• Korean Journal of Fisheries and Aquatic Sciences
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• v.30 no.3
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• pp.473-479
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• 1997

The yolk protein of spotted flounder, Verasper variegatus was purified by precipitation wit cold distilled water, followed by Sepharose CL-6B column chromatography. The purified protein was identified as vitellin by Ouchterlony's immunodiffusion test and immunoelectrophoresis. The purified vitellin from ovarian crude extracts has same antigenic determinants with the female specific serum protein, vitellogenin. The molecular weight of purified vitellin was estimated about 550 kD by gel filfration. The vitellin was composed of three major subunits with molecular weight of about 108, 85 and 31 kD, and two minor subunits. The vitellin was identified by western blot analysis using anti-vitellin antibody.

• The Korean Journal of Zoology
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• v.29 no.2
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• pp.86-106
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• 1986

For the comparative study of protein patterns of the sera of pregnant women, the protein in sera of normal male subjects, non-pregnant women, pregnant women and women delivered of children were analyzed by using the methods of SDS/polyacrylamide gel electrophoresis, two-dimensional electrophoresis and amino acid analysis. The results were as follows: 1. When the protein patterns of sera in normal male ranging from 10, 000 to 110, 000 daltons were compared to non-pregnant women by SDS/polyacrylamide gel electrohporesis, their protein patterns were same each other numerically but bands 3(22, 000 dalton) and 6(39, 000) were less in male than in non-pregnant women quantitatively. When the protein patterns in the pregnant women in which serum were collected two week intervals were compared with non-pregnant women, there was increased or decreased in several bands quantitavely. The protein patterns of sera in pregnant women were compared with those of non-pregnant women; band 3(22, 000) showed similar patterns each other until the 16th week but the quantity of protein was decreased continously from the 18th week to the third trimester of pregnancy. Contary, bands 4(24, 000), 9(69, 000), 10(70, 000), 12(80, 000), 14(86, 000), 15(91, 000) and 16(94, 000) were gradually increased in quantity from the begining of gestation, and band 7(51, 000) was increased until the 32th week of gestation only but somewhat decreased after this time. The quantities of bands 12(80, 000), 15(91, 000) and 16(94, 000) were relatively increased when the protein patterns of delivered women were compared with those of the third trimester of pregnancy. Women who were dilivered female children showed more increase in bands 4(24, 000), 7(51, 000) and 10(70, 000)than one who were delivered male chilren. 2. When the protein patterns of sera in normal males were compared with those of nonpregnant women by two-dimensional electrophoresis, three spots of spot a group were not appeared in the males and the spot c group in the males was less than in the non-pregnant women. In the pregnant women, albumin was significantly decreased during the 10-12 week of gestation but recovered after these times. And spot f(70, 000) was decreased in the 10th week of gestation but increased from this time. 3. Glutamic acid, lysine, aspartic acid, leucine and valine in pregnant women were large in quantity while methionine, isoleucine and glycine were small in quantity by amino acid analysis. The total amino acids were increased remarkably in the second trimester of pregnancy but began to decrease in the third trimester of pregnancy. As mentioned, this present paper deat with that proteins which consist of maternal serum were increased with specific period in pregnancy and that the change of characteristic protein patterns were identified in the serum protein of each trimester in the pregnant women. And furthermore, the study should be preformed for the sex-identification of a fetus in the early pregnancy.

• Korean Journal of Veterinary Research
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• v.31 no.4
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• pp.491-500
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• 1991

This study was conducted to detect the serum antibodies in the experimentally toxoplasma infected dogs and street dogs by use the of an enzyme-linked immunosorbent assay (ELISA). And this test was performed on the polystylene microplate by coating with the tachyzoites soluble antigen of T gondii (RH strain), incubated with sera diluted then, added with HPO-conjugated rabbit anti-dog IgG and o-phenylenediamine used as a substrate. Tachyzoites of T gondii harvested from mouse peritoneal cavity were purified by 30, 40 and 50% Percoll density gradient centrifugation and used as the source of antigen. The results obtained were summarized as follows; 1. The highest ratio of positive to negative (P/N ratio) was obtained at the level of $l{\mu}g/ml$ protein concentration of antigen with the 1/4000 dilution of the conjugate measured by checker-board titration. It was regarded as the optimum concentration of the antigen and conjugate. 2. Cut-off value in this IgG ELISA was 0.375 that was determined by mean absorbance (at 492nm) of IFA negative serum added with the dauble value of the standard deviation $(mean{\pm}2S.D.)$. 3. Serum ELISA IgG antibodies to T gondii in the exyerimentally infected dogs were detected firstly at the Week 3 after inoculation and the highest titer was recognized at the Week 4, 5 and 6 after inoculation. 4. Stability of the antigen absorbed in the microplates that were preserved at $4^{\circ}C$ and $-25^{\circ}C$ separately were prolonged up to 3 weeks and 10 weeks at $4^{\circ}C$ and $-25^{\circ}C$, respectively. However the reproducibility was not reliable after the preservation of 4 weeks and longer. 5. Positive rate of the specific antibodies in 312 test sera was 28.5% and there was no significant differences between the male (27.8%) and female (29.5%), respectively. 6. The IgG ELISA was proved to be a specific procedure for the detection of antibodies to canine toxoplasma infection and also evaluated as a screening test for the large scale of test samples in laboratory.

• Journal of Animal Science and Technology
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• v.63 no.2
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• pp.426-439
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• 2021

It is already well known that castration improves marbling quality but exact timing of castration is still highly debated in beef cattle production industry. After castration, blood hormonal changes occur in steer and objective of this study was to investigate the effects of growth hormone (GH) levels on adipocyte differentiation in stromal vascular cells (SVCs) and transdifferentiation into adipocytes in C2C12 myoblasts. Total GH concentrations were measured via enzyme-linked immunosorbent assay (ELISA) in 24 male calves and 4 female calves. Cell proliferation, cellular triglyceride (TG) accumulation, and the cell's lipolytic capability were measured in C2C12 myoblasts and SVCs. Myogenic, adipogenic, and brown adipocyte-specific gene expression was measured via real-time polymerase chain reaction (PCR) using SYBR green. Serum GH levels were the highest in late-castrated calves. Treatment with 5 ng/mL GH resulted in greater TG accumulation as well as increased CCAAT-enhancer-binding protein (C/EBP)α and peroxisome proliferator-activated receptor (PPAR)γ expression compared to that after treatment with 15 ng/mL GH. Treatment with 5 ng/mL GH also resulted in lower myogenin (myo)G and myoD expression compared to that after treatment with 15 ng/mL GH. The expression of bone morphogenetic protein (BMP) 7 after treatment with 5 ng/mL GH was higher than that after treatment with 15 ng/mL GH. But carcass characteristics data showed no significant difference between early and late castrated steers. Therefore, our results indicate that castration timing does not seem to be inevitable determinate of carcass qualities, particularly carcass weight and marbling score in Hanwoo beef cattle.

• Journal of Korean Neurosurgical Society
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• v.65 no.2
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• pp.204-214
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• 2022

Objective : Osteoporosis result from age-related decline in the number of osteoblast progenitors in the bone marrow. Probiotics have beneficial effects on the host, when administered in appropriate amounts. This study investigated the effects of probiotics expressing specific genes, especially the effects of genetically modified bone morphogenetic protein (BMP)-2-expressing Lactobacillus plantarum CJNU 3003 (LP) on ovariectomized rats. Methods : Twenty-eight female Wistar rats (250-300 g, 12 weeks old) were divided into four groups : the sham (control), the ovariectomy (OVX)-induced osteoporosis group (OVX), the OVX and LP (OVX/LP), OVX and genetically modified BMP-2-expressing LP (OVX/LP with BMP) groups. The three groups underwent bilateral OVX and two of these groups were administered two different types of LP via oral gavage daily. At 16 weeks post-OVX, blood was collected from the heart and the bilateral tibiae were extracted and were scanned by ex-vivo micro-computed tomography and stained with hematoxylin-and-eosin (H&E) and Masson's trichrome stain for pathological assessment. The serum levels of osteocalcin (OC), rat C-telopeptide of type I collagen (CTX-I), BMP-2, and receptor activator of nuclear factor-ĸB ligand (RANKL) were measured. Results : The 3D-micro-computed tomography images showed that the trabecular structure in the OVX/LP with BMP group was maintained compared with OVX and OVX/LP groups. No significant differences were detected in trabecular thickness (Tb.Th) between control and OVX/LP with BMP groups (p>0.05). Furthermore, a tendency toward increased BMD, trabecular bone volume, Tb.Th, and trabecular number and decreased trabecular separation was found in rats in the OVX/LP with BMP groups when compared with the OVX and OVX/LP groups (p>0.05). The H&E and Masson's trichrome stained sections showed a thicker trabecular bone in the OVX/LP with BMP group compared with the OVX and OVX/LP groups. There was no difference in serum levels of OC, CTX and RANKL control and OVX/LP with BMP groups (p>0.05). In contrast, significant differences were found in OC and CTX-1 levels between the OVX and OVX/LP with BMP groups (p<0.05). Conclusion : Our results showed that the expression of genetically modified BMP-2 showed inhibition effect for bone loss in a rat model of osteoporosis.

• Journal of Nutrition and Health
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• v.31 no.1
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• pp.46-61
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• 1998

This study was carried out to evaluate the effects of vitamin D nutritional status on bone mineral density of adults (21-49 years). To attain the aim, we measured bone mineral density (BMD) of the subjects at distal radius by single-photon absorptiometry (SPA). Serum level of 25-hydroxyvitamin D(25-(OH)D) , known to be the best indicator of indicator of vitamin D status in humans was analyzed . The factors affecting this vitamin D level were also investigated in autumn in 122 young adults. Serum level of 25-(OH)D was measured by high pressure liquid chromatography(HPLC) and biochemical variables, general health status, time spent outdoors, and dietary intakes of the subjects. BMD of the male subjects was significantly greater than that of female subjects. Weight, activity and total energy expediture (TEE) showed a positive correlation with distal BMD. The mean level of serum 25-(OH)D was 24.4$\pm$11.0 ng./ml and by sex, 26.0$\pm$6.8ng/ml for males and 23.3$\pm$12.3ng/ml for females , the level was significantly higher in male (p<0.01). there was significant correlation between BMD at distal-radius and s-25(OH)D levels (p<0.001). The serum level fo parathyroid hormone (PTH) showed a negative correlation with BMD(p<0.05), with the more obvious correlation in females. Vitamin D intake was estimated to be 3.75$\pm$2.19ug/day in average. Among the nutrients studied, protein ,fat, calcium , and vitamin D intake were positively correlated with distal BMD. When food frequencies were concerned , milk and dairy products showed a significant positive correlation with the BMD level, and driedfoods, eggs , fats and oils, and cereals also showed a positive correlation. Time spent outdoors was estimated to be about 70 minutes in average and positively correlated with the distal BMD level(p<0.01). During the day, the specific time between 12 :00pm and 2:00pm showed the most significant correlation with BMD (p<0.001). Multiple regression analysis with the variables showed that distal BMD could be fit 31.9% by the time spent outdoors a day, intake of Ca and vitamin D, and TEE. The standardized estimates were 0.344 for vitamin D intake, 0.284 for Ca intake 0.179 for the time spent outdoors a day and 0.273 for TEE. For males, s-25*OH)D level, TEE and time spent outdoors during a day showed a significant correlation. For females, intake of Ca and vitamin D could fit about 27.1% of the distal BMD.

• Development and Reproduction
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• v.9 no.2
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• pp.135-142
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• 2005

Vitellogenins(VTGs) are the precursor of egg-yolk proteins in most oviparous species from invertebrates to vertebrates. In oviparose vertebrates, VTGs are synthesized in the liver and transported through the blood to oocytes. In female fish, concentrations of plasma VTG increase rapidly at onset of vitellogenesis in the normal reproductive cycle. Male fishes also possess the gene for VTG, but plasma concentrations of the protein typically remain small, presumably due to low levels of endogenous estrogens. However, exposure of males to exogenous estrogenic mimics can result elevated. Therefore, the VTG in fish can be used as a useful biomarker for appropriate tools of endocrine disrupting compounds effects. In this studies, we prepared the test methods that can measure the plasma VTG level in the gobies that live in polluted area with mimic estrogen. For the purpose, we purified VTG of floating goby(Chaenogobius annularis) and prepared specific monoclonal and polyclonal antisera to yolk protein, then developed a sandwich competitive ELISA system for measurement of plasma VTG levels. Validation for the ELISA system using monoclonal and polyclonal antibodies against VTG was tested. The absorbance curve of serial dilutions of serum from vitellogenic female was paralleled to the standard curve of VTG, but normal male was not paralleled. The developed sandwich ELISA system was measured for VTG levels in plasma of common goby(Acanthogobius flaviman) and javeline goby(A. hasta) as well as in plasma of floating goby(C. annularis).

• Journal of Veterinary Science
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• v.21 no.6
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• pp.81.1-81.10
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• 2020

Background: Although previous in vivo studies explored urinary microRNA (miRNA), there is no agreement on nephrotoxicity-specific miRNA biomarkers. Objectives: In this study, we assessed whether urinary miRNAs could be employed as biomarkers for nephrotoxicity. Methods: For this, literature-based candidate miRNAs were identified by reviewing the previous studies. Female Sprague-Dawley rats received subcutaneous injections of a single dose or repeated doses (3 consecutive days) of gentamicin (GEN; 137 or 412 mg/kg). The expression of miRNAs was analyzed by real-time reverse transcription-polymerase chain reaction in 16 h pooled urine from GEN-treated rats. Results: GEN-induced acute kidney injury was confirmed by the presence of tubular necrosis. We identified let-7g-5p, miR-21-3p, 26b-3p, 192-5p, and 378a-3p significantly upregulated in the urine of GEN-treated rats with the appearance of the necrosis in proximal tubules. Specifically, miR-26-3p, 192-5p, and 378a-3p with highly expressed levels in urine of rats with GEN-induced acute tubular injury were considered to have sensitivities comparable to clinical biomarkers, such as blood urea nitrogen, serum creatinine, and urinary kidney injury molecule protein. Conclusions: These results indicated the potential involvement of urinary miRNAs in chemical-induced nephrotoxicity, suggesting that certain miRNAs could serve as biomarkers for acute nephrotoxicity.

• Journal of Veterinary Science
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• v.22 no.3
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• pp.44.1-44.15
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• 2021

Background: Intraoperative fluids are still poorly studied in veterinary medicine. In humans the dosage is associated with significant differences in postoperative outcomes. Objectives: The aim of this study is to verify the influence of three different fluid therapy rates in dogs undergoing video-assisted ovariohysterectomy. Methods: Twenty-four female dogs were distributed into three groups: G5, G10, and G20. Each group was given 5, 10, and 20 mL·kg^-1·h^-1 of Lactate Ringer, respectively. This study evaluated the following parameters: central venous pressure, arterial blood pressure, heart rate, respiratory rate, temperature, acid-base balance, and serum lactate levels. Additionally, this study evaluated the following urinary variables: urea, creatinine, protein to creatinine ratio, urine output, and urine specific gravity. The dogs were evaluated up to 26 h after the procedure. Results: All animals presented respiratory acidosis during the intraoperative period. The G5 group evidenced intraoperative oliguria (0.80 ± 0.38 mL·kg^-1·h^-1), differing from the G20 group (2.17 ± 0.52 mL·kg^-1·h^-1) (p = 0.001). Serum lactate was different between groups during extubation (p = 0.036), with higher values being recorded in the G5 group (2.19 ± 1.65 mmol/L). Animals from the G20 group presented more severe hypothermia at the end of the procedure (35.93 ± 0.61℃) (p = 0.032). Only the members of the G20 group presented mean potassium values below the reference for the species. Anion gap values were lower in the G20 group when compared to the G5 and G10 groups (p = 0.017). Conclusions: The use of lactated Ringer's solution at the rate of 10 mL·kg^-1·h^-1 seems to be beneficial in the elective laparoscopic procedures over the 5 or 20 mL·kg^-1·h^-1 rates of infusion.

• Journal of Veterinary Clinics
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• v.23 no.1
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• pp.72-76
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• 2006

A 1-year-old, female English cocker spaniel (ECS) dog was presented with 3-month history of vomiting and retaking of the vomitus, and chronic weight loss. The client had noticed mild abdominal distension 10 days before. The dog was diagnosed as chronic hepatitis with hepatic cirrhosis based on complete blood count (CBC), serum chemistry profiles, radiography, ascites assessment, bile acid evaluation, and liver biopsy through exploratory laparotomy and necropsy. CBC and serum chemistry profiles revealed mild anemia, slightly elevated hepatic enzymes (ALT and AST), increased creatinine kinase (CK), hyperammonemia, and hypoproteinemia with hypoalbuminemia. Ascites was transudate according to analysis of components. Bile acid assessment (fasting; $174.4{\mu}mol/L$ and postprandial; $198.4{\mu}mol/L$) showed strongly suspected hepatic insufficiency. On radiological findings, ascites was evident. Atrophied liver (especially left side lobes) and distended mesenteric vasculatures were observed by exploratory laparotomy. Histopathological examination of marginal lesion of left lateral lobe of liver by biopsy revealed the necrosis of hepatic cells, dilation of sinusoids, infiltration of neutrophils in sinusoids, and vacuolation of hepatic cytoplasm. The patient had been managed with careful low protein diet and specific supportive therapy (ursodeoxycholic acid, prednisolone, vitamine E, and interferon). Vomiting and ascites disappeared with medical management. The dog was monitored periodically by CBC, serum chemistry and radiographic examination. The dog survived more 18 months with medical therapy. After spontaneous death, necropsy and histopathologic examination were performed.