This experiment was carried out to study on changing phases of the concentrations of serum testosterone and metabolites in the various grwoing stages of male pigs. The eight males were used to obtain serial blood samples at a, pp.oximately 20kg body weight intervals from birth to 130kg body weight. The blood samples were taken from the jugular veins and serum was stored at -20$^{\circ}C$ until assay. Testosterone concentrations in the serum were analyzed by radioimmunoassay. The result obtained are as follows: 1. Serum testosterone concentrations were elevated at birth and were reached a maximum level between 50 and 70kg body weight, which was when sexual maturity was reached. 2. Calcium values did not vary a, pp.eciably with body weight, and ranged from 9.6${\pm}$0.6 to 11.9${\pm}$0.8mg/100$m\ell$. Potassium and sodium concentrations ranged from 38.5${\pm}$2.9 and 233.9${\pm}$2.1mg/100$m\ell$ to 64.2${\pm}$6.5 and 269.1${\pm}$9.5mg/100$m\ell$, respectively. Magnesium values dro, pp.d at birth and then rose to peak at 15kg of body weight. Iron concentrations was 0.12${\pm}$0.02mg/100$m\ell$ at birth, rose to 0.20${\pm}$0.04mg/100$m\ell$ at 15kg of body weight and then gradually increased to 0.29${\pm}$0.04mg/100$m\ell$ at 30kg of body weight. Serum zinc concentrations rose from a low of 56${\pm}$3.3mg/100$m\ell$ at birth to a high of 83.3${\pm}$3.4mcg/100$m\ell$ at 15kg of body weight. Co, pp.r values rose from a low of 25${\pm}$2.5mcg/100$m\ell$ at birth to a high of 183${\pm}$4.3mcg/100$m\ell$ at 15kg of body weight. 3. Serum cholesterol concenrtration did not vary a, pp.eciably with body weight, and ranged from 90.5${\pm}$6.0mg/100$m\ell$ to 95.0${\pm}$6.3mg/100$m\ell$. Glucose concentrations ranged from 80.5${\pm}$1.2mg/100$m\ell$ to 108.7${\pm}$8.4mg/100$m\ell$. Serum total protein rose from alow of 2.7${\pm}$0.8mg/100$m\ell$ at birth to a rapidly high of 4.3${\pm}$0.1mg/100$m\ell$ at 15kg of body weight and then gradually increased to 7.3${\pm}$0.4mg/100$m\ell$ at 130kg of body weight. Serum albumin values ranged from 0.5${\pm}$0.1$m\ell$ to 3.0${\pm}$0.3mg/100$m\ell$. 4. The total concentrations of essential/nonessential amino acid were 944.7mg/100$m\ell$ and 934.4mg/100$m\ell$ at birth, respectively. The values of essential/nonessential amino acid gradually rose from a low level at birth to a high level at 130kg of body weight. The total concentrations of essential/non-essential amino acid ratios remained from birth to 130kg of body weight.
During in vitro culture of mammalian oocytes and embryos, the cells are exposed to the risks that cause cell injury or death. Numerous studies have been reported that the cell injury may be induced by the action of free radicals generated by auto-oxidation. This study was undertaken to investigate the optimal culture condition system for in vitro culture of porcine embryos. We first evaluated the effect of culture media on the porcine embryo development. NCSU-23 and PZM-5, culture medium tested, were failed to produce significant difference on the rate of blastocyst formation. In NCSU-23, the developmental rate was slightly higher than that in PZM-5. During in vitro maturation (IVM), fertilizaton (IVF), and culture (IVC) under 5 or 20% oxygen ($O_2$), the rates of cleavage and development were insignificantly different from each other under our culture condition (20% $O_2$, in NCSU-23), the mean cell number per blastocyst was $40{\pm}10$. These results showed that medium and $O_2$ concentration had no significant effect on the development of porcine embryos.
This study was conducted to examine whether efficiency of oocyte production from superovulated prepubertal goats. Fifteen prepubertal and twenty adult goats, maintained in a pen under natural day length and fed hay ad libitum, were pretreated with progestagen implanted CIDR for 10 days. Superovulation treatment of the goats received twice daily intramuscular injection of a total of 70 mg FSH for 3 days from Day 8 of CIDR. All the gonadotrophin treated goats were injected with 10 mg $PGF_2{\alpha}$ on Day 8 and 400~600 IU hCG in the afternoon on Day 10. Oocytes were recovered by follicle aspiration or oviduct flushing at 35 to 40 h after hCG injection through mid-ventral incision. The in vivo matured oocytes was activated by ionomycin (5 min) and 6-DMAP (3.5~4 h). The activated oocytes were cultured in mSOF medium containing 0.8% BSA at 38.5$^{\circ}C$ in an atmosphere of 5% CO$_2$, 5% O$_2$, 90% N$_2$ for 7~8 days. There was no significant difference in the mean number of CL and in vivo matured and follicular oocytes recovered. But, quality of I + II grade follicular oocytes was lower (p<0.05) in the prepubertal goat (25.0%) than the adults (52.4%). The same results were also observed in the cleavage and blastocyst rate of activated oocytcs. The cleavage and blastocyst rate from prepubertal derived oocytes were lower (p<0.05) in the prepubertal goat (54.5%, 23.3%) than the adult goat (86.8%, 46.6%). Considering overall these results, we suggest that maturation of donor goats is a major factor affecting recovered oocytes quality and in vitro development of activated goat oocytes.
This study was performed to investigate the effect of 12-week weight control program including nutrition education, diet therapy and regular exercise on body weight, obesity degree, BMI and blood lipid levels among middle aged obese women. The 31 middle aged obese women participated in the weight control program for 12-week. The body weight, body composition, blood lipid levels and food habit were measured before and after the weight control program. Changes in body weight after 12 week weight control program showed the slight reduction from $67.4{\pm}10.8kg\;to\;65.1{\pm}9.2kg$. Obesity degree and BMI were decreased from $128.4{\pm}19.1%,\;27.8{\pm}3.9\;to\;123.4{\pm}15.7%,\;26.8{\pm}3.0$, respectively. Waist-hip ratio was also decreased from $0.98{\pm}10.16\;to\;0.94{\pm}0.04$. LBM was also slightly reduced from $39.9{\pm}5.6kg\;to\;37.0{\pm}8.5kg$ with the reduction of body fat. The levels of systolic blood pressure, total cholesterol and triglyceride were slightly decreased, HDL-chol level was significantly increased from $43.1{\pm}14.1mg/dL\;to\;50.4{\pm}16.5mg/dL$. Waist-hip ratio was also slightly decreased after 12 weeks. Food habit score was significantly increased after nutriton education and personal counseling about their eating behaviors and nutritional status.
Journal of the Korean Society for Marine Environment & Energy
/
v.11
no.2
/
pp.70-77
/
2008
In this study, to establish countermeasure from marine casualties as a basic study fur long-term prediction of topographical change around Jinudo in the Nakdong river estuary, spatio-temporal topographical change monitoring was carried out. Also, in order to estimate the deposition variations concerning SS (Suspended Solid) flux which moved at St.S1 during neap and spring tide, respectively. From the topographical monitoring, it was found that the annual mean ground level and deposition rate were 141 mm and 0.36 mm/day and all parts except the northern part of Jinudo had the active topographical changes and a tendency to annually deposit. From vertical distribution of SS net fluxes, $SS_{LH}$ (latitudinal SS net flux) during spring tide overall flows average 28 $kg/m^2/hr$ (eastward), and $SS_{LV}$ (longitudinal SS net flux) flows average 11.1 $kg/m^2/hr$ (northward). And, $SS_{LH}$ overall flows average 4.8 $kg/m^2/hr$ (eastward), and $SS_{LV}$ flows average 1.5 $kg/m^2/hr$ (northward) during neap tide similar with spring tide. The depth averaged values of the latitudinal and longitudinal SS net fluxes during spring tide were approximately 6 times higher than those during neap tide. As result of, it was considered that topographical change of southern part of Jinudo was affected by resuspension of bottom sediments due to strong current in bottom layer during flood flow.
Specific or non-specific cytolytic processes of free-living amoebae causing meningoencephalitls have been emphasized and the cytolytic ability related to hydrolases in Entantoeba sp. and Naegleria sp. has also been reported since the latter half of 1970's. However, no information on hydrolase activities in Acanthamoeba sp. is available. Hydrolases in Acanthamoeba culbertsoni, a pathogenic species of free-living amoebae, were assayed and compared with those in a non-pathogenic species, A. royreba. Pathogenicity of these two species was confirmed through experimental infection to BALB/c mice. Hydrolase activities and cytotoxic effects between pathogenic and non.pathogenic species were compared in the trophozoites cultured in CGV media and in CHO cell line, respectively. The results are summarized as follows: 1. The mice infected with A. culbertseni were all dead 15 days after nasal inoculation, and the mean survival time was 8.5 days. Also the mice infected with this pathogenic species manifested typical meningoencephalitis, whereas the mice infected with A. royreba did not. 2. Hydrolases detected both in the cell extracts and culture media were acid phosphatase, ${\beta}-N-acetyl$ galactosaminidase, ${\beta}-N-acetyl$ glucosaminidase, ${\alpha}-mannosidase$, neutral proteinase and acid proteinase, all of which were detected with remarkably higher rate in A. culbertsoni than in A. royreba. 3. A. cuzbertsoni revealed strong cytotoxicity for the target CHO cells, whereas A. royreba did not show any specific cytotoxicity. About 80% of the target cells mixed with A. culbertsoni were dead 48 hours after cultivation, and more than 95% of the target cells were dead 72 hours after cultivation. 4. Hydrolase activities in A. culbertsoni cultured with the target cell line were assayed according to the culture time. The activities of acid phosphatase, ${\beta}-N-acetyl$ galactosaminidase, ${\beta}-N-acetyl$ glucosaminidase, ${\alpha}-mannosidase$ and acid proteinase in this pathogenic amoeba were detected higher in amoeba extracts than in culture media up to 120 hours after cultivation, but after 120 hours of cultivation those activities were detected higher in culture media than in the amoeba Iysates. Neutral proteinase activity in A. culbertsoni increased more in EBSS medium than in the Iysate specimens although the activity in the extracts was generally steady according to the cultivation time. Summarizing the above results, it is concluded that there were differences in hydrolase activities between Pathogenic A. culbertsoni and non-pathogenic A. royreba, and that some hydrolase activities were detected remarkably higher in A. culbertsoni which revealed strong cytotoxicity to the target CHO cell line.
Choi, Hea Jin;Lee, Soo Young;Ma, Sang Hyuk;Kim, Jong Hyun;Hur, Jae Kyun;Kang, Jin-Han
Pediatric Infection and Vaccine
/
v.12
no.2
/
pp.186-194
/
2005
Purpose : Hepatitis A viral infections have been continued after re-emerging since mid 1990s in Korea. The incidence of this disease has been increased in young adults younger than 30 years of age since 2000. This study was performed to evaluate the prevalence of antibody to hepatitis A in Korea(two regions; Incheon and Changwon) in 2005, and was compared with the results of similar studies in mid 1990s. Methods : The study was conducted from January 2005 to June 2005, and consisted of 1,301 enrolled subjects, neonates to 50 years old, living in Incheon and Changwon in Korea. All sera were frozen and stored at $-70^{\circ}C$ until assayed. Anti-HAV IgG antibodies were measured by microparticle enzyme immunoassay(HAVAB, Abbott Lab., IL, USA). Results : The prevalence of anti-HAV IgG was 61.1% in infants younger than 1 year old, 30.5% in 1~5 years, 14.6% in 6~10 years, 1.7% in 11~15 years, 6.5% in 16~20 years, 36.6%in 21~30 years, 77.5% in 31~40 years, and 99.8% in 41~50 years. Statistical differences were not found between male and female, but there was statistical difference in 6~10 years old age group between the two areas. Conclusion : Our study indicate that the prevalence of antihepatitis A virus antibody has shifted from children to old adolescents and young adults. This result suggests that the risk of sudden outbreaks or increasing incidence of hepatitis A viral infections in young adults may be expected in our society. The preventive strategies of hepatitis A including vaccination should be prepared.
The present study was conducted to examine some factors affecting in vitro development and fecundity of embryos recloned with somatic cell nuclear transfer (SCNT). Fibroblast cells retrieved from the ear of a 3-week-old, cloned Korean goat (Jinsoonny) were used as karyoplast donors and serum-starvation was conducted in tissue culture medium (TCM)-199 supplemented with 0.5% FBS. Recipient oocytes were surgically collected by flushing the oviducts 35 h after hCG injection following FSH priming. The zonae pellucidae of the oocytes were partially perforated with a laser drill and a donor cell was transferred into an enucleated oocyte. The couplets were electrically fused and activated by ionomycin (5 min) and 6-DMAP (4 h). The reconstructed embryos were cultured in mSOF medium containing 0.8% BSA at $39^{\circ}C$ in an atmosphere of 5% $CO_2$, 5% $%O_2$, 90% $N_2$ for 12 to 15 h. Re-cloned embryos (2- to 4-cell stages) were surgically transferred into the oviducts of the recipients and pregnancy was subsequently diagnosed by progesterone assay and ultrasound on Days 21 and 63 of pregnancy. The fusion rate following 1st fusion pulse was higher (p<0.05) in 2nd cloning (65.9%) compared to 1st cloning (51.0%), but it was not different in the other groups. The rate of cleavage after fusion was significantly higher (p<0.05) in 1st (77.7%) than in 2nd cloning (56.0%). A total of 175 re-cloned embryos were transferred into 28 recipients. On day 21 and 60 after transfer, 11 (39.3%) and 4 recipients (17.4%) were pregnancy, respectively. In comparison of pregnancy rate by estrous synchronization, a total of 66 and 109 re-cloned embryos were transferred into 11 recipients in natural estrus and 17 recipients in induced estrus, respectively. Five (45.4%) and 2 recipients (18.2%) in natural estrus were pregnant on days 21 and 63 while 6 (35.3%) and 2 (11.8%) recipients in induced estrus were pregnant, respectively. These results show that recloning of goat can be achieved by SCNT and estrous synchronization between donor and recipient animals may be one of the major factors affecting success rate.
Heat shock protein 90 (Hsp90) is ATPase-directed molecular chaperon and affects survival of cancer cell. Inhibitory effect of Hsp90 by inducing cell cycle arrest and apoptosis in the cancer cell was reported. However, its role during oocyte maturation and early embryo development is very insufficient. In this study, we traced the effects of Hsp90 inhibitor, 17-allylamino-17-demethoxygeldanamycin (17-AAG), on meiotic maturation and early embryonic development in pigs. We also investigated several indicators of developmental potential, including structural integrity, gene expression (Hsp90-, cell cycle-, and apoptosis-related genes), and apoptosis, which are affected by 17-AAG. Then, we examined the roles of Hsp90 inhibitor on viability of primary cells in pigs. Porcine oocytes were cultured in the NCSU-23 medium with or without 17-AAG for 44 h. The proportion of GV arrested oocytes was significantly different between the 17-AAG treated and untreated group (78.2 vs 34.8%, p<0.05). After completion of meiotic maturation, the proportion of MII oocytes was lower in the 17-AAG treated group than in the control group (27.9 vs 71.0%, p<0.05). After IVF, the percentage of penetrated oocytes was significantly lower in the 17-AAG treated group (25.2%), resulting in lower normal pronucleus formation (2PN of 14.6%). Therefore, the inhibition of meiotic progression by Hsp90 inhibitor played a critical role in fertilization status. Porcine embryo were cultured in the PZM-3 medium with or without 17-AAG for 6 days. In result, significant differences in developmental potential were detected between the embryos that were cultured with or without 17-AAG. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) showed that the number of containing fragmented DNA at the blastocyst stage increased in the 17-AAG treated group compared with control (7.5 vs 4.4, respectively). Blastocysts that developed in the 17-AAG treated group had low structural integrity and high apoptotic nuclei than those of the untreated control, resulting in decrease the embryonic qualities of preimplantation porcine blastocysts. The mRNA expressions of cell cycle-related genes were down-regulated in the 17-AAG treated group compared with control. Also, the expression of the pro-apoptotic gene Bax increased in 17-AAG treated group, whereas expression of the anti-apoptotic gene Bel-XL decreased. However, the expression of ER stress-related genes did not changed by 17-AAG. Cultured pESF cells were treated with or without 17-AAG and used for MIT assay. The results showed that viability of pESF cells were decreased by treatment of 17-AAG ($2{\mu}M$) for 24 hr. These results indicated that 17-AAG decreased cell proliferation and increased cell death. Expression patterns Hsp90 complex genes (Hsp70 and p23), cell cycle-related genes (cdc2 and cdc25c) and apoptosis-related genes (Bax and Bcl-XL) were significantly changed by using RT-PCR analysis. The spliced form of pXbp-1 product (pXbp-1s) was detected in the tunicamycin (TM) treated cells, but it is not detected in 17-AAG treated cells. In conclusion, Hsp90 appears to play a direct role in porcine early embryo developmental competence including structural integrity of blastocysts. Also, these results indicate that Hsp90 is closely associated with cell cycle- and apoptosis-related genes expression in developing porcine embryos.
Chemical assessment of soil pollution with heavy metals was made by analyzing the changes in pH, ionic strength, cationic concentration and chemical species in the soil solution. Saturated pastes of the unpolluted soils were made by adding solutions containing Cu or Cd and the final Cu or Cd concentrations were in the range of 0 to 400 mg/kg. After equilibrating for 24 hours at $25^{\circ}C$, the soil solution was extracted from the saturated pastes by the vacuum extraction method and analyzed for pH, electrical conductivity, Cu, Cd, cations and inorganic ligands. Chemical species in soil solution were calculated by the GEOCHEM-PC program employing the input variables of pH, ionic strength(${\mu}$), molar concentrations of cations and ligands. Increasing Cu or Cd additions lowered pH of the soil solution but increased concentrations of Ca, Mg and K resulting in increases of ${\mu}$ of the soil solution. Effects of Cu on lowering pH and increasing ${\mu}$ were greater than those of Cd. Concentrations of Cu or Cd in soil solution were relatively very low as compared to those of additions, but increased linearly with increasing additions representing that concentrations of Cu were higher than those of Cd. At 400 mg/kg additions, concentrations of Cu were in the range of 0.51 to 11.70 mg/L but those of Cd were 34.4 to 88.5 mg/L. Major species of Ca, Mg and K were free ions and these species were equivalent to greater than 95 molar % of the existing respective molar concentrations. These cationic species were not changed by Cu or Cd additions. Major species of Cu in lower pH soils such as SiCL and SL were free $Cu^{2+}$ (>95 molar %), but those in LS having a higher pH were free $Cu^{2-}$ and Cu-hydroxide complex. At 100 mg Cu/kg treatment, $Cu^{2+}$ and Cu-hydroxide complex were equivalent to 73 and 22.4 molar %, respectively. These respective percentages were decreased and increased correspondingly with increasing Cu treatments. Major species of Cd in soil solution were free $Cd^{2+}$ and Cd-chloride complex, representing 79 to 85 molar % for $Cd^{2+}$ and 13 to 20% for Cd-chloride complex at 10 mg Cd/kg treatment. With increasing Cd additions to 400 mg/kg, $Cd^{2+}$ species decreased to $40{\sim}47%$ but Cd-chloride complexes increased to $53{\sim}60$ molar %. These results demonstrated that soil contamination with heavy metals caused an adverse effect on the plant nutritional aspects of soil solution by lowering pH, increasing cations temporarily, and increasing free metal concentrations and species enough to be phytotoxic.
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