• Clinical and Experimental Reproductive Medicine
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• v.30 no.1
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• pp.47-55
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• 2003

Objectives: Bok, Bcl-2-related ovarian killer, is a proapoptotic Bcl-2 family protein identified in the ovary based on its dimerization with the antiapoptotic protein Mcl-1. The present study examined the hormonal regulation and localization of Bok messenger RNA levels in the mouse ovary during the follicle development. Methods: The animals were implanted subcutaneously with Silastic brand capsules containing the synthetic estrogen, DES at $21{\sim}23$ days of age. Ovaries were collected $1{\sim}3$ days after implantation for RNA analysis and in situ hybridization. Some mice were removed capsule for $1{\sim}2$ days to induce ovarian follicle apoptosis. Ovaries were also collected from 26 day-old immature mice at various times after treatment with 10 IU PMSG. Some mice received a single intraperitoneal injection of 10 IU hCG to induce ovulation, and ovaries were obtained at different time intervals for Northern blot and in situ hybridization analysis, respectively. Results: Treatment of immature mice with diethylstilbestrol (DES) for $24{\sim}48$ h increased ovarian Bok mRNA levels. Bok mRNA was remained the same levels in mice removed DES for $24{\sim}48$ h to induce apoptosis. High signals of Bok mRNA after DES treatment were detected in granulosa cells of early antral follicles. Treatment of immature mice with PMSG for 12 h increased markedly ovarian Bok mRNA expression which was detected mainly in preantral and atretic follicles. Interestingly, low levels of Bok mRNA were also expressed in granulosa cells of preovulatory follicles. Treatment of PMSGprimed mice with hCG stimulated strongly ovarian Bok mRNA expression at $6{\sim}9$ h. At that time, Bok mRNA was expressed in granulosa cells of atretic and small growing follicles. Conclusion: These results demonstrate that Bok is one of proapoptotic Bcl-2 members expressed in early growing and atretic follicles during the ovarian follicular development. Gonadotropins induce a transient increase of Bok gene expression in granulosa cells of preantral and preovulatory follicles indicating some role in the ovulatory process.

• The Korean Journal of Cytopathology
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• v.7 no.2
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• pp.163-168
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• 1996

Abnormalities of p53 gene are common in lung cancers and are associated with immunologically detectable p53 protein. p53 immunoreactivity is uncommon in normal cells but is frequently seen in neoplasia. Therefore, assessment of p53 expression may assist in the cytological diagnosis of malignancy. The usefulness of p53 immunostaining as a marker of malignancy in the cytological analysis of bronchial brush specimens from the patients with lung cancers was investigated in this study. A total of 71 bronchial brush samples submitted for cytologic diagnosis were immunostained with D07, a monoclonal antibody to recombinant p53 protein. Resultant p53 data were correlated with cytologic diagnosis and clinical information. Of the 17 smears with a benign cytodiagnosis, all were p53 negative. Of the 40 cases with a malignant cytodiagnosis (histologically confirmed), 35 were p53 positive and 5 were negative. Of the 14 cases that were cytologically suspicious but nondiagnostic for malignancy, 11 were p53 positive, 9 of which were subsequently proved to be malignant by histologic examination, and the remaining 2 cases were tuberculosis clinically. Forty four of 51 histologically confirmed lung carcinomas were p53 positive, including 25 of 28 squamous cell carcinomas, 13 of 17 small cell carcinomas, 3 of 3 adenocarcinomas, and 3 of 3 large cell undifferentiated carcinomas. These results suggest that p53 immunostaining could be of value as a marker of malignancy in the cytologic examination of bronchial brush specimens. Furthermore, we have shown the possible clinical utility of p53 immunostaining in cytopathological diagnosis, that is, as a valuable adjunct to morphological assessment in the analysis of cytopathologically suspicious cases.

• Journal of Pharmacopuncture
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• v.2 no.1 s.2
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• pp.135-152
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• 1999

To investigate effects of Taraxaci herbal-acupuncture on Adjuvant Arthritis in rats, the edema rate, the number of WBC, the quantity of total protein, albumin and globulin in the blood serum and histological test of the muscular tissue were measured in the arthritis part. 1. After elicitating arthritis of Sprague Dawely(SD) rats by injection of Freund's complets adjuvant for 2 weeks, saline was injected for the Exp. I group and Taraxaci herbal-acupuncture was injected for the Exp. II group during 30days. Selected point was $ST_{35}$ in both the groups. And then the volume of the paw were checked. The volume of paw was $0.84{\pm}0.14mm$ in the Exp. I group and $0.38{\pm}0.17mm$ in the Exp. II group, the swelling of the paw was restricted significantly in the Exp. II group(p<0.05) 2. The number of WBC was $10.34{\pm}0.14(10^3/ml)$ in the normal group and $37.47{\pm}5.46(10^3/ml)$ in the Exp. I group. It was $20.39{\pm}4.23(10^3/ml)$ in the Exp. II group. This fact showed that the group Exp. II was more effective than the Exp. I group in the treatment of arthritis(p<0.05) 3. The content of the total protein in the blood serum was $6.14{\pm}0.43g/dl$ in the normal group, $7.95{\pm}0.94g/dl$ in the Exp. I group, and $6.38{\pm}1.75g/dl$ in the Exp. II group. This fact showed that the group Exp. II was more effective than the Exp. I group in the treatment of arthritis(p<0.05) 4. The contents of albumin in the blood serum was $2.94{\pm}0.13g/dl$ in the normal group, $2.01{\pm}0.48g/dl$ in the Exp. I group. and $2.71{\pm}0.34g/dl$ in the Exp. II group. This fact showed that the group Exp. II was more effective than the Exp. I group in the treatment of arthritis(p<0.05) 5. The contents of globulin in the blood serum was $3.19{\pm}0.48g/dl$ in the normal group, $4.70{\pm}1.26g/dl$ in the Exp. I group. and $3.67{\pm}0.56g/dl$ in the Exp. II group. There was no significance in the serum globulin between Exp. II group and Exp. I group from the stastical analysis 6. In histological finding, because of severe inflammatory reaction, remarkably irregular tissue and large amount of inflammatory cells were found in the Exp. I group. But the Exp. II group showed small amount of inflammatory cells, the refrained inflammatory state and even recovering state. From these results, it is showed Taraxaci herbal-acupuncture refrain inflammatory reaction and muscular tissue necrosis in SD rats paw were induced by Freund's complete adjuvant

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.2
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• pp.239-246
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• 2015

The present study was conducted to evaluate the effect of the dietary supplementation of Bacillus amyloliquefaciens-based direct-fed microbial (DFM) on growth performance, nutrient utilization, intestinal morphology and cecal microflora in broiler chickens. A total of two hundred and eighty eight 1-d-old Arbor Acres male broilers were randomly allocated to one of four experimental treatments in a completely randomized design. Each treatment was fed to eight replicate cages, with nine birds per cage. Dietary treatments were composed of an antibiotic-free basal diet (control), and the basal diet supplemented with either 15 mg/kg of virginiamycin as antibiotic growth promoter (AGP), 30 mg/kg of Bacillus amyloliquefaciens-based DFM (DFM 30) or 60 mg/kg of Bacillus amyloliquefaciens-based DFM (DFM 60). Experimental diets were fed in two phases: starter (d 1 to 21) and finisher (d 22 to 42). Growth performance, nutrient utilization, morphological parameters of the small intestine and cecal microbial populations were measured at the end of the starter (d 21) and finisher (d 42) phases. During the starter phase, DFM and virginiamycin supplementation improved the feed conversion ratio (FCR; p<0.01) compared with the control group. For the finisher phase and the overall experiment (d 1 to 42) broilers fed diets with the DFM had better body weight gain (BWG) and FCR than that of control (p<0.05). Supplementation of virginiamycin and DFM significantly increased the total tract apparent digestibility of crude protein (CP), dry matter (DM) and gross energy during both starter and finisher phases (p<0.05) compared with the control group. On d 21, villus height, crypt depth and villus height to crypt depth ratio of duodenum, jejunum, and ileum were significantly increased for the birds fed with the DFM diets as compared with the control group (p<0.05). The DFM 30, DFM 60, and AGP groups decreased the Escherichia coli population in cecum at d 21 and d 42 compared with control group (p<0.01). In addition, the population of Lactobacillus was increased in DFM 30 and DFM 60 groups as compared with control and AGP groups (p<0.01). It can be concluded that Bacillus amyloliquefaciens-based DFM could be an alternative to the use of AGPs in broilers diets based on plant protein.

• Journal of Life Science
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• v.20 no.12
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• pp.1777-1783
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• 2010

Plants generate reactive oxygen species (ROS) as a by-product of normal aerobic metabolism or when exposed to a variety of stress conditions, which can cause widespread damage to biological macromolecules. To protect themselves from oxidative stress, plant cells are equipped with a wide range of antioxidant proteins. However, the detailed reaction mechanisms of these are still unknown. Peroxiredoxins (Prxs) are ubiquitous thiol-containing antioxidants that reduce hydrogen peroxide with an N-terminal cysteine. The active-site cysteine of peroxiredoxins is selectively oxidized to cysteine sulfinic acid during catalysis, which leads to inactivation of peroxidase activity. This oxidation was thought to be irreversible. Recently identified small protein sulphiredoxin (Srx1), which is conserved in higher eukaryotes, reduces cysteine.sulphinic acid in yeast peroxiredoxin. Srx1 is highly induced by $H_2O_2$-treatment and the deletion of its gene causes decreased yeast tolerance to $H_2O_2$, which suggest its involvement in the metabolism of oxidants. Moreover, Srx1 is required for heat shock and oxidative stress induced functional, as well as conformational switch of yeast cytosolic peroxiredoxins. This change enhances protein stability and peroxidase activity, indicating that Srx1 plays a crucial role in peroxiredoxin stability and its regulation mechanism. Thus, the understanding of the molecular basis of Srx1 and its regulation is critical for revealing the mechanism of peroxiredoxin action. We postulate here that Srx1 is involved in dealing with oxidative stress via controlling peroxiredoxin recycling in Arabidopsis. This review article thus will be describing the functions of Prxs and Srx in Arabidopsis thaliana. There will be a special focus on the possible role of Srx1 in interacting with and reducing hyperoxidized Cys-sulphenic acid of Prxs.

• Parasites, Hosts and Diseases
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• v.51 no.6
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• pp.719-726
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• 2013

Mitochondrial genomes have been extensively studied for phylogenetic purposes and to investigate intra- and interspecific genetic variations. In recent years, numerous groups have undertaken sequencing of platyhelminth mitochondrial genomes. Haplorchis taichui (family Heterophyidae) is a trematode that infects humans and animals mainly in Asia, including the Mekong River basin. We sequenced and determined the organization of the complete mitochondrial genome of H. taichui. The mitochondrial genome is 15,130 bp long, containing 12 protein-coding genes, 2 ribosomal RNAs (rRNAs, a small and a large subunit), and 22 transfer RNAs (tRNAs). Like other trematodes, it does not encode the atp8 gene. All genes are transcribed from the same strand. The ATG initiation codon is used for 9 protein-coding genes, and GTG for the remaining 3 (nad1, nad4, and nad5). The mitochondrial genome of H. taichui has a single long non-coding region between trnE and trnG. H. taichui has evolved as being more closely related to Opisthorchiidae than other trematode groups with maximal support in the phylogenetic analysis. Our results could provide a resource for the comparative mitochondrial genome analysis of trematodes, and may yield genetic markers for molecular epidemiological investigations into intestinal flukes.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.11
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• pp.1445-1454
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• 2010

Two separate animal trials were conducted to evaluate the coincidence of dry matter intake (DMI) and average daily gain (ADG) predicted by the Cornell Net Carbohydrate and Protein System (CNCPS) and observed actually in crossbred growing bulls kept in a traditionally confined feeding system in China. In Trial 1, 45 growing Simmental${\times}$Mongolia crossbred F1 bulls were assigned to three treatments (T1-3) with 15 animals in each treatment. Trial 2 was conducted with 60 Limousin${\times}$Fuzhou crossbred F2 bulls allocated to 4 treatments (t1-4). All of the animals were confined in individual stalls. DMI and ADG for each bull were measured as a mean of each treatment. All of the data about animals, environment, management and feeds required by the CNCPS model were collected, and model predictions were generated for animals on each treatment. Subsequently, model-predicted DMI and ADG were compared with the actually recorded results. In the three treatments in Trial 1, 93.3, 80.0 and 73.3% of points fell within the range from -0.4 to 0.4 kg/d for DMI mean bias; similarly, in the four treatments in Trial 2, about 86.7, 73.3, 73.3 and 80.0% of points fell within the same range. These results indicate that the CNCPS model can accurately predict DMI of crossbred bulls in the traditionally confined feeding system in China. There were no significant differences between predicted and observed ADG for T1 (p = 0.06) and T2 (p = 0.09) in Trial 1, and for t1 (p = 0.07), t2 (p = 0.14) and t4 (p = 0.83) in Trial 2. However, significant differences between predicted and observed ADG values were observed for T3 in Trial 1 (p<0.01) and for t3 in Trial 2 (p = 0.04). By regression analysis, a statistically different value of intercept from zero for the regression equation of DMI (p<0.01) or an identical value of ADG (p = 0.06) were obtained, whereas the slopes were significantly different (p<0.01) from unity for both DMI and ADG. Additionally, small root mean square error (RMSE) values were obtained for the unbiased estimator of the two variances (DMI and ADG). Thus, the present results indicated that the CNCPS model can give acceptable estimates of DMI and ADG of crossbred growing bulls kept in a traditionally confined feeding system in China.

• Journal of Nutrition and Health
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• v.37 no.10
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• pp.917-927
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• 2004

This study was performed to examine component of variance in nutrient intakes and to estimate the level of accuracy with varying degree of precision in order to achieve estimates of usual nutrient intakes. Three-day dietary records including both weekdays and weekends were collected every 4 season over a I-year period from 36 males and 93 females aged 40 - 65 y. For each nutrient, we partitioned total intake variance into weekly (weekday vs weekend), seasonal, within- and between- individual variation as components of variance, using analysis of variance. It was found that major components of variance were within- and between-individual variation. Particularly, within-individual variation (57.2 - 87.1 %) was greater than between-individual variation (12.2 - 37.4%) for all nutrients. Weekly and seasonal variation contributed small components of variance for most nutrients. For protein, fat and carbohydrate, there were a little significant weekly variation (0.00 - 1.35%) in females but not in males. For some micronutrients, there were moderately significant seasonal variation (0.15 - 5.48%) in both sexes. Ratio of within- to between- individual variation ranged 1.4 (vitamin B$_2$) -4.5 (vitamin B$_1$) in males and 1.6 (carbohydrate) -2.9 (fat) in females. With total 12-day dietary records data, the maximum percentage deviation of observed intakes from usual (true) intakes ranged 12-37%. To estimate usual individual intakes within 20% of the true mean with 90% confidence level, 3 - 9 days of dietary survey were required for energy, protein, carbohydrate, phosphorus and iron, 13 - 19 days for fat and calcium, 25 - 29 days for vitamin A and vitamin C. Correlation coefficients between observed and true nutrient intakes were 0.71 - 0.91 for males, 0.81-0.91 for females. In conclusion, mean intakes of several nutrients can be reliably measured with the record method, using a limited number of days. Both nutrients of interest and the primary objectives should be taken account when planning method of assessment and number of replicates.

• Nutrition Research and Practice
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• v.11 no.3
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• pp.190-197
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• 2017

BACKGROUND/OBJECTIVES: Gallus gallus domesticus (GD) is a natural mutant breed of chicken in Korea with an atypical characterization of melanin in its tissue. This study investigated the effects of melanin extracts of GD on osteoblast differentiation and inhibition of osteoclast formation. MATERIALS/METHODS: The effects of the melanin extract of GD on human osteoblast MG-63 cell differentiation were examined by evaluating cell viability, osteoblast differentiation, and expression of osteoblast-specific transcription factors such as bone morphogenetic protein 2 (BMP-2), small mothers against decapentaplegic homologs 5 (SMAD5), runt-related transcription factor 2 (RUNX2), osteocalcin and type 1 collagen (COL-1) by reverse transcription-polymerase chain reaction and western blotting analysis. We investigated the inhibitory effect of melanin on the osteoclasts formation through tartrate-resistant acid phosphatase (TRAP) activity and TRAP stains in Raw 264.7 cell. RESULTS: The melanin extract of GD was not cytotoxic to MG-63 cells at concentrations of $50-250{\mu}g/mL$. Alkaline phosphatase (ALP) activity and bone mineralization of melanin extract-treated cells increased in a dose-dependent manner from 50 to $250{\mu}g/mL$ and were 149% and 129% at $250{\mu}g/mL$ concentration, respectively (P < 0.05). The levels of BMP-2, osteocalcin, and COL-1 gene expression were significantly upregulated by 1.72-, 4.44-, and 2.12-fold in melanin-treated cells than in the control cells (P < 0.05). The levels of RUNX2 and SMAD5 proteins were higher in melanin-treated cells than in control vehicle-treated cells. The melanin extract attenuated the formation of receptor activator of nuclear factor kappa-B ligand-induced TRAP-positive multinucleated RAW 264.7 cells by 22%, and was 77% cytotoxic to RAW 264.7 macrophages at a concentration of $500{\mu}g/mL$. CONCLUSIONS: This study provides evidence that the melanin extract promoted osteoblast differentiation by activating BMP/SMADs/RUNX2 signaling and regulating transcription of osteogenic genes such as ALP, type I collagen, and osteocalcin. These results suggest that the effective osteoblastic differentiation induced by melanin extract from GD makes it potentially useful in maintaining bone health.

• 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).