Menarche is a major biologic event associated with women's health through the lifespan. Social and economic deprivation is known to be related with delayed menarche. We explored the association between delayed menarche and deprivation index of the school area. Retrospective cohort study. A total of 187,024 female middle- and high-school students in the Korea Youth Risk Behavior Web-based Survey (KYRBS) from 2006 to 2015 were analyzed. Delayed menarche was defined as absence of menarche by 14 years of age. Area-level deprivation index where the school located were calculated using 2010 Korean Census data. We conducted regression analyses including body mass index (BMI), father's and mother's education, absence of one of the parents, year of survey and birth cohort (1988-1992, 1993-1997, 1998-2003) to calculate the odds ratio (OR) for delayed menarche. Delayed menarche occurred in 6.5% in the study population. The birth year-specific prevalence decreased from 8.3% (236/2,950) in girls born in 1988 to 3.8% (210/1,076) in those born in 1999. When the population was divided into 4 groups by deprivation index quartiles, late menarche occurred in 6.0% (least deprived), 6.1%, 6.5%, and 6.7% (most deprived) of each group. In the multivariable regression model, girls going to school in area with upper 3rd (1.11, 95% CI: 1.02, 1.21) and 4th quartiles (1.16, 95% CI: 1.07, 1.25) of deprivation index had higher risk for late menarche. This study suggests going to school located in more deprived area is associated with higher odds for delayed menarche. Future studies of timing of menarche would need to consider the potential effect of area-level deprivation.

We investigated the temperature-dependent locomotion of Caenorhabditis elegans by using the mobile phone-based microscope. We developed the customized imaging system with mini incubator and smartphone to effectively control the thermal stimulation for precisely observing the temperature-dependent locomotory behaviours of C. elegans. Using the mobile phone-based microscope, we successfully followed the long-term progress of specimens of C. elegans in real time as they hatched and explored their temperature-dependent locomotory behaviour. We are convinced that the mobile phone-based microscope is a useful device for real time and long-term observations of biological samples during incubation, and can make it possible to carry out live observations via wireless communications regardless of location. In addition, this microscope has the potential for widespread use owing to its low cost and compact design.

Background: Wnt/β-catenin signaling is important in development and differentiation of melanocytes. Objective: In this study, we evaluated the effects of ICG-001 on collagen expression in human normal skin- and keloid-derived fibroblasts. Methods: Fibroblasts were treated with Wnt/β-catenin signaling inhibitor, ICG-001, and then collagen expression level were checked. Results: To evaluate the effect of ICG-001 on type I collagen synthesis, we performed Western blot and RT-PCR. Treatment of normal fibroblasts with ICG-001 resulted in dramatic decrease of type I collagens in the dose- and time-dependent manners. Also, the secretion of procollagen was significantly reduced by ICG-001. The gel contraction was occurred in a time-dependent manner in control groups, while ICG-001 treatment resulted in significant inhibition of gel contraction. Moreover, ICG-001 markedly inhibited the type I collagen synthesis in keloid fibroblasts. Pretreatment with ICG-001 failed to inhibit TGF-β-induced phosphorylation of Smad2/3. We first confirmed whether ICG-001 actually inhibited Wnt/β-catenin signaling in dermal fibroblasts. Cells were transduced with TOPflash reporter adenovirus, then treated with ICG-001. Also, overexpression of β-catenin markedly diminished the inhibitory potential of ICG-001 on type I collagen production. Conclusions: In summary, ICG-001 inhibited the collagen production in normal and keloid fibroblasts. Our data suggest that ICG-001 can be applied as therapeutics for fibrotic skin diseases.

CFP1 is a component of histone H3 lysine 4 (H3K4) methyltransferase, SET1 complex and acts a epigenetic regulator. In this study, we investigated the role of CFP1 during development of the oocyte and fertilization using oocyte specific conditional knockout animal model. Experimental animal model. CFP1 conditional knockout (cKO) mice was generated and used for oocyte-specific deletion by mating with ZP3-cre transgenic mice. Fertility of CFP1 cKO female mice were monitored for 6 months. For mature oocytes collection, 4∼6 weeks old wild type or CFP1 cKO mice, were superovulated, and at 13.5∼14 h post hCG, ovulated oocytes were collected from oviduct. GV immature oocytes were recovered 44∼46h post-PMSG. Maturation rate were calculated by the first polar body formation. Fertilization ability was tested with IVF or ICSI. The pronucleus formation was observed 6h after IVF or ICSI. Oocytes were subjected intracellular calcium changes, GSH, MitoTracker Red CMXRos, or transmission electron microscopy. To examine the gene profiles from a single CFP1 cKO oocyte, single-cell RNA sequenceanalysis was performed by BGI technology. CFP1 cKO female mice were infertile. There is no significant difference in the number of GV immature oocytes between control and CFP1 cKO. Oocytes from CFP1 cKO showed higher percentages of abnormal maturation, such as big polar body, the second polar body, and intact chromosome and spindle in polar bodies. The number of ovulated oocytes from CFP1 cKO mouse were significantly low compared with control. Oocytes from CFP1 cKO showed insecure actin filament beneath plasma. Only 6% of oocytes from CFP1 cKO mice were fertilized and no further development was observed. Interestingly, there is no decondensation of sperm nucleus after IVF or ICSI. In addition, disruption of CFP1 in the oocyte disregulates the expression of numerous genes including Gstt1 which is one of factors involved in GSH metabolism. To examine the explanation of the sperm nucleus decondensation failure, we looked into the level of glutathione which is known to be essential for sperm head decondensation during fertilization. The oocytes from CFP1 cKO mice showed higher level of GSH compare to control. Mitochondria in ooplasm of CFP1 cKO formed aggregation and abnormal clusters beneath plasma membrane, whereas mitochondria from control oocytes distributed homogeneous in ooplasm with clustering. In the status of H3K4 methylation, CFP1 cKO contains significantly low H3K4me3, whereas the level of mono or di-methylation of H3K4 was similar. CFP1 is essential for oocyte maturation and sperm decondensation during fertilization via regulation of numerous gene expression which are involved in GSH metabolism and mitochondria distribution in the oocyte.

ObjectiveRecent information provided by genetically modified mouse model has suggested that PI3K-Akt-mTOR/PTEN signaling pathway controlled primordial follicle activation in vivo. Also, PPAR gamma agonist induced PTEN expression and inhibited Akt phosphorylation in vitro. Ovary specific PPAR gamma was predominantly expressed in ovarian tissue. The aim of this study was to investigate whether PPAR gamma modulator might induce the activation and growth of primordial follicles from 5day female mouse after birth.DesignAnimal study.Materials and MethodsOvaries were collected from 5days old B6D2F1 or ICR female mouse and cultured on six well plate and insert for 10 days in vitro. Ovaries were cultured in DMEM/F12 containing BSA, ITS-X, and ascorbic acid, with or without PPAR gamma modulator for 2 days and transferred PPAR gamma modulator free medium. To evaluate the expression of PPAR gamma, PTEN, and Akt1 on mouse ovary, we performed real-time-PCR, immunohistochemistry, and western blot. Also, end of culture, ovaries were fixed 10% neutral-buffered formalin and subjected of HE staining, immunohistochemistry against Ki-67, AMH, PTEN, Akt1 and FOXO-3a, and apoptosis assay with TUNEL. Also, western blot was performed to evaluate the changes of PTEN, Akt1.ResultsThe expression of PPAR gamma, PTEN, Akt1 and FOXO3a were evaluated in ovaries from 5, 10, 15, 20 days and 8 weeks old mouse by RT-PCR. PPAR gamma was expressed in ovaries from all the ages, it increased in ovary in 20 days explosively. PTEN and Akt1 were increased in ovary form 10 Days. In PPAR gamma antagonist treated ovaries, PTEN was decreased and Akt1 was activated. Also nuclear exclusion of FOXO3a was observed in oocytes of primordial follicles at 6hr treatment with PPAR gamma antagonist. AMH expression was observed in PPAR gamma antagonist treated ovaries, but was not in PPAR gamma agonist. After 12 days culture, increases of ovarian sizes of PPAR gamma antagonist treated groups were observed as compare to non-treated or PPAR gamma agonist treatment.ConclusionsFrom these result, PPAR gamma may participate primordial follicle activation and further development, possibly mediated in part of PI3K-PTEN signaling pathway in vitro. Generation of activated primordial follicle in cancer treatment patients, or other infertile women may allow fertility preservation. Also Production of a large number of oocytes may facilitate future derivation of embryonic stem cell for regenerative medicine. ObjectiveRecent information provided by genetically modified mouse model has suggested that PI3K-Akt-mTOR/PTEN signaling pathway controlled primordial follicle activation in vivo. Also, PPAR gamma agonist induced PTEN expression and inhibited Akt phosphorylation in vitro. Ovary specific PPAR gamma was predominantly expressed in ovarian tissue. The aim of this study was to investigate whether PPAR gamma modulator might induce the activation and growth of primordial follicles from 5day female mouse after birth. Recent information provided by genetically modified mouse model has suggested that PI3K-Akt-mTOR/PTEN signaling pathway controlled primordial follicle activation in vivo. Also, PPAR gamma agonist induced PTEN expression and inhibited Akt phosphorylation in vitro. Ovary specific PPAR gamma was predominantly expressed in ovarian tissue. The aim of this study was to investigate whether PPAR gamma modulator might induce the activation and growth of primordial follicles from 5day female mouse after birth. DesignAnimal study. Animal study. Materials and MethodsOvaries were collected from 5days old B6D2F1 or ICR female mouse and cultured on six well plate and insert for 10 days in vitro. Ovaries were cultured in DMEM/F12 containing BSA, ITS-X, and ascorbic acid, with or without PPAR gamma modulator for 2 days and transferred PPAR gamma modulator free medium. To evaluate the expression of PPAR gamma, PTEN, and Akt1 on mouse ovary, we performed real-time-PCR, immunohistochemistry, and western blot. Also, end of culture, ovaries were fixed 10% neutral-buffered formalin and subjected of HE staining, immunohistochemistry against Ki-67, AMH, PTEN, Akt1 and FOXO-3a, and apoptosis assay with TUNEL. Also, western blot was performed to evaluate the changes of PTEN, Akt1. Ovaries were collected from 5days old B6D2F1 or ICR female mouse and cultured on six well plate and insert for 10 days in vitro. Ovaries were cultured in DMEM/F12 containing BSA, ITS-X, and ascorbic acid, with or without PPAR gamma modulator for 2 days and transferred PPAR gamma modulator free medium. To evaluate the expression of PPAR gamma, PTEN, and Akt1 on mouse ovary, we performed real-time-PCR, immunohistochemistry, and western blot. Also, end of culture, ovaries were fixed 10% neutral-buffered formalin and subjected of HE staining, immunohistochemistry against Ki-67, AMH, PTEN, Akt1 and FOXO-3a, and apoptosis assay with TUNEL. Also, western blot was performed to evaluate the changes of PTEN, Akt1. ResultsThe expression of PPAR gamma, PTEN, Akt1 and FOXO3a were evaluated in ovaries from 5, 10, 15, 20 days and 8 weeks old mouse by RT-PCR. PPAR gamma was expressed in ovaries from all the ages, it increased in ovary in 20 days explosively. PTEN and Akt1 were increased in ovary form 10 Days. In PPAR gamma antagonist treated ovaries, PTEN was decreased and Akt1 was activated. Also nuclear exclusion of FOXO3a was observed in oocytes of primordial follicles at 6hr treatment with PPAR gamma antagonist. AMH expression was observed in PPAR gamma antagonist treated ovaries, but was not in PPAR gamma agonist. After 12 days culture, increases of ovarian sizes of PPAR gamma antagonist treated groups were observed as compare to non-treated or PPAR gamma agonist treatment. The expression of PPAR gamma, PTEN, Akt1 and FOXO3a were evaluated in ovaries from 5, 10, 15, 20 days and 8 weeks old mouse by RT-PCR. PPAR gamma was expressed in ovaries from all the ages, it increased in ovary in 20 days explosively. PTEN and Akt1 were increased in ovary form 10 Days. In PPAR gamma antagonist treated ovaries, PTEN was decreased and Akt1 was activated. Also nuclear exclusion of FOXO3a was observed in oocytes of primordial follicles at 6hr treatment with PPAR gamma antagonist. AMH expression was observed in PPAR gamma antagonist treated ovaries, but was not in PPAR gamma agonist. After 12 days culture, increases of ovarian sizes of PPAR gamma antagonist treated groups were observed as compare to non-treated or PPAR gamma agonist treatment. ConclusionsFrom these result, PPAR gamma may participate primordial follicle activation and further development, possibly mediated in part of PI3K-PTEN signaling pathway in vitro. Generation of activated primordial follicle in cancer treatment patients, or other infertile women may allow fertility preservation. Also Production of a large number of oocytes may facilitate future derivation of embryonic stem cell for regenerative medicine. From these result, PPAR gamma may participate primordial follicle activation and further development, possibly mediated in part of PI3K-PTEN signaling pathway in vitro. Generation of activated primordial follicle in cancer treatment patients, or other infertile women may allow fertility preservation. Also Production of a large number of oocytes may facilitate future derivation of embryonic stem cell for regenerative medicine.

While fate of ovarian failure after chemotherapy has been well established, the precise mechanism by which this occurs is less clear.Recent study has revealed that PTEN/PI3K/Akt signaling in oocytes is critically important for maintenance of the primordial follicle pool. Current study was designed to investigate role of this pathway in cisplatin induced primordial follicle depletion. This was an experimental study using animal model. Mouse was given daily intraperitoneal injection of 2mg/kg (Dong A Pharmaceutical, Korea) for 15days. Ovaries were collected at time point 0, 5, 10, 12, 15 days after the start of daily Cisplatin injection. Ovaries were fixed in PFA and paraffin embedded. Serially sectioned (10μm section) ovarian tissues were H&E stained for differential follicle count. Early stage follicle were counted according to previous definitions after staining with early stage ovarian follicle marker Lhx8. Ovary samples were used for immunohistochemistry and evaluated for apoptosis using Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Antibodies used in western blotting were PTEN, Akt, phospho-Akt, ERK, phospho-ERK, GSK3-β, phospho-GSK3-β and FOXO3a with visualization by enzymatic chemiluminescence. Quantification of the different follicle populations in each time point after Cisplatin injection showed that as cumulative dose of Cisplatin increase, there was significant increase in ratio of growing follicle versus dormant follicles. As with the TUNEL staining, apoptosis was only observed in granulosa cells of growing or mature follicle until day 10 and relatively few primordial follicle was stained compared to growing and mature follicle at day15. Analysis of the PI3K/PTEN/Akt/Foxo3 pathway demonstrated decreased PTEN as cumulative dose of cispaltin increases and activated pathway cascade resulting increase in cytoplasmic translocation of Foxo3 in cisplatin treated follicles. Cisplatin over activates the dormant primordial follicle in mice resulting loss of resting pool of ovarian follicles. Effect of the treatment showed activation of PTEN/Akt/PI3K/FOXO3 pathway thus increase in growing follicle pool while dormant follicles exhibit rapid depletion. Our finding suggests promising therapeutic target for fertility preservation during chemotherapy.

Background: There has been a paucity of data about the difference in gene expression between melasma lesional skin and normal adjacent one. Objective: Our aim was to identify novel genes involved in the pathogenesis of melasma. Methods: We performed a microarray analysis and confirmed the results on quantitative real-time polymerase chain reaction (qRT-PCR) in Korean women with melasma. Results: There were 334 genes whose degree of expression showed a significant difference between melasma lesional skin and normal adjacent one. Of these, five were confirmed on qRT-PCR. In melasma lesional skin, there were down-regulation of genes involved in the PPAR signaling pathway and up-regulation of genes involved in neuronal component and the functions of stratum corneum barrier. Conclusion: This result suggests that the pathogenesis of melasma might be associated with novel genes involved in the above signaling pathway in Korean women.

Seok, J. -H.; Kang, H.; Kim, T. -Y.; Yon, J. -H.; Oh, C. -S.; Yang, H. -S. Author Information

BACKGROUND:Melanocyte dendrites serve as the principal conduit for melanosome transfer. The dendrite formation requires actin polymerization mediated by Rho family GTPases including RhoA, Rac1 and Cdc42.OBJECTIVE:The aim of this study is to investigate and explore the involvement of p38 MAPK in melanocyte dendrite formation.METHODS:We transduced melanoma cells with adenovirus harboring the expression cassette for constitutive active form of MKK6, an upstream MAPKK for p38 MAPK.RESULTS:We investigated the effect of melanogenic inducers on melanocyte dendricity, using SK-mel-24 melanoma cells because that this cell line is refractory to several melanogenic inducers in terms of melanogenesis. TPA-induced the phosphorylation of p38 MAPK and the elongation of dendrite length, suggesting that MKK6 may be involved in this process. Overexpression of the constitutive active form of MKK6 resulted in significant elongation of dendrites in the melanoma cell line SK-mel-24. Moreover, overexpression of MKK6 ultimately led to the upregulation of Cdc42 and Rac1, suggesting that MKK6 acts as a crucial upstream signaling molecule for Rho family GTPases. When overexpressed in normal human epidermal melanocytes, MKK6 led also the increase of dendrite length.CONCLUSION:These results suggest that MKK6 is an authentic regulator for melanocytes dendricity, through the modulation of Rho family GTPases.

BACKGROUND:A callus is a local thickening of skin, characterized by accelerated keratinization and a reduced rate of desquamation. However, the mechanism of callus formation is not fully understood.OBJECTIVES:To evaluate the expression patterns, in callused skin, of genes that are implicated in keratinization and adhesion/desquamation.METHODS:Samples of skin from the dorsum of the foot (DF), centre of the plantar arch (CP) and anterior aspect of the heel (AH) were obtained from fresh cadavers, and protein and gene expression were determined by immunohistochemistry and reverse transcription-polymerase chain reaction, respectively.RESULTS:The stratum corneum in the DF showed a splitting phenotype by conventional haematoxylin and eosin staining, while the stratum corneum was normal in the AH. Cells of the stratum corneum in the AH were nonsquamous. Expression of cornification-related molecules including involucrin, filaggrin, caspase 14 and calcium-sensing receptor was higher in the AH. Similarly, expression of adhesive proteins such as corneodesmosin, desmoglein 1 and desmocollin 1 was increased in the AH. However, protease-activated receptor 2 expression was reduced in the stratum granulosum in the AH. The number of proliferating cells in the stratum basale was significantly increased in the AH, compared with the DF and CP.CONCLUSIONS:Our data suggest that calluses form as a result of hyperproliferation and incomplete differentiation of epidermal keratinocytes, and increased expression of adhesion molecules.

OBJECTIVE: We aimed to compare the outcomes of transfer cycles involving vitrified/warmed blastocysts and three different endometrial preparation methods.DESIGN: Retrospective study.MATERIALS AND METHODS: In all, 611 women with regular menstruation and normal ovarian function and who underwent vitrified/warmed blastocyst transfer cycles (648 cycles) between January 2007 and December 2009 were recruited. Based on the endometrial preparation method used, transfer cycles were divided into: group 1, natural cycle without human chorionic gonadotropin (hCG) (n = 310); group 2, natural cycle with hCG (n = 134); and group 3, artificial cycle with estrogen and progesterone supplementation (n = 204).RESULTS: Patients in three groups were similar in terms of demographic characteristics and reproductive history. The cycles leading to surplus embryo freezing were also found to be statistically comparable. Implantation rate, clinical pregnancy rates and ongoing pregnancy rates were significantly higher in group 1 than in group 3 (34.9% vs 24.2%: P =0.003, 41.9% vs 30.4%: p = 0.008 and 38.1% vs 27.5%: p =0.013 ,respectively). However, result between group 1and group 2 were comparable (34.9% vs 33.3%: p = NS, 41.9% vs 41.7%: p = NS, 38.0% vs 38.0%: p = NS, respectively). There were no significant difference between groups in the means of miscarriage rate (group 1: 8.0% vs group 2: 5.2% vs group 3: 6.4%). Other variables found to significantly affect the ongoing pregnancy rate upon multivariate analysis were age at transfer (OR: 0.938, 95% CI: 0.890–0.988, P = 0.016), number of embryos transferred (OR: 1.905, 95% CI: 1.348–2.692, P < 0.001), grade of transferred embryos (OR: 0.572, 95% CI: 0.391–0.839, P = 0.004), and maximal endometrial thickness (OR: 1.254, 95% CI: 1.143–1.376, P < 0.001).CONCLUSION: The present result suggest that natural cycles with or without hCG yield better outcomes than hormonally manipulated artificial cycles after the transfer of vitrified/warmed blastocysts. OBJECTIVE: We aimed to compare the outcomes of transfer cycles involving vitrified/warmed blastocysts and three different endometrial preparation methods. DESIGN: Retrospective study. MATERIALS AND METHODS: In all, 611 women with regular menstruation and normal ovarian function and who underwent vitrified/warmed blastocyst transfer cycles (648 cycles) between January 2007 and December 2009 were recruited. Based on the endometrial preparation method used, transfer cycles were divided into: group 1, natural cycle without human chorionic gonadotropin (hCG) (n = 310); group 2, natural cycle with hCG (n = 134); and group 3, artificial cycle with estrogen and progesterone supplementation (n = 204). RESULTS: Patients in three groups were similar in terms of demographic characteristics and reproductive history. The cycles leading to surplus embryo freezing were also found to be statistically comparable. Implantation rate, clinical pregnancy rates and ongoing pregnancy rates were significantly higher in group 1 than in group 3 (34.9% vs 24.2%: P =0.003, 41.9% vs 30.4%: p = 0.008 and 38.1% vs 27.5%: p =0.013 ,respectively). However, result between group 1and group 2 were comparable (34.9% vs 33.3%: p = NS, 41.9% vs 41.7%: p = NS, 38.0% vs 38.0%: p = NS, respectively). There were no significant difference between groups in the means of miscarriage rate (group 1: 8.0% vs group 2: 5.2% vs group 3: 6.4%). Other variables found to significantly affect the ongoing pregnancy rate upon multivariate analysis were age at transfer (OR: 0.938, 95% CI: 0.890–0.988, P = 0.016), number of embryos transferred (OR: 1.905, 95% CI: 1.348–2.692, P < 0.001), grade of transferred embryos (OR: 0.572, 95% CI: 0.391–0.839, P = 0.004), and maximal endometrial thickness (OR: 1.254, 95% CI: 1.143–1.376, P < 0.001). CONCLUSION: The present result suggest that natural cycles with or without hCG yield better outcomes than hormonally manipulated artificial cycles after the transfer of vitrified/warmed blastocysts.