2% CHX, 3% NaOCl, or 0.1% OCT. Dentin samples were then collected immediately for microbial analysis, including an analysis of colony-forming units (CFUs). Results: The MICs of each tested irrigant were 0.05% for CHX, 0.25% for NaOCl, and 0.0125% for OCT. All tested irrigants showed concentration-dependent increase in zones of inhibition, and 3% NaOCl showed the largest zone of inhibition amongst all tested irrigants (p < 0.05). There were no significant differences among the CFU measurements of 2% CHX, 3% NaOCl, and 0.1% OCT showing complete elimination of S. epidermidis in all samples. Conclusions: This study showed that OCT was comparable to or even more effective than CHX and NaOCl, demonstrating antimicrobial activity at low concentrations against S. epidermidis.

• Restorative Dentistry and Endodontics
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• v.44 no.2
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• pp.13.1-13.9
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• 2019

Objectives: This study evaluated the effect of dentin pretreatment with silver nanoparticles (SNPs) and chlorhexidine (CHX) on the microshear bond strength (${\mu}SBS$) durability of different adhesives to dentin. Materials and Methods: Occlusal surfaces of 120 human molars were ground to expose flat dentin surfaces. The specimens were randomly assigned to six groups (n = 20). Three groups (A, B, and C) were bonded with Adper Single Bond 2 (SB) and the other groups (D, E, and F) were bonded with Clearfil SE Bond (SEB). Dentin was pretreated with CHX in groups B and E, and with SNPs in groups C and F. The specimens were restored with Z250 composite. Half of the bonded surfaces in each group underwent ${\mu}SBS$ testing after 24 hours and the other half was tested after 6 months of water storage. Results: SNP application was associated with a higher ${\mu}SBS$ than was observed in the CHX and control groups for SEB after 24 hours (p < 0.05). A significantly lower ${\mu}SBS$ was observed when no dentin pretreatment was applied compared to dentin pretreatment with CHX and SNPs for SB after 24 hours (p < 0.05). The ${\mu}SBS$ values of the 6-month specimens were significantly lower than those obtained from the 24-hour specimens for all groups (p < 0.05). This decrease was much more pronounced when both adhesives were used without any dentin pretreatment (p < 0.05). Conclusions: SNPs and CHX reduced the degradation of resin-dentin bonds over a 6-month period for both adhesive systems.

• Journal of Embryo Transfer
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• v.16 no.2
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• pp.107-115
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• 2001

Development of effective activation protocols is of great importance for improving the success of cloning and subsequent transgenic. Three methods for oocyte activation, including 5$\mu$M ionomycin (5 min) alone, ionomycin + 1.9 mM 6-dimetylaminopurine (DMAP, 3 hrs) and ionomycin + 10 $\mu\textrm{g}$/ml cycloheximide (CHX, 3 hrs) were compared for their effects of pronuclei (PN) formation, development, developmental velocity and ploidy of parthenotes to IVF control in bovine. In group of ionomycin + DMAP, the oocytes having more 3PN were significantly (P<0.05) higher than in groups of ionomycin alone and of ionomycin + CHX (45.5% vs. 0 and 0%, respectively). Activation with the ionomycin alone, ionomycin + DMAP and ionomycin + CHX resulted in cleavage rates of 30, 85.5 and 57.9%, respectively. The blastocysts rate of parthenotes activated by ionomycin + DMAP treatment was significantly higher (12.3%. p<0.05) than those of other treated groups. Chromosome analysis shows that ionomycin + DMAP treatment greatly enhances the incidence of chromosomal abnormality of the parthenotes. From the results, we may conclude that DMAP treatment to the oocytes accelerates developmental velocity resulting in both the higher incidence of chromosome abnormality and of PN formation, and strongly suggest that CHX combined with ionomycin is better than DMAP for the purpose of successful nuclear transplantation. Developmental velocity of parthenotes activated by ionomycin + DMAP treatment was significantly (P<0.05) faster than others.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.15 no.9
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• pp.5644-5651
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• 2014

Propolis is an extremely safe natural antimicrobial substance that has been reported to have powerful antibacterial efficacy. The aim of this study was to evaluate the inhibitory effects of propolis against Candida albicans (C. albicans). Propolis was collected from the honey bee Apis mellifera. The strain of C. albicans was cultivated overnight in liquid media incubated at $37^{\circ}C$. The antimicrobial activity was investigated using phosphate buffered saline (PBS), 3% sodium hypochlorite (NaOCl), 0.1% chorhexidine (CHX), and propolis extracts ($5{\mu}l/ml$, $10{\mu}l/ml$). C. albicans were sensitive to 3% NaOCl, 0.1% CHX, and propolis ($5{\mu}l/ml$, $10{\mu}l/ml$) with zones of inhibition of 15, 14.5, 16, and 17 mm, respectively. The CFU of PBS, 3% NaOCl, 0.1% CHX, $5{\mu}l/ml$ and $10{\mu}l/ml$ of propolis led a 1, 7, 7, 5 and 7-log reduction. Among the groups tested, C. albicans was most sensitive to $10{\mu}l/ml$ of propolis, which showed the largest inhibition zones. Therefore, propolis can be a new antimicrobial therapy for oral mucosa disease in traditional medicine.

• Restorative Dentistry and Endodontics
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• v.35 no.4
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• pp.295-301
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• 2010

Recent studies demonstrated that the combination of chlorhexidine (CHX) and sodium hypochlorite (NaOCl) resulted in the formation of a precipitate, para-chloroaniline (PCA). Alexdidine (ALX) is a kind of biguanides like CHX, and has stronger detoxification effect against the bacterial virulence factors such as lipoteichoic acid and lipopolysacchardide compared with CHX. The purpose of this study was to determine whether PCA was formed after chemical interaction between ALX and NaOCl using mass spectrometry. Mass spectrometry was performed for the mixture of five different concentrations of ALX (1, 0.5, 0.25, 0.125, 0.0625%) and 4% NaOCl. Results showed that the peak of PCA was not detected in mixed solutions of ALX and NaOCl in mass spectrometry analysis. The color of mixed solution of ALX and NaOCl after chemical interaction was light yellow to white, but there wasn't any precipitate found.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.5
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• pp.718-724
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• 2007

Considerable attention has been focused on the cloning of mammalian embryos, as a consequence of poor development, in order to enhance the application of genetic engineering. Experiments were conducted to compare the developmental competence of parthenotes and reconstructed (NT) rabbit eggs with fetal fibroblasts (FFs) following various activation regimens. Oocytes and NT eggs were exposed to: electric stimulation (EST, Group 1) and EST followed by 6-dimethylaminopurine (DMAP, Group 2), cycloheximide (CHX, Group 3) or DMAP/CHX (Group 4). Pronuclear (PN) status, cleavage, blastocyst development and the ploidy were assessed. In parthenote groups 1, 2, 3 and 4, the PN formation differed significantly. And, the cleavage and blastocyst rates were 41.7 and 5%, 75.6 and 53.7%, 68 and 36%, 82.1 and 52.6%, respectively, among treatments. Polyploidy was observed in 17.2% of EST plus DMAP and 44.9% of EST plus DMAP/CHX groups. In SCNT groups (Group 1, 2, 3 and 4), the cleavage and blastocyst rates were 28.6 and 7.1%, 58.3 and 29.2%, 56.8 and 24.1%, 64.5 and 27.8%, respectively. The chromosomal composition differed significantly (p<0.05) among treatments. In Group 2 and 3, 53.8% and 81.8% of embryos revealed diploid chromosomal sets, respectively. However, in Group 4, 53.3% of embryos showed abnormal ploidy (mixoploid). Although DMAP or combination with DMAP/CHX resulted in higher in vitro development of rabbit SCNT embryos, higher incidence of chromosomal abnormality may induce problems related to fetal loss of at late stage of development.

• Proceedings of the KSAR Conference
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• 2001.10a
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• pp.62-62
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• 2001

Development of effective activation protocols is of great importance for improving the success of cloning and subsequent transgenic. Three methods for oocyte activation, including 5μM ionomycin (5 min) alone, ionomycin＋1.9 mM 6-dimetylaminopurine (DMAP, 3 hrs) and ionomycin＋10㎍/㎖ cycloheximide(CHX, 3 hrs) were compared for their effects of pronuclei(PN) formation, development, developmental velocity and ploidy of parthenotes to IVF control in bovine. In group of ionomycin＋DMAP, the oocytes having more 3 PN were significantly(P〈0.05) higher than in groups of ionomycin alone and of ionomycin＋CHX (45.5% vs. 0 and 0%, respectively). Activation with the ionomycin alone, ionomycin＋DMAP and ionomycin＋CHX resulted in cleavage rates of 30, 85.5 and 57.9%, respectively. The blastocysts rate of parthenotes activated by ionomycin＋DMAP treatment was significantly higher (12.3%, P〈0.05) than those of other treated groups. Chromosome analysis shows that ionomycin＋DMAP treatment greatly increases the incidence of chromosomal abnormality of the parthenotes. When compared the developmental velocity at 24 hrs after insemination and activation, 27% eggs in IVF control and 55% in DMAP treatment out of total cleaved eggs developed to 2-cell stage, respectively. Developmental velocity of parthenotes activated by ionomycin ＋DMAP treatment was significantly (P〈0.05) faster than others. From the results, we may conclude that DMAP treatment to the oocytes accelerates developmental velocity resulting in both the higher incidence of chromosome abnormality and of PN formation suggesting that CHX combined with ionomycin is suitable DMAP for the purpose of successful nuclear transplantation.