• Korean Journal of Animal Reproduction
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• v.25 no.4
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• pp.317-325
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• 2001

This study investigated the effect of ferrous sulfate (Fe$^{2+}$) and/or ascorbic acid (Asc) on fertilizing ability in vitro of frozen-thawed boar spermatozoa. Using chlortetracycline (CTC) fluorescence, the spermatozoa was treated in preincubation medium with control, Fe$^{2+}$(1 mM), Asc (0.5 mM) and Fe$^{2+}$Asc to assessed for acrosome reaction, and the oocyte penetration test to determine whether the Fe$^{2+}$ and/or Asc can promote the penetration ability in vitro. When frozen-thawed spermatozoa was washed with preincubation medium, there were significantly (P ＜ 0.05) more acrosome-reacted in medium with Fe$^{2+}$Asc (38%) than control (27%). The penetration rates were also significantly (P ＜ 0.05) higher in medium with Fe$^{2+}$Asc (76%) than control (55%). Next, the lipid peroxidation of sperm was evaluated on the basis of malondialdehyde production following same treatments. The addition of Fe$^{2+}$Asc to sperm suspension increases the formation of malondialdehyde. However, there were not significantly different under the all conditions. The sperm suspension were also treated with control, Fe$^{2+}$, Asc and Fe$^{2+}$/Asc and assayed for sulfhydry1(-SH) group content. In the Fe$^{2+}$/Asc group, sperm-SH group were higher than another groups. In spermatozoa treated with Fe$^{2+}$ and/or Asc, however, no changes in sperm -SH-groups were detected when compared to controls. In another experiment, the activity of sperm binding to zona pellucida was evaluated through binding to salt-stored porcine oocytes. In control and Asc treatment groups, sperm binding to zona pellucida were significantly (P ＜ 0.05) higher than in medium with Fe$^{2+}$. On the other hand, there is not a significant increase in binding to zona pellucida with spermatozoa treated by Fe$^{2+}$/Asc. In summary, the present study suggests that Fe$^{2+}$/Asc causes an enhancement in fertilizing ability that is associated with penetration rate increased without change of spermatozoa binding capacity to homologous zona pellucida.o homologous zona pellucida.

• Journal of Animal Science and Technology
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• v.46 no.6
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• pp.957-966
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• 2004

The objective of this study was to investigate the effects of BPA on reproductive characteristic and blood hematological and chemical values in mice. The male mice were intraperitoneally injected BPA in native control(no treatment), positive control(corn oil, 3$m\ell$/kg B.W), 0.05, 0.5 and 5.0mg BPA/kg B.W and female mice were injected BPA in control(corn oil, 3$m\ell$/kg B.W), 0.05, 0.5 and 5.0mg BPA/kg B.W with 5 times at 3 days interval for 14 days. The administration of BPA in male mice didn't affect the body and reproductive organ weight such as testis, epididymis, vesicular gland and coagulating gland. We found that the 5.0mg BPA group was significantly reduced the sperm concentration and increased the sperm abnormality compared to native, positive control and 0.05mg BPA groups(P<0.05), but any other effects were not found in sperm viability and motility in BPA treatment groups. The RBC, HB, HT, MCV, MCH, MCHC, albumin, BUN and total protein of blood hematological and chemical values in male were not different in all experimental groups(P>0.05). However, the value of WBC was slightly lower in BPA treatment groups than that of control groups and PLT was slightly higher in BPA groups than that of control groups, but not significantly defference among the experimental groups(P>0.05). In female mice, the effects of BPA on body weight didn’t affect in all experimental groups, but ovary weights in 0.5 and 5.0mg BPA groups were significantly increased compared to those in control and 0.05mg BPA group(P<0.05). The uterine weight in BPA groups was slightly higher than that of control group, but not significantly different in all experimental groups(P>0.05). The RBC, Hb, HT, MCV, MCH, MCHC, albumin and total protein of blood hematological and chemical values in female were not different in all experimental groups(P>0.05). The values of WBC and PLT in BPA groups were slightly higher than that of control, but not significantly different among the experimental groups. The concentration of BUN was the higher in BPA groups than that of control group. The histological evaluation of testis, ovary and uterus were not different in all experimental groups.

• Journal of Embryo Transfer
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• v.21 no.3
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• pp.255-262
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• 2006

The purpose of this study is to investigate the effects of vitrification in open pulled straws (OPS) methods on in vitro survival ability of porcine embryos. For in vitro maturation of immature oocytes, the porcine ovaries were collected from local slaughter-house. The cumulus-oocytes complexes were aspirated from 2 to 6 mm follicles. The collected oocytes were cultured for in vitro maturation in NCSU-23 medium with 5 mM hypotaurine, 0.57 mM cysteine, 10% porcine follicle fluid, 10 IU/ml PMSG and 10 IU/ml hCG for $21{\sim}22$ hrs. Then, the oocytes were more cultured $21{\sim}22$ hrs in vitro maturation in medium removed hormones. The frozen-thawed spermatozoa were washed by centrifugation 2 times for 10 min at 1,500 rpm in D-PBS with 5.56 mM glucose, 0.33 mM Na-pyruvate, 100 IU/ml penicillin, $100 {\mu}g/ml$ streptomycin and 4 mg/ml BSA. The fertilization medium used mTBM with 2 mM caffeine and 2 mg/ml BSA and adjusted to a pH of 7.2 to 7.4. The final concentration of spermatozoa was adjusted to $2.5{\times}10^6$cells/ml motile sperm during fertilization in vitro. At 8 hrs after insemination, the oocytes were transferred into NCSU-23 medium with 5.0 mM hypotaurine, 4 mg/ml BSA and 10 ng/ml EGF and cultured for 7 days. When the blastocysts of different stages were frozen-thawed by OPS methods, the proportions of embryos with normal morphology were significantly (p<0.05) higher in embryos frozen-thawed at expanded blastocyst stage (38.9%) than in early blastocyst stage (28.3%). On the other hand, the proportions of embryos damaged after frozen-thawing were significantly (p<0.05) higher in embryos frozen at early blastocyst stages than in expanded blastocyst stage. In another experiment, the normal embryos morphology after frozen- thawing were further cultured for 48 hrs. After culture, the proportions of embryos hatched were 6.7, 20.0 and 33.3% for embryos frozen-thawed at early blastocyst, mid-blastocyst and expanded blastocyst stages. These finding indicate the possible broader application for OPS methods, as frozen-thawed embryos may be accompanied by developmental stage according to requirements of the survival ability after freezing of blastocyst stage in the pig.

• Korean Journal of Poultry Science
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• v.40 no.3
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• pp.163-169
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• 2013

Cryopreservation of poultry semen has been reported, but preservation of female genetic material has not been possible because of the unique anatomical and physiological characteristics of the avian egg. Thus an alternative strategy for conservation of oviparous species of animals must be developed. Recent technological developments for producing germline chimeras by the transfer of primordial germ cells (PGCs) into recipient embryos has enabled the conservation and retrieval of chicken genetic resources in their complete form. In the present study, fertilized eggs were incubated for about 5.5 days to obtain embryos at stage 28. The whole embryo was collected from the germinal gonad using a fine glass micro pipette under a microscope. The PGCs were then purified using MACS method. Two commercially available cryoprotectants (A and B) were used to preserve the PGCs, and EG were used as a control. The average recovery rate of PGCs after thawing was 35.5% and 60.5% with the A and B treatments, respectively. There was no significant difference between B treatments and control, which showed an average recovery rate of 52.8%. However, the recovery rate obtained using A cryoprotectant (35.5%) was significantly lower than using treatment control and B. The average viability of the PGCs after thawing were 77.9% and 77.4% for cryoprotectants A and B, respectively, and the control were was 81.6%. There was no statistically significant difference between the two treatments and control. It was concluded that all of the available cryoprotectants examined in this study could be used for preservation of PGCs from embryos. Further experiments to produce germline chimera from PGCs preserved using this techniques are strongly recommended.

• Tuberculosis and Respiratory Diseases
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• v.43 no.2
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• pp.173-181
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• 1996

Background : Several studies have suggested that impaired mucociliary clearance plays a role in the pathophysiology of bronchial asthma. Cough productive of mucoid sputum is common, and mucous plugs in the airways are frequently observed. These clinical features are in keeping with the histologic lesions of asthma, which involve primarily the epithelial and mucous-producing structures of the conducting airways. Some studies have shown that the mucociliary clearance is impaired in adult asthma, but it has not been studied in childhood asthma. The objectives of this study were to examine whether the mucociliary clearance is impaired in childhood asthma and to estimate the degree of impairment in comparison with that of immotile cilia syndrome. Method : Thirteen children with mild stable asthma and eight patients with immotile cilia syndrome completed this study. Ten healthy children were recruited as a normal control group. The whole-lung mucociliary clearance was measured by the radioaerosol technique. Aerosols, tin colloid particles tagged with the radionuclide technetium-99m($^{99m}Tc$), were generated by means of nebulizer, and inhaled via a mouthpiece. The retention of radioactivity was measured at 30, 60, 90 and 120 minutes by gamma camera, and mucociliary clearance was calculated as percent retention at each time. Results: 1) In each subject, the percent retention decreased variably with the lapse of time. 2) The percent retention of radionuclide decreased at each time in order of normal control, bronchial asthma and immotile cilia syndrome and the percent retention of immotile cilia syndrome was significantly higher than that of normal control at each time(p<0.05). 3) At two hours, the percent retention of bronchial asthma($65.0{\pm}1.8$(SE)%) was significantly higher than that of the normal control($54.4{\pm}3.5%$, p<0.05), and significantly lower than that of immotile cilia syndrome($73.3{\pm}1.4%$, p<0.01). 4) When the percent retention was analyzed according to $PC_{20}$ in the children with bronchial asthma, they had no relationship with each other. Conclusion: Mucociliary clearance in the children with bronchial asthma was significantly lower than normal control. This finding indicates that impaired mucociliary clearance operates in childhood asthma as well, and suggests that it may be one contributing factor in the pathogenesis of asthma. The degree of impairment, however, was not so severe as immotile cilia syndrome.

• Tuberculosis and Respiratory Diseases
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• v.45 no.2
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• pp.311-321
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• 1998

Background: Mucoepidermoid carcinoma of the lung arises from submucosal gland of tracheobronchial tree. Histologically, the tumor is composed of mucin-secreting cells, squamous cells, and intermediated cells, which show no particular differentiating characteristics, in varying proportions. The tumor is divided into low grade and high grade depending on the proportion of cells, and the degree of the mitotic activity, cellular necrosis and nuclear pleomorphism. While favorable prognosis of low grade tumor, high grade tumor, which is very difficult to differentiate from adenosquamous carcinoma, has an aggressive clinical course. The tumor is rare, comprising 0.1 to 0.2% of primary lung cancers and 1 to 5% of bronchial adenomas. Method: A retrospective clinical study was done on 17 cases of mucoepidermoid carcinoma. The study investigated the clinical features, radiologic findings, bronchoscopic findings, histology and clinical courses. Results: Age ranged between second to seventh decade with a mean age of 42 years. Twelve out of 17 cases were male. Five out of 17 cases were smokers with a mean 11 pack-years. Common symptoms included dyspnea, cough, hemoptysis, and wheezing. Two out of 17 cases was asymptomatic. Atelectasis or mass was common radiologic finding. Plain chest radiography was normal in one patient whom the tumor was located in upper trachea. Bonchoscopy revealed exophytic mass in 12 cases and nodular infiltrations in 4 cases. One case having solitary pulmonary nodule in the right lower lung was normal on bronchoscopy. Histologically, ten out of 17 cases were low grade, and seven out of 17 cases were high grade. Among 10 patients with low grade tumor,9 patients were performed operation and have been alive without recurrence during a mean follow-up of 30 months. Two out of 7 patients with high grade tumor were performed pneumonectomy and have been alive during a follow-up of 3 and 8 months, respectively. Conclusion: Most of mucoepidermoid carcinoma is located at central airway and is presented symptoms by mucosal irirtation. Although atelectasis or mass is common radiologic finding. chest X -ray can be normal. The histologic grading and the extent of tumor are two most important factors for prognosis.

• Tuberculosis and Respiratory Diseases
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• v.52 no.1
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• pp.37-45
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• 2002

Background: Diffuse panbronchiolitis(DPB) is a chronic inflammatory lung disease that presents as coughing, copious sputum, exertional dyspnea, which progresses to bronchiectasis. The pathogenesis of bronchiectasis is controlled by inflammatory mediators, which are closely related to mucus hypersecretion, goblet cell dysplasia. In recent studies, the epidermal growth factor receptor(EGFR) system was reported to be associated with this process. It was hypothesized that a relationship exists between goblet cell dysplasia, EGFR expression, and inflammatory mediators produced by neutrophil. Method: Alcian blue/periodic acid -Schiff(AB/PAS) stain, MUC5AC, EGFR, CD16 immunohistochemical stain were examined to investigate a role for the EGFR system in a mucus hypersecretion in DPB using the lung biopsy specimens from 13 DPB patients and 6 controls. Results : In the DPB group, the AB/PAS and MUC5AC -stained areas were $8.31{\pm}3.36%$, $11.46{\pm}4.68%$, respectively. In the control group, the AB/PAS- and MUC5AC-stained areas were $50.5{\pm}5.77%$, $53.3%{\pm}6.67%$, which was significantly larger than in the DPB group (each comparison, p<0.05). The percentage of EGFR expression was $9.54{\pm}4.95%$ in the DPB group, but zero in of the control group. The extent of neutrophilic infiltration was $71.92{\pm}3.71$/5HPF in the DPB group and $45.0{\pm}5.73$/5HPF in the control group, which was statistically significant(p=0.002). Conclusion: The EGFR system is highly related to goblet cell dysplasia, mucus hypersecretion and neutrophilic inflammation in DPB.

• Journal of Embryo Transfer
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• v.31 no.1
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• pp.47-52
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• 2016

The objective of this study was to investigate the result of in vivo embryo collection and pregnancy rate after embryo transfer using sex-sorted sperm of Korean brindle cattle. Donor Korean brindle cattle superovulation treated by decreasing dose of FSH injection. Embryos were recovered on 7 days after the third artificial insemination. Control group semen straw used artificial insemination contained 20 million sperm. Sex-sorted semen straws contained 4 million sperm or 10 million sperm. As for the result of the recovery of the in vivo embryos derived from sex-sorted sperm, the number of transferable embryos was significantly highly recovered to be $6.20{\pm}2.28/donor$ from the control group and was significantly lowly recovered to be $1.57{\pm}1.72/donor$ from the group treated at a sperm concentration of $10{\times}10^6$ (p<0.05). The number of unfertilized embryo was $0.8{\pm}1.30/donor$ in control group which was significantly lower than the group treated at a sperm concentration of $4{\times}10^6$ (p<0.05). However, there was no significant difference in the number of undeveloped ova between control and treatment groups. Pregnancy rate after embryo transfer was shown to be 35.00% in control group and 12.50% in treatment group. The karyotype analysis of the calf derived from sex-sorted sperm resulted in a similar chromosomal distribution pattern (2n=60, XX) compared to those of common Korean native cattle.

• Development and Reproduction
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• v.8 no.2
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• pp.77-84
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• 2004

The purpose of this experiment was to determine the effects of di(2-ethylhexyl) phthalate(DEHP) on reproductive characteristic, blood hematological and chemical values in mice. The male mice were intraperitoneally injected DEHP in negative control(no treatment), positive control(corn oil, 3ml/kg B.W), 0.5, 1.0 and 10.0mg DEHP/kg B.W and the female mice were injected DEHP in control(corn oil, 3ml/kg B.W), 0.5, 1.0 and 10.0mg DEHP/kg B.W with 5 times for 15 days on 3 days interval. The administration of DEHP in male mice were not affect on body weight, epididymis, vesicular gland and coagulating gland weight. The testis weight were slightly higher in DEHP treatment groups than in control. The semen characteristics(sperm concentration, viability, motility and abnormality) of male mice were not difference in all experimental groups. The RBC, Hb, HT, MCV, MCH, MCHC< PLT, albumin, BUN and total protein of blood hematological and chemical values were not affect the administration of DEHP in mice. The WBC values in 10.0mg DEHP group was slightly difference in all experimental group(P>0.05). The histological evaluation of testis, ovary and affevt the reproductive characteristic, blood hematological and chemical values.

• Development and Reproduction
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• v.8 no.2
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• pp.91-97
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• 2004

The objective of this study was to assess that the effects of DEHP administration on reproductive characteristics and blood hematological and chemical values in pups born after DEHP administration in pregnant mice. DEHP was administrated to pregnant mice by intraperitoneally injection with 0, 0.5, 1.0 and 10.0mg/kg B.W, 5 times at 3 days interval from Day 1 to Day 16 in the gestation period. The body weight and reproductive organ weight(testis, epididymis and coagulating gland) in male pups on 45 day after birth was not affected in all experimental groups, but vesicular gland in DEHP groups was significantly lower than that of control group(P<0.05). The semen characteristics of male pups were not affected in DEHP treatment groups. The WBC, HB, HT, MCH and albumin values in male pups were not affected in all experimental groups, but RBC MCV, MCHC, PLT and total protein values were significantly different among the experimental groups(P<0.05). In female pups, the effects of DEHP administration were not affected the body and uterus weight, but the left ovary in 10.0mg DEHP group was significantly heavier than in control and 0.5mg DEHP group(P<0.05). The WBC, MCV, MCH, MCHC, PLT, albumin, BUN and total protein values in female were not different in all experimental groups. The RBC, HB and HT values were significantly different among the experimental gruop(P<0.05). The historical evaluation of testis in male pups that were grown to 45 days after birth was not different in all experimental groups. The ovary in female pups had many corpus luteum in 10.0mg DEHP group. The endometriosisi of uterus was significantly decreased in DEHP group. There results suggest that low concentration of DEHP administration in pup born after DEHP administration in pregnant mice was not affered on reproductive characteristic, but was affected on blood hematological and chemical values.