Vanadium is an essential trace element but has not been identified with a specific biogical role. To study the direct effects of vanadium on osteoblast, we incubated murin osteoblast-like (MC3T3-El) cells with various corcentration of vanadium oxide & sodium orthovanadate. This study was designed to investigate the effect of vanadium on DNA synthesis, alkaline phosphatase (ALP) activity, cAMP formation responsive to parathormone(PTH) and type I $\alpha$ 2 collagen ribonucleic acid (mRNA) level in murin osteoblast-like (MC3T3-El) cells. The cells were cultured in $\alpha-minimal$ essential medium$(\alpha-MEM)$ supplemented with $10\%$ fetal bovine serum (FBS) and then changed to $0.1\%$ FBS with various concenoation of vanadium oxide & sodium orthovanadate. Quiescent cultured MC3T3-El cells incubated for 24 hours with 2,5,10,15,20 ${\mu}M$ vanadium oxide incorporated $[^3H]Thymidine;$ every concentration showed increases in $[^3H]Thymidine$ incorporations dose dependant manner, the greatest response occurred at $20{\mu}M$. Quiescent cultured MC3T3-E1 cells incubated for 3days with 2,5,10,15,20 ${\mu}M$ vanadium oxide, for 2days with sodium orthovanadate and alkaline phosphatase was assayed with disodium phenyl phosphate as substrate. Vanadium oxide increased the alkaline phosphatase content in MC3T3-El cells at $2{\mu}M\;&\;6{\mu}M$ ; the greatest response occurred at $2{\mu}M$. But decreased at other content sodium orthovanadate increased alkaline phosphatase content in MC3T3-El cells at all concenoation ; the greatest response occurred at $4{\mu}M$. Quiescent cultured MC3T3-El cells incubated for 3days with $5,10{\mu}M$ vanadium oxide , with $5,8{\mu}M$ sodium orthovanadate and cAMP formation was measured by Radioimmunoassay(RIA). Vanadium oxide & sodium orthovanadate showed the tendency of inhibitory effects on cAMP responsiveness to PTH in MC3T3-El cells. Quiescent cultured MC3T3-El cells incubated for 24hours with $10,20{\mu}M$ vanadium oxide, with $5,10{\mu}M$ sodium orthovanadate and Type I $\alpha$ 2 collagen ribonucleic acid (mRNA) expression was studied by Nothern blot analysis. Northern blot analysis of vanadium oxide treated cells showed decreasing effects 0& sodium orthovanadate revealed increasing effects in type I $\alpha$ 2 collagen ribonucleic acid (mRNA) level.
Folate and Vitamin $B_{12}$ are essential nutrients important during pregnancy. This study was conducted to evaluate the folate and vitamin $B_{12}$ nutritional status of Korean pregnant women and to investigate the relationship between maternalumbilical cord serum folate and vitamin B12 levels and pregnancy outcomes. Dietary intakes of the pregnant women were estimated by 24 hour-recall (3 times). Serum folate and vitamin B12 levels in maternal blood and umbilical cord of 27 pregnant women at 1'st-, 2'nd-, 3'rd-trimester and delivery were measured by RIA (radioimmuno assay), respectively. Means of folate and vitamin $B_{12}$ intake were $283.53\pm58.01{\mu}g/day\;and\; 2.99\pm1.32mg/day$, respectively. Maternal mean serum folate levels of the trimester and delivery were $9.75\pm3.60ng/ml,\;10.46\pm4.63ng/ml,\;10.71\pm4.14ng/ml\;and\;15.05\pm7.04ng/ml$. Those maternal levels were significantly lower than that of umbilical cord blood $(23.99\pm9.42ng/ml)$. Serum vitamin $B_{12}$ levels of maternal trimester and delivery were $479.07\pm137.56 pg/ml,\;310.96\pm137.56pg/ml,\;308.22\pm74.65pg/ml,\;and\;295.67\pm93.36pg/ml$, which were significantly lower than those of umbilical cord blood $(500.13\pm185.60ng/ml)$. This finding indicates that the uptake of folate and vitamin $B_{12}$ in the fetus may be due to an active placental transport mechanism. Maternal serum level correlated positively with those of umbilical cord blood, showing that folate and vitamin $B_{12}$ concentration of umbilical cord blood might be affected by maternal status. There was no significant correlation between the serum folate levels in maternal-umbilical cord blood and the pregnancy outcomes. However, maternal vitamin $B_{12}$ level at l'st trimester was significant positive correlation between the gestational age except for birth weight and weight gain.
Endocrine disruptors (EDs) are exogenous chemicals which interfere several aspects of natural hormone properties. EDs with estrogenic activity have been recently reported to cause animal reproductive problems. This study was performed to investigate the effects of bisphenol A (BPA) on the mouse spermatogenesis in vivo. Male ICR mice were orally injected on a daily basis with low dose of BPA 20 mg/kg, high dose of BPA 200 mg/kg, or corn oil (vehicle control) for 7 days, and litter size and weights of body, testis, and cauda epididymis were measured. The level of serum testosterone and the expression of TGF- $\beta$$_1$ mRNA were also analyzed using RIA and RT-PCR, respectively. Also, morphological differences of testes after treatments were examined. Sperm concentration and level of serum testosterone showed a decreasing tendency detected as untreated >corn oil >low >high dose BPA treated mice, although there were no significant statistical differences. Interestingly, in mice treated with a high dose of BPA, partial disappearance of spermatozoa in seminiferous tubular lumen and the expression of TGF-$\beta$$_1$ mRNA were observed. Spermatogenesis was disrupted through TGF-$\beta$ system in the seminiferous tubules, resulting in no development of germ cells. Similarly, the litter size treated with a high dose of BPA was significantly different from that of untreated control group. In conclusion, these results that a high dose of BPA (200 mg/kg) acts as an endocrine disruptor during apermatogenesis in male mice md that there are BPA-specific lesions in the adult male reproductive tract might represent a permanently altered responsiveness to testosterone by BPA in the affected target tissue.
Purpose Assessment of Serum Thyroglobulin (sTg) value in total thyroidectomy patients having an ablation dose of radioactive iodine indicates remaining cancer or metastasis. Especially, sTg in patients on withdrawal thyroxine or thyrogen administration for radioiodine ablation is an important indicator to determine the direction of further treatment and prognosis. Current guidelines suggest measurement of sTg is performed at 72 hours after the last injection of thyrogen. and assumes that sTg reaches maximum serum levels at that time. The purpose of this study is to evaluate the variation of sTg measured after thyrogen administration. Materials and Methods We compared with sTg performed at 24hours(D0) and 72hours(D2) after the last injection of thyrogen. We reviewed D0 and D2 from 276 patients were divided them into three groups according to ablation dose of radioactive iodine, 5mCi(A group), 30~80mCi(B group) and 100~200mCi(C group). We used T-test for comparison between D0 and D2. sTg was measured in serum using immunoradiometric assay (Tg-plus RIA; BRAHMS, Berlin, Germany). Results There is no critical variation between D0 and D2 in A group(n=100)(P=0.32), The case of increase(D2>D0) is 45, no change(D2=D0) is 23, decrease(D2D0 is 91, D2=D0 is 28, D2D0 is 19, D2=D0 is 2. The biggest increase is 143.6 ng/mL from 98.4 to 242. Conclusion There was a significant difference in the group over 30mCi. and the case of D2>D0 is 45%, 58.7%, 90.5% for each group. therefore, D2 increased as the dose of radioactive iodine increased. Furthermore, the most sTg values of D0 and D2 are variation under 2.0 ng/mL, so reproducibility as well as sensitivity of sTg will be important at values below 2ng/mL.
This study was conducted to investigate the effects of streptozotocin-induced (STZ) diabetes on insulin-like growth factor-I (IGF-I), insulin-like growth factor binding proteins (IGFBPs), and IGF-I carrier proteins in serum, liver, and kidney. The levels of total and free IGF-I were measured by radioimmunoassay (RIA). The patterns of IGFBPs were determined by western ligand blotting (WLB) analysis. The profiles of IGF-I carrier proteins in serum were determined by column chromatography. The levels of total and free IGF-I in serum were lower in STZ-induced diabetic rat than normal rat (p<0.01). Similarly, the levels of total IGF-I in liver was lowered in STZ-induced diabetic rats. On the other hand, the levels of total IGF-I in kidney were increased in STZ-induced diabetic rats compared with normal rats (p<0.01). In serum and liver from STZ-induced diabetic rats, the amount of IGFBP-3 was decreased and the amount of IGFBP-2 was increased compared with normal rats. There was a not difference in amount of IGFBP-4 in serum between STZ-induced diabetic rats and normal rats. The serums of normal rats have higher 150kDa carrier proteins than in STZ-induced diabetic rats, whereas, most of 50kDa carrier proteins were found in STZ-induced diabetic rats. These results demonstrate that in STZ-induced diabetic rats, IGF-I/IGFBPs system that included functional bioactivity was changed in serum as well as tissues, and these changes may play an important role in pathogenesis of diabetes.
Insulin-like growth factor-I(IGF-I) plays an important role in the regulation of mammalian and poultry growth. IGF-I has many actions in different tissues, which include metabolic, mitogenic, and differentiative actions. IGF-I induces insulin-like effects - such as increased cell glucose uptake and glycogen sysnthesis, however several physiological actions of IGF-I may not have been identified yet. In order to investigate the effect on growth in broiler chicken treated with exogenous insulin-like growth factor-I, 30 chickens were injected $50{\mu}g$ reconbinant human IGF- I (rhIGF- I ) per kg body weight as experimental group and 30 ckickens saline subcutanously as control, 3 times according to ages from 2 to 35 days. We established radioimmunoassay method by which we can measure chicken IGF- I (cIGF- I ) as in rhIGF- I assay. The results obtained were as follows; 1) The dilution curve showed in parallelism between rhIGF- I and cIGF- I in the Sep-pak $C_{18}$ cartridge plasma extracts. 2) The body weight of broiler chicken were significantly increased at 31 days($1,176.50{\pm}99.79g$) and 35 days($1,252.84{\pm}125.21g$) of age in treatment groups, compared with control group($1,011.88{\pm}40.22g,\;1,111.32{\pm}153.67g$). The liver and kidney weights on 35 days$(35.24{\pm}5.18g,\;11.05{\pm}1.47g)$ were significantly higher in rhIGF- I treated group than control group($30.95{\pm}4.04g,\;10.01{\pm}1.60g$) 3) The plasma concentration of IGF- l and total protein in rhIGF- I treated group were $58.17{\pm}1.69ng/ml$, $3.75{\pm}0.62g/dl$ respectively compared with control group $45.70{\pm}1.64ng/ml$, $2.32{\pm}0.53g/dl$. The results suggest that exogenous rhIGF- I increased total body weight, liver and kidney weights in broiler chicken, and it may increase IGF- I and total protein concentration in serum.
Previously, we demonstrated that estradiol (E$_2$) was produced by medium sized follicles of Rana nigromaculata and Rana dybowskii in vitro. Present experiments were carried out to determine when the growing follicles have obtained the ability to produce E$_2$. Follicles in different growth stages were isolated and cultured for 6 hours in the presence or absence of frog pituitary homogenates (FPH, 0.1 pituitary/2m1) or various steroid precursors (200 ng/2m1). levels of progesterone (P$_4$), 17 -hydroxyprogesterone (17$\alpha$-OHP), androstenedione (AD), testosterone Cr) or E$_2$ in the medium were measured by RIA. The smallest follicles failed to produce steroids, whereas the smaller follicles produced considerable amounts of steroids (211 pg/follicle), and the medium sized follicles produced a large amounts of steroids (1653 pg/follicle) in response to FPH. Addition of pregnenolone (P5) resulted in a marked increase in P$_4$ but not in other steroids by the smallest follicles whereas the treatment resulted in a marked increase in P$_4$, 17$\alpha$-OHP, AD, T and E$_2$by the smaller and medium follicles. When the amounts of steroids are calculated on the basis of unit surface area (pg/mm$_2$), the ability of the smallest follicles to produce P$_4$ from P5 was similar to those of smaller and medium sized follicles. However, the smallest follicles failed to metabolize P$_4$ to other steroids whereas the smaller and medium follicles did. Taken together, the data suggest that the smallest follicles do not response to FPH in terms of steroid production but they have capacity to convert P5 to P$_4$ and that the smaller follicles have potential to produce E$_2$ although much less efficient than medium sized follicles.
This study was undertaken to define the varying responses of vascular smooth muscle to different wavelengths of ultraviolet radiation and to relate them to the changes in cyclic GMP contents. The ring preparations of rat thoracic aorta with intact or removed endothelium were irradiated with the ultraviolet or visible light (UVR) of wavelengths in step of 10 nm between 250 and 500 nm from xenon lamp of a spectrofluorometer, and the changes in vascular tension were recorded. For cyclic GMP assay, the preparations, pretreated with phenylephrine as in the tension experinents, were frozen after irradiation and homogenated in trichloroacetic acid. The supernatant was extracted with ether and the cyclic GMP contents were measured with radioimmunoassay. In the endothelium-intact preparations, biphasic responses, vasoconstriction (UVR-contraction) followed by vasodilatation (UVR-dilatation), were observed. The maximal UVR-contraction was observed at 320 nm, while the maximal vasodilatation was elicited at 420 nm. In the endothelium-removed rings, however, only vasodilatation was observed, with the maximal vasodilatation taking place at 370 nm. The cyclic GMP contents were not affected by the Irradiation with 320 nm for 30 sec or 1 min in the endothelium-intact preparations, while it was significantly increased by 380 and 420 nm. In the endothelium-removed preparations, UVR of 370 nm markedly increased the cyclic GMP contents. The present study indicates that the increase in cyclic GMP is closely related to vasodilatation induced by UVR of 420 nm in the endothelium-intact or 370 nm in the denuded preparations, whereas it is not involved in the vasoconstriction induced by UVR of 320 nm in the intact rings, and the mechanism leading to UVR-contraction remains to be clarified. These observations suggest that nitric oxide-cyclic GMP system is closely related to the UVR-dilatation in rat aortic preparation, while it is not involved in the UVR contraction.
The importance of thyroid hormones for the survival of rats in the cold is along-established fact. Hypothyroid animals are unable to survive in a cold environment. It was also reported that acute exposure of rats, guinea pigs and rabbits to cold produced an increased secretion of TSH and thereby thyroid hormone secretion within 10 to 30 min, but this increase of thyroid activity disappeared quite rapidly during warming. However, in human study no significant difference was found in the concentration of $T_4$, TSH and cortisol between summer and winter. But plasma $T_3$ concentration was increased significantly in winter in 56 adult men. On the other hand, it has been also known that catecholamines are important in the maintenance of body temperature of rat exposured to cold. Abundant evidences suggest that the sympathetic nervous system is involved in the activation of nonshivering thermogenesis and that thyroid hormone metabolism and secretion are influenced by catecholamines and consequently by the activity of the sympatheticadrenal system. Many of the metabolic effects of catecholamines are associated with an increase in the level of cAMP mediated through activation of adenylate cyclase which converts ATP to cAMP. Other studies have shown that thyroid hormones affect the amount of adenylate cyclase present in the adipose tissue. On the other hand. it was also reported that a particulate cAMP phosphodiesterase activity in fat cells was modulated by the action of thyroid hormones. The objective of the present study was to determine the interaction between thyroid activity and cyclic nucleotides during acute exposure to cold. Albino rats weighing around 200 g were used as the experimental animal. The room temperature group was kept at $25^{\circ}C$ and the cold-exposured group was kept at $4^{\circ}C$ for 1 week or 2 weeks. Each group was subdivided into three subgroups; control, KI, and MTU group. At the end of experiment the animals were etherized and blood was taken from abdominal aorta for $T_4,\;T_3$ and cyclic nucleotides. The determinations of $T_3,\;T_4$ and cyclic nucleotides were carried out with a radioimmunoassay(RIA) method. The results were summerized as followings. 1) A significant increase of thyroid weight was observed in rats exposured to cold for 2 weeks. Furthermore, in rats administered MTU while to exposure to cold the thyroid weight was also increased significantly. 2) After 2 weeks $T_3$ concentration in the plasma of cold-exposured rats was significantly increased in KI group and MTU group as well as in control group. On the contrary, after 2 weeks of cold exposure $T_4$ level was decreased in control group. 3) In the case of cyclic nucleotides, plasma cAMP was increased in the control group after 1 or 2 weeks of cold exposure. However, cAMP level in plasma was rather significantly decreased in KI group and MTU group as well as in control group.
Objectives: This study was designed to investigate the significance of serum SCC antigen, CA 19-9, CA 125 level and DNA microsatellite alterations (MSA) as prognostic factors and indicators for recurrences in the pre-treatment and post-treatment state, respectively in head and neck cancer patients. Materials and Methods: 120 patients who received curative treatment for head and neck cancer from 1995 to 2000 were followed up successfully, and were analyzed retrospectively. Thirty healthy subjects served as normal controls. Serum SCC Ag levels were measured by microparticle enzyme immunoassay technique via IMX SCC assay, CA 19-9 levels were measured by CA 19-9 RIA test kit, and CA 125 levels were measured by CA 125 IRMA kit. MSA were identified after PCR amplification. Heterozygosity was considered lost if the ratio of one allele was significantly decreased (>50%) in serum DNA compared with normal DNA from lymphocytes. Results: Preoperative tumor markers were higher in cancer patients than control, but not significant. Postoperative SCC Ag levels were lower than preoperative levels. The SCC Ag levels were remained low in no evidence of disease (NED) group, but increased in locoregional recurrence and distant metastasis group. CA 19-9 and CA 125 levels showed no correlation between levels and recurrences and were not decreased significantly after primary tumor removal. MSA were detected in five out of 21 cases, and highly detected in distant metastasis group. Conclusion: SCC Ag seems to be a helpful serum tumor marker for early detection of recurrence and distant metastasis of head and neck cancer after curative treatment. But, CA 19-9 and CA 125 were not reliable markers for head and neck tumors. MSA were not statistically significant because of the small number of study group. However they may be helpful for screening serum molecular markers for early detection of distant metastasis of head and neck cancers.
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