크롬 함유 페인트 분무작업자에서 발생한 비중격 천공

정희경,김두희,최병순,임현술,신용철,황보근 東國大學校醫學硏究所 1997 東國醫學 Vol.4 No.-

크롬은 산업에서 널리 사용되고 있으나 접촉성 피부염, 점막 자극, 비중격 천공, 천식, 폐암 등 건강에 영향을 미칠 수 있는데 크롬 및 그 화합물의 원자가, 수용성, 농도, 산성도, 폭로기간 및 경로에 따라 독성이 다르다. 약 5개월간 크롬에 폭로된 2명의 근로자에서 비중격 천공이 발생하여 35명 근로자의 작업조건과 건강상태를 조사하였다(페인트 분무 판넬부서 14명, 서스부서 16명, 기타 부서 5명). 공기 중 총크롬의 농도는 0.103-0.318 mg/m³이었고, 6가 크롬농도는 자동분무 중에 0.001-0.060 mg/m³, 수동분무 중에 0.029-0.226 mg/m³이었다. 모든 근로자에서 소변의 크롬농도는 50㎍/L를 넘지 않았으나, 혈액의 크롬농도는 3명에서 3㎍/dL를 초과하였다. 서스부서보다 판넬부서 근로자에서 비강의 미란이나 가피가 더 많이 발견되었다(교차비 7.0). 부서별로는 소변이나 혈액의 크롬농도가 차이가 없었고 비강소견에 따라서도 소변의 크롬농도가 차이가 없었으나, 혈액의 크롬농도는 비강의 미란이나 가피가 있는 경우 유의하게 높았다. 비중격 천공이 발생한 2명의 근로자에서 크롬에 의해 발생할 수 있는 폐나 비갑개, 간 및 신장의 이상소견과 천식은 발견되지 않앗다. 본 사업장에서 크롬에 의한 건강장애를 예방하기 위해서는 수동분무를 자동분무로 바꾸고 분무한 페인트가 완전히 건조한 후 다음 작업을 하여야 한다. 또한 크롬 미스트가 흡입되지 않도록 적절한 호흡기 보호구를 철저히 착용하여야 한다. After discovery, chromium has been widely used in industry. Health hazards of chromium and its compounds are contact dermatitis(allergic and irritant), mucosal irritation, nasal septal perforation, asthma, and lung cancer. Their toxicity depends on valency, solubility, concentration, PH, exposure duration and route. Nasal septal perforation was developed in two workers exposed to chromium for about 5 months, so we surveyed the working condition and health status of 35 workers(14 workers in paint spraying panel line, 16 in sus line and 5 in other line). Air concentration was as follows: total chromium 0.103-0.318 mg/m³, Cr(VI) 0.001-0.060 mg/m³in automatic spraying and 0.029-0.226 mg/m³in manual spraying. Urine chromium concentration was not over 50 ㎍/L in all workers, but blood chromium concentration was over 3 ㎍/dL in 5 workers. Erosion or crust of nasal cavity was detected more frequently in panel line workers than sus line workers(odds ratio=7.0). But blood or urine concentration of chromium was not different according to the job. Urine concentration of chromium was not different irrespective of nasal cavity finding, but blood concentration was high in the workers with erosion or crust of nasal cavity. Two workers with nasal septal perforation did not have any other diseases which can be developed by chromium exposure. To prevent health hazards of chromium in this factory, manual spraying process has to be changed to automatic process, the next process done after paint was absolutely dried, and respiratory protector should be weared to prevent inhalation of chromium mist effectively.

Production of Transgenic Male Piglet as Research Model for Alzheimer's Disease

Mi-Ryung Park,Kyong-Woon Kim,Jae-Seok Woo,Seongsoo Hwang,In-Sul Hwang1,Tae-Uk Kwak,Ji-Hyun Lim,Se-Pil Park 한국동물생명공학회(구 한국동물번식학회) 2017 발생공학 국제심포지엄 및 학술대회 Vol.2017 No.10

Alzheimer’s disease (AD) is a progressive neurodegenerative disease associated with memory loss and cognitive impairments. An AD transgenic (Tg) pig model would be useful for preclinical testing of therapeutic agents. In this study, we report the use of somatic cell nuclear transfer (SCNT) to produce transgenic pigs over-expressing the human AD related genes (APP, APPswedish, Presenilin 1, and Tau). Transgenic embryos were generated by SCNT of from ear fibroblasts expressing AD genes. A total of 1808 (average 258) SCNT embryos were transferred into 7 recipients. Pregnancy was successfully maintained in one recipient. We obtained 1 cloned male piglet from a surrogate gilts and the weight of piglet was 935 g. But, the male piglet died two days after birth. The piglet expressed AD related genes by PCR and western blot analysis. Transgenes were expressed in multiple tissues, and at especially high levels in brain. AD Tg pig might be very useful for studying the disease and for testing new therapeutics in preclinical studies of human AD.

Characterization of α-Gal Epitope in Cells and Tissues from Homozygous α-1,3-Galactosyltransferase Knockout Pigs

Hwang, In-Sul,Kwon, Dae-Jin,Kwak, Tae-Uk,Oh, Keon Bong,Ock, Sun-A,Chung, Hak-Jae,Im, Gi-Sun,Hwang, Seongsoo The Korean Society of Animal Reproduction 2015 Reproductive & developmental biology Vol.39 No.4

To overcome the hyperacute immune rejection during pig-to-non-human primates xenotranasplantation, we have produced and bred ${\alpha}$-1,3-galactosyltransferase knock-out ($GalT^{-/-}$) pigs. In this study, the somatic cells and tissues from the $GalT^{-/-}$ pigs were characterized by an analysis of the expression of Gal${\alpha}$-1,3-Gal (${\alpha}-Gal$) epitope. Briefly, ear fibroblast cell lines of 19 homozygous $GalT^{-/-}$ pigs were established and cryopreserved. The expression of ${\alpha}-Gal$ epitope in the cells was measured by fluorescence activated cell sorter (FACS) analysis using BS-I-B4 lectin. Also, the homozygous ($GalT^{-/-}$) cells and tissues samples were immunostained with BS-I-B4 lectin for analysis of ${\alpha}-Gal$ epitope expression. The results showed that the expression of ${\alpha}-Gal$ epitope in $GalT^{-/-}$ cells (0.2 %) were significantly (p<0.05) down-regulated to the range of cynomolgus monkey fibroblast (0.2 %) cells compared to heterozygous ($GalT^{-/+}$) (9.3 %) and wild type ($GalT^{+/+}$) (93.7 %) fibroblast cells. In the immunostaining results, while the expression of ${\alpha}-Gal$ epitope was detected a partly in $GalT^{-/+}$ cells and mostly in $GalT^{+/+}$ cells, it was almost not detected in the $GalT^{-/-}$ cells. Also, immunostaining results from various tissues of the $GalT^{-/-}$ pig showed that the expression of ${\alpha}-Gal$ epitope was not detectable, whereas various tissues from $GalT^{+/+}$ pig showed a strong expression of ${\alpha}-Gal$ epitope. Our results demonstrated that ${\alpha}-Gal$ epitope expressions from $GalT^{-/-}$ pigs were successfully knocked out to prevent hyperacute immune rejection for further study of xenotransplantation.

Effect of a short-term in vitro exposure time on the production of in vitro produced piglets

Hwang, In-Sul,Kwon, Dae-Jin,Kwak, Tae-Uk,Lee, Joo-Young,Hyung, Nam-Woong,Yang, Hyeon,Oh, Keon Bong,Ock, Sun-A,Park, Eung-Woo,Im, Gi-Sun,Hwang, Seongsoo The Korean Society of Embryo Transfer 2016 한국동물생명공학회지 Vol.31 No.2

Although piglets have been delivered by embryo transfer (ET) with in vitro produced (IVP) embryos and blastocysts, a success rate has still remained lower level. Unlike mouse, human, and bovine, it is difficult to a production of piglets by in vitro fertilization (IVF) because of an inappropriate in vitro culture (IVC) system in pig. Therefore, the present study was conducted to investigate whether minimized exposure time in IVC can improve the pregnancy and delivery rates of piglets. Immediately after IVM, the oocytes were denuded and co-incubated with freshly ejaculated boar semen for 3.5 to 4 hours at $38.5^{\circ}C$ under 5% $CO_2$ in air. To avoid long-term exposure to in vitro state, we emitted IVC step after IVF. After that the presumptive zygotes were transferred into both oviducts of the surrogate on the same day or 1 day after the onset of estrus. Pregnancy was diagnosed on day 28 after ET and then was checked regularly every month by ultrasound examination. The 3 out of 4 surrogates were determined as pregnant (75%) and a total of 5 piglets (2 females and 3 males) were delivered at $118.3{\pm}2.5$ days of pregnancy period. In conclusion, a short-term exposure time may be an important factor in the production of IVP-derived piglets. It can be apply to the in vitro production system of transgenic pig by IVF, cloning, and pronuclear microinjection methods.

Rescue of vitrified-warmed bovine oocytes with rho-associated coiled-coil kinase inhibitor.

Hwang, In-Sul,Hara, Hiromasa,Chung, Hak-Jae,Hirabayashi, Masumi,Hochi, Shinichi Society for the Study of Reproduction [etc.] 2013 BIOLOGY OF REPRODUCTION Vol.89 No.2

<P>Cryotolerance of matured bovine oocytes is not fully practical even though a promising vitrification procedure with a ultrarapid cooling rate was applied. The present study was conducted to investigate whether recovery culture of vitrified-warmed bovine oocytes with an inhibitor (Y-27632) of Rho-associated coiled-coil kinase (ROCK) can improve the developmental potential after in vitro fertilization (IVF) and in vitro culture. Immediately after warming, almost all oocytes appeared to be morphological normal. Treatment of the postwarming oocytes with 10 μM Y-27632 for 2 h resulted in the significantly higher oocyte survival rate before IVF as well as higher cleavage rate and blastocyst formation rate. Quality analysis of the resultant blastocysts in terms of total cell number and apoptotic cell ratio also showed the positive effect of the Y-27632 treatment. Time-dependent change in mitochondrial activity of the vitrified-warmed oocytes was not influenced by ROCK inhibition during the period of recovery culture. However, the ability of ooplasm to support single-aster formation was improved by the ROCK inhibition. Thus, inhibition of ROCK activity in vitrified-warmed bovine oocytes during a short-term recovery culture can lead to higher developmental competence, probably due to decreased apoptosis and normalized function of the microtubule-organizing center.</P>

Production of Cloned Korean Native Pig by Somatic Cell Nuclear Transfer

Hwang, In-Sul,Kwon, Dae-Jin,Oh, Keun Bong,Ock, Sun-A,Chung, Hak-Jae,Cho, In-Cheol,Lee, Jeong-Woong,Im, Gi-Sun,Hwang, Seongsoo The Korean Society of Developmental Biology 2015 발생과 생식 Vol.19 No.2

The Korean native pig (KNP) have been considered as animal models for animal biotechnology research because of their relatively small body size and their presumably highly inbred status due to the closed breeding program. However, little is reported about the use of KNP for animal biotechnology researches. This study was performed to establish the somatic cell nuclear transfer (SCNT) protocol for the production of swine leukocyte antigens (SLA) homotype-defined SCNT KNP. The ear fibroblast cells originated from KNP were cultured and used as donor cell. After thawing, the donor cells were cultured for 1 hour with 15 ${\mu}M$ roscovitine prior to the nuclear transfer. The numbers of reconstructed and parthenogenetic embryos transferred were $98{\pm}35.2$ and $145{\pm}11.2$, respectively. The pregnancy and delivery rate were 3/5 (60%) and 2/5 (40%). One healthy SLA homotype-defined SCNT KNP was successfully generated. The recipient-based individual cloning efficiency ranged from 0.65 to 1.08%. Taken together, it can be postulated that the methodological establishment of the production of SLA homotype-defined cloned KNP can be applied to the generation of transgenic cloned KNP as model animals for human disease and xenotransplantation researches.

Analysis of Semen Parameters in α1,3-Galactosyltransferase^-/- Boars

Hwang, In-Sul,Lee, Seung-Chan,Kim, Sung Woo,Kwon, Dae-Jin,Park, Mi-Ryung,Yang, Hyeon,Oh, Keon Bong,Ock, Sun-A,Woo, Jae-Seok,Im, Gi-Sun,Hwang, Seongsoo The Korean Society of Embryo Transfer 2017 한국동물생명공학회지 Vol.32 No.2

It is very difficult to get the information about semen quality analysis in transgenic pigs because of limited numbers and research facilities. Therefore, in the present study, we analyzed the semen quality of transgenic boars generated for xenotransplantation research. Briefly, the semen samples were collected from 5 homozygous ${\alpha}1,3$-Galactosyltransferase knock-out ($GalT^{-/-}$) transgenic boars and immediately transported to the laboratory. These semen samples were decupled with DPBS and conducted to analyze semen parameters by a computer-assisted semen analysis (CASA) system. The boar semen were examined all 12 parameters such as total motility (TM), curvilinear velocity (VCL), straight line velocity (VSL), average path velocity (VAP), and hyperactivated (HYP), etc. In results, among the 5 $GalT^{-/-}$ boars, three boars (#134, 144, and 170) showed normal range of semen parameters, but #199 and 171 boars showed abnormal ranges of semen parameters according to standard ranges of semen parameters. Unfortunately, #171 boar showed azoospermia symptom with rare sperm counts in the original semen. Conclusively, assessment of semen parameters by CASA system is useful to pre-screening of reproductively healthy boar prior to natural mating and artificial insemination for multiplication and breeding.

Effect of Cytochalasin B Treatment on the Improvement of Survival Rate in Vitrified Pig Oocyte

Hwang, In-Sul,Park, Mi-Ryung,Kwak, Tae-Uk,Park, Sang-Hyun,Lim, Ji-Hyun,Kim, Sung Woo,Hwang, Seongsoo The Korean Society of Developmental Biology 2018 발생과 생식 Vol.22 No.3

To improve survival rates of vitrified pig oocytes, the treatment of cytoskeletal stabilizer on an appropriate time is one of the possible approaches. However, the exact treatment timing and effect of cytoskeletal stabilizer such as cytochalasin B (CB) is not well known during oocyte vitrification procedures. Thus, the present study was conducted to determine optimal treatment timing of CB during vitrification and warming procedures. In experiment 1, the survival rates of the postwarming pig oocytes were analyzed by fluorescein diacetate (FDA) assays with 4 classifications. In results, post-warming oocytes showed significantly (p<0.05) decreased number of alive oocytes (31.8% vs. 86.4%) compared to fresh control. In detail, the significant difference (p<0.05) was found only in strong fluorescence (18.2% vs. 70.5%) not in intermediate fluorescence groups (13.6% vs. 15.9%). In experiment 2, CB was treated before (CB-Vitri) and after (Vitri-CB) vitrification. In results, group of Vitri-CB showed significantly (p<0.05) higher (91.6%) survival rates compared to group of CB-Vitri (83.7%), significantly (p<0.05) and comparable with group of Vitri Control (88.7%) by morphological inspection. In FDA assay results, group of Vitri-CB showed significantly (p<0.05) higher (44.2%) survival rates compared to groups of CB-Vitri (36.7%) and Vitri Control (35.1%). In conclusion, the increased survival rates of post-warming pig oocyte treated with Vitri-CB method are firstly described here. The main finding of present study is that the CB treatment during recovery could be helpful to refresh the post-warming pig oocyte resulting its improved survival rates.

Optimization of Major Culture Elements on Growth and Shikonin Production in the Lithospermum erythrorhizon Hairy Root Culture

Hwang, Ok-Jin,Kim, Yu-Jeong,Sung, Nak-Sul,Ahn, Jun-Cheul,Kim, Sik-Eung,Hwang, Baik The Korean Society of Medicinal Crop Science 2002 한국약용작물학회지 Vol.10 No.4

The effects of basal media, carbon, nitrogen, phosphate and some major macro elements on growth and shikonin production in Lithospermum erythrorhizon hairy root culture were studied. Among examined media, growth of hairy root cultured in B5 liquid medium was rapid, whereas shikonin production was high in MS liquid medium. Under B5 basal medium, sucrose concentration for optimal growth and shikonin production was 9% and 4% respectively. The growth and shikonin production on pH changes in B5 medium resulted little effect in pH 5.8 to pH 8.8 ranges, whereas growth was decreased dramatically in both above 8.8 and under 5.8. Nitrogen source and concentration effected on the growth and shikonin production. The highest growth rate was in B5 medium (50 mM $KNO_3$ and 1 mM $NaH_2PO_4)$, whereas the highest shikonin production was in the condition supplemented with 5 mM $KNO_3$ and 10 mM $NaH_2PO_4$.

The expression of α-gal epitope in skin derived from transgenic pigs for xenotransplantation

Seongsoo Hwang,In-Sul Hwang,Tae-Uk Kwak,Hyeon Yang,Mi-Ryung Park,Keon Bong Oh,Sun-A Ock,Jae-Seok Woo,Jeong-Woong Lee,Gi-Sun Im 한국실험동물학회 2017 한국실험동물학회 학술발표대회 논문집 Vol.2017 No.2