• 제목/요약/키워드: Negative Stain

검색결과 140건 처리시간 0.027초

누드 마우스에서 Poly(D,L-lactic-co-glycolic acid) (PLGA) 지지체 내 인체 지방줄기세포의 골성분화 (Osteogenic Differentiation of Human Adipose-derived Stem Cells within PLGA(Poly(D,L-lactic-co-glycolic acid)) Scaffold in the Nude Mouse)

  • 유결;조성돈;변준희;이종원
    • Archives of Plastic Surgery
    • /
    • 제34권2호
    • /
    • pp.141-148
    • /
    • 2007
  • Purpose: The object of this study was to evaluate the development of continuous osteogenic differentiation and bone formation after the subcutaneous implantation of the tissue-engineered bone, in vitro. Methods: Human adipose-derived stem cells were obtained by proteolytic digestion of liposuction aspirates. Adipose-derived stem cells were seeded in PLGA scaffolds after being labeled with PKH26 and cultured in osteogenic differentiation media for 1 month. The PLGA scaffolds with osteogenic stimulated adipose-derived stem cells were implanted in subcutaneous layer of four nude mice. Osteogenesis was assessed by RT-PCR for mRNA of osteopontin and bone sialoprotein(BSP), and immunohistochemistry for osteocalcin, and von Kossa staining for calcification of extracellular matrix at 1 and 2 months. Results: Implanted PLGA scaffold with adipose-derived stem cells were well vascularized, and PLGA scaffolds degraded and were substituted by host tissues. The mRNA of osteopontin and BSP was detected by RT-PCR in both osteogenic stimulation group and also osteocalcin was detected by immunohistochemistry at osteogenic stimulation 1 and 2 months, but no calcified extracellular deposit in von Kossa stain was found in all groups. Conclusion: In vivo, it could also maintain the characteristics of osteogenic differentiation that adipose-derived stem cells within PLGA scaffold after stimulation of osteogenic differentiation in vitro, but there were not normal bone formation in subcutaneous area. Another important factor to consider is in vivo, heterologous environment would have negative effect on bone formation as.[p1]

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

  • 최태열;김신규;김춘원;김기홍;황응수;차창룡;김광혁
    • 대한미생물학회지
    • /
    • 제22권3호
    • /
    • pp.197-208
    • /
    • 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.

  • PDF

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
    • /
    • 제41권6호
    • /
    • pp.367-370
    • /
    • 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
    • /
    • 제57권1호
    • /
    • pp.49-53
    • /
    • 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
    • /
    • 제32권5호
    • /
    • pp.575-581
    • /
    • 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
    • /
    • 제32권12호
    • /
    • pp.1553-1560
    • /
    • 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).

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

  • 박건;이종호;사영조;진웅;권종범;박재길;이선희;곽문섭
    • Journal of Chest Surgery
    • /
    • 제37권3호
    • /
    • pp.261-266
    • /
    • 2004
  • 식도암 환자에서 조기진단 및 수술적 치료 방법의 상당한 진전에도 불구하고 환자의 예후는 여전히 좋지 않다. p53 종양 억제유전자는 세포의 성장과 증식을 조절하는 것으로 알려져 있고 nm23 유전자는 설치류 흑색종에서 종양의 전이억제 효과가 있다고 알려져 있다. 이 실험은 p53과 nm23유전자 발현과 식도암 환자의 임상병리학적인 특징상의 관련성을 알아보고자 하였다. 가톨릭대학교 의과대학 부속 성모병원에서 수술한 식도암 환자 40명의 조직을 대상으로 하였고, p53 변이형 단백질과 nm23단백질을 면역화학적 염색을 시행하여 <10% 양성 종양세포 : negative ; 10∼30% 양성 종양세포: +; 30∼50% 양성 종양세포 : ++; >50% 양성 종양세포: +++의 4개의 군으로 분류하였고, 또한 종양의 침습 정도는 none, mild, moderate, severe로 분류하여 평가하였다. p53 변이형 단백질과 nm23 단백질의 과발현은 생존율 및 임상병리학적 특징과 연관성이 없었고, 또한 p53 및 nm23유전자 발현의 조합 분석에서도 유의한 상관관계를 발견하지 못하였다.

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)- (Studies on Glycolipids in Bacteria -Part I Occurrence of Glycolipids in Various Bacteria-)

  • 김교창
    • Applied Biological Chemistry
    • /
    • 제17권2호
    • /
    • pp.117-124
    • /
    • 1974
  • 23종(種)의 대표적(代表約)인 세균(細菌)에 대(對)하여 glucosamine 함량(含量)을 측정(測定)하여 당지질(糖脂質)의 분포(分布)를 조사(調査)하고 당지질(糖脂質)의 함량(含量)과 Gram염색(染色)과의 상관관계(相關關係)를 규명(究明)한바 다음과 같은 결과(結果)를 얻었다. 1. Gram음성균(陰性菌) 및 varieable균(菌)은 그 균체(菌體) 당지질중(糖脂質中) glucosamine함량(含量)이 최소(最少) Proteus vulgaris의 $0.04{\mu}g$에서 부터 최고(最高) Aerobacter aerogenes 의 $2.48{\mu}g$의 범위(範圍)로 분포(分布)되여 있었다. Gram positive균(菌) 및 Bacillus속중(屬中)의 Gram양성균(陽性菌)은 전부(全部) $0.02{\mu}g$ 이하(以下)의 함량(含量)을 갖고 있었다. Gram양성(湯性)인 Corynebacterium sepedonicum 및 Staphylococcus aureaus는 특(特)히 glucosamine함량(含量)이 최소(最少)였다. 이들보다 Gram음성균(陰性菌)은 양성균(湯性菌)과 명확(明確)히 구별(區別)할 수 있을만큼 glucosamine함량(含量)이 많았다. 2. Gram염색(染色)은 당지질(糖脂質) 추출전(抽出前)보다 추출후(抽出後)가 염색(染色)이 더 잘되는 경향(傾向)이였다. 3. Aerobacter aerogenes, Bacillus circulans, Pseudomonas fluorescens 및 Salmonella typhirurium의 4균주(菌株)에서 얻은 당지질(糖脂質)의 적외선(赤外線) spectra는 대체(大體)로 같았으며 다 OH-group C-O, C-O-C, $CH_2+CH_3$, amide 결합(結合), 지방산(脂肪酸) ester 결합(結合), ester carbonyl 결합(結合)의 존재(存在)를 확인(確認)하였다.

  • PDF

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

  • 이하균;권성준;백승삼
    • Journal of Gastric Cancer
    • /
    • 제4권3호
    • /
    • pp.169-175
    • /
    • 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.

  • PDF

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

  • 최현수;이만섭;박준봉;허익;권영혁
    • Journal of Periodontal and Implant Science
    • /
    • 제26권4호
    • /
    • pp.907-931
    • /
    • 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.

  • PDF