• Title/Summary/Keyword: reproductive medicine

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Production of a transgenic pig expressing 3D8 single chain variable fragment (scFv) and its evaluation of PRRS resistant (3D8 scFv 형질전환 돼지 개발 및 PRRS 저항성 평가)

  • Lee, Hwi-Cheul;Lee, Gunsup;Kim, Ji-Yoon;Yang, Hyeon;Lee, Bo Ram;Park, Mi-Ryung;Hwang, In-Sul;Lee, Poongyeon;Byun, Sung-June;Kim, Won-Il;Oh, Keon Bong
    • Korean Journal of Veterinary Service
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    • v.43 no.4
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    • pp.227-236
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    • 2020
  • In this study, we have developed 3D8 scFv transgenic pig (TG) by microinjection of fertilized one-cell pig zygotes (2.17%). The effect of 3D8 scFv TG on porcine reproductive and respiratory syndrome virus (PRRSV) resistance were evaluated through PRRSV VR2332 (1×103 TCID50/mL) challenge and transmission experiments. As a result, the average daily weight gain (ADWG) of TG increased compared to the wild type pigs (WT) in PRRSV challenge groups and the serum viremia levels of the TG was significantly lower than of WT on the 7 day and 21 day after infection, meaning that the viral shedding was suppressed by 3D8 scFv expression. These results suggest that the expression of 3D8 scFv in pig could suppress spreading of infected virus to pigs sharing a room.

Alternative Immunossays

  • Barnard, G.J.R.;Kim, J.B.;Collins, W.P.
    • Korean Journal of Animal Reproduction
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    • v.9 no.2
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    • pp.133-139
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    • 1985
  • An immunoassay may be defined as an analytical procedure involving the competitive reaction between a limiting concentration of specific antibody and two populations of antigen, one of which is labelled or immobillized. The advent of immunoassay has revolutionised our knowledge of reproductive physiology and the practice of veterinary and clinical medicine. Radioimmunoassay (RIA) was the first of these methods to be developed, which meausred the analyte with good sensitivity, accuracy and precision (1,2). The essential components of RIA are:-(i) a limited concentration of antibodies, (ii) a reference preparation, and (iii) an antigen labelled with a radioisotope (usually tritium or iodine-125). Most procedures invelove isolating the antibody-bound fraction and measuring the amount of labelled antigen. Good facilities are available for scintilltion counting, data reduction nd statistical analysis. RIA is undergoing refinement through:-(i) the introduction of new techniques to separate the antibody-bound and free fractions which minimize the misclassification of labelled antigen into these compartments, and the amount of non-specfic binding. (3), (ii) the development of non-extration for the measurement of haptens (4), (iii) the determination of a, pp.rent free (i.e. non-protein bound) analytes (5), and (iv) the use of monoclonal antibodies(6). In 1968, Miles and Hales introduced in important new type of immunoassay which they termed immunora-diometric assay (IRMA) based on t도 use of isotopically labelled specific antibodies(7) in a move from limited to excess reagent systems. The concept of two-site IRMAs (with a capture antibody on a solid-phase, and a second labelled antibody to a different antigenic determinant of the analyte) has enabled the development of more sensitive and less-time consuming methods for the measurement of protein hormones ovar wide concentration of analyte (8). The increasing use of isotopic methos for diverse a, pp.ications has exposed several problems. For example, the radioactive half-life and radiolysis of the labelled reagent limits assay sensitivity and imposes a time limit on the usefulness of a kit. In addition, the potential health hazards associated with the use and disposal of radioactive cmpounds and the solvents and photofluors necessary for liquid scientillation counting are incompatable with the development of extra-laboratory tests. To date, the most practical alternative labels to radioisotopes, for the measurement of analytes in a concentration > 1 ng/ml, are erythrocytes, polystyrene particiles, gold sols, dyes and enzymes or cofactors with a visual or colorimetric end-point(9). Increased sensitivity to<1 pg/ml may be obtained with fluorescent and chemiluminescent labels, or enzymes with a fluorometric, chemiluminometric or bioluminometric end-point. The sensitivity of any immunoassay or immunometric assay depends on the affinity of the antibody-antigen reaction, the specific activity of the label, the precision with which the reagents are manipulated and the nonspecific background signal (10). The sensitivity of a limited reagent system for the measurement of haptens or proteins is mainly dependent upon the affinity of the antibodies and the smalleest amount of reagent that may be manipulated. Consequently, it is difficult in practice to improve on the sensitivity obtained with iodine-125 as the label. Conversely, with excess reagent systems for the measurement of proteins it is theoretically possible to increase assay sensitivity at least 1000 fold with alternative luminescent labels. To date, a 10-fold improvement has been achieved, and attempts are being made to reduce the influence of other variables on the specific signal from the immunoreaction.

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Expression of Antisense Mouse Obese Gene in Transgenic Mice (형질전환 생쥐에서 Antisense 비만유전자의 발현)

  • Kwon, B.S.;Hong, K.H.;Jahng, J.W.;Lee, H.T.;Chung, K.S.
    • Korean Journal of Animal Reproduction
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    • v.24 no.4
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    • pp.419-428
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    • 2000
  • Leptin, the product of obese (ob) gene, is an adipocyte-derived satiety factor that plays a major role in the regulation of food intake, energy homeostasis, body weight, reproductive physiology and neuropeptide secretion. The present study was designed to generate transgenic mice expressing antisense mouse ob (mob) gene. Total RNA was extracted from the adipose tissues of mouse, then reverse transcription was performed. The 303 and 635 bp fragments of anti I and II cDNAs were amplified from mob cDNAs by PCR. The two mob cDNAs were reversely ligated into between adipose tissue specific aP2 promote and SV40 poly(A) site. Transgenic mice carrying two different kinds of antisense mob transgenes were generated by DNA microinjection into pronucleus. Total 14 transgenic mice were born, and the 4 and 5 founder lines of the transgenic mice with anti I and II transgenes were respectively established. Antisense mRNA expression was detected in transgenic F$_1$ mice by RT-PCR analysis. This result suggests that the transgenic mice expressing antisense mob mRNA may be useful as an animal disease model to be obesity caused by decreased amount of leptin secretion.

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Assessment of In vitro Antioxidant, Antidiabetic and Cytotoxic Activities of Sphaeranthus africanus Extracts

  • Tran Thi Huyen;Julien Dujardin;Nguyen Thi Thu Huong;Chung Thi My Duyen;Nguyen Hoang Minh;Ha Quang Thanh;Dao Tran Mong;Ly Hai Trieu;Nguyen Mai Truc Tien;Mai Thanh Chung;Nguyen Nhat Minh;Nguyen Thi Ngoc Dan;Huynh Loi
    • Natural Product Sciences
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    • v.29 no.2
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    • pp.98-103
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    • 2023
  • Sphaeranthus africanus is commonly used as a traditional remedy for sore throats and pain treatment in Vietnam. The aerial parts have been studied for its anti-inflammatory and anti-proliferative properties. However, the antioxidant and antidiabetic potential of the plant has not been explored. In this work, hydrophilic extracts of the plant's aerial parts were prepared in order to investigate its antioxidant and anti-diabetic properties. Also, the cytotoxicity of the root was evaluated and compared to that of the aerial parts. All of the extracts inhibited lipid peroxidation with IC50 values ranging from 2.05 to 3.56 ㎍/mL, indicating substantial antioxidant activity. At an IC50 value of 4.80 ㎍/mL, the 50% ethanol extract exhibited the most potent inhibition of α-glucosidase. The cytotoxic activity of root extracts is 2 to 5-fold less than that of the aerial parts. Nevertheless, dichloromethane and ethyl acetate extracts of the root demonstrated a selective effect on leukemia cells, with no harm towards the normal HEK-293 cell line. This work provides a scientific support for the antioxidant and antidiabetic activity of the plant. Hence, it may find a promising material for the development of novel antioxidant and antidiabetic agents. More research can be conducted on the phytochemistry and anticancer activities of the plant's root.

Enhanced Drug Carriage Efficiency of Curcumin-Loaded PLGA Nanoparticles in Combating Diabetic Nephropathy via Mitigation of Renal Apoptosis

  • Asmita Samadder;Banani Bhattacharjee;Sudatta Dey;Arnob Chakrovorty;Rishita Dey;Priyanka Sow;Debojyoti Tarafdar;Maharaj Biswas;Sisir Nandi
    • Journal of Pharmacopuncture
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    • v.27 no.1
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    • pp.1-13
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    • 2024
  • Background: Diabetic nephropathy (DN) is one of the major complications of chronic hyperglycaemia affecting normal kidney functioning. The ayurvedic medicine curcumin (CUR) is pharmaceutically accepted for its vast biological effects. Objectives: The Curcuma-derived diferuloylmethane compound CUR, loaded on Poly (lactide-co-glycolic) acid (PLGA) nanoparticles was utilized to combat DN-induced renal apoptosis by selectively targeting and modulating Bcl2. Methods: Upon in silico molecular docking and screening study CUR was selected as the core phytocompound for nanoparticle formulation. PLGA-nano-encapsulated-curcumin (NCUR) were synthesized following standard solvent displacement method. The NCUR were characterized for shape, size and other physico-chemical properties by Atomic Force Microscopy (AFM), Dynamic Light Scattering (DLS) and Fourier-Transform Infrared (FTIR) Spectroscopy studies. For in vivo validation of nephro-protective effects, Mus musculus were pre-treated with CUR at a dose of 50 mg/kg b.w. and NCUR at a dose of 25 mg/kg b.w. (dose 1), 12.5 mg/kg b.w (dose 2) followed by alloxan administration (100 mg/kg b.w) and serum glucose levels, histopathology and immunofluorescence study were conducted. Results: The in silico study revealed a strong affinity of CUR towards Bcl2 (dock score -10.94 Kcal/mol). The synthesized NCUR were of even shape, devoid of cracks and holes with mean size of ~80 nm having -7.53 mV zeta potential. Dose 1 efficiently improved serum glucose levels, tissue-specific expression of Bcl2 and reduced glomerular space and glomerular sclerosis in comparison to hyperglycaemic group. Conclusion: This study essentially validates the potential of NCUR to inhibit DN by reducing blood glucose level and mitigating glomerular apoptosis by selectively promoting Bcl2 protein expression in kidney tissue.

Captive breeding of endangered betta fish, Betta rubra, under laboratory conditions

  • Agus Priyadi;Asep Permana;Eni Kusrini;Erma Primanita Hayuningtyas;Bastiar Nur;Lukman;Josie South;Sawung Cindelaras;Sulasy Rohmy;Rendy Ginanjar;Muhamad Yamin;Djamhuriyah S Said;Tutik Kadarini;Darmawan Setia Budi
    • Fisheries and Aquatic Sciences
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    • v.27 no.4
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    • pp.213-224
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    • 2024
  • Betta rubra, classified as endangered fish species by the International Union for Conservation of Nature (IUCN), has been successfully bred and raised in captivity for two generations under laboratory conditions. This study aimed to provide comprehensive information on the captive breeding of B. rubra, focusing on various parameters crucial for ex-situ conservation and domestication. The research involved breeding trials, embryo and larvae observation, first feeding experiments, larva and fry rearing trials, and the evaluation of growth and reproduction in two generations. The study revealed that the female B. rubra, with an average total length of 5.17 ± 0.15 cm and weight of 1.61 ± 0.06 g, produced an average of 73.67 ± 7.09 eggs, 34.33 ± 5.13 total larvae, and exhibited a hatching rate of 46.67 ± 5.77%. The embryogenesis process commenced on the day of spawning (dps) and continued until the eggs hatched at 6 dps. Larvae development and yolk absorption occurred from 0 to 6 days post-hatching (dph). The study also examined the impact of different initial feeding options, with chopped Tubifex resulting in the most significant in- crease (p < 0.05) in length. The growth pattern of B. rubra larvae showed slow initial growth during the first seven days, followed by a rapid exponential growth phase from day 8 to day 39. Two generations of B. rubra (G1 and G2) were successfully bred in captivity, with G2 showing a better tendency for growth in length and weight compared to G1. Notably, there were no significant differences (p > 0.05) in reproductive success between the wild-origin broodstock (G0), G1, or G2. This research contributes valuable insights into the captive breeding of B. rubra and its early life stages, offering critical information for the conservation and sustainable management of this endangered species. Further research is needed to explore the long-term effects of domestication on behavior, physiology, and phenotypic diversity.

Viability Assessment of Fresh and Frozen-thawed Dog Spermatozoa by Flow Cytometry (Flow Cytometry에 의한 개 신선정액과 동결정액의 생존성 분석)

  • Hong Y. M.;Kim Y. J.;Yu I.;Ji D. B.;Kim M. S.
    • Reproductive and Developmental Biology
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    • v.28 no.3
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    • pp.167-172
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    • 2004
  • This study was performed to examine the correlations among dog sperm viabilities evaluated by flow cytometry, by microscopic evaluation (ME), by carbo-xifluorescein diacetate and propidium iodide (CFDA/PI) and by hypoosmotic swelling (HOS) test. Semen were collected from 5 dogs ranging in age from 2 to 4 years. Each ejaculate was divided into 3 aliquots and different proportions of freeze-killed cells were added to each aliquot (1:0, 1:1 and 1:3). In the other experiment, semen was extended with Sweden extender containing 5% glycerol and equex STM paste, and frozen using liquid nitrogen vapor. Fresh and frozen-thawed dog sperm viability were assessed by flow cytometry using PI staining method. The accuracy of flow cytometry was evaluated by comparing with other classic assessments, microscopic evaluation, epifluorescence microscopic analysis using CFDA/PI, and HOS test. High correlations of sperm viabilities were found among flow cytometry, epifluorescence evaluation, HOS test (p<0.01) in fresh semen. Especially, sperm viability assessed by HOS test was highly correlated with viability by flow cytometry in all the ratios of live and dead spermatozoa, 1:0, 1:1 and 1:3 (p<0.01). The viability evaluated by ME were significantly correlated with that by flow cytometry in ratios of 1:0 and 1:3 (p<0.05) however, there was no significance in ratio of 1:1. The viability evaluated by C/p were highly correlated with that by flow cytometry in ratio of 1:0 and 1:1 (p<0.01) and significantly correlated in ratio of 1:3 (p<0.05). In frozen-thawed spermatozoa, the viability determined by HOS test was considerably correlated with that by flow cytometry (p<0.01). There was significant correlation between the viabilities by ME and by flow cytometry (p<0.05). But the viability evaluated by CFDA/PI was not correlated with viability by flow cytometry. The result from this study validate the use of flow cytometry as a precise method for assessing the viability of fresh and frozen-thawed dog spermatozoa.

The Health Status of Rural Farming Women (농촌여성(農村女性)의 건강실태(健康實態)에 관한 연구(硏究))

  • Park, Jung-Eun
    • Journal of agricultural medicine and community health
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    • v.15 no.2
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    • pp.97-106
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    • 1990
  • 1. Background Women's health and their involvement in health care are essential to health for everyone. If they are ignorant, malnourished or over-worked, the health &-their families as well as their own health will suffer. Women's health depends on broad considerations beyond medicine. Among other things, it depends upon their work in farming. their subordination to their families, their accepted roles, and poor hygiene with poorly equipped housing and environmental sanitation. 2. Objectives and Contents a. The health status of rural women : physical and mental complaints, experience of pesticides intoxication, Farmer's syndrome, experiences of reproductive health problems. b. participation in and attitudes towards housework and farming c. accessibility of medical care d. status of maternal health : fertility, family planning practice. induced abortion, and maternal care 3. Research method A nationwide field survey, based on stratified random sampling, was conducted during July, 1986. Revised Cornell Medical index(68 out of 195 items). Kawagai's Farmers Syndrome Scale, and self-developed structured questionnaires were used to rural farming wives(n=2.028). aged between 26-55. 4. Characteristics of the respondents mean age : 40.2 marital status : 90.8% married mean no. of household : 4.9 average years of education : 4.7 yrs. average income of household : \235,000 average years of residence in rural area : 36.4 yrs average Working hours(household and farming) : 11 hrs. 23 min 5. Health Status of rural women a. The average number of physical and mental symptoms were 12.4, 4.7, and the rate of complaints were 22.1%, 38.8% each. revealing complaints of mental symptomes higher than physical ones. b. 65.4% of rural women complained of more than 4 symptoms out of 9, indicating farmer's syndrome. 11.9 % experienced pesticide overdue syndrome c. 57.6% of respondents experienced women-specific health problems. d. Age and education of respondents were the variables which affect on the level of their health 6. Utilization of medical services a. The number of symptoms and complaints of respondents were dependent on the distance to where the health-care service is given b. Drug store was the most commonly utilized due to low price and the distance to reach. while nurse practitioners were well utilized when there were nurse practitioner's office in their villages. c. Rural women were internalized their subordination to husbands and children, revealing they are positive(93%) in health-care demand for-them but negative(30%) for themselves d. 33.0% of respondents were habitual drug users, 4.5% were smokers and 32.3% were alcohol drinkers. and 86.3% experienced induced-abortion. But most of them(77.6%) knew that those had negative effects on health. 7. Maternal Health Care a. Practice rate of contraception was 48.1% : female users were 90.9% in permanent and 89.6% in temporary contraception b. Induced abortions were taken mostly at hospital(86.3%), while health centers(4.7%), midwiferies(4.3%). and others(4.5%) including drug stores were listed a few. The repeated numbers of induced abortion seemed affected on the increasing numbers of symptoms and complaints. c. The first pre-natal check-up during first trimester was 41.8%, safe delivery rate was 15.6%, post-natal check-up during two months after delivery. Rural women had no enough rest after delivery revealing average days of rest from home work and farming 8.3 and 17.2. d. 86.6% practised breast feeding, showing younger and more educated mothers depending on artificial milk 8. Recommendations a. To lessen the multiple role over burden housing and sanitary conditions should be improved, and are needed farming machiner es for women and training on the use of them b. Health education should begin at primary school including health behavior and living environment. c. Women should be encouraged to become policy-makers as well as administrators in the field of women specific health affairs. d. Women's health indicators should be developed and women's health surveillance system too.

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A study on the concentration of trace metal elements in whole blood of the women in reproductive ages in urban and rural area (도시 및 농촌지역 가임연령 여성들의 혈중 미량금속원소의 함량에 관한 조사)

  • Jun, Jin-Ho
    • Journal of Preventive Medicine and Public Health
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    • v.17 no.1
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    • pp.95-106
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    • 1984
  • To acquire the essential basic data to the establishment of control measure for the hazardous health effect that could be caused by harmful metals, the author measured the concentrations of trace metals in whole blood of women of $20{\sim}39$ years old living in urban and rural area using atomic absorption spectrophotometer. The summarized results were as follows; 1. The mean concentration of zinc in whole blood was $10.69{\pm}8.07{\mu}g/ml$ in rural area. The frequency distribution by zinc concentration level was nearly L-type and the cumulative frequency distribution was showed bimodal type in both area. 2. The mean iron concentration in whole blood was $323.09{\pm}87.15{\mu}g/ml$ and $322.07{\pm}104.74{\mu}g/ml$ in urban and rural area, respectively. The frequency distribution was similar to normal distribution type in both area, but the cumulative distribution was unimodal type in urban area and bimodal type in rural area. 3. The mean magnesium concentration was $41.08{\pm}19.58{\mu}g/ml$ and $40.28{\pm}16.82{\mu}g/ml$ in the area, respectively. The frequency distribution type had skewness to the right and the cumulative frequency distribution was unimodal type in both area. 4. The mean copper concentration was $1.417{\pm}0.761{\mu}g/ml$ and $1.375{\pm}0.743{\mu}g/ml$ in the area, respectively. The frequency distribution type had skewness to the right and the cumulative frequency distribution was bimodal type in both area. 5. The mean manganese concentration was $0.079{\pm}0.039{\mu}g/ml$ and $0.07{\pm}0.058{\mu}g/ml$ in the area, respectively. The frequency distribution type had skewness to the right in both area but slight irregular in rural area and the cumulative distribution was unimodal and bimodal type in urban and rural area, respectively. 6. The mean cadmium concentration in whole blood was $0.031{\pm}0.026{\mu}g/ml$ in urban and $0.028{\pm}0.023{\mu}g/ml$ in rural area. The frequency distribution type had skewness to the right and cumulative frequency distribution was bimodal type in both area.

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A Preliminary Epidemiologic Study on Korean Veterans Exposed to Herbicides in Vietnam War (파월국군장병의 고엽제 위해에 관한 예비적 역학조사)

  • Kim, Joung-Soon;Lee, Hyun-Sul;Lee, Hong-Bok;Lee, Won-Young;Park, Young-Joo;Kim, Sung-Soo
    • Journal of Preventive Medicine and Public Health
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    • v.27 no.4 s.48
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    • pp.711-734
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    • 1994
  • Among chemical agents in herbicides, dioxin (2, 3, 7, 8-tetrachloro dibenzo-$\rho$-dioxin : TCDD), a chemical contaminant in herbicides sprayed during the Vietnam War has been known to be the major agent causing toxic effects. Approximately 320,000 korean soldiers participated the Vietnam War from 1964 to 1974. Although the potential hazards of the herbicides among Korean veterans exposed were implicated, the problem had not been a public issue until 1991 when Korean veterans were informed U.S. companies, the herbicides manufacturer payed fund, from which a trust fund for New Zealand and Australian Class members were established in 1985. After a series of appeals and demonstration by the Korean Veterans demanding medical care and compensation for their serious health damages, a bill of medical care and compensation for herbicides victims was promulgated in March 1993 and become effective from May 1993, This study was carried out with two major objectives : the first to understand the health problems caused from the herbicides by reviewing literatures published, and the second to examine the nature and extent of health impacts among Korean veterans exposed and to develop valid study methods for the major study by interviewing and reviewing records on a part of veterans (638 persons) registered and completed medical examination in Seoul Veterans Administration Hospital from June to October 1993. The results obtained are as followings: 1. The literature review of 107 papers revealed that 1) Dioxin is teratogenic, carcinogenic and affects almost all organs including nervous, endocrine, and reproductive systems in animal experiments. 2) The diseases showing evidence of causal association were Hodgkin's disease, non-Hodgkin's disease, lung cancer, lymphoma, soft tissue sarcoma, chloroacne and polyneuropathy when judged on the basis of consistency in study results and biological plausibility. 2. Interview and medical record review study on 638 veterans, though limited validity owing to lack of control group, crude estimates of dioxin exposure levels (no biomarkers measurable), and uncertainty of diagnosis, showed that: 1) Most of the study subject's were in their 40's of age and had been dispatched to Vietnam during the period from $1965{\sim}1970$ around one year. 2) Most frequently complained symptoms in medical examination were motor weakness (32%), sensory abnormalities in extremities (23%), skin diseases (22%), and pain in extremities (20%) whereas in Interview they were more frequent in order of skin problem (44%), motor weakness (38%), sensory abnormalities and pain in extremities(17% and 19% each). Kappa indices on the same category of complaints between two sources of information were variable and relatively low. 3) On medical examination, only a part of the 638 subjects had initial impression (442 pts) and final diagnosis (218 pts) suggesting decision making on diagnosis appeared to be difficult even with all available modern medical technologies: in initial impression disorders from peripheral and central neuropathy were predominant whereas in final diagnosis various types of skin disorder were most frequent 4) When dose-response relationship between several conditions (from questionnaire) and arbitrary exposure scores were examined by CMH linear trend test, spontaneous abortion, sexual problems and health problem of offsprings showed statistically significant linear trends. However, pregnancy, accident and suicidal attempts did not show any relationship in this study capacity. 5) Among complaints, psychosis and neurosis (anxiety, phobia) in interview study, and memory disorder and psychosis in medical record study revealed linear trend. 6) Skin disorder was the only condition showing linear trend in initial impression and none in final diagnosis on medical examination. Even though objective to select out dioxin-related disease or group of diseases from this study was not achieved the research experiences provided firm basis for developing various methodological approaches. 3. From this preliminary study we concluded that a larger scale major epidemiologic study on health impacts of herbicides among Korean veterans exposed is not only Indispensible but also well designed study with more valid exposure information and diagnosis may be able to establish causal relationship between certain groups of diseases and exposure to the herbicides among Korean veterans.

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