Purpose : Recent reports pointed out that gonadotropin releasing hormone analogue (GnRHa) therapy alone is not so promising for improving adult height in precocious puberty. So, that we studied the growth promoting effect of combined therapy with GnRHa and growth hormone (GH) in early pubertal girls. Methods : Twenty three early pubertal girls ($9.73{\pm}1.59yr$) with predicted adult heights (PAH) below-2 standard deviation score (SDS) were included. They were divided into two groups as follows; Group I before menarche (n=19) and Group II after menarche (n=4). After combined therapy, various growth parameters were compared between two groups and between the before and after therapy. Results : Between the two groups before therapy, chronologic age (CA), growth velocity (GV), body mass index (BMI), target height (TH), PAH and serum insulin-like growth factor binding protein-3 were not different, but BA, height and difference between bone age (BA) and CA were significantly higher and insulin-like growth factor-1 (IGF-1) was marginally higher in group II. After therapy, BA still remained higher in group II, but other parameters were not different. In both groups, after therapy, the difference between BA and CA, the ratio of BA over CA, and GV were significantly decreased, but PAH, height SDS and BMI were significantly increased. Regarding IGF-1 level, a significant increase was noted in group I, but not in group II. Conclusion : With combined therapy of GnRHa and GH, PAH in early pubertal girls might be improved significantly and even approach TH. Among them, those who were before menarche might have greater potential for the height gain than those after menarche in view of IGF-1 changes during therapy.
Although some phytoes rogens might have beneficiary rather than adverse effects, most endocrine disrupting compounds(EDCs) are considered to be harmful to human and wildlife health through interfering the endocrine system. Previously we found that prepubertal exposure to genistein(GS), a well-known isoflavone phytoestrogen, could activate the reproductive system of immature female rats resulting precocious puberty. Interestingly, di(2-ethyl hexyl) phthalate(DEHP) exposure brought inverse result, a delayed puberty, in the same experimental regimen. In this study, we examined whether prepubertal exposure to GS or DEHP affect the gene expressions of estrogen receptors($ER\;{\alpha}$ and $ER\;{\beta}$) and LH receptor(LHR) which represent the maturational status of ovary and uterus in immature rats. GS (100 mg/kg/day) was administered daily from postnatal day 25 to the day when the first vaginal opening(VO) was observed, and the animals were sacrificed on the next day(day 32). Similarly, DEHP(l00 mg/kg/day) was administered daily from postnatal day 25 through the day when the first V.O. in control group was observed, and the animals were sacrificed on the next day(day 36). To determine the transcriptional changes in the hormone receptors, total RNAs were extracted from ovary and uterus and were applied to semi-quantitative reverse transcription polymerase chain reaction(RT-PCR). In the GS group, the transcriptional activities of $ER\;{\alpha}$, $ER\;{\beta}$ and LHR in uterus and LHR in ovary were significantly increased when compared to those of control group. In the DEHP group, the transcriptional activities of all the hormone receptors measured were significantly lowered when compared to those of control group. These alteration of the reproductive hormone receptor expressions in ovary and uterus might be represent the phenotypic aspects(secondary sexual characteristics) such as tissue weights and reproductive hormone levels during perinatal period in immature female rats.
Parabens were commonly used for preventing the growth of microorganisms as preservatives in the pharmaceutical, cosmetic and food industry. Also, parabens are known endocrine disruptors because of their estrogenic effects on human. Parabens affect the endocrine system and show adverse effect such as, genital malformations, precocious puberty and testicular cancer in young children, infants and fetuses. In this study, we developed analytical method for four parabens (methyl paraben, ethyl paraben, propyl paraben, butyl paraben) in human breast milk which frequently consumed by newborn baby. The analytes were extracted using liquid-liquid extraction (LLE) after enzyme hydrolysis with protease and lipase, then quantitative analysis was performed by liquid chromatography tandem mass spectrometry (LC-MS/MS). The method validation results were as follows; the linearity of calibration curves were excellent with coefficient of determinations (r2) higher than 0.999, the limit of detections (LODs) were 0.019~0.044 ng/mL, the accuracies were 85.3~105.9% and the precisions were lower than 10%. The average concentration ± standard deviation of parabens in ten human breast milk sample were MP 0.660 ± 0.519 ng/mL, EP 1.631 ± 2.081 ng/mL and PP 0.326 ± 0.320 ng/mL, and BP was not detected.
Kim, Boyoung;Lee, Joohee;Kim, Seonyeob;Lee, Eunji;Choi, Chorong;Kho, Younglim
Journal of Environmental Health Sciences
/
v.42
no.1
/
pp.53-60
/
2016
Objectives: Preservatives are commonly used in pharmaceuticals, cosmetics and other products to extend the expiration date and prevent the growth of microorganisms. Preservatives are generally effective in controlling mold and inhibiting yeast growth, and against a wide range of bacterial attacks as well. They also adversely affect the quality of sperm and cause precocious puberty in children. This study was performed to analyze seven preservatives used in pharmaceuticals and personal care products. Methods: Five kinds of pharmaceuticals and personal care products (PPCPs) were examined for analysis with a high performance liquid chromatography-diode array detector. Each sample was homogenized and the targeted compounds were extracted with methanol. The suspended particulate was removed by syringe filter. Next, the sample was injected into an HPLC system. The separation of the seven preservatives was achieved with a C18 column and gradient mode. The accuracies were between 73% and 120% and precision was lower than 11.58% (RSD). Results: All of the calibration curves showed good linearity with a coefficient of determination ($r^2$) over 0.999. Among the PPCP samples, the detection rate of preservatives was 32.5% for pharmaceuticals, 44.8% for toothpaste, 76.9% for mouthwash, 40.0% for body lotion and 56.0% for wet tissues. The average concentrations of the preservatives in PPCPs were BA 1141.0 mg/kg, MP 709.8 mg/kg, EP 624.9 mg/kg, PP 216.9 mg/kg, BP 167.8 mg/kg, and TCS 538.2 mg/kg. The most frequently detected preservatives in pharmaceuticals and personal care products were BA, MP and PP. The concentrations of preservatives exceeded Korean regulatory standards in 11 samples of medicines, three of mouthwash and two of body lotion. Conclusion: We found that most of the PPCP samples contained various preservatives. It is necessary to identify which preservatives were used and to determine the level of preservatives in PPCPs and to assess the health risk to susceptible populations such as children.
In this paper, we develop a cost effective and disposable voltammetric sensing platform involving screen-printed carbon electrode (SPCE) modified with the nanocomposites composed of multi-walled carbon nanotubes, polyelectrolyte, and tyrosinase for bisphenol A. This is known as an endocrine disruptor which is also related to chronic diseases such as obesity, diabetes, cardiovascular and female reproductive diseases, precocious puberty, and infertility. A negatively charged oxidized multi-walled carbon nanotubes (MWCNTs) wrapped with a positively charged polyelectrolyte, e.g., polydiallyldimethylammonium, was first wrapped with a negatively charged tyrosinae layer via electrostatic interaction and assembled onto oxygen plasma treated SPCE. The nanocomposite modified SPCE was then immersed into different concentrations of bisphenol A for a given time where the tyrosinase reacted with OH group in the bisphenol A to produce the product, 4,4'-isopropylidenebis(1,2-benzoquinone). Cyclic and differential pulse voltammetries at the potential of -0.08 V vs. Ag/AgCl was employed and peak current changes responsible to the reduction of 4,4'-isopropylidenebis(1,2-benzoquinone) were measured which linearly increased with respect to the bisphenol A concentration. In addition, the SPCE based sensor showed excellent selectivity toward an interferent agent, bisphenol S, which has a very similar structure. Finally, the sensor was applied to the analysis of bisphenol A present in an environmental sample solution prepared in our laboratory.
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 consist 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-t63I 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 agonist-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.
Some phytoestrogens in soy and red wine, for example, might have beneficiary rather than adverse effects. In particular, dietary soy intake seems to be highly correlated with protection of breast cancer, osteoporosis and cardiovascular disorders. However, questions persist on the potential adverse effects of the main soy constituent genistein (GS) on female reproductive physiology. Previously we found that prepubertal exposure to GS could activate the reproductive system of immature female rats leading to precocious puberty onset, and intracerebroventricularly (ICV) injected GS could directly activate hypothalamic kisspeptin-GnRH neuronal circuits in adult female rats. The present study was performed to examine the hypothalamus-specific GS effects in prepubertal female rats and which subtype of estrogen receptor is mediated in this GS effect. Prepubertal female rats (PND 30) were anaesthetized, treated with single dose of GS (3.4 ${\mu}g$/animal), and sacrificed at 2 hrs post-injection. To determine the transcriptional changes of reproductive hormone-related genes in hypothalamus, total RNAs were extracted and applied to the semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). ICV infusion of GS significantly lowered the transcriptional activities of mTOR (1:$0.361{\pm}0.058$ AU, p<0.001) but increased that of GAD67 (1:$1.285{\pm}0.099$ AU, p<0.05), which are known to act as an upstream modulator of kisspeptin and GnRH neuronal activities in the hypothalamus, respectively. GS administration enhanced significantly the mRNA levels of KiSS-1(1:$1.458{\pm}0.078$ AU, p<0.001), and exerted no effect on the mRNA level of kisspeptin receptor GPR-54 (1:$1.29{\pm}0.08$ AU). GnRH gene expression was significantly decreased in GS-treated group compared to control group (1:$0.379{\pm}0.196$ AU, p<0.05). There was no difference in the mRNA level of $ER{\alpha}$ in the GS-treated group compare to control group (1:$1.180{\pm}0.390$ AU, Fig. 3A). However, icv infusion of GS significantly increased the transcriptional activities of $ER{\beta}$ (1:$4.209{\pm}0.796$ AU, p<0.01, Fig. 3B). Taken together, the present study indicated that the acute exposure to GS could directly alter the hypothalamic GnRH modulating system in prepubertal female rats. Our study strongly suggested the involvement of $ER{\beta}$ pathway in GS's hypothalamus-specific action, and this idea is consistent with the GS's well-known $ER{\beta}$-mediated protective action in breast cancer.
Lee, Hae Yeon;Seo, Han Kyung;Jang, Yi Sun;Kim, Hee Jeoung
The Korean Journal of Nuclear Medicine Technology
/
v.21
no.2
/
pp.44-48
/
2017
Purpose Estradiol (E2) is a steroid hormone mainly produced in women and is a useful indicator for diagnosis of gynecological diseases, menstrual cycle, menopause, and precocious puberty. E2 measurement is performed by diluting the $^{125}I$ radioactive tracer and tracer buffer in the kit. However, It was not precisely specified when the period of tracer is available after activating. The purpose of this study was to determine the appropriate dilution time based on the measurement value with dilution time. Materials and Methods From December 2016 to February 2017, 60 E2 samples with concentrations ranging from 8 to 4577 pg/mL were divided into low, medium, and high concentrations. Dilution of the $^{125}I$ tracer was performed on a 230 RPM agitator for 30 minutes, 1 hour 30 minutes, and 2 hours 30 minutes, respectively. 24 hour dilution was gently shaken and refrigerated. To verify the difference and significance of the results according to the dilution time, a test of normality was performed using SPSS 18.0 and analyzed by Kruskal-Wallis test. The measured value according to the dilution time was compared with the interquartile range of the absolute error. Results The results of Kruskal-Wallis test were not significant (P>0.05). Measurement results are showed as interquartile range of absolute error. At low concentration, it is 0.052 between 1 hour 30 minutes and 2 hours 30 minutes, and 0.105 between 30 minutes and 1 hour 30 minutes. At medium concentration, 0.062 between 30 minutes and 1 hour 30 minutes, and 0.038 between 1 hour 30 minutes and 2 hours 30 minutes. At high concentration, it is 0.029 between 1 hour 30 minutes and 2 hours 30 minutes, and 0.06 between 2 hours 30 minutes and 24 hours. Conclusion There were no statistically significant differences. However, the change in the measured value is the smallest between 1 hour and 30 minutes to 2 hours and 30 minutes. Therefore, we recommend diluting time between 1 hour 30 minutes and 2 hours 30 minutes.
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.
Journal of the Korean Applied Science and Technology
/
v.37
no.3
/
pp.594-603
/
2020
The aim of this study was to investigate the issue in obese children with a high probability of leading to precocious puberty for they have more advanced skeletal maturation compared to normal children in order to contribute to the health of obese children by identifying the relationship between physique and physical fitness according to their skeletal maturation. A total of 243 obese children between the ages of 10-13 were enrolled as subjects, and the skeletal maturation, which symbolize indicators of biological maturation, were evaluated by using the TW3 method after hand-wrist radiographs, thus classified the skeletal maturation level into retarded group(n=70), normal group(n=128), and advanced group(n=45). A stadiometer and InBody 270 (Biospace, Korea) were used to measure 3 components in physique. A total of 7 components in physical fitness, which included muscular strength, muscular endurance, flexibility, power, cardiovascular endurance, balance, agility, were measured as well. Descriptive statistics and one-way variable analysis (ANOVA) were conducted for data processing using SPSS 25.0, and Duncan's multiple-range method was used for post hoc analysis; and it was considered significant at the level of P< .05. Analysis results of this study may be summarized as follow. First, height and weight in physique factors according to the skeletal maturation between the retarded, normal, and advanced groups in males and females were highly significant. Second, physical fitness comparison according to the skeletal maturation in males were statistically significant in muscular strength, power, and agility whereas muscular strength and balance were statistically significant in females.
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