• Title/Summary/Keyword: micromethod

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A Micromethod for Rapid and Simple Isolation of Genomic DNA from Small Scale Culture of Bifidobacterium (소량의 Bifidobacterium 배양액에서 genomic DNA 추출을 위한 신속/간단한 방법)

  • Jeakal, Soo;Park, Hee-Kyung;Song, Ji-Eun;Heo, Tae-Ryeon;So, Jae-Seong
    • Microbiology and Biotechnology Letters
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    • v.23 no.6
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    • pp.781-783
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    • 1995
  • A method is described for the rapid and simple isolation of genomic DNA from 3 ml culture of Bifidobacterium. The method is expected to be used in gene manipulation of Bifidobacterium spp. The isolated DNA using this method is shown to be an excellent substrate for restriction endonuclease digestion and ligation with T4 DNA ligase.

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Thyroid Stimulating Immunoglobulin Bioassay Using Cultured Human Thyroid Cells; A Simplified Micromethod (갑상선질환에서 갑상선 자극면역글로불린측정의 의의에 관한 연구 -Micro법 갑상선세포배양에 의한 측정의 기본적 검토-)

  • Lee, Myung-Chul;Chung, June-Key;Cho, Bo-Youn;Koh, Chang-Soon;Lee, Mun-Ho;Ahn, Il-Min;Ahn, Hee-Kwon
    • The Korean Journal of Nuclear Medicine
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    • v.19 no.1
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    • pp.95-102
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    • 1985
  • The activation of adenylate cyclase of human thyrocytes in primary cell culture and the release of c-AMP into the medium are used to detect b-TSH and TSAb in sera of patients with autoimmune thyroid disease. Sera of patients are used directly as a part of cell culture without immunoglobulin precipitation. In the above TSI bioassay, TSAb pooled serum show c-AMP concentration between that of 1mU/ml and 10 mU/ml b-TSH but normal control pooled serum doesn't show any detectable c-AMP response. Ninety fiye percent of untreated Graves' patients shows TSAb activity above normal range, 20% of Hashimoto's and 36% of euthyroid Graves' patients show detectable TSAb activity.

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A Rapid Serotyping of Hydrophobic Strains of Mycobacterium acrofulaceum by Fluorescent Anti-Complement Technique (형광항보체법(螢光抗補體법)에 의한 Mycobacterium scrofulaceum 소수성균주(疏水性菌株)의 형별동정(型別同定))

  • Choi, Chul-Soon;Kim, Yong-Jae;Yang, Yong-Tae
    • The Journal of the Korean Society for Microbiology
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    • v.15 no.1
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    • pp.9-17
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    • 1980
  • In the ecology and epidemiologic studies on various serotypes of atypical mycobacteria(AM), Schaefer's bacterial agglutination test(BA) provided the basis of the serologic procedures. Recently, attempts have been made to modify and to simplify the Schaefer's BA such as a slide agglutination test(Engel & Beerwald, 1970), a "simplified" BA(Reznikov & Leggo, 1972), an agglutination inhibition test(Richards & Eacret, 1972) and "micromethod"(Thoen et al., 1975). The BA, however, was not widely applied as a routine laboratory test mainly because it requires much times and labors to perform and partley because it is not applicable to hydrophobic strains either often encountered in the isolation of AM in the clinical bacteriology or stock strains maintained in the laboratory. On the contrary, fluorescent antibody technique with mycobacteria may have advantages over the BA because it is far more simpler in serologic procedures and is applicable to all strains of mycobacteria regardless of smooth or rough types of cultures. At the present, it is well known that the type-specific antigens are lacking on the surface of rough type of AM compared to that on smooth type of strain, but the antigenicity on the surface of the hydrophobic strains of AM which resulted from a series of subculture and the strain in the laboratory for 3 to 6 months has not been clarified. In this study, an attempt to serotype the hydrophobic strains of M. scrofulaceum serotype 41, 42 and 43 by fluorescent anti-complement(FAC) technique was made. The FAC technique with mycobacteria was also described in detail. In the summary, the complement fixing antibody titres of reference sera to smooth types of homologous serotype was highest, but the antibody titres of reference sera to hydrophobic strains of serotypes, 41, 42 and 43 gave two-to 8-folds lower than those to smooth type of strains. Although the sensitivity of type-specific antigens on the hydrophobic strains to reference sera was much lower, using the two units of reference sera determined by titration with hydrophobic strains, three serotypes, i. e., 41, 42 and 43 were specifically differentiated one another by FAC technique. This result indicated that the hydrophobic strains which were maintained in the laboratory at least for 6 months still retain type-specific antigen detectable by FAC technique.

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C-분염(分染)에 의한 사람의 염색체 다형성에 관한 연구

  • Choi, Soo-Kyung;Paik, Yong-Kyun;Lee, Hyung-Hoan
    • Clinical and Experimental Reproductive Medicine
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    • v.16 no.1
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    • pp.41-55
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    • 1989
  • Lymphocyte chromosome preparations obtained by the micromethod (Arakaki and Sparkes, 1963) from 234 our patients (165 females and 69 males) were analysed by C-, NOR-and GC-bandings for chromosome heteromorphisms. The centromeric regions of chromosomes 1,9,16 and the long arm of the Y chromosomes were tested for C heteromorphism. Minor variations found in this study such as inv(9), prominant short arms and large satellites of acrocentrics were also examined by appropriate banding techniques. Of the 234 probands, a total of 125 different C-variants were detected, and the average frequency of the variants per individual was estimated to be 0.53. The observed variations were as follows : 99 qh variants, 5 pericentric inversions of chromosome 9, and 21 satellite and/or short arm variants.

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The Study on the Ferrokinetics and Acquired Immunity in Repeated Hookworm Infections (구충성빈혈(鉤蟲性貧血)에 관(關)한 연구(硏究))

  • Lee, Mun-Ho;Lee, Pyl-Ung
    • The Korean Journal of Nuclear Medicine
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    • v.1 no.2
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    • pp.61-74
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    • 1967
  • In order to confirm whether acquired immunity or resistance can be developed by the repeated hookworm infections, the 150 mature actively moving filariform ancylostoma duodenale larvae obtained from the severe hookworm anemia patients were orally given to 8 healthy volunteers in three divided doses, 50 in each, at 5 day interval. Also the hematological changes as well as several ferrokinetics using $^{59}Fe$ were done and were compared with 10 controls. The clinical symptoms and signs were checked every day for the first 3 weeks and then twice weekly until the end of the experiment. The appearance of the ova in the stool was examined by the formalin ether method and the ova was counted by the Stoll's method. The following laboratory tests were done: 1) Red blood cell count, venous blood hematocrit(micromethod), hemoglobin count (cyanomethemoglobin method) were checked every 5 to 7 day interval. 2) Plasma iron concentration (Barkan's modified method) was determined every 2 to 3 week interval. 3) Radioisotope studies: a) Ferrokinetics: Huff et al and Bothwell's method were applied. Erythropoietic Index (% of normal)=$\frac{Subject's\;turnover/100ml\;whole\;blood{\times}100}{Average\;normal\;turnover/100ml\;whole\;blood}$ of the gastrointestinal absorption of iron: Radioiron($^{59}Fe$) balance b) Quantitative measurement method was applied. c) Determination of the plasma erythropoietin activity: Fried's method was applied. Following were the results: 1) The serum iron level was lower. The red cell volume was decreased, but with relative increase of plasma volume. 2) The plasma iron disappearance time was accelerated and the plasma iron turnover rate was decreased. The red cell iron turnover rate was markedly increased, while all of the red cell iron concentration, circulating red cell iron. plasma iron pool were decreased. The daily iron pool turnover and red cell renewal rate were increased. 3) The erythropoietic index, erythropoietin activity and intestinal absorption of iron($^{59}Fe$) were markedly increased. 4) The infectivity was $9.8{\pm}1.31%$ which was lower than that observed in the single infection. 5) From these observations, it is concluded that the hookworm anemia is essentially iron deficieny in its origin and some immunity acquisition is possible with repeated infections.

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Evaluation of Acid Steatocrit in Korean Infants (한국인 영아에서의 Acid Steatocrit 표준치에 관한 연구)

  • Kim, Kyo-Sun;Park, Ji-He;Sim, Jae-Geon
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.4 no.1
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    • pp.77-82
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    • 2001
  • Purpose: The fecal acid steatocrit is an improved steatocrit method for the evaluation of fecal fat. The present study was set up in order to define the normal range of acid steatocrit values during the first 3 months of life. Methods: Fecal acid steatocrit values were determined in 78 healthy full term and in 21 healthy prematurely born infants between May 1998 and April 2000. The acid steatocrit method was performed in these babies during the first 3 months of life. Results: Steatorrhea occurs during the first month and then decreases, as shown by the fall in the acid steatocrit curve from 1st to 3rd month in our subject. Very high acid steatocrit results (above 90%) were found in all full term and premature infants during the first month of age. Acid steatocrit results of human milk-fed infants were significantly lower than those of formula-fed infants (p=0.0018). Conclusion: We conclude that high acid steatocrit results during the first 1 month of age can be due to physiologic steatorrhea. The acid steatocrit micromethod can be used for the evaluation of milk fat absorption in infants and monitoring steatorrhea instead of other more cumbersome methods.

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Studies on the Simplified Hemagglutination Reaction to Pasteurella multocida and the Stabilization of Erythrocytes (Pasteurella multocida에 대(對)한 간이적혈구(簡易赤血球) 응집반응(凝集反應)과 적혈구(赤血球)의 안정화(安定化)에 관(關)한 연구(硏究))

  • Lee, Hak Cheul;Chung, Yung Gun;Kim, Kyo-Joon
    • Korean Journal of Veterinary Research
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    • v.10 no.1
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    • pp.11-23
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    • 1970
  • Recently Carter(1952) reported the capsule antigens of Pasteurella multocida could be divided into four serological types A,B,C and D by means of precipitation tests. Subsequently he showed that the most sensitive for identification of these types involved the use of capsule substance adsorbed by erythrocytes in hemagglutination test. It may be somewhat difficult to conduct the hemagglutination test in small laboratory, because relatively large amounts of antisera and erythrocytes of the human O type are required for the test. A simple method for serological typing of P. multocida was the slide agglutination test employed by Little et al. (1943) and Namioka et al. (1962), but this method is still in controversy. The author tried adapting Carter's hemagglutination method to the slide method so called "micromethod technique", and studied on the stabilization of erythrocytes for use of slide hemagglutination to P. multocida although many invesigators reported the stabilization of erythrocytes. The results obtained are summarized as follows: 1. A simplified method (slide method) for capsule typing of the organism was developed by adapting Carter's hemagglutination reaction(tube method). Antibody-containing serum can be diluted serially on Boerner's microtest slide with capillary or serological pipetts with a considerable accuracy. The slide reaction can be carried out with case on the slide by adding $0.05m{\ell}$ of antigen-sensitized erythrocytes suspension diluted to one percent on $0.05m{\ell}$ of serially diluted antibody-containing sera, and the final result can be read after 60 minutes at the room temperature ($15^{\circ}C$). 2. It is difficult to determine superiority of inferiority between the slide method and the tube method on the pattern of the reaction of hemagglutination. 3. The pH range of 6.6 to 8.3 is optimal for the slide hemagglutination reaction. 4. The antigen-sensitization against erythrocytes at $37^{\circ}C$ is optimal for the slide hemagglutination. 5. Both the doses and concentration of antigen do not influence the antigen-adsorbing capacity of erythrocytes. 6. The reduction of antigen-sensitizing hours does not influence the antigen-adsorbing capacity of erythrocytes even 30 minutes. 7. The tannic acid treatment against formalinized and non-formalinized erythrocytes showed no effect on the reaction of hemagglutination. 8. The erythrocytes preserved at $4^{\circ}C$ in the ACD solution do not decrease the reactivity on the reaction of hemagglutination for 60 days, while they begin slight hemolysis 30 days after preserving. 9. The stable preparation of erythrocytes can be obtained by treating the cells at $37^{\circ}C$ for 20 hours with from 4 to 8 percent of formalin in saline or buffer. These cells can be preserved at $4^{\circ}C$ for more than 8 months experimented without hemolysis. With low concentration of formalin, the cells were not sufficiently stabilized resulting in the hemolysis after short period of preservation at $4^{\circ}C$. 10. The erythrocytes treated with 16 percent of formalin remain constantly or increase the reactivity for the reaction of hemagglutination. On the contrary, the cells treated with I to 8 percent of formalin decrease the reactivity. 11. There is no difference between nontreated fresh erythrocytes and the erythrocytes preserved in the ACD solution on the reactivity against the hemagglutination, and the erythrocytes treated with 16 percent of formalin showed the reactivity of higher level than that of the above two kinds of erythrocytes. 12. There is no difference between the saline and the isotonic buffer solution on the reaction of hemagglutination.

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