• Clinical and Experimental Pediatrics
• /
• v.55 no.9
• /
• pp.337-343
• /
• 2012

Purpose: Leptin has been considered a link between metabolic state and reproductive activity. Defective reproductive function can occur in leptin-deficient and leptin-excessive conditions. The aim of this study was to examine the effects of centrally injected leptin on the hypothalamic KiSS-1 system in relation to gonadotropin-releasing hormone (GnRH) action in the initial stage of puberty. Methods: Leptin (1 ${\mu}g$) was injected directly into the ventricle of pubertal female mice. The resultant gene expressions of hypothalamic GnRH and KiSS-1 and pituitary LH, 2 and 4 hours after injection, were compared with those of saline-injected control mice. The changes in the gene expressions after blocking the GnRH action were also analyzed. Results: The basal expression levels of KiSS-1, GnRH, and LH were significantly higher in the pubertal mice than in the prepubertal mice. The 1-${\mu}g$ leptin dose significantly decreased the mRNA expression levels of KiSS-1, GnRH, and LH in the pubertal mice. A GnRH antagonist significantly increased the KiSS-1 and GnRH mRNA expression levels, and the additional leptin injection decreased the gene expression levels compared with those in the control group. Conclusion: The excess leptin might have suppressed the central reproductive axis in the pubertal mice by inhibiting the KiSS-1 expression, and this mechanism is independent of the GnRH-LH-estradiol feedback loop.

• Clinical and Experimental Pediatrics
• /
• v.63 no.4
• /
• pp.146-150
• /
• 2020

Background: The incidence of precocious puberty has increased throughout the 20th century. The association between precocious puberty and endocrine disrupting chemicals including agricultural pesticides has been a subject of global study, but human data are lacking. Purpose: We investigated the relationship between agricultural pesticides and the development of precocious puberty. Methods: We enrolled 60 female subjects at Severance Children's Hospital from December 2015 to January 2017. Of them, 30 were diagnosed with precocious puberty, while the other 30 prepubertal girls were enrolled as normal controls. We investigated their clinical characteristics and analyzed the urinary levels of 320 different agricultural pesticides. Results: Agricultural pesticide was detected in one of 30 patients with precocious puberty (3.3%) versus 2 of 30 girls in the normal control group (3.3% vs. 6.7%, P=0.554). Dinotefuran, a neonicotinoid-class insecticide, was detected in the samples of all 3 positive subjects. Conclusion: Our results showed no relationship between agricultural pesticides and the development of precocious puberty. Larger sample sizes and robustly controlled variables are necessary to further investigate this topic.

• Journal of the Korean Academy of Child and Adolescent Psychiatry
• /
• v.24 no.3
• /
• pp.151-156
• /
• 2013

Objectives : It has been reported that higher percentage of B cells react with monoclonal D8/17 antibody in patients with rheumatic fever, childhood onset obsessive-compulsive disorder, Tourette's disorder, or prepubertal anorexia nervosa. The purpose of this study is to replicate the previous studies in a Korean young population with tic disorder and to identify any relationship between D8/17 and clinical symptoms. Methods : The binding of D8/17 to B cells was determined in patients with tic disorder (N=21) and healthy controls (N=9) by Fluorescence-Activated Cell Sorter analysis. Results : In the sample examined by this study, the average percentage of B cells expressing D8/17 in tic disorder was 2.05%; healthy controls was 3.15%. No statistically significant differences were found in the mean percentages of D8/17 between the two groups. Conclusion : The expression of D8/17 in B cells was very low in this study. No subjects with tic disorder or healthy controls was above 12% in D8/17 positive proportion. Further studies, including higher number of patients and control group members, should be performed.

• Asian-Australasian Journal of Animal Sciences
• /
• v.12 no.6
• /
• pp.862-867
• /
• 1999

The present study was undertaken to determine the effect of administration of exogenous progesterone early in gestation on uterine levels of IGF-I mRNA and on embryo survival at day 25 of gestation in the pig. Forty-one prepubertal gilts were induced into oestrus with PG600 and artificially inseminated at their subsequent naturally occurring oestrus. Gilts were then randomly assigned to one of three groups. Gilts in the two treatment groups were injected intramuscularly with 50 mg of progesterone either from day 2 to 14 (N=14) or from day 4 to 14 (N=15) after breeding while those in the control group (N=12) were given corn oil (0.5 ml) from day 2 to 14. Between days 25 and 28 of gestation, gilts were slaughtered and reproductive tracts were recovered. Endometrial tissue (1 g) was collected and analysed for IGF-I mRNA levels using a reverse transcription-polymerase chain reaction, Progesterone treatment, starting either on day 2 or 4 after breeding, neither significantly increased embryo survival rate by day 25 of gestation nor altered IGF-I mRNA levels in uterine tissue. However, across all samples, the IGF-I mRNA level in the uterus was highly correlated with embryo survival rate (r=0.8193, p<0.01), supporting the involvement of IGF-I in the regulation of porcine embryo development.

• The Korean Journal of Pain
• /
• v.9 no.1
• /
• pp.94-97
• /
• 1996

Two hundred and eighty eight patients suffering from excessive sweating of palms, soles and axillae etc., visited our Neuro-Pain clinic from November 1991 to March 1996. The sex ratio was 1:1.2. the third decade of age was the major age group. the onset time of hyperhidrosis was prepubertal period (in 95.1% of them). the provocative factors fo excessive sweating were tension and stress from interpersonal relationship. they had the family history (30.9%) and the past history treated with herb medication (56.9%), medicine (30.6%), operation (1.4%), and no treatment (39.6%). We treated 113 patients by sympathetic ganglion block with pure alcohol. the average times of thoracic sympathetic ganglion block were 2.1 (left), 2.4 (right) and those of lumbar sympathetic ganglion block were 1.2 (left), 1.6 (right). Average admission period was 14.7 days. Recurrence rare was 7.1%. Most longstanding effective period was 45 months. We conclude from our results that sympathetic ganglion block is one of the most effective treatments for hyperhidrosis owing to its simple technique and low recurrence rate.

• Asian-Australasian Journal of Animal Sciences
• /
• v.15 no.10
• /
• pp.1403-1411
• /
• 2002

The aims of this study were, firstly, to analyze the biochemical compositions of serum and follicular fluid (FF) from prepubertal gilts after PMSG (1,000 IU) treatment. The concentrations of total proteins, lipids, cholesterol, glucose and sex hormones (progesterone, $P_4$; estradiol-$17{\beta}$, $E_2$; testosterone, T) were measured. Secondary, the effects of porcine FF (pFF) addition (40% and 100%) in IVM media and different culture conditions [Exp. 1: mBMOC-2+20% porcine serum (PS), fresh IVM medium, filtered IVMconditioned medium, or rabbit oviducts; Exp. 2: mBMOC-2+20%PS or stepwise medium replacement procedures (SMRP) cocultured with or without cumulus cells] on the in vitro development (IVD) of porcine oocytes were also examined. Results showed that no significant differences were found in total protein levels between serum and pFF from different sizes (large, >7 mm; medium, ~5-7 mm; small, <3-5 mm) of follicles (75-85 and 49-90 mg/dl; p>0.05). Total lipid concentrations remained constant in serum (395-472 mg/dl), and reduced significantly in the pFF from large follicles (287 mg/dl) at 132 h after PMSG treatment when compared to those at other time points (441-480 mg/dl). Basal cholesterol levels in serum and pFF at 12 h were similar (153-161 mg/dl), but increased at 36 h (186-197 mg/dl). Basal P4 and E2 levels in serum (0.1 ng/ml and 5.5 pg/ml) were low, but increased from 0.34 ng/ml and 12.13 pg/ml at 24 h to 0.81 ng/ml and 61.70 pg/ml at 98 h, respectively, after PMSG treatment (p<0.05). P4 levels increased linearly in pFF from large follicles during 12 through 132 h (138-1,288 ng/ml). A similar increase was also observed in $E_2$ levels (22-730 pg/ml) before 60 h post PMSG treatment, and then dropped afterwards (730-121 pg/ml). The development of the oocytes fertilized in 40% pFF-medium was greater than that in 100% pFF-medium group without gonaodtropin addition (31% vs 10%, p<0.05). However, both were lower than those in mBMOC-2+20%PS and in rabbit oviducts (p<0.05). When cocultured with cumulus cell monolayers, a greater cleavage rate was observed in the group cultured in filtered IVM-conditioned medium than the SMRP group (36% vs 18%, p<0.05). A similar phenomenon was also observed in the culture without cumulus cell monolayers (33% vs 19%, p<0.05). It is concluded that neither the fresh IVM nor filtered IVM-conditioned medium has positive effect on the IVD of oocytes. Coculture with cumulus cell monolayers and the SMRP were not beneficial to the development of IVF pig oocytes.

• Reproductive and Developmental Biology
• /
• v.29 no.2
• /
• pp.127-131
• /
• 2005

The objective of this study was to examine the effect of EGF on meiotic maturation and pronuclear (PN) formation of porcine oocytes. Prepubertal gilt cumulus-oocyte-complexes (COCs) aspirated from $2\~6mm$ follicles of abbatoir ovaries were matured in TCM199 containing 0.1mg/ml cysteine, $0.5{\mu}/ml$ FSH and LH, and EGF (0, 5, 10, 20, 40 ng/ml) for 22 hr at $39^{\circ}C$ in a humidified atmosphere of $5\%$ $CO_2$ in air. They were then cultured for an additional 22hr without hormones. In Experiment 1, to examine the nuclear maturation at 44hr of culture, the expanded cumulus cells were removed by vortexing for 1 min in 3 mg/ml hyaluronidase. The oocytes were fixed in acetic acid: methanol (1:3, v/v) at least for 48 hr and stained with $1\%$ orcein solution for 5 min. Nuclear status was classified as germinal vesicle (GV), germinal vesicle breakdown (GVBD), prophase-metaphase I (PI-MI), and PII-MII under microscope. In Experiment 2, to investigate PN formation, oocytes were fertilized with Percoll-treated freshly ejaculated sperm $(1\times10^5\; cells/ml)$ in mTBM with $0.3\%$ BSA and 2mM caffeine for 5hr, and cultured in NCSU-23 medium with $0.4\%$ BSA. At 6hr of culture, the embryos were fixed in $3.7\%$ formaldehyde for 48hr and stained with 10ug/ml propidium iodide for 30 min. PN status was classified as no or one PN (unfertilized), 2 PN (normal fertilized) and $\geq3$ PN (polyspermy). Differences between groups were analyzed using one-way ANOVA after arc-sine transformation of the proportional data. The rate of oocytes that had reached to PII-MII were significantly (P<0.05) higher in all groups added EGF than that of non-treated group $(67\%)$, but it did not differ among the all added groups $(86\%,\;85\%,\;79\%\;and\;81\%$, in 5, 10, 20 and 40 ng/ml EGF, respectively). No differences on the incidence of 2PN were observed in all treated groups $(25\%,\;30\%,\;33\%,\;29\%\;and\;29\%$, in 0, 5, 10, 20 and 40 ng/ml EGF, respectively), however, in non-treated group, polyspermy tended to be increased ($66\%\;vs\;. 58\%,\;54\%,\;52\%\;and\;55\%$, 0 vs. 5, 10, 20, 40 ng/ml EGF, respectively). These results suggest that EGF can be effectively used as an additive for enhancing oocyte maturation and reducing the incidence of polyspermy in pig.

• Asian-Australasian Journal of Animal Sciences
• /
• v.31 no.2
• /
• pp.189-197
• /
• 2018

Objective: Hemicastration is a unilateral orchiectomy to remove an injured testis, which can induce hormonal changes and compensatory hypertrophy of the remaining testis, and may influence spermatogenesis. However, the underlying molecular mechanisms are poorly understood. Here, we investigated the impact of hemicastration on remaining testicular function. Methods: Prepubertal mice (age 24 days) were hemicastrated, and their growth was monitored until they reached physical maturity (age 72 days). Subsequently, we determined testis DNA methylation patterns using reduced representation bisulfite sequencing of normal and hemicastrated mice. Moreover, we profiled the testicular gene expression patterns by RNA sequencing (RNA-seq) to examine whether methylation changes affected gene expression in hemicastrated mice. Results: Hemicastration did not significantly affect growth or testosterone (p>0.05) compared with control. The genome-wide DNA methylation pattern of remaining testis suggested that substantial genes harbored differentially methylated regions (1,139) in gene bodies, which were enriched in process of protein binding and cell adhesion. Moreover, RNA-seq results indicated that 46 differentially expressed genes (DEGs) involved in meiotic cell cycle, synaptonemal complex assembly and spermatogenesis were upregulated in the hemicastration group, while 197 DEGs were downregulated, which were related to arachidonic acid metabolism. Integrative analysis revealed that proteasome 26S subunit ATPase 3 interacting protein gene, which encodes a protein crucial for homologous recombination in spermatocytes, exhibited promoter hypomethylation and higher expression level in hemicastrated mice. Conclusion: Global profiling of DNA methylation and gene expression demonstrated that hemicastration-induced compensatory response maintained normal growth and testicular morphological structure in mice.

• Journal of Radiation Protection and Research
• /
• v.24 no.2
• /
• pp.87-92
• /
• 1999

The present study was carried out to evaluate the morphological changes in the degenerating primordial follicles induced by $\gamma$-radiation. The prepubertal female mice of three weeks old ICR strain were whole-body irradiated with a dose of $LD_{80(30)}$ (8.3 Gy). The ovaries were collected at 0 h, 3 h, 6 h, and 12 h post-irradiation. The largest cross sections were prepared with histological semi-thin sections and then observed microscopically. The ratio of normal to atretic follicles was reduced significantly after 6th post-irradiation. At 6 h post-irradiation, the number of degenerated primordial follicles increased. Germinal vesicles disappeared, and lipid droplets increased. No more ooplasmic membranes were seen. Granulosa cells became round in shape, and apoptotic cells started to appear. The ratio of normal to atretic follicles in the control group was 62.50%. The ratio decreased with time after irradiation. The ratio decreased down to 51.61 %, 48.97 %, 11.11 %, and 7.14 % at 0 h, 3 h, 6 h, and 12 h, respectively. Taken together, ionizing radiation acutely induced the degeneration of primordial follicles. The patterns of degeneration are 1) apoptosis of one or more granulosa cells with relatively intact oocyte, 2) apoptosis of oocyte with intact follicle cells, or 3) apoptotic degenerations of both cells. The Present study can provide morphological clues for the identification of degenerating primordial follicles.

• Toxicological Research
• /
• v.27 no.2
• /
• pp.61-70
• /
• 2011

Brominated flame retardants (BFRs) are present in many consumer products ranging from fabrics to plastics and electronics. Wide use of flame retardants can pose an environmental hazard, which makes it important to determine the mechanism of their toxicity. In the present study, dose-dependent toxicity of tetrabromobisphenol A (TBBPA), a flame retardant, was examined in male prepubertal rats (postnatal day 18) treated orally with TBBPA at 0, 125, 250 or 500 mg/kg for 30 days. There were no differences in body weight gain between the control and TBBPA-treated groups. However, absolute and relative liver weights were significantly increased in high dose of TBBPA-treated groups. TBBPA treatment led to significant induction of CYP2B1 and constitutive androstane receptor (CAR) expression in the liver. In addition, serum thyroxin (T4) concentration was significantly reduced in the TBBPA treated group. These results indicate that repeated exposure to TBBPA induces drug-metabolising enzymes in rats through the CAR signaling pathway. In particular, TBBPA efficiently produced reactive oxygen species (ROS) through CYP2B1 induction in rats. We measured 8-hydroxy-2'-deoxyguanosine (8-OHdG), a biomarker of DNA oxidative damage, in the kidney, liver and testes of rats following TBBPA treatment. As expected, TBBPA strongly induced the production of 8-OHdG in the testis and kidney. These observations suggest that TBBPA-induced target organ toxicity may be due to ROS produced by metabolism of TBBPA in Sprague-Dawley rats.