• The Journal of Korean Physical Therapy
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• v.17 no.2
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• pp.22-37
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• 2005

This study was performed to investigate LBP(low-back pain) prevalence rate and its related factors inphysical therapists. A self-administered questionnaire survey was conducted to 522 physical therapists in Daegu and Gyeongsangbuk. The LBP prevalence rate for recent 6 months was $61.7\%$. According to age, the prevalence rate was highest in the group aged 24 or less for both male and female physical therapistis. For male physical therapists , it was higher in smokers and drivers, but for female ones, it was not in a significant relationship with smoking and driving. According to work experience, the prevalence rates of those who had been working for less than one year were the highest, and those who had been working at hospitals and clinics were $69.1\%$, significantly high. According to therapy techinque, the prevalence rate related ti PNF was $71.6\%$, the highest and to occupational therapy was $34.3\%$, the lowest. According to working environment for six months, in both male and female physical therapists. LBP prevalence rate was in a statistically significant relationship with work hours during the day, night and weekend duties, repetitive works, motions that bend or twist the waist, the number of patients per day, the number of time to help patients per day, the number of time to carry medical equipment per day and the length of time to work standing. The rate was also higher when physical therapists fekt nire stress from their work. According to the result of multiple logistic regression analysis, male was in a significant relationship with work experience and motions that bend or twist the waist and female was in a significant relationship with work hours during the night and weekend duties, the number of time to help patients per day, motions that bena or twist the waist, actions taken to protect the waist. As for the developmental pattem of LBP in the group of physical therapists with LBP for six moths, $15.7\%$ of them had LBP for first time, $42.3\%$ had recurred LBP, and $42.0\%$ had chronic LBP. As for the causes of LBP, $51.7\%$ said that it was because they worked in the same posture for a long time, and as for how to treat LBP, 48.8% said that they treated themselvs or got help from their co-wokers. The results presented above suggest that physical therapists are exposed to high risk of LBP caused form occupational activities. Therefire, it is necessary to improve working environment to reduce the occurrence of LBP and to develop education programs for preventing the occurrence and recurrence of LBP.

• Development and Reproduction
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• v.15 no.2
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• pp.77-85
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• 2011

To investigate the influence of density and feeding frequency (food amount) on growth, cannibal dynamics and survival of puffer Takifugu obscurus (TL $25.0{\pm}0.5$ mm, BW $0.53{\pm}0.03$ g) during early life history, we conducted one experiment composed with 6 density groups set up as 1.43 inds./${\ell}$, 2.14 inds./${\ell}$, 2.86 inds./${\ell}$, 3.57 inds./${\ell}$, 4.29 inds./${\ell}$ and 7.14 inds./${\ell}$. The experiment was triplicately performed in 1 ton FRP tank for 30 days. Daily food intake (DFI), feed efficiency (FE), growth, cannibal frequency, and survival rates were examined. Although not find any significant relation of density to DFI and FE, we found that growth decreased significantly with the increasing of density under 3.57 inds./${\ell}$, but significantly increased with the increasing of density over 4.29 inds./${\ell}$. Cannibal frequency and daily mortality rate was significantly increased with increasing density and decreasing feeding frequency, inducing the most poor survival rate in the highest density group and low feeding frequency. Therefore, we concluded that the density and the feeding frequencies affect sibling cannibal dynamics and survival of puffer stock.

• Reproductive and Developmental Biology
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• v.35 no.3
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• pp.313-317
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• 2011

This study was conducted to examine the effect of oocyte donor age and micromanipulation medium on the development of mouse cloned embryos receiving cumulus cells. Mouse oocytes were obtained from 6 to 11 week-old mice BDF1 female mice(experiment 1) and cumulus cells were used as donor cells. Micromanipulation procedures for nuclear transfer(NT) were performed in FHM, M2 or Hepes-buffered TCM199(TCM199) medium(experiment 2). After nuclear transfer, the reconstructed oocytes were activated by 10 mM $SrCl_2$ in Ca-free CZB medium in the presence of 5 II ${\mu}$g/ml cytochalasin B for 5 h and cultured in KSOM medium for 4 days. In experiment 1, the survival rate of oocytes after injection of cumulus cells were significantly(p<0.05) lower in oocytes from 6~7 week-old mice(53.3%) than in oocytes from 8~9(80.9%) and 10~11 week-old mice(77.1%). In experiment 2, the survival rate of oocytes after cell injection were significantly(p<0.05) higher in FHM and M2 medium(71.7% and 76.9%) than in TCM199 medium(51.2%). The activation rates of cloned embryos were not different among the micromanipulation media. However, the embryos developed to blastocyst stage were significantly(p<0.05) higher in FHM medium(13.9%) than in M2 and TCM199 medium(0.0% and 0.0%). In conclusion, the present study suggest that oocytes from above 8 week-old mice are superior to oocytes from 6~7 week-old mice as a source of recipient cytoplasm and FHM is superior to M2 and TCM199 as a micromanipulation medium for mouse somatic cell cloning.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.61-61
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• 2001

The main goal of this study was to produce transgenic piglets by the method of injection of sperm-mediated exogenous DNA. Spermatozoa (1$\times$106 sperm of final concentration) obtained from caudal epididymis were mixed with pBC1-hEPO (20 ng/${mu}ell$) or pcDNA3 LAC Z (20 ng/${mu}ell$), and followed by electroporation (500 V, 25 ㎌). Matured oocytes having the first polar body and dense cytoplasm were selected and centrifuged at 12,000g for 6 min. After sperm injection, the oocytes were activated electrically (1.7 ㎸/cm, 30 $\mu$ sec, single pulse) in 0.3 M mannitol solution. Eggs injected sperm were cultured in NCSU 23 medium (0.4% BSA) at 39$^{\circ}C$, 5% $CO_2$ in air for 192 h. This study were comprised 3 experiments. Experiment 1 compared the developmental efficiencies between the sperm-injected oocytes (Group 1) and further activated electrically (Group 2). Experiment 2 compared the expression of pcDNA3 LAC Z in the embryos produced by Group 1 and Group 2. Finally, experiment 3 carried out transfer of embryos (1-8 cell stage) transfected with pBC1 -hEPO into surrogate recipients synchronized by injection of combination of PG600 with hCG. The rates of cleavage and development into blastocyst stage in Group 2 were significantly higher than those of Group 1 (71.3% and 28.1% vs. 43.3% and 10.3%, respectively, p<0.05). Thirty (24.2%) out of 124 embryos analyzed in Group 2 were positive by X-gal. Similarly, in Group 1, 16.3% (8/49) were positive. After transfer of 789 embryos to 7 recipient gilts, three out of them examined by ultrasound became pregnant. One recipient is in day 50 pregnancy. On day 54 of gestation, two were carried out uterotomy in order to confirm the pregnancy One had 7 and another had 2 fetuses. We conclude that injection of sperm-mediated gene transfer will be used as a valuable tool for the production of transgenic piglets.

• Reproductive and Developmental Biology
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• v.30 no.2
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• pp.87-92
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• 2006

The possibility of producing transgenic embryos expressing the green fluorescence protein (GFP) gene have been evaluated after transfer of exogenous gene into the porcine zygote cytoplasm using the intracytoplasm sperm injection (ICSI) as gene delivery method. For DNA binding to sperm heads, 0.05% Triton X-100 or Lipofectin was used. After injection of the sperm bound to DNA by means of Lipofectin or Triton X-100 triturate, the blastocyst formation rates on day 6 were not significantly different from that of ICSI only group (18.8, 19.2 and 25.3%). In terms of GFP expression, more embryos were in GFP form in Triton X-100 group than in Lipofectin group (40.6 vs 36.4%), while percentage of non-mosaic embryos expressing the GFP gene in all blastomere was higher (P<0.05) in Lipofectin group than in Triton X-100 group (4.2 vs 0.9%). ICSI embryos derived from sperm treated with Lipofectin/DNA complex was transferred into 3 recipients and were collected by uterine flushing on days 5, 7 and 15 after embryo transfer, and then GFP expression was observed by a fluorescence microscopy. Over 26% of the collected embryos were normally expressed GFP gene. These results suggest that foreign gene transfer method with DNA bound sperm caused minimal damage to structure of oocytes that can result to full development of porcine embryos. This was confirmed in this study when the embryos that were transferred after ISCI of DNA bound sperm had a normal development and gene expression until preimplantation.

• Korean Journal of Animal Reproduction
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• v.17 no.2
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• pp.149-157
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• 1993

A new technique was established to maximize the numbers of follicular oocytes recovered from the ovaries obtained at the slaughter house. And their further developmental capacity was demonstrated. There recovery techniques were used; aspiration (ASP, control), slicing (SLC) and slicing combining aspiration (ASP+SLC). Recovered oocytes were cultured in TCM 199+15% FCS+gonadotrophins in an atmosphere of 5% CO$_2$ in air at 39$^{\circ}C$ for 24 h. The nuclear maturation was detemined with chromo-some configuration by rapid staining. And cytoplasmic maturation was examined by the formation of female pronuclei with parthenogenetic activation of the matured oocyte after 18 h of co-culture with granulosa cell monolayer. Total 1,641 bovine follicular oocytes recovered from 245 ovaries. The number of oocytcs per ovary was 1.87 in ASP, 11.05 in SLC and 7.88 in ASP+SLC, respectively. SLC would yield 5.9 folds increase, compared with ASP. The rate of maturation were 92.9% in ASP, 79.1% in SLC and 71.7% in ASP+SLC, respectively. Although the maturation rate in ASP was the highest, metaphase II oocytes per ovary in SLC was 5 times higher than that of ASP. The rates of pronuclei formation upon ethanol activation were 75% in ASP, 67% in SLC and 62.5% in ASP+SLC, respectively. The results demonstrate that it should be possible to maximize the number of the follicular oocyte from the ovary for mass production of bovine embryos. Thus the established technique may provide efficient supply of bovine embryos for biochemical and molecular study of early bovine embryos.

• Reproductive and Developmental Biology
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• v.33 no.1
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• pp.35-40
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• 2009

The objectives of this study was to evaluate the efficiency of the bacteria eliminated sperm by percoll gradient method on sperm quality and embryo cleavage in vitro in pig. The semen of miniature pig collected by gloved-hand method pre-warmed ($37^{\circ}C$) in thermos bottle, and separated by 65% percoll. Analysis of sperm ability was estimated by examining viability, capacitation and acrosome reaction using chlortetracycline (CTC) and the abnormality. Also, fertility of sperm was monitored with cleavage rate of embryo after IVF using separated and un-separated sperm by percoll. The result, viability of separated sperm was significantly(p<0.05) higher($83.6{\pm}$2.0 vs $59.0{\pm}4.4%$) than un-separated sperm. The results of CTC analysis showed the percentage of F- and B-patterned separated sperm was higher in separated that than un-separated sperm. On the contrary, the percentage of AR-patterned form unseparaed sperm was significantly(p<0.05) higher($13.6{\pm}0.8$ vs $8.1{\pm}0.6%$) than separated sperm. Also, abnormality of un-separated sperm was significantly(p<0.05) higher($2.2{\pm}0.4$ vs $16.8{\pm}2.8%$) than separated sperm. However, the cleavage rates of embryo using separated sperm by percoll and un-separated sperm had not significantly difference on 2 cell stage(9.25 vs 11.88%), 4 cell stage(26.76 vs 24.51%) and >4 cell stage(63.99 vs 63.61%) at 48h of IVF. Therefore, the sperm separated by percoll method showed improvement in sperm quality than un-separated sperm in miniature pig.

• Reproductive and Developmental Biology
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• v.32 no.2
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• pp.117-121
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• 2008

Our previous study showed that transgenic (TG) pigs harboring human EPO (hEPO) gene have been shown to have reproductive disorders, including low pregnancy rates, irregular estrus cycle and low little size. To investigate these reasons, we assessed estrus behavior (standing response) and plasma $17{\beta}$-estradiol ($E_2$) level, which partly reflect reproductive function, during the estrus cycles after synchronization and superovulation by hormone treatments. Then, we analysed blood composition and expression of hEPO gene in TG pigs. Pigs were injected with PG600. After 10 days, pigs were fed with Regumate porcine for 6 days. Blood samples were collected from jugular vein. Analysis of blood composition and $E_2$ level were measured by Hemavet 950 and $E_2$ ELISA kit, respectively. And, the expression of hEPO gene in reproductive organs was quantitated by real-time RT-PCR. The percentage of estrus behavior in TG was significantly decreased. Hematocrit (HCT), hemoglobin (Hb) concentration and red blood cell (RBC) number were significantly higher in TG than wild type (WT). On the other hand, high expression of hEPO gene in TG was observed in the mammary gland as well as in the uterus. Moreover, plasma $E_2$ level was significantly higher in TG than WT. These results suggest that nonspecific expression of hEPO gene in the other organs of TG may affect blood composition and plasma $E_2$ level, thereby causing reproductive disorders.

• Journal of Animal Science and Technology
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• v.49 no.2
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• pp.287-294
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• 2007

The present study was to conducted the sexing efficiency and accuracy of bovine embryo by LAMP (Loop-mediated isothermal amplification) method, the development of the biopsied embryos into re- reformation and the freezability of these blastocysts by slow-freezing and vitrification. In vivo embryos were superovaluted with gonadotropin(Antorin R-10) for 4 days combined with progestrone releasing intravaginal(CIDR) insertion in Hanwoo donors, and in vitro embryos were used blastocyst embryos at Day 7 or Day 8 after post-insemination in vitro. The biopsy of bovine embryo was carried out in a 80μl drop with Ca2+-Mg2+ free D-PBS and the viability of biopsied embryos were evaluated in IVMD medium at over 12 h culture time in 5% CO2 incubator.For embryo sexing, about five or seven blastomeres were isolated from in vitro and in vivo embryos at blastocysts with microblade. and were then subjected to LAMP. The survivability of biopsied embryos were no difference in the development rate to re-formation of blastocysts between in vivo and in vitro embryos(100% and 90% respectively). The rates of sexed embryos were compared according to two groups, the female rate was lower than that the male in the in vivo and in vitro embryos(46% vs, 54% and 40% vs, 60%, respectively). However, there were no difference in the overall sexing ratio between the two groups. The survivability of frozen-thawed sexed embryos were lower in the in vitro than in vivo embryos in the slow-freezing(Group 1) and vitrification method(Group 2), (41.7% vs. 58.8% and 57.1% vs, 77.8. respectively).

• Clinical and Experimental Reproductive Medicine
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• v.30 no.2
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• pp.111-118
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• 2003

Objectives: The development of an useful method for obtaining metaphase chromosomes from a biopsied blastomere would allow differentiation between embryos with balanced and normal chromosome complements in the preimplantation genetic diagnosis for chromosomal translocations. This study was performed to evaluate the effects of microtubule depolymerizing agents (MTDAs) on the blastomeres of mouse and human preimplantation embryos, and to establish an effective method for obtaining metaphase chromosomes of biopsied blastomeres in human early embryos. Materials and Methods: Early embryos (2-4 cell stage) from superovulated mice (ICR strain) were collected and treated with single or mixture MTDAs, such as vinblastine, nocodazole and colcemid. After the treatment of MTDAs for 16 hours, the metaphase aquisition (MA) rates were evaluated by the observation of chromosome status with bis-benzimide or DAPI staining. The optimal condition from the above experiment was applied to human embryos, which were developed from abnormal fertilization (3-pronuclei). Fluorescence in-situ hybridization (FISH) with whole chromosome probes was conducted on the human metaphase chromosomes by the MTDAs. Results: In mouse embryos, the effective concentrations of each MTDAs for obtaining metaphase chromosomes were $1.0{\mu}M$ of vinblastine (20.3%), $5.0{\mu}M$ of nocodazole (28.1%) and $1.0{\mu}M$ colcemid (55.6%), respectively. The highest MA rate (91.2%) in the mouse embryos was obtained by a mixture of vinblastine ($1.0{\mu}M$) and nocodazole ($1.0{\mu}M$). In the human embryos, the metaphase chromosomes of blastomeres were obtained in 44 of 113 blastomeres (38.9%) by treatment of the mixture of vinblastine and nocodazole. FISH signals of the metaphase chromosomes were successfully observed in human individual blastomeres. Conclusions: The treatment of a mixture MTDAs for obtaining metaphase chromosomes was an efficient method, and the MA rate was above 90% in the mouse embryos. However, only a relatively small proportions of the blastomeres yielded metaphase chromosomes by the MTDAs in the human embryos. The inconsistent effects of MTDAs may be related to the variation of different species and the poor developmental potency of abnormally fertilized human embryos. We should develop more reliable and efficient methods for obtaining the metaphase chromosomes in the biopsied blastomeres of human preimplantation embryos.