• Korean Journal of Animal Reproduction
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• v.23 no.1
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• pp.53-61
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• 1999

The present study was designed to investigate the effects of gonadotrophins on in vitro growth and maturation in mouse preantral follicles. Ovaries were removed from 12-day-old ICR mice. Follicles were dissociated enzymetically in Leibovitz L-15 medium containing 1 mg/$m\ell$ collagenase and 0.2 mg/$m\ell$ DNase I. The follicles were cultured on Transwell-COL membrane inserts in six well cluster dishes for 10 days. The culture medium was $\alpha$MEM medium supplemented with 5% fetal bovine serum and FSH or HMG. After 10 days of growth in vitro, follicles were allowed to mature for 18~20 hr in medium supplemented with 1.5 IU/$m\ell$ hCG. The oocytes were then denuded of their cumulus cells and assessed maturation status. Concentrations of oestradiol and progesterone were measured with a radioimmunoassay. Oocyte diameter was determined with an ocular micrometer. The survival and Metaphase II rates of oocytes were significantly higher in FSH treatment groups than in control group (P＜0.001), but there were no differences among the groups of treated FSH concentration. The survival and Metaphase II rates of oocytes in HMG treatment group (60.9 and 40.6%) were higher than in FSH treatment group (76.6 and 48.2%) and control group (49.2 and 7.1%). The survival and Metaphase II rates of oocytes on both FSH and LH treatment groups were no differences among the ratios of FSH and LH. Diameter of oocyte was no differences among the treatment groups, but smaller than compared to in vivo grown oocyte. Through the entire culture period, secretions of oestradiol and progesterone were significantly less in control group than in HMG and FSH treatment groups. These results suggest that gonadotrophins playa key role in in vitro culture of mouse preantral follicles. Especially, addition of FSH and LH should be more effective than FSH alone.

• Korean Journal of Animal Reproduction
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• v.21 no.1
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• pp.47-52
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• 1997

The objective of this study was to investigate the in vitro / in vivo embryonic development after vitrification of mouse zygotes and the chrom osomal normality of delivered live young after embryo transfer. Mouse IVF zygotes were cryopreserved by vitrification using vitrification solution, EFS40 (40% ethylene glyc이, 30% Ficoll a and 0.3 M sucrose in phosphate buffer saline c containing 10% FBS ) . After mouse zygotes were exposed to EFS40 at 25"C for 30 sec., they were immediately plunged into LN$_2$. Vitrified thawed mouse zygotes were cultured upto bIastocysts in M16 for 4 days. The rates of in vitro development were 71.5% under this condition. Cultured blastocysts were transferred to recipients (3 day of pseudopregnant) on one or both uterus horns (6-8 embryos per a uterus horn). And all recipients were allowed to produce litters. The results obtained in these experiments were summarized as follows: The pregnancy rates and in vivo survival rates, live fetus rates, for vitrified zygotes (80.0, 39.6%) were not significantly difference in those of control zygotes (77.8%, 50.0%). Also, all of live-born young mice were chromosomally normal (n=40). This results suggested that proposed rapid vitrification procedures can be effectively use in 1-cell mouse zygotes cryopreservation.

• Journal of Animal Science and Technology
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• v.45 no.1
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• pp.13-22
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• 2003

Growth Hormone (GH) gene is a member of gene family through the evolutionary process from a small common ancestral gene by a series of gene duplications. The role of the GH in growth and performance controls has been extensively studied in human, mice and livestock. Many researchers have considered GH as a strong candidate gene for evaluation of genetic polymorphisms that could be associated with economic traits in cattle. We report here a novel missense mutation within the exon 5 of the bovine Growth Hormone (bGH) gene. We could amplified 522 bp fragments from eight unrelated Hanwoo cattle by PCR, then, subsequently cloned and sequenced. An Msp I RFLP corresponding to a C to T transition was observed at position 2258 nt. From this result, we could predict a missense mutation (Arg to Trp) at codon 166 in a highly conserved region among many mammals. Codominant Mendelian segregation of the two alleles, Msp I (+) and Msp I (－), was observed in two full-sib F2 families (n = 32, African taurine Bos taurus ${\times}$ African zebu Bos indicus) and eight half-sib Hanwoo families. For the availability of genetic marker, we have performed PCR-RFLP with a large number of individual animals from 15 different cattle breeds (European and Asian taurines, and African indicines). Consideration of breed frequencies of Msp I (－) allele in relation to breed type and their geographic origins, shows higher frequencies in humped breeds or Asian cattle breeds than in humpless or European breeds. This result indicates that the missense mutation can be contributed the functional significance such as the signal transduction through the receptor binding, also may be used as a marker for selection of the economic traits in Hanwoo.

• Development and Reproduction
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• v.3 no.1
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• pp.95-100
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• 1999

Growth differentiation factor-9 (GDF-9) is a member of the transforming growth factor $\beta$ (TGF-$\beta$) superfamily. It has been known that GDF-9 is a growth factor having a crucial role in normal folliculogenesis and its expression is oocyte-specific. The present study was aimed to elucidate the expression of GDF-9 mRNA in the mouse primordial follicles as well as in the other developmental stages. The semiquantitative analysis of GDF-9 mRNA expression was conducted. Total RNA was extracted from the ICR mice ovaries at gestational day 19, postnatal day 1, day 10, day 21, and day 28, and RT-PCR was performed to measure GDF-9 and $\beta$-actin mRNA levels. Level of GDF-9 mRNA were normalized against the level of $\beta$-actin mRNA, and compared among different stages. GDF-9 mRNA was detected in all samples including the fetal ovaries that mainly consists of primordial follicles. The highest level of mRNA was observed in ovaries obtained at day 10 that mainly consists of growing follicles. The present result suggests that GDF-9 may play an important role in the early stage of folliculogenesis.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.23 no.5
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• pp.345-352
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• 1990

This study was investigated on the antitumor of protein-polysaccharide fraction(PPF) extracted from seaweeds such as sea-staghorn and laver toward sarcoma-180 cells. In the PPF extracted from these sewaweeds, the polysaccharide contents of sea-staghorn and laver were $62.26\%$ and $65.78\%$, respectively. The highest levels of polysaccharides found in seaweeds was fructose. The major amino acids were aspartic acid, glutamic acid, glycine and cystein. The solid tumor growth inhibition showed the highest level of $53.30\%$ when 50mg/kg sea-staghorn was administrated. The life prolongation effect was $17.35\%$ at 50 mg/kg of laver. In the effects of immunologic activity, when 100mg/kg sea-staghorn was administrated, the number of circulating leucocyte showed the highest level of $82.23\%$ but decreased leucocyte for prolonged times. The number of total peritoneal exudate cells of the sea-staghorn administerated group was increased significantly in comparison with the control group. The hematobiolgoical analysis of the experimental group was similar with that of the control group. This experiments indicated that hemeastasis still maintained nor-mal state and not showed any harmful effects in normal mice.

• Journal of Life Science
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• v.19 no.11
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• pp.1672-1678
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• 2009

The purified antifungal substances produced by Bacillus amyloliquefaciens IUB158-03 was positive to ninhydrin but negative to aniline, suggesting that the antifungal substance could be a peptide. FAB-MS, UV adsorption spectrum, and amino acid composition analysis revealed that the molecular weight of the antifungal substance was 1042 and that maximal adsorption was at 220 nm and 277 nm. The antifungal substance was composed of $Asn_3$, $Gln_2$, $Ser_1$, $Gly_1$, and $Tyr_1$. The composition and structural characteristics of antifungal substance were analysed by $^1H$-NMR spectrum, $^1H$-COSY, HMQC, which revealed that the compound belongs to the iturin A family. Temperature and pH had little effect on the stability of the antifungal substance in the ranges of $-70{\sim}121^{\circ}C$ and pH 6.0~10.0, respectively. It showed strong antibiotic activity against fungi. An in vitro cytotoxicity test using NIH3T3 cell showed that the antifungal substance does not have cytotoxicity. The number of circulating leukocytes and the hematobiological analysis of the mice administered with the antifungal substances was similar to those of the control group, indicating no cytotoxicity in vivo. Therefore, the antifungal substances extracted from culture broth of Bacillus amyloliquefaciens IUB158-03 have future potential as biocontrol agents against plant diseases caused by fungi.

• Journal of Digital Convergence
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• v.18 no.3
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• pp.267-273
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• 2020

To determine the biological effects of exercise on hippocampus in mice brain exposed to radiofrequency radiaton (RF), the expression of AMPKα, p-AMPKα, ERK1/2, p-ERK1/2, p38, and p-p38 protein in the mouse exposed to RF were investigated in the hippocampal tissues, Western blot method was used to compare the protein expression levels for each molecule. Significant increases in protein expression of individual and phosphorylated molecules were observed in the spontaneous exercise group, and the expression of these molecules was notably decreased in the RF exposure and spontaneous exercise group. This study shows that neuroplasticity can be increased by exercise in hippocampus that is responsible for memory, but memory and cognitive function may be affected by exposure to RF. We may expect clinically interesting results on dementia or Alzheimer disease if we proceed further investigation on the effect of RF.

• Clinical and Experimental Reproductive Medicine
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• v.26 no.1
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• pp.103-109
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• 1999

Objective: Heat shock protein 70-2 (Hsp70-2) gene knockout mice are found to have premeiotic arrest at the primary spermatocyte stage with a complete absence of spermatids and spermatozoa. This observation led to the hypothesis that hspA2 may be disrupted in human testes with abnormal spermatogenesis. To test this hypothesis, we studied the mRNA expression of hspA2 in infertile men with azoospermia. Design: The mRNA expression were analyzed by competitive RT-PCR among testes with normal spermatogenesis, pachytene spermatocyte arrest, and sertoli-cell only syndrome. Materials and methods: Testicular biopsy was performed in men with azoospermia (n=15). Specimens were subdivided into three groups: (group 1) normal spermatogenesis (n=5), (group 2) spermatocyte arrest (n=5), (group 3) Sertoli-cell only syndrome (n=5). Total RNA was extracted by Trizol reagent. Total extracted RNA was reverse transcribed into cDNA and amplified by PCR using specific primers for hspA2 target cDNAs. A competitive cDNA fragment was constructed by deleting a defined fragment from the target cDNA sequence, and then coamplified with the target cDNA for competitive PCR. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene was used as an internal control. Results: On Competitive RT-PCR analyses for hspA2 mRNA, significant amount of hspA2 expression was observed in group 1, whereas a constitutively low level of hspA2 was expressed in groups 2 and 3. Conclusion(s): The study demonstrates that the hspA2 gene expression is down-regulated in human testes with abnormal spermatogenesis, which in turn suggests that hspA2 gene may play a specific role during meiosis in human testes.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.18 no.1
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• pp.81-98
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• 1992

Hairless mice (Skh:HR-1) exposed to single doses (0.5, 1.0 and 3. OMED) of UV-B radiation were displayed remarkable changes of barrier function and surface morphology. Trans- epidermal water loss (TEWL) as an index of barrier function was measured by evaporimeter, and wrinkle density (WD) as an index of morphological alteration was measured by image analyzer. Significant changes of TEWL were not observed in the control and 0. SMED group, but 1.OMED and 3. OMED groups noted significant difference. TEWL of 3. OMED group was rapidly increased to the 3rd day and decreased until the 14th day when it reached nearly to normal level, Time-courses of TBWL for 1. OMEB and 3. OMED groups displayed similar pattern, but different only in the magnitude. WD were significantly decreased during the 3rd-5th day in all of the irradiated groups and then increased during the last period to the 14th day, but did not recover the normal level at the 14th day. Time-courses of WD for all groups exhibited similarity, and were entirely dependent on the exposed doses. We also observed histological changes which included hyperplasia, sunburn cell (SBC) formation, accumulation of polymorphonuclear leukocyte (PMNs), and loss of collagen of UVB- exposed hairless mouse skin. Changes of TEWL and WD are helpful in understanding of epidermal and dermal damages by single exposure of UVB.

• The Korean Journal of Zoology
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• v.18 no.4
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• pp.163-172
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• 1975

This work was undertaken to elucidate some aspects of mechanisms underlying the increased deoxycytidineuria following irradiation in the mice, by observing Dische-positive substances liberated from thissues, the activity of CdR-aminohydrolase of tissues and the CdR excreted in the urine at various times after single whole-body exposure to 400 and 800 R of X-rays. The activity of CdR-aminohydrolase declined markedly at 1 hour in the small intestine and liver, followed by a gradual rise reaching a maximum at 3 days after irradiation. In the case of the spleen and blood, however, only a trace of activity was observed in the control and irradiated animals. The amount of Dische-positive substance liberated from the small intestine postirradiation was elevated from 3 to 12 hours, showing a maximum during 6 to 9 hours after irradiation. On the contrary, the activity of the enzyme in the liver, spleen and kidney was less than one twentieth that of the small intestine, suggesting a prediction that these organs are not attributable to the increased deoxycytidineuria. A maximum deoxycytidineuria was exhibited at 9-12 hours period, attributed a large amount of CdR to the small intestine, which might correlate with the change in the CdR-aminohydrolase activity. Radiation-induced CdR seems to be liberated from the small intestine into the blood when the CdR-aminohydrolase activity declines abruptly. Then, the CdR is rapidly subjected to a filtration in the kidney without undergoing a further degradation pathway in the blood.