• Title/Summary/Keyword: Isolation system

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TNF-α stimulated IL-8 and IL-10 expression in monocytes from patients with chronic granulomatous disease (만성육아종질환 환자 단핵구에서 TNF-α 자극에 의한 IL-8과 IL-10의 발현 양상)

  • Shin, Kyung-Sue
    • Clinical and Experimental Pediatrics
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    • v.51 no.10
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    • pp.1096-1101
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    • 2008
  • Purpose : Patients with chronic granulomatous disease (CGD) have genetic mutations in a component of the NADPH oxidase enzyme that is necessary for the generation of the superoxide anion. The profound defect in innate immunity is reflected by the patients susceptibility to catalase-positive bacteria and fungi. In addition, CGD patients display signs of persistent inflammation, which is not associated only with deficient superoxide anion production. The aim of this study was to elucidate the cytokine responses in CGD patients after $TNF-{\alpha}$ stimulation. Methods : Heparinized blood samples were collected from 8 CGD patients and 10 healthy volunteers. Monocytes ($1{\times}10^6cell/well$) isolated by the magnet cell isolation system were incubated with a constant amount of $TNF-{\alpha}$ (10 ng/mL) at $37^{\circ}C$ for 6 h. Incubated cells were harvested at 60-min intervals for IL-8 and IL-10 mRNA analysis, and the supernatant was collected at the same intervals to determine IL-8 and IL-10 expression. Monocytes from healthy volunteers were also incubated with antioxidants followed by $TNF-{\alpha}$ stimulation for IL-8 and IL-10 expression. Results : In CGD patients, a high expression of IL-8 together with a significantly higher IL-10 expression than in the healthy controls was seen after $TNF-{\alpha}$ stimulation. Moreover, normal monocytes treated with antioxidants exhibited increased IL-8 responses. Conclusion : The absence of phagocyte-derived reactive oxidants in CGD might be associated with a dysregulated production of pro- and antiinflammatory cytokines. Additional research related to reactive oxidants is needed to clarify the role of cytokines in CGD patients.

Evaluation of Selective Media for Isolation of Staphylococcus aureus from Agricultural Products (농산물 중 Staphylococcus aureus의 분리를 위한 선택배지 평가)

  • Kim, Se-Ri;Lee, Seo-Hyun;Seo, Min-Kyoung;Kim, Won-Il;Park, Kyeong-Hun;Yun, Hye-Jeong;Yoon, Yo-Han;Yoo, Soon-Young;Ryu, Kyoung-Yul;Yun, Jong-Chul;Kim, Byung-Seok
    • Journal of Food Hygiene and Safety
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    • v.27 no.2
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    • pp.169-175
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    • 2012
  • Five kinds of selective media, such as mannitol salt agar (MSA), Baird-Parker agar (BPA), Baird-Parker supplemented with rabbit plasma fibrinogen (BPA+RPF), CHROMagar Staphylococcus aureus (CSA), and Petrifilm Staph Express count system (Petrifilm), were compared to recommend the optimum selective media for isolation of Staphylococcus aureus from agricultural products. Seventy four target and non target bacteria were inoculated on five selective media to analyze sensitivity and specificity. In the recovery test of injured S. aureus cells, S. aureus was exposed to acid (1% lactic acid for 10 min), heat ($60^{\circ}C$ for 90s), and cold ($-20^{\circ}C$ for 1h) conditions. And artificially contaminated agricultural products (iceberg lettuce, green pepper, and cherry tomato) was enumerated on five selective media. The sensitivity of BPA+RPF, CSA, Petrifilm, MSA, and BPA were 100%, 100%, 100%, 90.5%, 90.5%, respectively. In addition, the specificity of BPA+RPF, CSA, MSA, BPA and Petrifilm were 100%, 100%, 84.6%, 75.0%, 67.3%, respectively. However, no difference among five selective media was observed in recovery on injured S. aureus cell and enumeration from agricultural products. This results suggest that BPA+RPF and CSA are the optimum media for detection of S. aureus from agricultural products.

Ethnography of Caring Experience for the Senile Dementia (노인성 치매 환자의 돌봄경험에 대한 문화기술지)

  • 김귀분;이경희
    • Journal of Korean Academy of Nursing
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    • v.28 no.4
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    • pp.1047-1059
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    • 1998
  • Senile Dementia is one of the dispositional mental disorder which has been known to the world since Hippocratic age. It has become a wide-spread social problem all over the world because of chronic disease processes and the demands of dependent care for several years as well as improbability of treatment of it at the causal level. Essentially, life styles of the older generation differ from those of the younger generation. While the fomer is used to the patriarchal system and the spirit of filial piet and respect, the latter is pragmatized and individualized under the effects of the Western material civilization. These differences between the two generations cause conflict between family members. In particular, the pain and conflict of care-givers who take care of a totally dependent dementia patient not only is inciting to the collapse of the family union, but is expanding into a serious social problem. According to this practical difficulty, this study has tried to compare dementia care-givers' experiences inter-culturally and to help set up more proper nursing interventions, describing and explaining them through ethnographies by participant observation and in-depth interviews that enable seeing them in a more close, honest and certain way. It also tries to provide a theoetical model of nusing care for dementia patients which is proper to Korean culture. This study is composed of 12 participants (4 males, 8 females) whose ages range from 37-71 years. The relations of patients are 5 spouses(3 husbands, 2 wives), 4 daughters-in-law, 2 daughters, and 1 son-in-law. The following are the care-givers' meaning of experiences that results of the study shows. The first is "psychological conflict". It contains the minds of getting angry, reproaching, being driven to dispair, blaming oneself, giving up lives, and being afraid, hopeless, and resigned. The second is "physical, social and psychological pressure" . At this stage, care-givers are shown to be under stress of both body and soul for the lack of freedom and tiredness. They also feel constraint because they hardly cope with the care and live through others' eyes. The third is "isolation". It makes the relationship of patient care-giver to be estranged, without understanding each other. They, also, experience indifference such as being upset and left alone. The forth is "acceptance" They gradually have compassion, bear up and then adapt themselves to the circumstances they are in. The fifth is "love". Now they learn to reward the other with love. It is also shown that this stage contains the process of winning others' recognition. The final is "hope". In this stage they really want situations to go smoothly and hope everything will be O.K. These consequences enable us to summarize the principles of cue experience such as, in the early stage, negative response such as physical·psychological confusion, pain and conflict are primary. Then the stage of acceptance emerges. It is an initial positive response phase when care-givers may admit their situations. As time passes by a positive response stage emerges. At last they have love and hope. Three stages we noted above : however, there are never consistent situations. Rather it gradually comes into the stage of acceptance, repeating continuous conflict, pressure and isolation. If any interest and understanding of families or the support of surrounding society lack, it will again be converted to negative responses sooner or later. Otherwise, positive responses like hope and love can be encouraged if the family and the surroundings give active aids and understanding. After all, the principles of dementia care experiences neither stay at any stage, nor develop from negative stages to positive stages steadily. They are cycling systems in which negative responses and positive responses are constantly being converted. I would like to suggest the following based on the above conclusions : First, the systematic and planned education of dementia should be performed in order to enhance public relations. Second, a special medical treatment center which deals with dementia, under government's charge, should be managed. Third, the various studies approaching dementia care experiences result in the development of more reasonable and useful nursing guidelines.

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Epidemiological Studies of Avian Reovirus Infection in Broilers in Korea (국내 육계에서의 조류 레오바이러스 감염에 대한 역학 조사)

  • Kim, J.M.;Kim, M.J.;Song, J.S.;Mo, I.P.
    • Korean Journal of Poultry Science
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    • v.35 no.1
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    • pp.85-99
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    • 2008
  • Avian reovirus (ARV) is a causative agent of viral arthritis/tenosynovitis, and malabsorption syndrome in broiler. The characteristics of malabsorption syndrome caused by ARV are diarrhea, poor feed conversion and stunting. Therefore, ARV infection has been recognized as one of the most important disease in the poultry industry because of economical losses. However, few study of ARV infection in broiler industry has been conducted in Korea. To evaluate the presence of ARV infection in broiler farms, epidemiological survey such as serological test and virus isolation has been conducted. For the serological survey using ELISA method, we selected five broiler farms which were located at different area and had a history of growth retardation, lameness, diarrhea and poor feathering. From these farms serum samples were collected at 1 day, 14 days and market age. All these farms had no history of vaccination against ARV. In addition to serological survey, we tried to isolate ARV from birds of designated farms at market age and collected feces and tissue samples such as cecal tonsil, intestine and liver. We were identified ARV by RT-PCR and transmissible electron microscopy. The samples were inoculated into 9-day-old embryonated eggs via the chorioallantoic membrane to observe the pock formation. For the pathogenicity test of ARV isolates, we inoculated with the isolates to the right footpad of 3-week-old SPF chicks and observed clinical signs and pathological changes for 14 days after challenge. Most broilers sampled for serological survey have maternal antibodies which were widely distributed at 1 day and decreased by 14 days. However, at the market age several broiler farms showed fairly high antibody titer against ARV. This increase of antibody titer at market age means the possible infection of ARV during the grow-out period. Among total 15 samples for the isolation of ARV. 2 samples were positive by RT-PCR and finally identified as a ARV. We inoculated these isolates in the SPF birds and observed that the antibody titer was increased from 7 days after challenge. However, we did not find any clinical signs both control and challenge groups. Based on the above results, it is clear that the ARV infection has been circulated in the broiler industry and caused significant economic losses. Further study is needed to evaluate the virulence of the isolates in the digestive system of broiler and the molecular characteristics of isolates.

An Analysis of Elements in Yen-Ben Street That Form a Sense of Place as an Ethnic Enclave (소수민족집단체류지역(Ethnic Enclave)으로서의 옌볜거리의 장소성 형성 요인 분석)

  • Han, Sung-Mi;Im, Seung-Bin
    • Journal of the Korean Institute of Landscape Architecture
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    • v.36 no.6
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    • pp.81-90
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    • 2009
  • This study seeks those elements that form a sense of place in Yen-Ben Street, which represents a typical ethnic enclave in Seoul, to provide a basic resource in the creation of an urban landscape that can provide a positive space for cultural diversity. The results of the study can be summarized as follows: First, the element of a physical environment that develops a sense of place was in fact the poor dwellings that correspond to the economic condition of Korean Chinese. While this element has a negative cognition to outsiders, Korean Chinese feel positively toward it. Secondly, signboards were a physical element of sense of place which retains cultural identity as a means of communication inside the community. Thirdly, it was found that activities such as shopping, recreation, and the exchange of information that are found in the pursuit of daily life act as an essential element in the formation of a sense of place even more than architectural elements. Fourthly, the appropriation of space by Korean Chinese and the isolation from the surroundings were obvious. This isolation is perceived as a negative sense of place formation to outsiders in Yen-Ben Street. Fifthly, the aspects of cultural dualism, mingling the concepts of home country, language, writing, and food have also affected the formation of a sense of place in the area. Sixthly, transience was a prominent phenomenon of Yen-Ben Street and is strengthened by illegal immigration. Although transience causes negative impacts such as in a lack of concern for the residential environment, it acts as a positive factor in the sense of place by mitigating uneasiness, and strengthening insider ties and cooperation.

Establishment of the High-Throughput Hair Roots' DNA Isolation System and Verification of Its Appicability for Hanwoo Traceability Using the 11 Microsatellite Makes (대량 모근 시료 DNA 분리 체계 확립과 11 microsatellite maker를 사용하는 한우 생산이력제로의 적용가능성 검증)

  • Lim, Hyun-Tae;Lee, Sang-Ho;Yoo, Chae-Kyoung;Sun, Du-Won;Cho, In-Cheol;Yoon, Du-Hak;Yang, Dae-Young;Cheong, Il-Cheong;Lee, Jung-Gyu;Jeon, Jin-Tae
    • Journal of agriculture & life science
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    • v.44 no.6
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    • pp.91-99
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    • 2010
  • We used a multiplex PCR primer set composed of 11 microsatellite (MS) markers and two sexing markers for gender detection. Genomic DNA extracted from hair roots of 3,510 Hanwoo were genotyped. Based on the 11MS markers, no animals had identical genotypes(TGLA227, BM2113, TGLA53, ETF10, SPS115, TGLA122, ETH3, ETH225, BM1824 and INRA23). The expected probability of identity among genotypes of random individuals (PI), the probability of identity among genotypes from random half-sibs ($PI_{half-sibs}$) and among genotypes of random individuals, and the probability of identity among genotypes from random sibs ($PI_{sibs}$) were estimated as $1.31{\times}10^{-23}$, $2.52{\times}10^{-16}$and $1.09{\times}10^{-6}$, respectively using the API-CALC program, version 1.0. We successfully completed the genotype analysis of 3,510 Hanwoo with a 3.93% genotyping failure rate. It was revealed that extracting DNA from the hair root was a time-efficient and cost-effective method to collect specimens for DNA isolation from live animals. This method also minimized stress for the animals during specimen collection. Among the hair roots from the back, belly, upper tail and lower tail, 5~13 hair roots of the lower tail led to the best genotype analysis results. Finally, we established a 96-well-format method of DNA preparation applicable for high- throughput genotype analysis.

Isolation of Polysaccharides Modulating Mouse’s Intestinal Immune System from Peels of Citrus unshiu (귤피로부터 분리한 마우스의 장관면역 활성 다당류의 검색)

  • Yang, Hyun-Seuk;Yu, Kwang-Won;Choi, Yang-Mun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.9
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    • pp.1476-1485
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    • 2004
  • Of solvent-extracts prepared from the 90 kinds of Korean traditional tea and rice gruel plants, cold-water extract from peels of Citrus unshiu (CUI-0) showed the most potent intestinal immune system modulating activity through Peyer’s patch whereas other extracts did not have the activity except for cold-water extracts of Laminaria japonica, Polygonatum japonicum, Poncirus trifoliata, and hot-water extracts of Gardenia jasminoides, Lycium chinense having intermediate activity. CUI-0 was further fractionated into MeOH-soluble fraction (CUI-1), MeOH insoluble and EtOH-soluble fraction (CUI-2), and crude polysaccharide fraction (CUI-3). Among these fractions, CUI-3 showed the most potent stimulating activity for the proliferation of bone marrow cells mediated by Peyer’s patch cells, and contained arabinose, galacturonic acid, galactose, glucose, glucuronic acid and rhamnose (molar ratio; 1.00:0.53:0.45:0.28:0.28:0.19) as the major sugars, and a small quantity of protein (9.4%). In treatments of CUI-3 with pronase and periodate (NaIO₄), the intestinal immune system modulating activity of CUI-3 was significantly reduced, and the activity of CUI-3 was affected by periodate oxidation particularly. The potently active carbohydrate-rich fraction, CUI-3IIb-3-2 was further purified by anion-exchange chromatography on DEAE-Sepharose FF, Sepharose CL-6B and Sephacryl S-200. CUI-3IIb-3-2 was eluted as a single peak on HPLC and its molecular weight was estimated to be 18,000 Da. CUI-3IIb-3-2 was consisted mainly of arabinose, galactose, rhamnose, galacturonic acid and glucuronic acid (molar ratio;1.00:0.54:0.28:1.45:0.63) in addition to a small amount of proteins (3.2%). In addition, CUI-3IIb-3-2 showed the activity only through Peyer’s patch cells, but this fraction did not directly stimulate proliferation of bone marrow cells. It may be concluded that intestinal immune system modulating activity of peels from C. unshiu is caused by pectic polysaccharides having a polygalacturonan moiety with neutral sugars such as arabinose and galactose.

A Survey of Staphylococcus aureus Contamination and Antibiotic Susceptibility in Retail Meat (국내유통 축산물에서의 Staphylococcus aureus 오염도 및 항생제 감수성 조사)

  • Yang, Jung-Im;Lee, Sun-Min;Lee, Gil;Lee, Hwan-Ju;Kim, Min-Kyu;Jeong, Eun-Jeong;Cha, Yong-Jun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.4
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    • pp.528-533
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    • 2008
  • Staphylococcus aureus is a pathogene of major concern in livestock products. This study was conducted to test imported and domestic meat sold by retail stores for the presence S. aureus. In addition, the antibiotic susceptibility of any S. aureus found was also evaluated. The overall isolation rate of S. aureus was 20.2% (13.9% in pork and 33.8% in beef) in retail meats. The percentage of imported meats found to contain S. aureus (33.3% in pork and 40.4% in beef) was higher than that of domestic meat (13.0% in pork and 14.7% in beef). In addition, the detection rate of S. aureus was higher in raw material meat than in ready to cook packaged meat. When the antibiotic susceptibility of S. aureus isolated from the meat products was evaluated, ampicillin was found to be the highest (76.5%), followed by penicillin (75.3%), tetracycline (27.1%) and erythromycin (21.2%). Penicillin and tetracycline resistant were detected in 55.6% and 13.3% of the beef isolates, respectively, and 97.5% and 42.5% of the pork isolates, respectively. The tetracycline and erythromycin resistant plasmids of the isolated strain were transferred into S. aureus DPRMM2429 by the filter mating method and the frequencies of transfer was found to be $1.1{\times}10^{-5}{\sim}1.9{\times}10^{-9}$ and $1.2{\times}10^{-5}{\sim}4.0{\times}10^{-8}$ respectively.

Antioxidative and Hepatoprotective Effects of Injinho-Tang and Osumogwa-Tang (인진호탕(茵蔯蒿湯)과 오수목과탕(吳茱木瓜湯)의 항산화(抗酸化) 및 간세포(肝細胞) 보호효과(保護效果))

  • Lee, Sang-Hyun;Kim, Young-Bok
    • Herbal Formula Science
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    • v.16 no.1
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    • pp.117-130
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    • 2008
  • Korean traditional medicine has been used for the treatment of the various diseases based on both oriental medicinal theory and clinical trials. Thus, the prescriptions of Korean traditional medicine would be useful for the development of new therapeutics. This research focuses on the fundamental study in Korean traditional prescriptions for the development of new hepatoprotective agents. We found two prescriptions. Injinho-Tang and Osumogwa-Tang, showed the significant DPPH free radical scavenging and hepatoprotective effect, respectively. It is well-known that free radical scavenging effect is related to the prevention of various pathological events including liver injury. This paper deals with hepatoprotective effects on tacrine-induced cytotoxicity in Hep G2 cells, free radicals scavenging on both DPPH and superoxide of above two prescriptions. Hot water extract of Injinho-Tang did not show the significant hepatoprotective effect on tacrine-induced cytotoxicity in Hep G2 cells, however, it shows the significant scavenging effects for both DPPH and superoxide radicals. On the other hand, all of the hot water extracts of constituent herbal drugs in Injinho-Tang exhibited the promising protective effect on tacrine-induced cytotoxicity in Hep G2 cells. Of these, water extract of Rhei Rhizoma showed the most prominent effect on tacrine-induced cytotoxicity in Hep G2 cells. Bioassay-guided fractionation of Rhei Rhizoma extract has furnished four compounds, and their chemical structures have been identified by comparison of their spectral data with those of literature as chrysophanol (1), emodin (2), 3,5-dihydroxy-4'- methoxystilbene (3), and rhapontigenin (4), respectively. Among the isolated compounds, compounds 2-4 revealed the significant hepatoprotective effect in vitro when their $EC_{50}$ values compare with that of silybin, as a positive control. It also exhibited that emodin possessed the most hepatoprotective effect among these active compounds. In case of Osumogwa-Tang, its hot water extract showed the moderate protective effect on tacrine-induced cytotoxicity in Hep G2 cells. Hot water extract of Chaenomelis Fructus, one of the constituent herbal drug of this prescription, exhibited the significant hepatoprotective effect with $EC_{50}$ value of $7.8{\pm}0.1\;{\mu}g/ml$, however, it showed strong cytotoxicity in Hep G2 cells above the concentration of $25\;{\mu}g/ml$. It was revealed that both hot water extract of Evodiae Fructus and its butanol soluble fraction showed the moderate hepatoprotective effect but concentration-dependent activity in Hep G2 assay system. Two quinolone alkaloids, evocarpine and dihydroevocarpine, also tested for their hepatoprotective effects on tacrine-induced cytotoxicity in Hep G2 cells, however, these two compounds derived from the Evodiae Fructus as the major constituents did not show in vitro hepatoprotective effect. From these results, it would be necessary to further isolation of its hepatoprotective compounds from the butanol soluble fraction of the hot water extract of Evodiae Fructus.

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Adenyl Cyclase Activity in Cold-acclimatized Animals (한냉적응이 Adenyl Cyclase Activity에 미치는 영향)

  • Kang, Bok-Soon;Lee, Sang-Ho;Kang, Doo-Hee
    • The Korean Journal of Physiology
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    • v.8 no.2
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    • pp.67-74
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    • 1974
  • The object of this research is aimed to determine the activity of adenyl cyclase in both skeletal muscle sarcolemma and fat cell ghost of epididymal adipose tissue isolated from rats exposed to cold for various length of time in an attempt to evaluate whether the tissue sensitivity to catecholamine is increased when rats are exposed to cold for long periods of time Methods: a)Animals: Albino rats ranging in weight from 150 to 200 gm were used throughout this study. For experimental purposes, the rats are divided into two groups: experimental animals were place4 in a cold room at $4^{\circ}C$, controls being kept at $25^{\circ}C$. At the end of 2, 4, 6, 12, and 16 weeks. exposure to cold the rats were used to measure the adenyl cyclase activity. b) Isolation of plasma membrane from skeletal muscle and adipose tissue: The Plasma membrane of skeletal muscle from hind limbs of rats are prepared by the method employed by Rosenthal et at. and fat cell ghost of epididymal adipose tissue of rats by the method employed by Rodbell. c) Adenyl cyclase assay: Adenyl cyclase activity were measured by the method employed by Marinetti et al. Briefly, plasma membrane was incubated with $3^H-ATP$, various amount of noradrenaline and other incubation mixture at $37^{\circ}C$ for 20 minutes. After stopping the enzyme reaction by immersion in boiling water, carrier 3',5'-AMP was added to the system as a marker and $100\;{\mu}1$ aliquots of incubation mixture were pipetted on $20{\time}20$ Whatman No. 3 MM filter paper for one dimensional chromatography. The cyclic AMP spots were cut off and placed in counting vials containing 10ml of Bray's scintillation cocktail. Radioactivity was determined with a Packard Tri-Carb liquid scintillation counter. The enzyme activity is expressed as nanomoles of cyclic AMP produced per mg of membrane per hour. Result: 1. Average adenyl cyclase activity in the plasma membrane of skeletal muscle before and after noradrenaline administration was significantly higher in the cold-exposed rats as compared to the control. Continuous exposure to cold Produced an increased adenyl cyclase activity before and after noradrenaline administration. Adenyl cyclase activity reached peak levels at the 6 weeks exposure to told and level of adenyl cyclase activity remained high. Noradrenaline administration to the incubation medium induced a significant increase in adenyl cyclase activity and the degree of stimulation were proportional to the hormonal concentration But the rate of inclement in adenyl cyclase activity by noradreasline was the same in both groups. 2. Adenyl cyclase activity in fat cell ghost between cold exposed and control rats showed no significant differences before and after noradreualine administration. In summary, it can be concluded that cold adaptation give rise an increased activity of adenyl cyclase in plasma membrane of skeletal muscle in rats.

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