• Korean Journal of Animal Reproduction
• /
• v.22 no.4
• /
• pp.425-435
• /
• 1998

To improve the efficiency of nuclear transplantation in bovine, in this study the development in vitro of nuclear transferred (NT) embryos was compared by different activation regimens of the enucleated oocytes. The effect of developmental stage and culture system of donor nuclei on fusion and development in vitro of NT embryos were also evaluated. Oocytes were collected from Hanwoo ovaries obtained from slaughterhouse and matured in Ham's F-10 supplemented with hormones. After 20~22 h maturation, the oocytes were vortexed to be free from cumulus cells and subsequently their nucleus and the first polar body were removed. Enucleated oocytes were divided into 3 groups for activation; the oocytes of group I were activated with ionomycin for 5 min and subsequently incubated in 6-dimetylarninopurine (DMAP) for 4 h, Those of group II were treated with DMAP for 4 h at 39 h after onset of in vitro maturation (IVM) and those of group III were kept in room temperature ($25^{\circ}C$) for 3 h at 39 h after onset of IVM. After in vitro fertilization (IVF) the embryos for muclear donor were cultured either by group culture (20 embryos /50 ${mu}ell$ drop) or individually (1 embryo /50 ${mu}ell$ drop) for 4 day and 5 day. At day 4 and 5 after IVF, blastomeres were separated in calcium-magnesium free medium, and then classified into small (day 5: $\leq$ 38 ${\mu}{\textrm}{m}$, day 4: $\leq$ 46 ${\mu}{\textrm}{m}$) and large (day 5 : $\geq$ 38 ${\mu}{\textrm}{m}$, day 4 ; $\geq$ 46 ${\mu}{\textrm}{m}$). The separated blastomeres were replaced into enucleated and activated recipient cytoplasm. The blastomere-oocyte complexes were fused by electrically. The NT embryos were cultured in TCM-199 containing 10% FCS in 39$^{\circ}C$, 5% $CO_2$ incubator for 7 day. The results obtained were summarized as follows; There were no differences in fusion and development to blastocyst between groups as group I (68%, 10%), group II (75%, 14%) and group III (73%, 9%), respectively. However, the cell number in blastocyst of NT embryos in group III were significantly fewer than in the other groups (P＜0.05). No differences in fusion and development to blastocyst were found between individual or group cultured and between small or large blastomeres of day 4 and day 5 donor embryos. From these results, it was concluded that the combination of ionomycin and DMAP, or treatment of DMAP at 39 h after onset of IVM were useful for the efficient of production of NT bovine embryos, and the individual cultured embryos could be simply used as donor nuclei for NT bovine embryo.

• Tuberculosis and Respiratory Diseases
• /
• v.48 no.4
• /
• pp.448-463
• /
• 2000

Background : Pulmonary infiltrate is a frequent cause of morbidity and mortality in patients with leukemia. It is often hard to obtain a reliable diagnosis by clinical and radiologic findings alone. The aim of this study was to evaluate diagnostic and therapeutic benefits of invasive procedures for new lung infiltrates in leukemia. Methods : Patients with leukemia who developed new lung infiltrates from December 1994 to March 1999 were included in this study. These patients were classified into the empirical group who received empirical therapy only and into the invasive group who underwent bronchoscopy or surgical lung biopsy for the diagnostic purpose of new lung infiltrates. A retrospective chart review was done to find the etiologies of new lung infiltrates, the yield of invasive procedures, outcome as well as predicting factors for survival. Results : 1) One hundred-two episodes of new lung infiltrates developed in 90 patients with leukemia. Invasive procedures were performed in 44 episodes while 58 episodes were treated with empirical therapy only. 2) Invasive procedures yielded a specific diagnosis in 72.7%(32/44), of which 78.1% had infectious etiology. Therapeutic plan was changed in 52.3%(23/44) of patients after invasive procedures. None of them showed procedure-related mortality. 3) The overall survival rate was 62.7%(64/102). Survival rate in the invasive group (79.5%) was significantly better than that in the empirical group (50.0%) (p=0.002). 4) Upon multivariate analysis, the performance of invasive procedures, no need for mechanical ventilation and achievement of complete remission of leukemia after induction chemotherapy were the independent predicting factors for survival in patients with leukemia and new lung infiltrates. Conclusion : Bronchoscopy and surgical lung biopsy are useful in the diagnosis of new lung infiltrates in patients with leukemia. However, survival benefits of invasive procedures should be considered together with disease status of leukemia and severity of respiratory compromise.