Up-to-date artificial insemination (AI) using frozen sperm consider as the most widely using technology for improvement of Korean Native Cow (Hanwoo) embryo production. However, it is time consuming, required at least 15~20 years to make more than 6 generations, and their offspring number is limited. To overcome such limitations, superovulation and in vitro fertilization have been developed. For superovulation, the number of produced embryos are not enough for commercialization and donor cows need rest period. This led to use of slaughterhouse ovary for in vitro fertilization, but it is impossible to repeat the collection from the same individual and it only can improve the genetic merits of offspring for one generation. Production of embryos using Ovum Pick-Up (OPU) technique, where oocytes can be repeatedly collected from living elite donor, might overcome these limitations. In this study, we investigated the possibility of using OPU technique from donors at different age and different session periods for mass-embryo-production. Oocytes were collected from 26 donor cows twice per week, 3 - 4 months per year, between 2013 and 2016. Results showed that, the average number of embryo produced in first year used donor was significantly higher than that in second year used donor (3.89 ± 2.85 vs 3.29 ± 2.70), however, there was no significant difference between third year used donor (3.51 ± 3.32) and other groups. Taken together, our data showed that repeated using of donor up to three years is possible for in vitro embryo mass-production. Moreover, OPU can be used as suitable embryo producing technique for livestock breed improvement.

Biofilms of oral microbes can cause various diseases in the oral cavity, such as dental caries, periodontitis and mucosal disease. Electrolyzed water generated by an electric current passed via water using a metal electrode has an antimicrobial effect on pathogenic bacteria which cause food poisoning. This study investigated the antimicrobial activity of electrolyzed waters using various metal electrodes on the floatage and biofilms of oral microbes. The electrolyzed water was generated by passing electric current using copper, silver and platinum electrodes. The electrolyzed water has a neutral pH. Streptococcus mutans, Porphyromonas gingivalis and Tannerella forsythia were cultured, and were used to form a biofilm using specific media. The floatage and biofilm of the microbes were then treated with the electrolyzed water. The electrolyzed water using platinum electrode (EWP) exhibited strong antimicrobial activity against the floatage and biofilm of the oral microbes. However, the electrolyzed water using copper and silver electrodes had no effect. The EWP disrupted the biofilm of oral microbes, except the S. mutans biofilm. Comparing the different electrolyzed waters that we created the platinum electrode generated water may be an ideal candidate for prevention of dental caries and periodontitis.

There are estimated to be about 700 species of bacteria in the oral cavity. Based on epidemiological investigations, some of these strains have been proposed as the pathogens responsible for oral diseases such as dental caries, gingivitis and periodontitis. Since electrolyzed hydrogen-rich water has been shown to have beneficial effects on human im- munity, its use has increased. In our study, the antibacterial activity of hydrogen-rich water for oralagainst bacteria asso- ciated with oral disease was evaluated. The bacterial strains Streptococcus mutans, Fusobacterium nucleatum, Porphy- romonas gingivalis and Tannerella forsythia were cultured in specific growth medium. S. mutans, F. nucleatum and P. gingivalis were soaked to thein both hydrogen water and tap water for 30 sec and then inoculated onto mitis-sali- varius agar and brain heart infusion agar including supple- mented withvitamin K and hemin, respectively. The num- bers of bacterial colonies were then measured after cultiva- tion for 48 hours. In the case of T. forsythia, which does not grow well on agar plates, inoculations into modified new oral spirochete (NOS) broth were performed and growth curve analysis was undertaken every day with a spectrophotometer. Hydrogen water showed antibacterial activity against all four bacterial strains in comparison with tap-water. We conclude from this that hydrogen water may have a positive impact on oral hygiene by helping to remove cariogenic bacteria and periodontopathogens.

MspTL is the major surface protein of Treponema lecithinolyticum associated with periodontitis and endodontic infections. Our recent investigation revealed that MspTL induces proinflammatory cytokines and intercellular adhesion molecule 1 in THP-1 cells and periodontal ligament cells. In this study we conducted oligonucleotide microarray analysis to investigate the global transcriptional regulation in THP-1 cells stimulated with purified recombinant MspTL. MspTL upregulated the expression of 90 genes in THP-1 cells at least four fold, and the functions of these genes were categorized into adhesion, apoptosis/antiapoptosis, cell cycle/growth/differentiation, chemotaxis, cytoskeleton organization, immune response, molecular metabolism, proteolysis, signaling, and transcription. The majority of the modified genes are known to be NF-κB-responsive and interferon-stimulated genes (ISGs). The expression of 12 selected genes was confirmed by real-time RT-PCR. Because prostaglandin E2 (PGE2) is an important inflammatory mediator and Cox-2 was found to be induced by MspTL in the microarray analysis, we determined the level of PGE2 in the culture supernatants of MspTL-treated cells and found that MspTL significantly increased PGE2. Our results provide insight into the gene regulation of host cells in response to MspTL, and may contribute to the understanding of the molecular mechanism in periodontitis.

Treponema denticola is the best studied oral spirochete and numerous studies have shown that it is strongly associated with periodontitis and expresses several putative virulence factors. In this study, we report on a surface protein of T. denticola, Td92, which is homologous to Tp92 of Treponema pallidum, an agent of syphilis. Immunofluorescence assay and immunogold labeling with anti-Td92 Ab revealed that Td92 had surface-exposed epitopes. And Td92 was capable of binding to fibronectin and KB cells, an oral epithelial cell line. In addition, Td92 could enter the KB cells. These results indicate that Td92 is a fibronectin-binding protein which can bind to and internalize into the host cells, facilitating the virulence of T. denticola.

Treponema maltophilum, a Group IV oral spirochete, is associated with periodontitis and endodontic infections. In this study we analyzed the functional role of the major surface protein of this organism (MspA) in human gingival fibroblasts (HGFs). The full-length gene encoding MspA was cloned and expressed in Escherichia coli by using the expression vector pQE-30. The recombinant protein (rMspA) was purified by affinity chromatography with nickel-nitrilotriacetic acid agarose and possible contamination of E. coli endotoxin in rMspA was removed by using polymyxin B-agarose. rMspA significantly induced the expression of pro inflammatory cytokines like IL-6 and IL-8 and intercellular adhesion molecule (ICAM)-1 in HGFs, when analyzed by reverse transcription-PCR, flow cytometry, and enzyme-linked immunosorbent assay. Our results indicate that MspA of T. maltophilum may play an important role in amplifying the local immune response by upregulating the expression of proinflammatory cytokines and ICAM-1.

Newly hatched black porgy larvae (Acanthopagrus schlegeli) swam to the surface, with the mouth and anus still closed and were 1.90–2.11 mm (mean, 2.0 mm) in total length (TL). The larvae were 2.71–2.94 mm TL (mean, 2.82 mm) on day 2 after hatching. At this time, about two-thirds of the yolk was absorbed, the bladder and intestines had formed, and the mouth and anus were open. Total length was 4.32–4.66 mm (mean, 4.45 mm) at the post-larval stage on days 5–6 after hatching, and the yolk and oil globule were almost absorbed. The end of the notochord began to flex, and 6–8 caudal fin rays were visible. The larvae were 15.37–16.1 mm TL (mean, 15.83 mm) at the juvenile stage on days 30–32 after hatching, and the number of rays in all fins was completely revealed.