• Journal of Microbiology and Biotechnology
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• 제9권3호
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• pp.311-317
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• 1999

Human T-cell leukemia virus Type-I (HTLV-I) is etiologically associated with rare adult T-cell leukemia, a malignant T-cell disorder. cDNAs encoding p24 (gag), gp21(env), and pXII of HTLV-I were amplified by polymerase chain reaction (PCR) using the genomic DNA extracted from HUT102 cell line as a template. The amplified cDNAs were cloned into the Escherichia coli expression vectors and over-expression of the recombinant proteins were achieved by adding IPTG into the culture media in order to induce the promoter. The molecular weights of the recombinant p24, gp21, and pXII, determined by SDS-PAGE, were found to be approximately 28 kDa, 23 kDa, and 15 kDa, respectively. Reactivity of the recombinant proteins with human sera was tested by the immunoblot assay. The gp21 and p24 reacted against the sera obtained from HTLV-I-infected individuals but not against the sera obtained from normal persons. These results suggest that the recombinant proteins expressed in E. coli were recognized by antibodies in sera from HTLV-I infected patients. These recombinant proteins would be applicable for detecting the presence of antibodies against HTLV-I in human blood samples.

• Journal of Microbiology and Biotechnology
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• 제20권10호
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• pp.1457-1462
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• 2010

Three types of serotypically atypical Shigella flexneri isolates were collected between 2007 and 2008 from Korean patients at the Korea National Institute of Health (NIH). These atypical isolates were characterized and compared with serologically typical S. flexneri. The first grouping of 11 atypical isolates displayed agglutination only with polyB antiserum and exhibited no reaction with any typing or grouping sera (PolyB:un). The second group of 3 isolates displayed reactions with typing sera IV, but also did not bind with any grouping sera (IV:un). The third group of 14 isolates exhibited a plural agglutination pattern, reacting with typing sera II, and two grouping sera (II:(3)4,7(8)). Amongst these atypical isolates, isolates belonging to IV:un and II:(3)4,7(8) exhibited greater antibiotic resistance, in particular to ampicillin, streptomycin, and trimethoprim-sulfamethoxazole, than typical S. flexneri strains. Furthermore, all II:(3)4,7(8) strains harbored integrons. This study suggests that these multiple antibiotic-resistant atypical S. flexneri are new subserotypes of S. flexneri that await further serological classification.

• Restorative Dentistry and Endodontics
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• 제19권2호
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• pp.485-498
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• 1994

Porphyromonas endodontalis is a black-pigmented anaerobic Gram-negative rod which is associated with endodontal infections and this microorganism possesses a potential for pathogenicity. The purpose of this study was to compare the membrane components of Porphyromonas endodontalis and Porphyromonas gingivalis and to study the immune reaction patterns of Porphyromonas endodontalis with patients with periapical lesion. Porphyromonas endodontalis (ATCC 35406), Porphyromonas gingivals serotypea (381), serotype b(W50), serotype c(A7A1-28) were cultured in anaerobic condition. Rabbit antisera were prepared by intravenous injection of formalized whole cells and human sera were obtained from patients and dental students. Indirect immunofluorescence method was used to study on the cross reaction between Porphyromonas endodontalis and Porphyromonas gingivalis serotype a, b, c antigen. Total membrane protein profiles of Porphyromonas endodontalis antigen were studied by sodium dodecyl sulfate polyacrylamide gel electrophoresis and the reactivity of antigenic components of Porphyromonas endodontalis against sera of patients and rabbit anti-Porphyromonas endodontalis antisera were assessed by Immunoblotting method. The following results were obtained : 1. Antigens of Porphyromonas endodontalis has multiple antigenic components, and both patients with periapical lesion and normal healthy individual showed immune response to this. 2. Patients group and healthy individual group showed a diversity of immune reaction pattern but they showed immune response against 43kd protein. 3. Patients with periapical lesion showed more diverse immune response than healthy individual and in some patients, much more bands appeared to lower molecular weight protein. 4. According to indirect immunofluorescence and Immunoblotting study, Porphyromonas endodontalis did not share common antigen with Porphyromonas gingivalis serotype a, b, c.

• Journal of Microbiology
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• 제42권2호
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• pp.117-125
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• 2004

Propionibacterium acnes (P. acnes) plays an important role in the disease pathogenesis of acne vulgaris, a disorder of pilosebaceous follicles, seen primarily in the adolescent age group. In the present study, the presence of antibodies against P. acnes (MTCC1951) were detected in acne patient (n=50) and disease free controls (n=25) using dot-ELISA and Western blot assay. The ability of P. acnes to induce pro-inflammatory cytokines by human peripheral blood mononuclear cells (PBMCs), obtained from acne patients and healthy subjects, were also analysed. The patients (n=26) who were culture positive for skin swab culture, were found to have a more advanced disease and higher antibody titres (1:4000 to >1:16000) compared to the P. acnes negative patients (n=24) and normal controls (n=25). An analysis of patients' sera by western blot assay recognized a number of antigenic components of P. acnes, rang-ing from 29 to 205 kDa. The major reactive component was an approximately 96 kDa polypeptide, which was recognised in 92％ (24 of 26) of the patients sera. Further, the P. acnes culture supernatant, crude cell lysate and heat killed P. acnes whole cells, obtained from 72-h incubation culture, were observed to be able to induce significant amounts of IL-8 and tumor necrosis factor alpha (TNF-${\alpha}$) by the PBMCs in both the healthy subjects and patients, as analysed by cytokine-ELISA. The levels of cytokines were significantly higher in the patients than the healthy subjects. A major 96 kDa polypep-tide reactant was eluted from the gel and was found to cause dose dependent stimulation of the pro-ductions of IL-8 and TNF-${\alpha}$. Thus, the above results suggest that both humoral and pro-inflammatory responses play major roles in the pathogenesis of acne.

• Parasites, Hosts and Diseases
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• 제50권3호
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• pp.191-197
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• 2012

Seroepidemiological changes of Toxoplasma gondii infection among the residents of the islands of Gangwha-gun, Incheon for 2 years were surveyed and evaluated by ELISA using a crude extract antigen. In 2010, sera of 919 adult residents in Gyodong-myeon and 313 adults in Samsan-myeon were collected and checked for IgG antibody titers, which showed 14.5% (133 sera) and 19.8% (62 sera) positive rates, respectively. In 2011, sera of 955 adults in Gyodong-myeon and 341 adults in Samsan-myeon were examined, which showed an increase of positive rates to 23.8% (227 sera) and 31.7% (108 sera), respectively. Totally, the seroprevalence of the first year was 15.8% and it increased rapidly to 25.8% in the second year. The positive rates of both sexes increased simultaneously with the significant ratio of males to females by 1.7-2.2 fold (P<0.05). In both myeons, 661 sera were collected every year and showed changes in optical density (OD) in 177 sera; newly found as positives in 73 persons (11.0%), negative conversion in 10 persons (1.5%), and maintained or increased in 94 persons (14.2%). This rapid increase in the prevalence of toxoplasmosis in Gangwha islands may be due to in part peculiar changes in the toxoplasmic environment of the islands and presumably the consumption of the pork bred domestically within the islands or imported from high endemic nations. It is necessary to find out symptomatic toxoplasmic patients and confirm the risk factors for further infection in the islands of Gangwha-gun.

• 대한의생명과학회지
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• 제16권4호
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• pp.311-316
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• 2010

We studied a comparison of the concentration of biochemical markers in sera of patients hospitalized with high fever (n=296) in Jeonbuk province during the last 2 years (2008 to 2009). The patients were divided into three patient groups of viral hemorrhagic fever (VHF) patient group tested positive for Hantavirus (n=53), leptospirosis (LEP) patient group tested positive for Leptospira interrogans (n=137) and scrub typhus (TSU) patient group tested positive for Orientia tsutsugamushi (n=106). We analyzed the concentration of ALP, AST, ALT, blood urea nitrogen, creatinine and glucose and compared the mean levels of them to normal range, the first sample and last sample. The frequencies of abnormal patient elevated above the upper limit of normal for ALP, AST and ALT were 18~43.4%, 78~97% and 62.3~92.7% in patient groups, and 24.5~47.4% (total protein) and 13.2~50.0% (albumin) of patients in patient groups had decreased below the lower limit of normal. The patients showed higher abnormal levels of glucose in patient groups were 58.5% (viral hemorrhagic fever patient group), 66.4% (leptospirosis patient group), 71.7% (scrub typhus patient group) and 66.9% (total patient group). There were significant difference between the first sample and the last sample in the mean levels of AST (decreased 22.2% in viral hemorrhagic fever patient group, 30.2% in leptospirosis patient group, 20.4% in scrub typhus patient group and 24.1% in total patient group), BUN (43.0% in viral hemorrhagic fever patient group, 41.6% in leptospirosis patient group, 47.4% in scrub typhus patient group and 43.0% in total patient group) and glucose (20.2% viral hemorrhagic fever patient group, 17.9% in leptospirosis patient group, 18.6% in scrub typhus patient group and 18.9% in total patient group) in the first sample and the last sample. According to these results, those diseases may cause liver damage and have high concentration of ALP, AST, ALT and glucose in blood even though the patients get out of the hospital.

• Asian Pacific Journal of Cancer Prevention
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• 제16권8호
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• pp.3131-3135
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• 2015

Objective: To investigate whether the expression of serum soluble neural cell adhesion molecule (sNCAM) is associated with hepatic encephalopathy (HE) in hepatocelular carcinoma (HCC) patients. Materials and Methods: The Oncomine Cancer Microarray database was used to determine the clinical relevance of NCAM expression in different kinds of human cancers. Sera from 75 HCC cases enrolled in this study were assessed for expression of sNCAM by enzyme linked immunosorbent assay (ELISA). Results: Dependent on the Oncomine Cancer Microarray database analysis, NCAM was down regulated in 10 different kinds of cancer, like bladder cancer, brain and central nervous system cancer, while up-regulated in lung cancer, uterine corpus leiomyoma and sarcoma, compared to normal groups. Puzzlingly, NCAM expression demonstrated no significant difference between normal and HCC groups. However, we found by quantitative ELISA that the level of sNCAM in sera from HCC patients with HE ($347.4{\pm}151.9ng/ml$) was significantly more up-regulated than that in HCC patients without HE ($260.3{\pm}104.2ng/ml$), the p-value being 0.008. sNCAM may be an important risk factor of HE in HCC patients, the correlation coefficients was 0.278 (P<0.05) on rank correlation analysis. Conclusions: This study highlights that up-regulated level of serum sNCAM is associated with HE in HCC patients and suggests that the high expression can be used as an indicator.

• 대한의생명과학회지
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• 제13권4호
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• pp.287-291
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• 2007

The factors and mechanisms by infection of Oriental Tsutsugamushi caused disease are not well understood. The onset of tsutsugamushi disease is characterized by chilliness, fever, malaise, headache and generalized aching. Infection of tsutsugamushi is the cause of impairment of function of a major organ often complicate the picture and immediately change the prognosis for the worse. Tsutsugamushi disease is reported that this disease is characterized by the histopathogenesis of liver, kidney, heart, and lung, but the variation of biochemical components in serum of tsutsugamushi disease patient are not clear. We analyzed total protein (TP), albumin (AL), aspartic aminotranferase (AST), alanine aminotransferase (ALT), alkaline phosphotase (ALP), urea nitrogen (UN), creatinine (CRE), glucose (GLD), cholesterol (CHOL) and total bilirubin (TB) in sera of patients with tsutsugamushi disease. In comparison with reference, total protein and albumin were abnormally decreased in 19.6% and 39.2% of patients, respectively. AST, ALT, ALP, creatinine, UN, glucose, cholesterol and total bilirubin were abnormally increased in 94.1 %, 72.5%, 25.5%, 15.7%, 9.8%, 62.7%, 25.5% and 6.0% of patients, respectively. The patients showed abnormal relative rate of protein electrophoretic fractions to total protein in serum compared to them of reference were 43.1% (albumin), 12.9% ($\alpha_1$-globulin), 58.8% ($\alpha_2$-globulin), 60.8% ($\beta$-globulin) and 70.6% ($\gamma$-globulin), respectively. These data suggest that infection of Oriental Tsutsugamushi causes impairment of function of a major organ and abnormal serum protein electrophoresis fractions to tsutsugamushi patients.

• Molecular & Cellular Toxicology
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• 제2권1호
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• pp.67-73
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• 2006

In previous studies, it has been discovered that cancer cells not only overexpress regulatory subunit I (Rl)/protein kinase type I (PKA-I) but also secrete outside the cell an extracellular form of PKA (ECPKA) and that the ECPKA secretion detected in patients' serum is obviously greater than that found in non-cancer patients or healthy subjects. We now found that ECPKA elicits the formation of serum autoantibodies that can serve as a cancer diagnostic and prognostic marker. To measure the presence of anti-ECPKA autoantibody in the human sera, basic methodology for ECPKA assay was established an enzyme-linked immunosorbent assay (ELISA). We obtained serum samples from 199 patients with different types of cancer, and also obtained 31 serum samples to compare with ECPKA concentrations from non-cancer patients and 119 normal volunteers. Compared with normal or non-cancer patient sera, we found that the frequency of anti-ECPKA autoantibody was significantly higher in cancer patients (88%) than in those without cancer (17%). Furthermore the presence of anti-ECPKA autoantibodies in the serum of cancer patients was highly correlated with the site of metastasis. The immunoassay developed for anti-ECPKA antibodies is highly sensitive and specific. Therefore, this discovery of an autoantibody-based cancer diagnostic may have serious clinical application and may become an important advance over current technology.

• Parasites, Hosts and Diseases
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• 제26권3호
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• pp.163-168
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• 1988

스파르가눔증을 혈청학적으로 진단하기 위하여 효소면역측정법을 실시할 경우, 그 민감도와 특이도가 높다고 이미 보고되었다. 실제로 조직기생충 감염이 의심되는 환자에 대하여 특이 IgG항체가를 일상적으로 검사하면 중추신경계 환자중에서 뇌스파르가눔증은 드물지 않게 발견할 수 있다. 그러나 혈청학적 진단에 이용되는 스파르가눔 조항원(조항원)은 조충증(조충증), 유구낭미충증이나 포충증 환자의 혈청과 비특이적인 교차반응을 일으키는 경우가 있다. 이러한 교차반응의 문제를 해결하려면 우선 스파르가눔 항원의 구성 단백질이 환차혈청과 반응하는 양상을 알아야 할 필요가 있다. 이 연구에서는 외과적으로 스파르가눔증을 확인한 환자 15명에서 얻은 혈청이 스파가눔의 생리식염수 추출액의 항원단백질중 어느 분획과 반응하는지 검토하였다. 그리고 뇌유구낭미충증 환자 24명(그중 8명은 효소면역측정법으로 교차반응이 있었음)의 혈청과 교차반응을 일으키는 단백질 분획을 관찰하였다. 스파르가눔충체의 생리식염수 추출액을 10∼15% linear gradient gel에서 SDS-폴리아크릴아마이드 전기영동을 실시하여 단백질대 30개를 구별할 수 있었다. 분리한 단백질(대)를 nitrocellulose 종이에 옮긴후 스파르가눔증 환자 혈청 및 conjugate와 차례로 반응시키고 발색반응을 일으킨 결과 (효소면역 전기영동이적법, immunoblot), 스파르가눔 항원단백질중 29kDa 및 36kDa가 가장 많이 또 가장 진하게 반응한 것이어서 민감하고 항원성이 높은 단백질 분획이라고 판단하였다. 그리고 위의 단백질 이외에도 158kDa, 130kDa, 107kDa, 78kDa, 72kDa, 52kDa, 23kDa, 21kDa, 15kDa 및 6kDa 등에도 반응이 있었다. 유구낭충증 환자 24명의 혈청중 스파르가눔 항원에 혈청학적 교차반응이 있었던 혈청은 36kDa 및 29kDa 단백질에 반응하고 있어 이 단백질이 스파르가눔의 종 특이(종특이)단백질 항원은 아닌 것으로 판단되나 그 성질에 대해서는 앞으로 더 추구할 가치가 있다고 생각한다.