An improved antibody-coated sensor system based on quartz crystal microbalance was developed for the detection of Salmonella spp. An antibody against Salmonella common structural antigen was immobilized onto one gold electrode of the piezoelectric quartz crystal surface by various immobilization procedures. The best results in sensitivity and stability were obtained with the thin layers of protein A and 3,3'-dithiopropionimidate 2HCl (DTBP), a homobifunctional thiol-cleavable crosslinker. After the addition of a S. typhimurium suspension into a reaction cell with 0.1 M sodium phosphate buffer, pH 7.2, the resonant frequency owing to S. typhimurium adsorption decreased conspicuously. The antibody-immobilized crystals prepared by the gold-protein A complex formation and DTBP thiolation showed the frequency shifts of 80 and 283 Hz, respectively. The time required for maximum frequency shift was about 30-60 min. The antibody-coated crystal could be reused for 6-8 consecutive assays.

Anti-phospholipid antihodies (aPL) have important roles in various pregnancy complications such as recurrent miscarrige, growth retardation, placental abruption and stillbirth. However, their biological actions on preimplantation development of oocytes are still unclear. In this study, we investigated whether either aPL containing sera or phospholipids could affect in vitro fertilization and development of mouse oocytes. Sera used in this study were collected from three patients and the presence of aPL in the sera was confirmed by enzymatic-linked immunosorbent assay. When mouse oocytes were cultured in a serum-free, Chatot, Ziomek and Bavister (CZB) medium (Experiment 1), addition of aPL-containing sera (10%) to CZB medium did not. significantly (P>0.05) influence sperm penetration of oocytes. However, development to the blastocyst stage was significantly (P<0.05) inhibited by serum addition, and formation of morulae (16-23% vs. 58%) and blastocysts (0-4% vs. 21%) was markedly reduced compared with no addition (Experiment 2). In Experiment 3, pronuclear stage embryos were cultured for 96 h in GZB medium supplemented with 1 g /ml phosphatidyl ethanolamine, 1 g/ml phosphatidyl inositol or 1 g /ml phosphatidyl choline. No increase in embryo development was found after addition of the phospholipids to CZB medium. These results suggest that 1) aPL have an inhibitory role in preimplantation development of mouse embryos, and that 2) the action of aPL may be related to a specific phospholid (s) rather than the tested phospholipids in the present study.

The envelope of the rnannnalian oocyte plays crucial roles in sperm-oocyte interactions by providing sperm receptors, inducing acrosome reaction and preventing polyspermy. Understanding of properties of the zona pellucida (ZP) is essential for the artificial control of fertility in mammals. This study was carried out to produce and characterize monoclonal antibodies(MAbs) to porcine ZP proteins. Approximately 8,000 ZPs were obtained from follicular oocytes and dissolved in 40l of double distilled water. Following immunization through foot-pad injections of Balb /c mice with a ZP solution, the popliteal lymph nodes were recovered at 2 weeks after the last injection. Hybridoma cell lines were established by fusing lymph node cells with P3X63 myeloma cells through selection using HAT medium and screening by immunofluorescence(IF) microscopy on the isolated ZP. Secreted MAbs were found to consist k chains and different heavy chains as evidenced by isotyping. Some of the MAbs demonstrated high specificity to the ZP in IF. The Mabs also showed positive cross reactivity with hamster and mouse eggs, while negative with bovine eggs. The results implicate that the MAbs can be used not only for identification of functional regions of the ZP, but also for elucidation of mechanisms involved in fertilization of mammals. The MAbs will provide basic information on biochemical anatomy of the ZP as well as can be candidates for the future contraceptive vaccines.

The present study was performed to investigate the effects of caffeine and heparin on capacitation and acrosome reaction of bovine spermatozoa, effects of antisperm antibodies on acrosome reaction of bovine spermatozoa. The rates of acrosome reaction in control group, caffeine treated group, heparin treated group, caffeine-heparin complex treated group were 40.3, 54.3, 63.3, 72.3%, respectively and there were significant differences among the groups(p<0.01), especially higher in caffeine-heparin complex treated group than the others. The rates of acrosome reaction of antisperm antibodies serum supplemented groups(5, 10 and 20%) were 60.4, 48.9 and 37.1%, respectively and there were significant differences among the groups(p<0.0l), and the more increases in serum concentrations, the more decreases in acrosome reaction, but this phenomenon was not seen in fetal calf serum supplemented group and heifer serum group. When the serum concentration was 5%, the rates of acrosome reactions were significantly lower in fetal calf serum supplemented group than heifer serum group and in antisperm antibodies serum group(p<0.01), and there were no significant differences between heifer serum group and antisperm antibodies serum group(p<0.01). When the serum conecntrations were 10%, 20%, the rates of acrosome reactions were significantly lower in antisperm antibodies serum supplemented group than in fetal calf serum group and in geifer serum group(p<0.01), and there were no significant differences between fetal calf serum group and heifer serum group(p<0.01). These results indicate that caffeine-heparin complex treatment is very effective for inducing acrosome reaction of bovine spermatozoa and that antisperm antibodies block acrosome reaction.

식이성 항원에 특이적인 IgG 및 IgA가 모유 중에 어느 정도 함유되어 있는지를 측정하고, 모체가 섭취한 식이 내용과 이들 특이 항체가 어떠한 상관성을 갖고 있는지를 조사한 결과는 다음과 같았다. 대상 임산부의 모유에 함유된 식이성 항원에 특이적인 항체 수준은 초유에서 가장 높게 나타났으며, 이행유로부터 성숙유 사이에서의 수준은 거의 변화하지 않았다. 측정된 우유단백질 및 계란단백질에 대해서는 특이 항체가에 커다란 차이는 없었으나 밀단백질인 gliadin에 대해서는 IgG, IgA 모두 높은 항체가를 유지하였다. 한 종류의 식이성 항원에 대해 높은 항체가를 나타내는 경우는 다른 특이 항체도 높은 수준을 나타내었으며 이러한 경향은 IgG에서 더욱 뚜렷하게 나타났다 모유 중에 함유된 특이 항체 수준은 임산부의 열량 및 단백질 섭취량에 의해 영향을 받지 않는 것으로 나타났다. 이들 특이 항체 수준이 모체가 섭취한 식이 단백질에 의해 영향을 받는지를 24시간 회상법 및 식품섭취 빈도 조사에 의해 살펴보았다. 24시간 회상법에 의해서는 특이 항체와 식이내용 중 그 특이항체를 유도할 수 있는 특정 식품 단백질 섭취량 사이에서의 상관성을 살펴볼 수 없었다. 식품섭취 빈도조사에서는 IgG의 성우 우유의 섭취빈도는 우유 유래의 특이 항체가에 영향을 주지 못한 것으로 보이나 알류섭취량과 OVA항체 수준 사이에서는 유의적인 상관관계를 볼 수 있었고, 육류의 섭취빈도는 α-La을 제외한 모든 항체수준과 영향을 준 것으로 나타났다. IgA의 경우는 육류의 섭취빈도가 항-BSA항체의 유도에 매우 유의적으로 작용한 것으로 나타났다.

In order to establish the enzyme linked immunosorbent assay(ELISA) of sterigmatocystin produced by Aspergillus versicolor, we experimented and obtained following results. Two of three rabbits which had been immunized with sterigmatocystin-hemiacetal-BSA produced antibodies against sterigmatocystin at 15 weeks. The produced antibodies were specific for sterigmatocystin and sterigmatocystin-hemiacetal but didn't cross react with other sterigmatocystin analogues in a significant degree. DMF : 4% KCl (18:2) mixed solution was most effective to dissolve sterigmatocystin. For the preparation of sample solution to determine sterigmatocystin by ELISA, sample was extracted with CHCl₃ and dried, then the dried sample was redissolved with 100 ul DMF+4% KCl mixture. 10-1,000 ng/ml level. of standard sterigmatocystin could be applied to the established ELISA. When artifically contaminated rice were assayed by the ELISA, the average recovery of sterigmatocystin spiked to 25-500 ng/g was 109% (97-116%), and mean interwell coefficient of variation was 21% (11-28%).