• Title/Summary/Keyword: Negative Stain

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Production of Monoclonal Antibody to Chlamydia Trachomatis (Chlamydia trachomatis 진단에 유용한 단세포군 항체 생산에 관한 연구)

  • Choi, Tae-Yeal;Kim, Think-You;Kim, Choon-Won;Kim, Ki-Hong;Hwang, Eung-Soo;Cha, Chang-Yong;Kim, Kwang-Hyuk
    • The Journal of the Korean Society for Microbiology
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    • v.22 no.3
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    • pp.197-208
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    • 1987
  • Chlamydia trachomatis has now shown that this interesting intracellular parasite is a cause of nongonococcal urethritis, infantile pneumonia, pelvic inflammatory disease and epididymitis, in addition to lymphogranuloma venerum and inclusion conjunctivitis. There are several diagnostic methods for C. trachomatis, but the method using monoclonal antibody is the most sensitive and specific. The hybride cell were prepared by fusion of myeloma cell($P_3X_{63}\;Ag_8{\cdot}V_{653}$) of mouse and lymphocyte of mouse(BALB/c) that were immunized with formalin killed C. trachomatis serotype D. The cell mixtures after fusion were dispensed into 640 wells of the 96 well culture plates and continuously cultured in HAT medium for 2 weeks. The supernatants of culture media in 83(13%) wells were reacted with C. trachomatis, which were determined by enzyme-linked immunosorbent assay in 96 well microplate. The clones that secreted antibody to C. trachomatis were cloned by limiting dilution. Only six monoclones secreted antibody to C. trachomatis. The antibody titer of ascitic fluid that collected from same BALB/c mice bearing hybridoma cells was above 1:100,000. These monoclonal antibodies that were IgG reacted with elementary and reticulate bodies of all serotypes(Ba, D, E, F, G, H, J and LGV type-I) using ELISA and indirect immunofluorescence stain, but there were no cross reaction with other bacteria(coagulase negative Staphylococcus, Proteus and E. coli). We concluded these six monoclones secreted the same monoclonal antibody to C. trachomatis. The sensitivity and specificity of the monoclonal antibody compared with Microtrak(confirmatory test of C. trachomatis, Syva) was 100%, respectively.

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Correlation of Clinical and Immunohistochemical Diagnosis in Patients with Pituitary Adenomas

  • Park, Sung-Ku;Jung, Shin;Jung, Tae-Young;Kim, In-Young;Kim, Soo-Han;Kang, Sam-Suk
    • Journal of Korean Neurosurgical Society
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    • v.41 no.6
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    • pp.367-370
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    • 2007
  • Objective: Pituitary adenomas are common neurological lesions believed to account for 10% to 15% of all primary brain tumors. There can be diagnostic confusion due to discordance of the preoperative endocrine and the postoperative immunohistochemical diagnosis. In this study, the rate of discordance between preoperative and postoperative findings and their clinical implications were investigated. Methods: From March 2005 to March 2006, 26 patients who underwent surgery for a pituitary adenoma were enrolled in this study. The preoperative pituitary hormone level and postoperative immunohistochemical results were compared and analyzed. Results: The median age of the patients was 38 years [range 15-66 years]. The male to female ratio was 8 to 18. The endocrine evaluation showed 16 hormonally-active and 10 hormonally-inactive adenomas. The immunohistochemical findings showed : 13 prolactin-positive, 1 GH-positive, 1 FSH-positive, 8 pleurihormone-positive and 3 stain-negative adenomas. The percentage of discordance observed between the preoperative endocrine and postoperative immunohistochemical diagnosis was 54%. Nine of 10 endocrine non-functioning adenomas showed : 3 PRL positive, 1 GH positive, 2 PRL+GH positive, 1 TSH+FSH positive, 1 FSH+ACTH+PRL positive and 1 FSH+LH+PRL positive adenomas by immunohistochemistry. Three endocrine PRL+GH secreting adenomas showed 2 PRL positive and 1 FSH+GH positive by immunohistochemistry. One endocrine PRL secreting and 1 GH secreting adenoma showed 1 PRL+ TSH positive and 1 GH+PRL positive by immunohistochemistry, respectively. The diagnosis of the other 12 pituitary adenomas showed concordance. Conclusion : The results of this study showed 54% discordance rate between the preoperative endocrine and postoperative immunohistochemical diagnosis for pituitary adenomas.

Effect of Temperature on the Killing of Opisthorchis viverrini Eggs In Vitro

  • Boueroy, Parichart;Duenngai, Kunyarat;Eamudomkarn, Chatanun;Sripan, Panupan;Boonmars, Thidarut;Pumhirunroj, Benjamabhorn;Artchayasawat, Atchara;Songsri, Jiraporn;Chomphumee, Kanpicha;Rattanasuwan, Panaratana;Laummaunwai, Porntip;Khueangchiangkhwang, Sukhonthip;Boonjaraspinyo, Sirintip
    • Parasites, Hosts and Diseases
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    • v.57 no.1
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    • pp.49-53
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    • 2019
  • Contaminated liver fluke egg in the environment has led to the high prevalence of human opisthorchiasis associated with cholangiocarcinoma in Southeast Asia. To find the effective lessening methods of Opisthorchis viverrini eggs in the contaminated environment, we investigated the temperature conditions for killing of these trematode eggs in vitro. Numerous O. viverrini eggs were obtained in the proximal part of uteri of adult worms from experimental hamsters. Mature eggs with miracidium were allocated by experimental groups (2 control: positive and negative and 4 treatment: 50, 60, 70, and $80^{\circ}C$) with 0.85% saline, and treated by the experimental plan. Eggs in each experimental groups were observed under the confocal microscope after stain with Propidium Iodide (PI) to evaluate the effect of temperatures. Eggs in 70 and $80^{\circ}C$ groups were all killed after over 10 min heated. Majority of eggs in $60^{\circ}C$ (10, 15, and 30 min heated), 70 and $80^{\circ}C$ (5 min heated) groups were inactivated. However in $50^{\circ}C$ group, below half of eggs were to be killed in all time lapse (10, 15 and 30 min). In order to prevent O. viverrini infection and cholangiocarcinoma, direct treatment of sewage by heating at 70 or $80^{\circ}C$ at least 10 min is essential. Therefore, treatment of O. viverrini eggs at a high temperature is a potential method for controlling egg contamination in sewage.

Caenimonas aquaedulcis sp. nov., Isolated from Freshwater of Daechung Reservoir during Microcystis Bloom

  • Le, Ve Van;Ko, So-Ra;Lee, Sang-Ah;Kang, Mingyeong;Oh, Hee-Mock;Ahn, Chi-Yong
    • Journal of Microbiology and Biotechnology
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    • v.32 no.5
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    • pp.575-581
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    • 2022
  • A Gram-stain-negative, white-coloured, and rod-shaped bacterium, strain DR4-4T, was isolated from Daechung Reservoir, Republic of Korea, during Microcystis bloom. Strain DR4-4T was most closely related to Caenimonas terrae SGM1-15T and Caenimonas koreensis EMB320T with 98.1% 16S rRNA gene sequence similarities. The average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values between strain DR4-4T and closely related type strains were below 79.46% and 22.30%, respectively. The genomic DNA G+C content was 67.5%. The major cellular fatty acids (≥10% of the total) were identified as C16:0, cyclo C17:0, summed feature 3 (C16:1ω7c and/or C16:1ω6c), and summed feature 8 (C18:1ω7c and/or C18:1ω6c). Strain DR4-4T possessed phosphatidylethanolamine, diphosphatidylglycerol, and phosphatidylglycerol as the main polar lipids and Q-8 as the respiratory quinone. The polyamine profile was composed of putrescine, cadaverine, and spermidine. The results of polyphasic characterization indicated that the isolated strain DR4-4T represents a novel species within the genus Caenimonas, for which the name Caenimonas aquaedulcis sp. nov. is proposed. The type strain is DR4-4T (=KCTC 82470T =JCM 34453T).

Mucilaginibacter aquariorum sp. nov., Isolated from Fresh Water

  • Ve Van Le;So-Ra Ko;Mingyeong Kang;Hee-Mock Oh;Chi-Yong Ahn
    • Journal of Microbiology and Biotechnology
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    • v.32 no.12
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    • pp.1553-1560
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    • 2022
  • A Gram-stain-negative, rod-shaped bacterial strain, JC4T, was isolated from a freshwater sample and determined the taxonomic position. Initial identification based on 16S rRNA gene sequences revealed that strain JC4T is affiliated to the genus Mucilaginibacter with a sequence similarity of 97.97% to Mucilaginibacter rigui WPCB133T. The average nucleotide identity and digital DNA-DNA hybridization values between strain JC4T and Mucilaginibacter species were estimated below 80.92% and 23.9%, respectively. Strain JC4T contained summed feature 3 (C16:1 ω7c and/or C16:1 ω6c) and iso-C15:0 as predominant cellular fatty acids. The dominant polar lipids were identified as phosphatidylethanolamine, one unidentified aminophospholipid, one unidentified phospholipid, and two unidentified lipids. The respiratory quinone was MK-7. The genomic DNA G+C content of strain JC4T was determined to be 42.44%. The above polyphasic evidences support that strain JC4T represents a novel species of the genus Mucilaginibacter, for which the name Mucilaginibacter aquariorum sp. nov. is proposed. The type strain is JC4T (= KCTC 92230T = LMG 32715T).

Clinical Significance of p53 Gene and nm23 Gene Expression in Esophageal Cancer (식도암 조직에시 p53 및 nm23 유전자 발현의 임상적 의의)

  • Park, Kuhn;Lee, Jong-Ho;Sa, Young-Jo;Jin, Ung;Kwon, Jong-Bum;Park, Jae-Gil;Lee, Sun-He;Kwak, Moon-Sub
    • Journal of Chest Surgery
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    • v.37 no.3
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    • pp.261-266
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    • 2004
  • Although significant progress has been made in the surgical treatment of esophageal carcinoma as well as in the detection of early stage esophageal carcinoma by diagnostic techniques, the prognosis of the esophageal carcinoma patients remain poor. The p53 gene product is known to regulate cell growth and proliferation. And the nm23 gene was identified originally as an anti-metastatic influence whose expression was correlated inversely with tumor metastatic potential in murine melanoma cell lines. This experiment was intended to know the relationship among the p53 and nm23 gene expression versus clinicopahologic characteristics of the esophageal cancer. Total 40 cases were collected from patients who had undergone esophagectomy at St. Mary's Hospital, Catholic university of Korea. Immunohistochemical stain for p53 mutant-type protein and nm23 protein was graded as <10% positive tumor cells: negative; 10∼30% positive tumor cells: + ; 30∼50% positive tumor cells: ++, and >50% positive tumor cells: +++. The tumor invasion was grades as none:- ; mild:+ ; moderate:++ ; severe: +++. Overexpression of p53 protein and nm23 was not associated with the survival and cliniocopathologic characteristics of the esophageal cancer. Moreover, the combination analysis of p53 and nm23 revealed that there was no relationship between the gene expression and the clinicopatholic characteristics of the esophageal cancer.

Studies on Glycolipids in Bacteria -Part I Occurrence of Glycolipids in Various Bacteria- (세균(細菌)의 당지질(糖脂質)에 관(關)한 연구(硏究) -제1보(第一報) 세균(細菌)에 있어서의 당지질(糖脂質)의 분포(分布)-)

  • Kim, Kyo-Chang
    • Applied Biological Chemistry
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    • v.17 no.2
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    • pp.117-124
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    • 1974
  • The 23 representative bacteria were studied for the glucosamine contents which represent the glycolipid content of the cell wall. The distribution of glycolipid in various bacteria was examined and the relationship between the glycolipid contents and the Gram stain was elucidated. The results were as follows: 1. The contents of glucosamine in the glycolipid of Gram negative and variable bacterial cell wall were large ranging from the least 0.04 ${\mu}g$ of Proteus vulgaris to the largest 2.48 ${\mu}g$ of Aerobacter aerogenes. The Gram positive bacteria and only those Gram positive among Bacilli contained less than 0.02 ${\mu}g$ of glucosamine contents. The least glucosamine containing Gram positive bacteria were Corynebacterium sepedonicum and Staphylococcus aureus. It could generally be said that the Gram negative and variable bacteria contain the higher content of glucosamine in the cell wall than the positives. 2. The bacteria were better stained by the Gram solution after the extraction of glycolipid from the cell wall than those without extraction. 3. The four infrared spectra of glycolipids obtained from Aerobacter aerogenes, Bacillus circulans, Pseudomonas fluorescens, and Salmonella typhirurium showed all the similar characteristics. All showed the existence of groups; OH, C-O, C-O-C, $CH_2+CH_3$, amide band, fatty acid ester band and ester carbonyl bond.

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Clinicopathological Significance of p53 and HSP27 in Gastric-cancer Patients (위암 환자에서 p53과 HSP27의 임상병리학적 의의)

  • Lee, Ha-Gyoon;Kwon, Sung-Joon;Baek, Seung-Sam
    • Journal of Gastric Cancer
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    • v.4 no.3
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    • pp.169-175
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    • 2004
  • Purpose: The tumor suppressor gene p53 has been shown to be a factor in the carcinogenesis or progression of gastric cancer. The mutant p53 has been reported to cause a higher risk of lymph-node metastasis. Futhermore, mutation of the p53 has been linked to a poor prognosis for gastric cancer. The heat shock protein-27 (HSP27), a stress protein, has also been reported to be a poor prognostic factor in ovarian and breast cancers. However, in gastric-cancer patients, controversies exist as to its influence on the prognosis. In the present study, we used an immunohistochemical stain to observe the effects of p53 and HSP27 on the clinicopathological factors and on the prognosis for gastric-cancer patients. Materials and Methods: To evaluate the significance of p53 and HSP27 in gastric cancer patients, we analyzed 212 cases of gastric cancer (stage I.IV). Tissue samples of 212 patients were stained immunohistochemically for the mutant p53 protein and for HSP27. The correlations between protein expression and the clinicopathological factors were investigated. Results: The overall expression rates for p53 and HSP27 were $36.9\%\;and\;27.8\%$, respectively. p53 and HSP27 were correlated to each other because the HSP27 expression rate was higher in the p53-positive group (P=0.046). Statistically, the p53 and the HSP27 expression rates were significantly increased in the case of tumor invasiveness, lymphatic metastasis and vessel involvement. Therefore, they play a role in cancer progression. The 5-year survival rates of the p53-positive and the p53-negative groups were $62.8\%\;and\;60.1\%$, respectively (P=0.793) while the 5-year survival rates for the HSP27-positive and HSP27-negative groups were $54.2\%\;and\;63.1\%$, respectively (P=0.090). Conclusion: p53 and HSP27 were correlated to each other in our immunohistochemical study of gastric carcinomas and they were not independent prognostic factors in gastric- cancer patients. However, further studies are needed to determine their prognostic values for gastric-cancer patients.

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THE EFFECTS OF THE BIOGLASS AND THE NATURAL CORAL ON HEALING PROCESS OF THE ALVEOLAR BONE DEFECTS (생체유리 및 천연산호 이식재가 성견 치조골 결손부의 재생에 미치는 영향)

  • Choi, Hyun-Soo;Lee, Man-Sup;Park, Joon-Bong;Herr, Yeek;Kwon, Young-Hyuk
    • Journal of Periodontal and Implant Science
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    • v.26 no.4
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    • pp.907-931
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    • 1996
  • The purpose of this study was to study of the effects of the bioglass and the natural coral on healing process of the alveolar bone defects. Three adult dogs aged 1 to 2 years were used in this study. Experimental alveolar bone defects were created surgically with surgical bur and bone chisel at the furcation area of the buccal surface of the right and left mandibular 3rd, 4th premolars. Twelve experimental alveolar bone defects were devided into four groups according to the type of graft materials. The groups were as follows : 1. flap operation with root planing & curettage(Negative control group) 2. flap operation with autogenous bone(Positive control group) 3. flap operation with bioglass(BG group) 4. flap operation with natural coral(NC group) At 2, 4, and 8 weeks, the dogs were serially sacrificed and specimens were prepared with Hematoxylin-Eosin stain for light microscopic evaluation. The results of this study were as follows : 1. The defect areas were filled with granulation tissue at two weeks in negative control group. But in other groups, the appearance of connective tissues around graft materials were formed more densely and the response of inflammation by graft materials itself was not found. 2. In every control and experimental groups at two weeks, there was seen the accumulation of the formation of new bone trabeculae at the bottom of defects and gradually expanded toward the graft materials and in autogenous group there was slightly seen the formation of new cementum. 3. There was seen the erosion of central portion of bioglass particles at two weeks in BG group, and the erosion of the central portion was developed more progressively and was filled with bone-like tissues at eight weeks. 4. The natural coral particles were encapsulated by densely connective tissues and seen the formation of new bone tissues at four weeks and developed more new bone and cementum formation at eight weeks. From the results of this study, the bioglass and the natural coral may be biocompatible and have a weak adverse reaction to the periodontal tissues.

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Two cases of Listeria monocytogenes isolation from celebrospinal fluid (뇌척수액에서의 Listeria monocytogenes 분리 2예 보고)

  • Chong, Yun-Sop;Kim, Yoon-Chung;Kim, Byung-Soo;Yi, Kui-Nyung;Lee, Sam-Uel Y.
    • The Journal of the Korean Society for Microbiology
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    • v.13 no.1
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    • pp.1-5
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    • 1978
  • Listeria monocytogenes infection was considered a rather rare disease and occurs mostly either in newborn babies or in young children. However, there has been increasing reports of this infection in elderly person with various underlying disease. Recently we have experienced two cases of Listeria meningitis; one in a 4-year-old male with an acute lymphoblastic leukemia, and the other in a 43-year-old female with a breast cancer. Both were on various chemotherapeutic agents for their primary diseases when the organism, L. monocytogenes was found in their celebospinal fluid(CSF). The degree of CSF pleocytosis were quite different by cases. The former case showed a marked increase, $3,350/mm^3$, and the latter slight, $410/mm^3$, Both showed a slight decrease of CSF glucose ranging 39 to 43mg/100ml. It seems that a routine CSF analysis bears a limitted value in the diagnosis or Listeria meningitis. A direct smear of CSF with Gram's stain revealed gram-positive bacilli in one case, but none in the other. Bacterial culture of CSF yielded plenty colonies in one case, but a few in the other. It seems that isolation of L. monocytogenes must not be considered very easy, and a negative direct smear does not necessarily mean a negative culture. The two isolates we obtained showed the typical cultural and biochemical characteristics of L. monocytogenes and were found to belong to serotypes 1b and 4b. It was our experience that the identification of this organism was not very much matter because of its distinct characteristics, but the most important matter was how to think of the possibility of this organism at the begining. The two isolates were both susceptible to cephalothin, chloramphenicol, erythromycin, tetracycline and gentamicin; intermediate to ampicillin, penicillin and kanamycin; and resistant to cloxacillin.

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