• Clinical and Experimental Reproductive Medicine
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• v.30 no.3
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• pp.207-215
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• 2003

Objective: To compare the pregnancy outcomes after in vitro fertilization with intracytoplasmic sperm injection (IVF-ICSI) between obstrucvtive and non-obstrucvtive azoospermia. Methods: From January 1994 to December 2002, 524 patients with obstructive azoospermia (886 cycles) and 163 patients with non-obstructive azoospermia (277 cycles) were included in this study. Microsurgical epididymal sperm aspiration (MESA) or testicular sperm extraction (TESE) in obstructive azoospermia and TESE in non-obstructive azoospermia were perfomed to retrieve sperm, which was used for ICSI and then fertilized embryos were transferred. The results of ICSI - fertlization rate (FR), clinical pregnancy rate (CPR), clinical abortion rate (CAR) and delivery rate (DR) - were statistically analysed in obstructive versus non-obstructive azoospermia. Results: There were no differences in the number of retrieved oocytes, injected oocytes for ICSI and oocyte maturation rate. FR was significantly higher in obstructive than non-obstructive azoospermia (71.7% vs. 61.1%, p<0.001). There was no difference in CPR per embryo transfer cycle. After pregnancy was established, however, CAR was significantly higher in non-obstructive than obstructive azoospermia (25.6% vs. 12.5%, p=0.004). DR per clinical pregnancy cycle was significantly higher in obstructive than non-obstructive azoospermia (78.0% vs. 64.4%, p=0.012). In the karyotype ananlysis of abortus, abnormal karyotypes were found in 75.0% (6/8) of obstructive and 55.6% (5/9) of non-obstructive azoospermia. Conclusion: Our data show significantly higher FR in obstructive than non-obstructive azoospermia. Though there was no differrence in CPR, CAR was significantly higher in non-obstructive than obstructive azoospermia. The abortion may be related to the abnormal karyotype of embryo, but further investigations are necessary to elucidate the cause of clinical abortion in azoospermia.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.17 no.3
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• pp.206-218
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• 1984

The gonadal development and gametogenesis of shad, Konosirus, punctatus (TEMMINCK et SCHLEGEL) were studied by comparing with various quantitative indices, such as seasonal changes of gonadosomatic index, fatness, egg-diameter composition, first maturing size, and by comparing with histological changes of gonad and gonadotrophs(GTH) in pituitary. The materials were monthly sampled from Dadaepo at the estuary of the Nakdong river in Korea from September, 1982 to October, 1983. The ovary of shad is a pair of sac-shaped organs revered with a fibromuscular capsule and consisting of numerous sacs. The type of testicular structure is lobular type with development of germ cells, mesenchymal tissue on the lobuli. The gonadosomatic index (GSI) is rather low till March, but increases in April and reaches to peak in June in females and May in males. And it suddenly falls in July. The gonads become active on the increase of water temperature and spawning season ends before high water temperature. After spawning, the small oocytes continue to remain as they are untill the growing period next year. The reproductive cycle includes the successive stages of growing from March to April, mature from April to May, ripe and spawning in June, and recovery and resting from July to February next year. In egg-diameter composition of an ovary taken in the spawning season, 2-3 modes were recognized with some batches shown in an ovary. An individual shad spawns twice or more in a month-spawning season. The individual spawning interval is estimated to be ten days or less. Changes of fatness are corelated with those of water temperature that affect on the condition of feeding, but less corelated with spawning. The percentage of mature of female and male fish, are $50\%$ in 17.0-18.0 cm and $100\%$ in 18.0-19.0 cm. GTH cells are activated from growing period and decrease their activity at pre-spawning season with peak activity for mature period.

• The Korean Journal of Malacology
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• v.17 no.1
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• pp.45-55
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• 2001

Gametogenes, reproductive cycle, first sexual maturity(biological minimum size), sex ratio and larval development of the marsh clam Corbicula japonica were investigated monthly by histological observations. Samples were collected in brackish water of Gokgang stream, Kyungsangbuk-Do, Korea, from August 1997 to July 1998. Sexuality of Corbicula japonica is dioecious and the species are an oviparous clam. The gonads are irregularly arranged from the sub-region of mid-intestinal gland in visceral cavity to reticular connective tissue of foot. The ovary is composed of a number of ovarian sac which are branched arborescent. Oogonia actively proliferate along the germinal epithelium of ovarian sac, in which young oocytes are growing. The testis is composed of a number of testicular tubules, and the epithelium of the tubule has function of germinal epithelium, along which spermatogonia actively proliferate. A great number of undifferentiated mesenchymal tissue and eosinophilic granular cells are abundantly distributed between developing oocytes and spermatocytes in the early developmental stages. With the further development of the ovary and testis these tissue and cells gradually disappear. Then the undifferentiated mesenchymal tissue and eosinophilic granular cells are considered to be related to the growing of the oocytes and spermatocytes. The spawning period is from July to September, and the main spawning occur between July and August when seawater temperatures reach above 22$^{\circ}C$. The reproductive cycle of this species can be divided into five successive stages; early active (February to April), late active (May to July), ripe (June to September), partially spawned (July to September), degenerative (September to October) and resting stage (October to February). Percentages of first sexual maturity of female and male clams ranging in length from 10 mm to 12 mm are over 50% and 100% for clams over 16.0 mm in shell length. Fertilized eggs or Corbicula japonica were 80-90 ${\mu}{\textrm}{m}$ in diameter. In the early embryonic development of C. japonica, the appearance of polar body, trochophore and D-shaped veliger were observed around 40 min., 27 hours and 4 days after spawning, respectively, at a water temperature of 26.5-28.$0^{\circ}C$. The size of larvae of early umbo stage was about 185-210 ${\mu}{\textrm}{m}$ in shell length, 160-180 ${\mu}{\textrm}{m}$ in shell height around 7 days after fertilization. The correlation of relative growth between the culture day (D) and shell length (SL) was expressed by the following simple formula from D-shaped veliger to metamorphosing stage; SL = 13.300D + 209.36($r^2$= 0.9078).

• Journal of the Korean Society of Radiology
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• v.12 no.1
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• pp.9-16
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• 2018

Hangover after drinking is different from person to person symptoms and degree, but usually thirst, fatigue, headache, general boredom, gastrointestinal disorder, vomiting, diarrhea, deficiency of vitamin appears. This hanging phenomenon is caused by the action of precursors such as ethyl acetate and acetaldehyde, which are the by products of fermentation contained in alcohol and alcohol accumulated in hepatocytes and body. In order to solve the hangover phenomenon, the same Origin as polysaccharide Polydeoxyribonucleotide, which is a nucleic acid-sugar-phosphate complex, which is a semen or testicular extract in salmon extract, and a water soluble salmon extract powder having the same structure and lower price than Polydeoxyribonucleotide And D-Glucuronic acid and N-Acetyl glucosamine. It has excellent biocompatibility, viscoelasticity and moisturizing power. It has effect on reduction of body water loss and skin moisture content in hangover phenomenon. It is antioxidant and skin moisturizing effect Hyaluronic acid was irradiated with gamma rays, and the composition was prepared by using the salmon extract powder and the main raw material. The ethanol degradation, the acetaldehyde reduction amount, the blood acetaldehyde concentration and the acetic acid concentration were measured to evaluate the alcoholysis effect, Skin moisture evaporation rate To examine the evaluation unit water content of the skin was improved determine whether the antioxidant and provide skin moisturizing effect. The addition of ethanol extracts of salmon extracts showed a decrease of 5 to 7 times compared with no addition, and a decrease of 3 to 5 times of acetaldehyde. In addition, the change of acetaldehyde concentration and acetic acid concentration in blood showed a rapid decrease compared to the no - added control group. In addition, when the raw material of hyaluronic acid was used, skin moisture content was high and skin moisture evaporation amount was decreased. Therefore, hyaluronic acid, which is a polysaccharide polymer, has excellent viscoelasticity and moisturizing ability, It is considered to provide antioxidant and skin moisturizing effect. Therefore, it can be said that the composition containing salmon extract powder and hyaluronic acid as a main ingredient is effective for the hangover phenomenon which occurs after drinking.

• Journal of Aquaculture
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• v.20 no.4
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• pp.219-225
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• 2007

This study investigated the correlationship between artificial maturation season and reproduction coefficient of cultured eel Anguilla japonica from May (spring) to next January (winter). The brood stock, female eels ($400{\sim}600\;g$) were artificially matured by weekly intramuscular injections of salmon pituitary extracts (SPE, 20 mg/fish) to induce a completion of vitellogenesis. After completion of vitellogenesis, final oocyte maturation and ovulation was induced by injection of $17{\alpha}$, $20{\beta}-dihydroxyprogesterone$ (DHP) at about $2\;{\mu}g/g$ body weight. Most fish ovulated $15{\sim}18\;h$ following the DHP injection. The ovulated fish were induced to natural spawning or artificial fertilization by the dry method. Males ($200{\sim}350\;g$) were received weekly intramuscular injections of human chorionic gonadotropin (HCG) at a dosage of 1 IU/g body weight to induce testicular maturation and spermiation. Seasonal reproduction coefficient which includes the rate of ovulation, buoyancy, fertilization and hatching of eggs in the artificially matured eel during spring to summer ($May{\sim}July$) were significantly higher than the other season, while there were no significant difference among spring and summer (P<0.05). Furthermore, the number of eggs spawned and larvae hatched in the artificially maturated eel during spring to summer ($May{\sim}July$) were significantly higher than the other season, while there were no significant difference in spring and summer (P<0.05). These results indicate that artificial maturation by hormone treatment of A. japonica was successful only during spring to summer, which is the maturation period in the wild stock in nature. Consequently, it is possible to determine the period of artificially induced sexual maturity by the reproduction coefficient which includes the rate of ovulation, buoyancy, fertilization and hatching of eggs in the cultured eel A. japonica.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.22 no.5
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• pp.266-280
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• 1989

Gonadal maturation of the Korean pomfrets, Pampus echinogaster (Basilewsky) and Pampus argenteus (Euphrasen) were histologically investigated based on the samples captured in the East China Sea from January 1987 to December 1988. Gonadosomatic index (GSI) of P. echinogaster began to increase from March, and reached maximum between May and July. It began to decrease from July and reached mini-mum between August and February. P. argenteus had a similar cycle, however, P. argenteus has higher values in April than P. echinogaster. Hepatosomatic index (HSI) were positively related to GSI. HIS of P. echinogaster and P. argenteus reached maximum in $April\~July$ and $April\~August$, respectively, Fatness coefficient of two Pampus species were low in the summer, and high in the winter. Ovary is of saccular structure, and testis is of lobular structure. From February, the early oocyte (ca. $100\mu$ in diameter grows) rapidly at the germinal epithelium of ovarian sacs. From March to April the oocytes grew up to cu $400\~500\mu$ in diameter. At this stage, the yolk globules are accumulated rapidly in the cytoplasmic layer. From May, the oocytes roached ca. $650\~850\mu$ in diameter, and they are spawned in $May\~July$. After spawning the residual follicles and remained ripe eggs degenerate. From February, spermatogonia grows into spermatocyte on the epithelium of the testicular lobuli. From May, spermatozoa appeared and spawning occurs. After spawning, the epithelium is thickened and the remained spermatozoa degenerate. Annual reproductive cycle of two Pampus species could be divided into four successive stages: Growing stage ($March\~April$), Mature stage ($April\~May$), Ripe and spent stage ($June\~July$) and Recovery and resting stage ($August\~January$). Absolute fecundity of P. echinogaster was $9,441\~135,294$, and that of P. argenteus was $50,678\~221,894$. Absolute fecundity of two Pampus species were positively related to body length and total weight. Relative fecundity was positively related to body length, while it was reversely related to total weight. The increasing rate of absolute fecundity of P. echinogaster was lower than P. argenteus. In P. echinogaster half of female and male reached first maturity at body length of $15.0\~$17.9cm and $12.0\~14.9cm$, respectively. All of females and males reached first maturity at body length of $18.0\~20.9cm$ and $21.0\~23.9cm, respectively. In P. argenteus all of females and males reached first maturity at body length of 18.6cm and 16.7cm$, respectively.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.29 no.1
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• pp.35-43
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• 1996

The appearance of the primordial germ cells (PGC's), early gonadogenesis, sex differentiation and sex ratio of the embryo in the viviparous teleost, Ditrema temmincki were investigated by using photomicroscopy. The PGC's were first observed in the fibrous mesenchymal tissue located between the early alimentary tract and the dorsal body wall in the late embryonic development stage. During the period from the hatching to the individual total length (TL) of 4.0 mm the PGC's were evenly distributed in the fibrous mesenchymal tissue between alimentary tract and body wall. But the period of TL 5.0 mm mesenchymal tissue separated from the dorsal body wall, the PGC's moved to the posterior mesenchymal tissue and formed the primitive gonad. During the early gonadogenesis, differentiation of the testis and ovary were distinguished from the arrangement of the germ cells and somatic cells. Gonad of embryo in TL 10.0 mm can be separated into the ovary and testis by external morphology. The testis had a separated form which was consisted with two lobes, and the ovary had a fused form in half-posterior part. In the testicular differentiation of the embryo, histological pattern of the seminiferous tubule appeared when TL of the embryo was to be 25.0 mm, for the seminal vesicle was formed In the individual TL of 30.0mm. The testis of the embryo with TL of 45.0 mm was similar to that of the adult fish in the external and internal structures. In the ovarian differentiation, formation of the ovigerous folds and the ovarian cavity were clearly observed when the TL reached to 30.0 mm. The ovary from the individual with TL of 60.0 mm was differentiated into a similar ovary as seen in the adult fish in the external and internal structure. Right before parturition the total length of the individual was approximately 63.0 mm of which the individual embryo has an ovary containing the oocytes in the chromatin nucleolus stage, or a testis containing the spermatogonia, respectively. And the embryonic sex ratio of female to male was 1.65 : 1. Ditrema temmincki is dioecism and the pattern of sex differentiation is belonged to a differentiation type.

• Korean Journal of Animal Reproduction
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• v.24 no.4
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• pp.357-364
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• 2000

Members of the glycoprotein family, which includes CG, LH, FSH and TSH, comprise two noncovalently linked $\alpha$- and $\beta$-subunits. Equine chorionic gonadotropin (eCG), known as PMSG, has a number of interesting and unique characteristics since it appears to be a single molecule that possesses both LH- and FSH-like activities in other species than the horse. This dual activity of eCG in heterologous species is of fundamental interest to the study of the structure-function relationships of gonadotropins and their receptors. CG and LH $\beta$ genes are different in primates. In horse, however, a single gene encodes both eCG and eLH $\beta$ -subunits. The subunit mRNA levels seem to be independently regulated and their imbalance may account for differences in the quantities of $\alpha$ - and $\beta$-subunits in the placenta and pituitary. The dual activities of eCG could be separated by removal of the N-linked oligosaccharide on the $\alpha$-subunit Asn 56 or CTP-associated O-linked oligosaccharides. The tethered-eCG was efficiently secreted and showed similar LH-like activity to the dimeric eCG. Interestingly, the FSH-like activity of the tethered-eCG was increased markedly in comparison with the native and wild type eCG. These results also suggest that this molecular can implay particular models of FSH-like activity not LH-like activity in the eCG/indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion. A single-chain analog can also be constructed to include additional hormone-specific bioactive generating potentially efficacious compounds that have only FSH-like activity. The LH/CG receptor (LH/CGR), a membrane glycoprotein that is present on testicular Leydig cells and ovarian theca, granulosa, luteal, and interstitial cells, plays a pivotal role in the regulation of gonadal development and function in males as well as in nonpregnant and pregnant females. The LH/CGR is a member of the family of G protein-coupled receptors and its structure is predicted to of a large extracellular domain connected to a bundle of seven membrane-spanning a-helices. The LH/CGR phosphorylation can be induced with a phorbol ester, but not with a calcium ionophore. The truncated form of LHR also was down-regulated normally in response to hCG stimulation. In contrast, the cell lines expressing LHR-t631 or LHR-628, the two phosphorylation-negative receptor mutant, showed a delay in the early phase of hCG-induced desensitization, a complete loss of PMA-induced desensitization, and an increase in the rate of hCG-induced receptor down-regulation. These results clearly show that residues 632~653 in the C-terminal tail of the LHR are involved in PMA-induced desensitization, hCG-induced desensitization, and hCG-induced down-regulation. Recently, constitutively activating mutations of the receptor have been identified that are associated with familial male-precocious puberty. Cells expressing LHR-D556Y bind hCG with normal affinity, exhibit a 25-fold increase in basal cAMP and respond to hCG with a normal increase in cAMP accumulation. This mutation enhances the internalization of the free and agoinst-occupied receptors ~2- and ~17- fold, respectively. We conclude that the state of activation of the LHR can modulate its basal and/or agonist-stimulated internalization. Since the internalization of hCG is involved in the termination of hCG actions, we suggest that the lack of responsiveness detected in cells expressing LHR-L435R is due to the fast rate of internalization of the bound hCG. This statement is supported by the finding that hCG responsiveness is restored when the cells are lysed and signal transduction is measured in a subcellular fraction (membranes) that cannot internalize the bound hormone.

• Toxicological Research
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• v.33 no.2
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• pp.107-118
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• 2017

Although alternative test methods based on the 3Rs (Replacement, Reduction, Refinement) are being developed to replace animal testing in reproductive and developmental toxicology, they are still in an early stage. Consequently, we aimed to develop alternative test methods in male animals using mouse spermatogonial stem cells (mSSCs). Here, we modified the OECD TG 489 and optimized the in vitro comet assay in our previous study. This study aimed to verify the validity of in vitro tests involving mSSCs by comparing their results with those of in vivo tests using C57BL/6 mice by gavage. We selected hydroxyurea (HU), which is known to chemically induce male reproductive toxicity. The 50% inhibitory concentration ($IC_{50}$) value of HU was 0.9 mM, as determined by the MTT assay. In the in vitro comet assay, % tail DNA and Olive tail moment (OTM) after HU administration increased significantly, compared to the control. Annexin V, PI staining and TUNEL assays showed that HU caused apoptosis in mSSCs. In order to compare in vitro tests with in vivo tests, the same substances were administered to male C57BL/6 mice. Reproductive toxicity was observed at 25, 50, 100, and 200 mg/kg/day as measured by clinical measures of reduction in sperm motility and testicular weight. The comet assay, DCFH-DA assay, H&E staining, and TUNEL assay were also performed. The results of the test with C57BL/6 mice were similar to those with mSSCs for HU treatment. Finally, linear regression analysis showed a strong positive correlation between results of in vitro tests and those of in vivo. In conclusion, the present study is the first to demonstrate the effect of HU-induced DNA damage, ROS formation, and apoptosis in mSSCs. Further, the results of the current study suggest that mSSCs could be a useful model to predict male reproductive toxicity.

• Korean Journal of Veterinary Research
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• v.45 no.3
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• pp.325-334
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• 2005

Changes in the rabbit Leydig cell from birth to adulthood were studied in New Zealand white rabbits of 1, 7, 21, 35, 49, 70, 105, 147, 196, and 252 days (n = 8 rabbits per group) of age. The objectives of this study were to understand the fate of the fetal Leydig cells, to determine the changes in serum testosterone levels, and leutenizing hormone-stimulated testosterone production per testis in vitro, and to quantify adult Leydig cells by number and average volume with age. Testes of rabbits were fixed by whole body perfusion using a fixative containing 2.5% glutaraldehyde in cacodylate buffer, processed and embedded in Epon-araldite. Using $1{\mu}m$ sections stained with methylene blue-azure II, qualitative and quantitative (stereological) morphological studies were performed. Testosterone levels in the incubation medium of luteinizing hormone-stimulated (100 ng/ml) testosterone secretion per testis in vitro, and in serum were determined by radioimmunoassay. The average volume of a testis of 1-day-old rabbits was determined as $0.0073cm^3$ and the parameter increased linearly from birth to 252 days ($3.93cm^3$). The volume density of the seminiferous tubules increased with age from 33.76% at day 1 to 88.2% at day 252. The volume density of the interstitium represents 66.24% of the testicular parenchyma at day 1. This proportion progressively diminished during development to reach a value of 11.8% at day 252. The volume density of Leydig cells increased almost linearly from birth (0.001%) to 252 days (2.62%). Leydig cell mass per testis increases from 0.0012 mg to 0.25 mg between days 1 and 35, from 2.66 mg to 44.3 mg between days 49 and 105 and from 65.42 mg and 102.9 mg between days 147 and 252. The absolute numbers of adult Leydig cells per testis increased linearly from birth to 252 days. The average volume of adult Leydig cell on days 1, 7, 21 and 35 was not significantly different; a gradual and continued increase was observed thereafter, reaching a 3-fold increase at 196 and 252 days. Serum testosterone concentrations were not significantly different at day 1 compared days 7, 21, 35. Significant increases were observed at days 49 and 70. Values at days 70 and 105 and days 147, 196, and 252 were not significantly different. LH-stimulated testosterone production per testis in vitro was significantly different at day 1 compared days 7, 21, 35. Significant increases were observed at days 49 and 70. Hormonal values at days 105, 147, 196, and 252 were not significantly different. These data suggested Leydig cell developmental phase can be classified: a neonatal phase (1-7 days), a prepubertal phase (14-49 days) and an adult phase (70-252 days). Immature and mature adult Leydig cells, initially detected at days 7 and 49, respectively, and mature adult Leydig cells were abundant Leydig cell type according to the number and absolute volume per testis form day 49 onwards.