• 제목/요약/키워드: Enzymeimmunoassay

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Enzymeimmunoassay for the Plasma Vitellogenin and Early Determination of Ovarian Maturation in Red Seabream, Pagrus major (참돔(Pagrus major)의 혈장 난황단백전구체에 대한 효소면역측정법과 난소성숙의 조기판정)

  • Han Chang-Haa;Yang Mun-Ho;Paek Jae-Min;Lim Sang-Koo;Kim Kwang-Hyun
    • Journal of Aquaculture
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    • v.8 no.1
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    • pp.1-19
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    • 1995
  • In red seabream, Pagrus major the female specific protein in the vitellogenic female serum was identified by Ouchterlony's immunodiffusion test and immunoelectrophoresis. The female specific serum protein might be vitellogenin based on the results of the immunological analysis for the male and vitellogenic female sera and crude egg extracts. Also, it was identified by the immunodiffusion test that the purified yolk protein from ovarian egg extracts has antigenic identities shared with the female specific serum protein. To study the relationship between the maturational stages of gonad and plasma levels of vitellogenin, these were measured from the late resting period (January) to the vitellogenic preiod (April) by the modified enzymeimmunoassay (EIA) using antiserum against yolk protein. The level of plasma vitellogenin began to increase in February (previtellogenesis stage) and continuously increased with the ovarian growth during the vitellogenesis period (March to April). The plasma vitellogenin levels were significantly different between the females and the males in February. Validation for the modified EIA system. was tested .The absorbance curve of serial dilutions of serum from the vitellogenic female was paralleled to the standard curve of yolk protein; $109\pm5.6\%$ recovery was achieved by the modified EIA. And the intraassay coefficients of variation were less than 10% within the concentration ranging from 31.3 ng/ml to 1,000 ng/ml. These findings suggest that the sex determination in adult red seabreams could be possible by using the modified EIA as early as in February.

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Effect of Suckling Systems on Serum Oxytocin and Cortisol Concentrations and Behavior to a Novel Object in Beef Calves

  • Chen, Siyu;Tanaka, Shigefumi;Ogura, Shin-ichiro;Roh, Sanggun;Sato, Shusuke
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.11
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    • pp.1662-1668
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    • 2015
  • We investigated differences between effects of natural- and bucket-suckling methods on basal serum oxytocin (OT) and cortisol concentrations, and the effect of OT concentration on affiliative and investigative behavior of calves to a novel object. Ten Japanese Black calves, balanced with birth order, were allocated evenly to natural-suckling (NS) and bucket suckling (BS) groups. Blood samples were collected at the ages of 1 and 2 months (1 week after weaning) calves, and serum OT and cortisol concentrations were measured using enzyme-linked immunosorbent assay and enzymeimmunoassay tests, respectively. Each calf at the age of 2 months (2 weeks after weaning) was released into an open-field with a calf decoy, and its investigative and affiliative behaviors were recorded for 20 minutes. In 1-month-old calves, the basal serum OT concentration ($25.5{\pm}4.9$ [mean${\pm}$standard deviation, pg/mL]) of NS was significantly higher than that of BS ($16.9{\pm}6.7$) (p<0.05), whereas the basal cortisol concentration ($5.8{\pm}2.5$ [mean${\pm}$standard deviation, ng/mL]) of NS was significantly lower than that in BS ($10.0{\pm}2.8$) (p<0.05). Additionally, a negative correlation was noted between serum OT and cortisol concentrations in 1-month-old calves (p = 0.06). Further, the higher serum OT concentration the calves had at 1 month old, the more investigative the calves were at 2 months old but not affiliative in the open-field with a calf decoy. Thus, we concluded that the natural suckling method from a dam elevates the basal serum OT concentration in calves, and high serum OT concentrations induce investigative behavior and attenuate cortisol concentrations.

Evaluation of the Usefulness of GDH & Toxin Test for the Diagnosis of Clostridioides difficile in a Tertiary Hospital in Seoul

  • Joo, Ho-Joong;Kim, Sang-Ha;Kwon, Pil-Seung;Ryu, Jae-Ki;Yook, Keun-Dol;Yu, Young-Bin;Kim, Young-Kwon
    • Biomedical Science Letters
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    • v.26 no.3
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    • pp.210-216
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    • 2020
  • The purpose of this study was to evaluate GDH & Toxin (GDT) tests for the identification of the presence of Clostridioides difficile (C. difficile) as well as to detect whether any toxin was present in the feces of patients suspected of diarrhea associated with C. difficile. Data related to the results of toxin and culture (TC) tests and GDT tests conducted on patients with diarrhea and suspected CDI between January 2017 and august 2018, positive test rates, patient ages and sexes, whether the patients were hospitalized, and turnaround time (TAT) were analyzed retrospectively. Of the 7,554 total tests conducted for CDI diagnosis, 1,010 TC tests (14.9%) were positive, while 92 GDT tests (12.0%) were positive. Of these positive cases, 815 (80.7%) identified through TC test and 80 (87%) identified through GDT test were inpatients. also, among the patients with positive test results, 497 (49.2%) diagnosed through TC test and 45 (48.9%) diagnosed through GDT test were aged 61 years or older. The total time required to complete a TC test was 83.6 hours, while the time required for a GDT test was 11.2 hours, equating to an approximately three-day difference between the two tests. The detection of toxin-producing C. difficile is important in CDI diagnosis, but the commonly used Enzymeimmunoassay (EIA) toxin tests with low sensitivity result in delayed CDI diagnosis time. Therefore, primary screening tests for CDI diagnosis using the GDT method and secondary tests using additional methods are considered most effective.

THE CONCENTRATIONS OF PROSTAGLANDIN E2, 6-KETO-PROSTAGLANDIN F1α, AND LEUKOTRIENE B4 IN PULPAL AND PERIAPICAL LESIONS (치수 및 치근단병소에서 Prostaglandin E2, 6-keto-Prostaglandin F1α, Leukotriene B4의 분포에 관한 연구)

  • Shon, Won-Jun;Baek, Seung-Ho;Lim, Sung-Sam
    • Restorative Dentistry and Endodontics
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    • v.25 no.2
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    • pp.193-201
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    • 2000
  • Prostaglandins (PGs) and Leukotrienes (LTs) have been implicated in the genesis of pulpal and periapical inflammation. In this study, the relationships among $PGE_2$, 6-keto-PG $F_1{\alpha}$ (a stable metabolite of $PGI_2$) and $LTB_4$ concentrations in inflamed pulp and periapical lesions were discussed. Pulp tissue were obtained in routine endodontic treatment and periapical lesions in periapical surgery after clinical diagnoses were made. These specimens were divided into four groups as normal pulp group (Control group), acute pulpitis group, chronic pulpitis group, and periapical lesion group. Pulp tissue and periapical lesions were stored in liquid nitrogen. The concentration of $PGE_2$, $PGI_2$ and $LTB_4$ were measured with ELISA. The data were analyzed by one-way ANOVA. Significantly higher levels of $PGE_2$, 6-keto-PG $F_1{\alpha}$ a and $LTB_4$ were found in acute pulpitis group than chronic pulpitis group and periapical lesion group(p<0.05). Periapical lesion group showed significantly higher mean concentrations of $PGE_2$ and $LTB_4$ than chronic pulpitis group. In control and chronic pulpitis group, significant higher levels of $PGI_2$ than $PGE_2$ and $LTB_4$ were found. These results suggested that the high levels of $PGE_2$ and $LTB_4$ in periapical lesions may be due to rich endothelium., fibroblast and lymphocyte known as the main producers of $PGE_2$ and $LTB_4$. $PGI_2$ may be thought to one of the most abundant PGs in normal pulp tissue.

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EFFECT OF OXYGEN TENSION ON THE ACTIVITY AND FUNCTION OF THE CELLS DERIVED FROM HUMAN PERIODONTAL LIGAMENT (산소 장력이 치주인대에서 유래한 세포의 활성과 기능에 미치는 영향)

  • Sa, Muyng-Hee;Yang, Won-Sik
    • The korean journal of orthodontics
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    • v.27 no.3 s.62
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    • pp.503-513
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    • 1997
  • This study was undertaken to investigate the effect of oxygen tension on the activity and function of the cells derived from human periodontal ligament by measuring cell activity, total protein synthesis, collagen synthesis, $IL-1{\beta},\;IL-6,\;TNF-{\alpha}$ Human periodontal ligament fibroblasts were collected from premolars extracted for orthodontic treatment and incubated in the environment of $37^{\circ}C,\;5\%\;CO_2,\100\%$ humidity. After the fifth to sixth passage they were used for the experiment. Gaspack system to which $0.2{\mu}m$ Millipore filter was attached was connected to mixed-gas tanks. The mixed gases were composed of $10\%\;O_2,\;5\%\;CO_2,\;85\%\;N_2$ in hyoxic group or $90\%\;O_2,\;5\%\;CO_2,\;5\%\;N_2$ in hyperoxic group and $5\%\;CO_2,\;95\%$ air for control. After incubation in $37^{\circ}C$ for 2, 4, 6 days, cell activity was determined by tetrazolium(MTT) assay and total protein synthesis was assayed using sulforhodamine B(SRB). And measurement of 4-hydroxyproline was performed to assess collagen synthesis md $IL-1{\beta},\;IL-6,\;and\;TNF-{\alpha}$ were measured by enzymeimmunoassay. The results were as follows. 1. The cell activity and total protein synthesis in hypoxic group were a little higher than or almost the same with those in control group. 2. In hyperoxic group, the cell activity was lower than that in control group and total protein synthesis was decreased. 3. Collagen synthesis was significantly decreased initially in both hypoxic and hyperoxic group and increased nearly to the level of control group as the duration of cell incubation was longer 4. As a result of enzymeimmunoassay, the amount of cytokines was $IL-6,\;TNF-{\alpha}\;and\;IL-1{\beta}$ in order. 5. $IL-6,\;TNF-{\alpha}\;and\;IL-1{\beta}$ were increased more rapidly in both hypoxic and hyperoxic group than in control group as the duration of cell incubation was longer. 6. There were more $IL-6\;and\;TNF-{\alpha}$ in hyperoxic group than in control group after 6 days, and there were more $IL-6\;and\;TNF-{\alpha}$ after 6 days than after 2 or 4 days in hyperoxic group. These results suggested that oxygen tension might modulate the production of extracellular matrix and cytokines in the cells derived from human periodontal ligament.

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Ultrasensitive Enzymeimmunoassay for Testosterone in Human Saliva (사람 타액내 Testosterone의 초감도 효소면역측정법)

  • 윤용달;전은현;이창주;도병록;이준영
    • Development and Reproduction
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    • v.4 no.1
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    • pp.115-123
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    • 2000
  • A few enzymeimmunoassay (EfA) for testosterone (T) have been reported but was not suitable for all biological samples. The present study was designed to develop a rapid, ultrasensitive EIA and to apply this technique for study the physiological changes of T in biological samples. Saliva samples were collected at 06:00~09:00 hour during one menstrual cycle from 18 normally menstruating women and on 09:00~10:00 hour from 20 normal men. The present study shows an established EIA for testosterone, using horseradish peroxidase (HRP), which was covalently bonded to testosterone-3-carboxymethyloxime (T-3-CMO). One batch of T-antisera was also covalently linked to microcrystalline cellulose particles by a mixed anhydride method in order to facilitate separation of bound and free steroids. The established EIA was validated in terms of sensitivity, accuracy, specificity, precisions etc., comparing with conventional radioimmunoassay. The sensitivity of the established EIA was less than 25 pg/tube. The correlation coefficients between the expected T-values and observed T-values measured by EIA or RIA were r=0.985 and r=0.941 respectively. The cross reactivity of antiserum in EIA was a little higher than that of RIA, especially by 5 ${\alpha}$-DHT. The intra- and inter-assay precisions of the present EIA were similar to those of RIA. The present study also demonstrates that the normal T-values in saliva of Korean male & female samples are 265.65${\pm}$15.80 pmol/l and 109.74${\pm}$ 12.01 pmol/l, respectively. The present EIA seems to be established and suitable for use in the endocrinological studies. The advantages of this EIA system also might make the present T-EIA an ideal procedure for use in a routine assay of ordinary laboratory with a conventional spectrophotometer.

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EFFECT OF VARIOUS CYTOKINES ON THE PRODUCTION OF PROSTAGLANDIN $E_2$, LEUKOTRIENE $B_4$ AND COLLAGENASE IN HUMAN PERIODONTAL LIGAMENT FIBROBLASTS IN VITRO (수종의 cytokine이 사람 치주인대 섬유아세포의 prostaglandine $E_2$, leukotriene $B_4$ 및 collagenase 생산에 미치는 영향)

  • Kim, Jung-Ho;Suhr, Cheong-Hoon
    • The korean journal of orthodontics
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    • v.24 no.4 s.47
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    • pp.871-883
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    • 1994
  • This experiment was designed to study possible roles of $interleukin-1\beta$, interleukin-6 and tumor necrosis $factor-\alpha$ in bone remodeling by measuring their effects on $PGE_2,\; LTB_4$ and collagenase production when they were administered to human periodontal ligament fibroblasts. Human periodontal ligament fibroblasts were collected from first premolars extracted for orthodontic treatment. They were incubated in the environment of $37^{\circ}C,\;5\%\;Co^2,\;and\;100\%$ humidity. They were treated with $0.25\%$ trypsin-EDTA solution and centrifuged. PDL cells in the fifth to seventh passage were used for the experiment. Cells were seeded onto the culture dishes and when they were successfully attached, human recombinant $interleukin-1\beta$, interleukin-6, and tumor necrosis $factor-\alpha$ were administered, alone or in combination. They were incubated for 4, 8 and 24 hours and the levels of $PGE_2,\;LTB_4$ and collagenase released into the culture media were assessed by enzymeimmunoassay and collagenase activity assay. The conclusions are as follows: 1. $IL-1\beta\;and\;TNF-\alpha$ were very active in stimulating the production of $PGE_2$ and collagenase by human periodontal ligament fibroblasts, while IL-6 increased $LTB_4$ production. 2. $IL-1\beta$ significantly increased $PGE_2$, but $LTB_4$ Production was not increased. $IL-1\beta$ is thought to act mainly via the cyclooxygenase pathway of arachidonic acid metabolism. 3. IL-6 tended to inhibit $IL-1\beta$ in the production of $PGE_2$ and collagense whereas IL-6 and $TNF-\alpha$ showed auditive effect in the level of $PGE_2$. The above cytokines increased the release of at least one of $PGE_2,\;LTB_4$ and collagenase. It suggests that cytokines are involved in bone remodeling process by stimulating PDL fibroblasts to produce various bone-resorptive agents. The roles of cytokines in bone remodeling as a whole would need further study.

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Monitoring the Reproductive Status of Dairy Cows by Urinary Pregnanediol Glucuronide

  • Yang, C.J.;Wu, L.S.;Liu, S.H.;Lin, J.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.4
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    • pp.460-466
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    • 2004
  • This study was undertaken with the aim to establish a reliable radioimmunoassay (RIA) system for urinary pregnanediol glucuronide (PdG) and to employ it for monitoring the reproductive status of dairy cows. Urine and blood samples were collected from the Holstein cows both pregnant and non-pregnant. The samples were then investigated for evaluating the relationship between progesterone ($P_{4}$) in blood and PdG in urine adjusted with or without urinary creatinine basis. Biweekly urine collection was employed for three cows in estrous and those artificially inseminated, while urine from pregnant cows was collected on a monthly basis. P_{4}$ and PdG levels were measured by enzymeimmunoassay (EIA) and RIA techniques, respectively. Our results indicated the sensitivity of PdG for RIA being 35 pg/tube and the recovery rate of 100%. Urinary creatinine concentrations also fluctuated within a day, but change at midday was not noteworthy. Regardless of the time of urination the change in concentrations of PdG was relatively smaller and did not vary significantly. The urinary PdG concentration showed periodic changes as that with serum P_{4}$ levels during the cow's estrus cycle. The correlation coefficient rose when creatinine level in urine was adjusted but the change was also not significant. The concentrations of PdG during the luteal phase were detected between 8.2 and 17.4 ng/ml, three to five times higher than that in the follicular phase. The concentration of PdG from pregnant cows (21 days after conception) was three to four times higher than in the nonpregnant cows. Our finding suggests that the determination of urinary PdG could be reliably employed for early pregnancy detection. The urinary PdG level continued to raise until 30 days pre-partum while the concentration reached its peak at 30 ng/ml, after which it started to fall 18 to 30 days before parturition and finally fell to its nadir value one week after parturition. As the correlation coefficient between the urinary PdG and serum P_{4}$ was higher than that corrected by urinary creatinine it can be suggested that the adjustment is not needed. The concentrations of urinary PdG could be maintained stably for 2 days in urine samples stored at room temperature and extended to 8 days when the samples were pretreated by boiling for 30 minutes. In conclusion urinary PdG concentration even without the need for creatinine basis adjustment can be used directly for monitoring the reproductive status of dairy cows.

Detection of Matrix Metanoproteinase -1, -2, -3 in pulpal and periapical pathosis (치수 및 치근단 병소에서 기질금속단백분해효소(matrix metalloproteinase) -1, -2, -3의 분포에 관한 연구)

  • Su-Jung Shin;Seung-Ho Baek;Sung-Sam Lim
    • Restorative Dentistry and Endodontics
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    • v.26 no.3
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    • pp.191-199
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    • 2001
  • 목적 - 기질금속단백분해효소(matrix metalloproteinase)는 조직의 염증 및 치유과정에서 숙주세포에서 생성, 분비되어 세포외기질(extracellular matrix)의 분해에 작용한다. 다양한 염증반응에 기질금속단백분해효소가 중요한 역할을 하는것으로 보고되고 있으나 치수 및 치근단 질환에서의 그 역할은 거의 알려져 있지 않은 상태이다. 본 연구에서는 염증이 있는 사람의 치수 및 치근단 조직을 채취하여 Enzymeimmunoassay 및 면역조직화학적 검색을 통해 제1형, 2형, 3형 기질금속단백분해효소의 수준 및 그 분포를 측정하여 치수 및 치근단 병소에서 이 효소의 작용을 알아보는 것을 목적으로 한다. 방법 - 연구재료는 근관치료를 위해 서울대학교 병원 치과 진료부 보존과에 내원한 환자를 대상으로 34개의 치아에서 통상의 근관치료 중 발수한 치수조직과 치근단 수술중 얻은 치근단 병소(n=10)를 이용하였다. 치수는 발수 전에 임상진단을 통해 급성 치수염(n=12), 만성 치수염(n=12), 정상 치수(n=10)로 구분하고 정상치수로 진단된 것을 대조군으로 설정하였다. 채취된 표본은 둘로 나누어 절반은 30분 이내에 5$\mu\textrm{m}$ 두께로 동결절단을 시행하여 조직표본을 제작하였고 deep freezer에 보관하였다가 헤마톡실린-에오신 염색 및 면역조직화학적 검색을 시행하였다. 나머지 조직은 ELISA를 위해 액체 질소에 보관하였다. ELISA를 시행하기전 표본의 단백질 정량을 시행하여 모든 표본의 단백질 양을 50mg/$\mu\textrm{l}$로 일치시키고 Amersham사의 ELISA kit를 사용하여 제1형, 2형, 3형의 기질금속단백분해효소의 양을 측정하였으며 그 결과를 Mann-Whitney U test를 사용하여 각 군간의 통계학적 유의성을 검증하였다. 결과 1. ELISA의 결과 제1형 기질금속단백분해효소의 농도는 모든 실험군에서 대조군보다 유의성있게 높게 나타났다.(p<.05). 또한 급성치수염군의 제1형 기질금속단백분해효소의 농도가 다른 실험군보다 유의성있게 높았다(p<.05). 2. 제2형 기질금속단백분해효소의 경우 급성치수염군과 대조군에서만 유의성있는 차이를 보였다(p<.05). 3. 제3형 기질금속단백분해효소의 경우 급성치수염군에서 대조군이나 만성치수염군보다 유의성 있는 높은 수치를 보였다(p<.05). 4. 면역조직화학검색 결과 염증성 치수에 존재하는 급성 및 만성염증세포 주위로 기질금속단백분해효소에 대한 면역 반응이 존재하였으며 주로 제1형과 제3형 기질금속단백분해효소의 경우 대식세포 및 림파구 주위로 강한 발색제의 침윤양상이 관찰되었다. 5. 치근단병소의 면역조직화학적 검색 결과 만성염증 세포 주변으로 미약한 발색제의 침윤양상이 관찰되었다.

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Effects of Cigarette Smoking on Clinical Symptoms and Level of Serum Prolactin in Schizophrenic Patients (흡연이 정신분열병 환자의 임상 증상 및 혈청 Prolactin에 미치는 영향)

  • Woo, Haing-Won;Lim, Weon-Jeong;Yun, Kyu-Wol
    • Korean Journal of Psychosomatic Medicine
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    • v.5 no.2
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    • pp.143-150
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    • 1997
  • This study was performed to evaluate the characteristics of smoking behavior and the effects of smoking on clinical symptoms and level of serum prolactin in schizophrenic patients. Methods : 76 male schizophrenic patients answered the questionnaire about the characteristics of smoking patterns. And patients were assessed by brief psychiatric rating scale(BPRS), positive and negative syndrome scle(PANSS), Hamilton rating scale for depression(HAM-D), assessment for involuntary movement scale(AIMS) and symptom checklist 90 R(SCL-90-R). Serum prolactin levels were measured by enzymeimmunoassay. Results: 1) The frequences of drinking coffee were significantly higher in smokers. The reasons for smoking were to relieve tension, to avoid boredom, due to habit and to do with friends. 80.1% of smokers tried quitting, but smoking was relapsed due to craving and withdrawal symptoms. 2) No significant difference was seen in mean neuropeltic doses, scores of PANSS and AIMS. But as for BPRS, scores of anxiety/depression subscale were significanly lower in smokers. Scores of HAM-D and scores of interpersonal sensitivity and phobia among SCL-90-R were significantly lower. 3) Levels of serum prolactin were significanlty lower in smokers. Conclusion : These findings suggest that in schisophrenia smoking relieves anxiety and depression subjectively But decreased prolactin levels may suggest that the possibility of increased dopamine in CNS.

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