In this study, we analyzed signal transduction by equine follicle-stimulating hormone receptor (eFSHR) on stimulation with recombinant eelFSHβ/α (rec-eelFSHβ/α), natural porcine FSH (pFSH), and natural human FSH (hFSH). cAMP stimulation in CHO-K1 cells expressing eFSHR was determined upon exposure to different doses (0-1450 ng/mL) of these hormones. The EC50 value of rec-eelFSHβ/α was 53.35 ng/mL. The Rmax values of rec-eelFSHβ/α and pFSH were 28.12 and 2.88 ng/mL, respectively. The activity of rec-eelFSHβ/α was much higher than that of natural pFSH. However, signal transduction in CHO PathHunter Parental cells expressing eFSHR was not enhanced by stimulation with natural hFSH. Thus, rec-eelFSHβ/α was completely active in cells expressing eFSHR. However, natural hFSH did not invoke a signal response in cells expressing eFSHR. Particularly, natural pFSH was weakly active in the same cells. These results showed that eelFSHβ/α has potent activity in cells expressing eFSHR. Thus, rec-eelFSHβ/α may efficiently bind to eFSHR, where as natural hFSH does not bind to eFSHR.

In all mammalian species, progesterone is essential to both the preparation for, and maintenance of, pregnancy. The 20α-hydroxysteroid dehydrogenase (20α-HSD) enzyme predominantly converts progesterone into its biologically inactive form 20α-hydroxyprogesterone, thereby regulating its activity. Thus, to directly assess sexual maturation in the MediKinetics micropig®, we analyzed the concentration of the steroid hormones progesterone and estradiol during the estrous cycle.Our results show that the progesterone level exhibited by the analyzed micorpig® was low at the beginning of the estrous cycle, and then abruptly increased to 30.32±10.0 ng/mL and 46.37±11.0 ng/mL by days 9 and 11 of the cycle, respectively. It reached the highest level 55.87±3.5 ng/mL on day 13 of the estrous cycle, before decreasing to 46.58±13.1 ng/mL and 10.0±7.6 ng/mL by days 15 and 17 of the cycle, respectively. In contrast, the estradiol level was shown to be highest (27.13±11.2 ng/mL) at the initiation of the estrous cycle, after which point it decreased to 13.29±6.5 ng/mL and 10.94±5.9 ng/mL by days 4 and 5 of the estrous cycle, respectively. By day 17 of the estrous cycle, the estradiol level decreased to 4.13±7.6 ng/mL.We anticipate that these results will provide useful information to enable the study of human ovulation and reproductive physiology using the MediKinetics micoripig® as a model system. We recommend further investigation to elucidate the functional mechanisms underlying the regulation of sexual maturation in the MediKinetics micropig®.

The purpose of this study was to compare the quantitative and qualitative assessment of dietary intake between patients with metabolic syndrome (MetS) and healthy subjects and to investigate dietary factors related to MetS. Anthropometric measurements, blood analysis, and dietary intake as assessed by 24-hour recall were conducted in MetS patients (n=15) and healthy subjects (n=25). In order to assess the quantity and quality of dietary intake, daily nutrient intake, nutrient density, nutrient intake to dietary reference intake (DRI), nutrient adequacy ratio (NAR), food intake, dietary diversity score (DDS), and dietary variety score (DVS) were analyzed. The statistical differences between MetS patients and controls were analyzed using the SAS software program. Daily energy intake and food intake were not significantly different between the two groups (2,154.3 kcal vs. 1,872.9 kcal; 1,280.0 g vs. 1,261.6 g). There were also no significant differences in daily nutrient intake, nutrient intake ratio to DRI, NAR, or DVS between the MetS group and the control group. However, daily intake of eggs and milk in MetS patients was significantly lower than in the control group (9.0 g/day vs. 30.3 g/day, p<0.05; 0 g/day vs. 49.7 g/day, p<0.05). These results indicate that low intake of eggs and dairy products may be related to the development of MetS.

In this study, we conducted an oral glucose tolerance test (OGTT) so as to compare antidiabetic activities of general potatoes, purple-flesh potatoes, and potato pigments in rats at various concentration levels. After allowing the rats to abstain from food for 12 hours, 10%/20% general potato, purple-flesh potato, and potato extract was orally administered to rats at 100 and 500 mg/kg concentrations. The blood glucose level was measured after an hour. Then, immediately, 1.5 g/kg of sucrose was administered through the abdominal cavity and the blood glucose measured after 30, 60, 120, and 180 minutes. 20% purple-flesh potato group and 10% general potato group, both 100 and 500 mg/kg, showed a significant concentration-dependent decrease in blood glucose levels after 30 minutes. The 100 mg/kg potato pigment group also showed a statistically significant decrease after 30 minutes. In conclusion, administration of 10% general potato, 20% purple-flesh potato, and potato pigment can reduce blood glucose level in an OGTT using rats.

The purpose of present study was to analyze mineral contents in various tissues and investigate theirs relation with bone mineral density (BMD) in rats. Fifteen Sprague-Dawley rats were fed standard diet for 4 weeks. Body weight gain, feed intake, and feed efficiency ratio were 41.00 g/week, 171.15 g/week, and 0.24 respectively. Among 12 minerals in serum, Ca is the highest with 6.86 mg/dl. Serum Mg, Se, and Cu were 2.52 mg/dl, 0.23 mg/dl and 0.22 mg/dl respectively. Mg contents in liver, spleen, and kidney were 246.36 μg/g, 105.01 μg/g, and 273.38 μg/g respectively. Tibia contents of Ca, Mg, Zn, Fe and V were 194.91 mg/g, 23.10 mg/g, 0.60 mg/g, 0.35 mg/g and 0.14 mg/g respectively. BMDs of right tibia and spine were 122.04 mg/cm2 and 153.61 mg/cm2. There were significantly positive correlations between tibia BMD and Se (p<0.05), tibia BMD and V (p<0.01), spinal BMD and V(p<0.05), respectively. It's expected that these results are used as a reference data in following study to elucidate physiological function of minerals.

The large extracellular domain of glycoprotein hormone receptors is a unique feature within the G protein-coupled receptors (GPCRs) family. After interaction with the hormone, the receptor becomes coupled to Gs, which, in turn stimulates adenylyl cyclase and the production of cAMP. Potential phosphorylation sites exist in the C-terminal region of GPCRs. The experiments described herein represent attempts to determine the functions of the eel follicle-stimulating hormone receptor (eelFSHR). We constructed a mutant of eelFSHR, in which the C-terminal cytoplasmic tail was truncated at residue 614 (eelFSHR-t614). The eelFSHR-t614 lacked all potential phosphorylation sites present in the C-terminal region of eelFSHR. In order to obtain the eelFSHR ligand, we produced recombinant follicle-stimulating hormone (rec-eelFSHβ/α) in the CHO-suspension cells. The expression level was 2-3 times higher than that of the transient expression of eelFSH in attached CHO-K1 cells. The molecular weight of the rec-eelFSHβ/α protein was identified to be approximately 34 kDa. The cells expressing eelFSHR-t614 showed an increase in agonist-induced cAMP responsiveness. The maximal cAMP responses of cells expressing eelFSHR-t614 were lower than those of cells expressing eelFSHR-wild type (eelFSHR-WT). The EC50 following C-terminal deletion in CHO-K1 cells was approximately 60.4% of that of eelFSHR-WT. The maximal response in eelFSHR-t614 cells was also drastically lower than that of eelFSHR-WT. We also found similar results in PathHunter Parental cells expressing β-arrestin. Thus, these data provide evidence that the truncation of the C-terminal cytoplasmic tail phosphorylation sites in the eelFSHR greatly decreased cAMP responsiveness and maximal response in both CHO-K1 cells and Path-Hunter Parental cells expressing β-arrestin.

Previous studies showed that recombinant equine chorionic gonadotropin (rec-eCGβ/α) exhibits both folliclestimulating hormone (FSH) and luteinizing hormone (LH)-like activities in rat LHR- and FSHR-expressing cells. In this study, we analyzed signal transduction by eelFSHR and eelLHR upon stimulation with rec-eCGβ/α and native eCG. The cyclic adenosine monophosphate (cAMP) stimulation in CHO-K1 cells expressing eelLHR was determined upon exposure to different doses (0–1,450 ng/mL) of rec-eCGβ/α and native eCG. The EC50 values of rec-eCGβ/α and native eCG were 172.4 and 786.6 ng/mL, respectively. The activity of rec-eCGβ/α was higher than that of native eCG. However, signal transduction in the CHO PathHunter Parental cells expressing eelFSHR was not enhanced by stimulation with both agonist rec-eCGβ/α and native eCG. We concluded that rec-eCGβ/α and native eCG were completely active in cells expressing eelLHR, similar to the activity in the mammalian cells expressing LHRs. However, rec-eCGβ/α and native eCG did not invoke any signaling response in the cells expressing eelFSHR. These results suggest that eCG has a potent activity in cells expressing eelLHR. Thus, we also suggest that rec-eCGβ/α can induce eel maturation by administering gonadotropic reagents (LH), such as salmon pituitary extract.

Equine chorionic gonadotropin (eCG) is a unique molecule that elicits the response characteristics of both follicle- stimulating hormone (FSH) and luteinizing hormone (LH) in other species. Previous studies from this laboratory had demonstrated that recombinant eCG (rec-eCG) from Chinese hamster ovary (CHO-K1) cells exhibited both FSH- and LH-like activity in rat granulosa and Leydig cells. In this study, we analyzed receptor internalization through rec-eCGs, wild type eCG (eCGβ/α) and mutant eCG (eCGβ/αΔ56) with an N-linked oligosaccharide at Asn56 of the α-subunit. Both the rec-eCGs were obtained from CHO-K1 cells. The agonist activation of receptors was analyzed by measuring stimulation time and concentrations of rec-eCGs. Internalization values in the stably selected rat follicle-stimulating hormone receptor (rFSHR) and rat luteinizing/ chorionic gonadotropin receptor (rLH/CGR) were highest at 50 min after stimulation with 10 ng of rec-eCGβ/α. The dose-dependent response was highest when 10 ng of rec-eCGβ/α was used. The deglycosylated eCGβ/αΔ56 mutant did not enhance the agonist-stimulated internalization. We concluded that the state of activation of rFSHR and rLH/CGR could be modulated through agonist-stimulated internalization. Our results suggested that the eLH/CGRs are mostly internalized within 60 min by agonist-stimulation by rec-eCG. We also suggested that the lack of responsiveness of the deglycosylated eCGβ/ αΔ56 was likely because the site of glycosylation played a pivotal role in agonist-stimulated internalization in cells expressing rFSHR and rLH/CGR.