• Korean Journal of Environmental Biology
• /
• v.20 no.3
• /
• pp.224-231
• /
• 2002

Photoperiod (length of light per day) is a major factor in regulating reproductive function in golden hamsters. The information of photoperiod is transmitted to the reproductive endocrine system by melatonin. Thus the effects of melatonin aye investigated in male golden hamsters exposed to photoperiods. Paired testicular weights were markedly reduced in the animals housed in short photoperiod $(SP,\le{12\;hours\;day^{-1})$ and injected with melatonin in the evening, but not in long photoperiod $(LP,\le{12.5}\;hours\;day^{-1})$ and injected with melatonin in the morning. The histological examination of regressed testes showed reduction of tubular lumen diameter including the numbers of cells and Leydig cell number. The mean values of both follicle stimulating hormone (FSH) and luteinizing hormone (LH) were also lowered in the sexually inactive animals than in the sexually active animals. Melatonin receptor was identified by reverse-transcription polymerase chain reaction (RT-PCR) and its expression was examined in various tissues to scrutinize the action site of melatonin. It turned out 309 nucleotides and was definitely expressed in hypothalamus and pituitary including spleen, retina, and epididymis. And gonadotropin releasing hormone (GnRH) gene, which is a key element in regulating reproduction, was identified by RT-PCR but the expression of GnRH was not modified by the treatment of melatonin. Taken together, photoperiod via melatonin indirectly affects reproductive endocrine system, possibly through the release of GnRH, not the synthesis of GnRH.

• Proceedings of the KSAR Conference
• /
• 2003.06a
• /
• pp.67-67
• /
• 2003

Early development of porcine oocytes fertilized in vitro was examined in different culture conditions. Porcine ovaries were collected from local slaughter-house. 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 cystein, 10% porcine follicle fluid, 10 IU/ml PMSG and 10 IU/ml hCG for 42~44 hrs. The frozen-thawed spermatozoa were washed by centrifigation 2 times at 1, 500 rpm in D-PBS with 5.56 mM glucose, 0.33 mM Na-pyruvate, 100 IU/ml penicillin, 1$\mu\textrm{g}$/ml streptomycin and 1ng/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 8hrs h after insemination, the oocytes were transferred into NCSU-23 medium with 5.0 mM hypotaurine and 4 mg/ml BSA and cultured for 7 days. In first experiment, the mean numbers of oocytes collected from 20 ovaries were 674.4 oocytes, and 4.1(27.6), 12.5(84.0), 25.4(171.6) and 57.9%(390.8) for A, B, C and D grade in morphological classification. In the second experiment, when culture medium was supplemented with various concentrations of EGF, the proportions of oocytes cleaved were 56.9, 55.7, 61.9 and 54.7% for 0, 5, 10 and 20ng/ml EGF. The higher proportions(15.1%) of oocytes developed to morular stage were obtained at concentration of 10ng/ml than 0 and 5ng/ml EGF (P<0.05). However, the proportions of embryos developed to blastocyst stage were not significantly different among concentrations of EGF. In another experiment, when the medium supplemented with catalase was used, the proportions of oocytes cleaved were higher in the concentration of 0 unit (56.5%, 61/108) than 100 and 1, 000 unit/ml of catalase (P<0.05). Although the developmental capacity of embryos was improved by medium with 0 unit/ml compared with 100, 500 and 1, 000 units/ml of catalase in oocytes developed to morula and blastocyst stages, were not significantly different among concentrations of catalase.

• Journal of Pharmacopuncture
• /
• v.18 no.4
• /
• pp.20-25
• /
• 2015

Objectives: Recently, thread-embedding therapy (TET) has been widely applied in Korean medicine for cosmetic purposes such as reducing skin wrinkles. An inserted thread was reported to have induced continuous stimulation, followed by support for connective tissue regeneration. However, the potential role of TET in hair-growth has not yet been reported. Methods: We designed this study to evaluate whether TET has a hair-growth-promoting effect. C57 black 6 (C57BL/6) mice were divided into three groups: normal saline-treated, minoxidil-treated, and thread-embedded groups. Normal saline or 5% minoxidil was topically sprayed on the dorsal skin of the mice once a day for 16 days. Medical threads were embedded into the dorsal skin of the mice in a single application. Hair growth activity was evaluated by using dermoscopic and microscopic observations. Sections of the dorsal skin were stained with hematoxylin and eosin. Expressions of bromodeoxyuridine (BrdU), proliferating cell nuclear antigen (PCNA), fibroblast growth factor-7 (FGF-7), and fibroblast growth factor-5 (FGF-5) were detected by using immunohistochemical staining. A reverse transcription-polymerase chain reaction (RT-PCR) analysis was adopted to measure the messenger RNA (mRNA) expressions of FGF-7 and FGF-5. Results: TET enhanced anagen development in the hair follicles of C57BL/6 mice. The expressions of BrdU and PCNA, both of which imply active cellular proliferation, were increased by using TET. Moreover, TET increased the expression of FGF-7, an anagen-inducing growth factor, while decreasing the expression of FGF-5, an anagen-cessation growth factor, both at the protein and the mRNA levels. Conclusion: TET enhanced hair re-growth in C57BL/6 mice. TET regulated the expressions of anagen-associated growth factors and activated the proliferation of hair follicular cells in depilated skin lesions. Considering its long-lasting effect, TET may be a good alternative therapeutic for the treatment of alopecia.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.25 no.5
• /
• pp.432-442
• /
• 1992

To identify the histological site of synthesis of yolk protein precursor, vitellogenin, by immunocytochemical method in the freshwater crab Eriocheir japonicus, we purified the yolk protein, vitellin, from crude egg extracts, and prepared the anti-rabbit serum against vitellin. Then, the site of vitellogenin synthesis was demonstrated by immunotytochemical method with PAP(peroxidase-antiperoxidase) reaction using the rabbit antiserum aganist vitellin. Female specific serum protein was identified in female serum by immunoelectrophoresis and Ouchterlony's immunodiffusion test for mature male and female sera. Based on the immunoelectrophoresis and Ouchterlony's diffusion test for mature male and female sera and crude egg extracts using antiserum against vitellogenic female serum absorbed with male serum, the female specific serum protein was identified as vitellogenin, detected in female serum only. The major yolk protein, vitellin, was purified from the crude egg extracts by DEAE-cellulose ion exchange chromatography, followed by sepharose CL-4B gel filteration chromatography. The molecular weight of vitellin was estimated to be about 245,000 dalton by sepharose CL-4B gel filteration chromatography. from the results of immunological analysis for vitellin, it was found that the vitellin antiserum contained the antibody against vitellogenin. In the results of immunocytochemical reaction by PAP method with the rabbit antiserum against vitellin, the vitellogenic oocytes and the hepatopancreas of mature female showed positive PAP reaction, but not in follicle cells and previtellogenic oocytes nf ovary, muscle of female and mature male hepatopancreas. Therefore, it showed that the hepatopancreas of mature female is the site of vitellogenic synthesis.

• The Korean Journal of Malacology
• /
• v.25 no.2
• /
• pp.145-151
• /
• 2009

The ultrastructural changes in germ cells during oogenesis of the melania snail, Semisulcospira libertina libertina have been investigated by light and electron microscopy. The ovary is located on the surface of the hepatopancreas in the spiral posterior region. The ovary exhibited greenish color in the gonadal mature season. The ovary was composed of a number of oogenic follicles. Oogenesis was divided into five stages with histological features: (1) oogonia, (2) previtellogenic, (3) initial vitellogenic, (4) active vitellogenic, and (5) mature stages. Oogonia were oval in shape, $4-6\;{\mu}m$ in diameter, and had a large nucleus. Previtellogenic oocytes were about $20\;{\mu}m$ in diameter and the cytoplasm reacted with hematoxylin in H-E satin. Initial vitellogenic stage, oocytes were $60-80\;{\mu}m$ in diameter, and small yolk granules of low electron density are scattered in the cytoplasm. Oocytes in the initial vitellogenic stage were connected with ovarian follicle by egg stalk. Active vitellogenic oocyte were $100-120\;{\mu}m$ in diameter. Electron density, size and quantity of yolk granules that are distributed in the cytoplasm have increased from the previous stage. Result of TEM observations, the oocyte contains well-developed Golgi complex, endoplasmic reticula and tubular mitochondria in the cytoplasm. Cytoplasm of mature oocyte was filled with proteinaceous yolk globules of high electron density. In this stage, the length of microvilli in the egg envelope was approximately $1.1\;{\mu}m$.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.22 no.2
• /
• pp.357-364
• /
• 2008

Mylabris is the dried body of the chinese blister beetle. The species used in medicine are Mylabris phalerata and M. cichorii. In recent studies, it has been found that Mylabris possesses antitumor properties, increases the number of leukocytes, and has irritant effects on the urinary organs. The crude extracts of Mylabris have been noted for their highly irritant action and other traditional uses of Mylabris include treatment of poor local blood circulation. The active constituent of Mylabris is cantharidin. The chemical is notable for its vesicant properties, but with severe side effects such as nephrotoxicity. This experiment examined the effect of extracts and fractions, obtained from Mylabris phalerata Pall. on hair growth activity of the C57BL/6N mice after topical application to skin. First, we examined the effect of an extracts, obtained from the alcohol extracts of dried Mylabris phalerata Pall. on hair growth activity of the C57BL/6N mice after topical application to skin. Second, we examined on hair growth activity of the cantharidin fraction of Mylabris phalerata Pall. compared to the control and 1% minoxidil groups. Third, we investigated the number of hair follicle and mast cells after topical application of extracts of Mylabris phalerata Pall. to skin for 16 days. The results were as follows: Hair growth effect from the extracts of Mylabris phalerata Pall.(0.312%) was observed in 80% of mice whose hair had been removed in 13 days. Hair growth effect from the extract of Mylabris phalerata Pall.(0.312 and 0.625%) and 1% minoxidil group was observed in 100% of mice whose hair had been clipped in 20 days. Hair growth effect from the cantharidin fraction(0.5%) and water fraction(0.5%) of Mylabris phalerata Pall. was observed in 100% of mice whose hair had been clipped in 24 days. The hair growth effect on the cantharidin fraction(0.125%) was observed to be strong compared with the minoxidil(3%) group, commercial hair growth agents, in mice whose hair had been clipped in 19 days. In the spontaneous alopecia mice model, the hair growth effect from the cantharidin fraction (0.125%) was observed to be strong as compared with the states before the 13 days experiment. These experiments suggest that extracts and fractions of Mylabris phalerata Pall. may stimulate the topical hair growth activity in low doses.

• Journal of Life Science
• /
• v.28 no.5
• /
• pp.532-539
• /
• 2018

Hair graying is processed by loss of melanin production caused by the decrease of activity and number of melanocyte and the accumulation of hydrogen peroxide ($H_2O_2$) in the hair follicle with increase of age. The purpose of this study was to investigate the effect the Black oryzasativa ethanolic extract (BLEE) on the melanin production. In this study BLEE at $8{\mu}g/ml$ or more showed a significant 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity and reduction power. BLEE at $16{\mu}g/ml$ or more showed promoted tyrosinase activity and melanin production. In addition BLEE scavenged intracellular $H_2O_2$ in 2',7'-dichlorodihydrofluorescein (DCF) fluorescence assay in B16F1 cells. However, Western blot analyses displayed that BLEE decreased the expression level of catalase, but no effect on the expression level of tyrosinase, tyrosinase associated protein-1 (TRP-1), tyrosinase associated protein-2 (TRP-2) and microphthalmia-associated transcription factor (MITF) transcription factor involved in melanogenesis. Thus, the promotive effect of BLEE on melanin production is attributed to the increase of tyrosinase activity and the reduction of intracellular $H_2O_2$ level. In conclusion, BLEE played a key role in in promoting melanin production, which suggests that the BLEE could be applied as a potential functional material in the development of hair care cosmetics related to the promotion of melanin production for the growth of black hair.

• The Korean Journal of Malacology
• /
• v.20 no.1
• /
• pp.55-63
• /
• 2004

Gonadosomatic index, condition index and reproductive cycle with the gonadal development of the female Octopus ocellatus were investigated by histological observations and morphometric data, from January to December, 2000. And changes in biochemical components of the ovary and the trunk tissues including the digestive organ associated with gonadal development were studied by biochemical analysis from January to October, 2001. The specimens were collected at the coastal waters of Buan, Jeollabuk-do, Korea, from January 2000 to October 2001. O. ocellatus is a dioecious organism. The gonad of O. ocellatus locates medially in posterior region of the body. Morphology of the ovary shows round and oval in shape, the average diameter and external colour of ripe ovary was 32 mm and semitransparent light brown in colour. As the ovary was getting mature, transparent elongated eggs covered with chorion were present in the ovarian cavity. Monthly changes in the gonadosomatic index (GSI) showed a similar pattern with those of the condition index. The GSI and condition index began to increase in March and reached the maximum in April. And then, their values decreased from May and reached the minimum in September. Reproductive cycle of O. ocellatus can be categorized into five successive stages: early developing stage (September to December), late developing stage (November to March), ripe stage (March to May), partially spawned stage (April to June), and degenerative/resting stage (June to October). Follicle cells attached to an oocyte were involved in vitellogenesis in the cytoplasm of the vitellogeneic oocyte and formation of chorion (secondary egg membrane) of the ovarian eggs. Spawning occurred between April and June. The spawning period was once a year and the peak took place between May and June. This species belongs to semelparity. According to changes in biochemical contents of the ovary and the digestive organ, monthly variations of moisture, total protein, total lipid and glycogen contents (%) in the ovary showed a negative correlationship with those of the trunk tissues including the digestive organ. Accordingly, it is assumed that the ovary only may be received nutrient supply (total lipid content) for gonadal development from the trunk tissues including the digestive organ (r = -0.55, p < 0.05).

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
• /
• v.17 no.3
• /
• pp.38-60
• /
• 2004

We made an experiment if the extracts of DanGuiBoHyulTangGami-Bang(DBTG) and 15 kinds of the medical herbs used the materials of DBTG were effective on the hair formation palpation and the falling out of hair, and came to the following conclusions. 1. The extracts of Paeonia lactiflora, Cuscuta chinensis and Angelica tenuissima of DBTG consisted of the 15 kinds of the medical herbs kept the activity of 5${\alpha}$-reductase type Ⅱ from being active 75.3$\%$, 63.8$\%$. 75.5$\%$. 2. The hair growth index, 1.6(control group 0.8) of the extracts of DBTG bas a little effect on the hair growth palpation and that of Rubus coreanus 1.8(control group 0.4) was the most effective one of the medical herbs, and Paeonia lactiflora 2.3(control group 1.7) and Vitex rotundifolia 2.3(control group 1.5) showed the effect on hair formation palpation. 3. The hair growth period couldn't be extended by DBTG in this experimental stage. 4. The 15 kinds of constitution medicines of DBTG didn't have effects in dermal papilla cells DNA increase, IGF- I, KGF, HGF the revelation of a gene heredity, the protein synthesis of the hair follicle tissues. 5. All of the 15 kinds of constitution medicines of DBTG didn't have the antibacterial activity in Paper disc rule. 6. The results from the test of a radical scavenging activity of the 15 kinds of constitution medicines of DBTG showed that the extracts of Paeoria lactiflora, Scutellaria baicalensis, Rubus coreanus have the superior antioxidant activity in the concentration of 0.01$\%$ and 0.001$\%$ 7. In the formation controlled experiment, Vitex rotundifolia (70.6$\%$), Scutellaria baicalensis (47.1$\%$, Saposhnikovia (44.8$\%$) of the 15 kinds of constitution medicines of DBTG in the 50㎍／㎖ concentration controled NO forming and Vitex rotundifolia (12.7$\%$) controled NO forming in the 5㎍／㎖ concentration in order. 8. MTT(lC／50) of the extracts of Rehmannia glutinosa, Paeonia lactiflora, Scutellaria baicalensis, Lycium chinense, Rubus coreanus of the 15 kinds of constitution medicines of DBIG was more than 500㎍／㎖ and had the least cell virulence.

• The Korean Journal of Zoology
• /
• v.18 no.2
• /
• pp.87-101
• /
• 1975

Electron microscopic studies on the ultrastructure of the mouse oocyte were made to investigate the inhibition of germinal vesicle breakdown by dibutyryl cAMP. The nuclear membrane of the dibutyryl cAMP-treated oocyte is characterized by a decreased degree of folding, maintains the normal double membrane structure, and shows an increased occurrence of the nuclear pore. It is suggested that these may be related to the suppression of the maturation of oocytes at the germinal vesicle. Mitochondria in the control cell were shown to be spread evenly throughout the cytoplasm and structurally underdeveloped or transitionary having little cristae development. On the contrary, mitochondria in the treated oocyte were found to be localized mainly around the nucleus and to show a greater extent of cristae development. The oocyte treated with dibutyryl cAMP appears to have fewer and structurally simpler lysosomes as compared to the control. The Golgi complex in the control oocyte exhibits the typical granular and lamellar structure, whereas that in the treated cell is poorly developed. Many multivesicular bodies, tonofilaments, and free ribosomes were observed in the control as well as in treated cells. The microvilli become structurally irregular, and a development of the perivitelline space is apparent in the treated oocyte. It is concluded that there is no basic difference in the ultrastructure between the oocytes treated with dibutyryl cAMP for 24 hours in the medium and those collected directly from the follicle. However, the finding that dibutyryl cAMP induces a development of more pores along the nuclear membrane strongly suggests the possibility that this compound inhibits the maturation of oocytes by influencing the permeability of the nuclear membrane.