• Korean Journal of Veterinary Research
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• v.41 no.4
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• pp.535-542
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• 2001

A multiplex PCR technique was developed for detecting specifically each Mycobacterium bovis, M. tuberculosis, M. avium and M. avium subsp, paratuberculosis, respectively, using clinical samples of field cattle. To apply this novel technique to clinical specimens, blood sample was obtained from live cows comprising 11 intradermal tuberculin test (ITT)-positive and 17 ITT-negative and tested by multiplex PCR. Positive results were obtained from 15 cows by the multiplex PCR, showing that 4 (23.5%) of the 17 ITT-negative cows were multiplex PCR positive. The multiplex PCR results also showed that among the 15 positive cows, 7 (46.7%) were infected with M. bovis, 1 (6.7%) with M. tuberculosis and 7 (46.7%) with M. avium. The sensitivity and specificity of multiplex PCR in comparison with those of ITT were 100% and 76.5%. The correlation between the multiplex PCR and ITT assays with blood samples was considered excellent, 85.7% agreement and ${\kappa}=0.72$. The results obtained, using reference mycobacterial strains and typed clinical samples, show that the multiplex PCR method may be a rapid, sensitive, and specific tool for the differential identification of various mycobacterial strains in a single-step assay. Therefore, multiplex PCR assay is a useful tool for early diagnosis of tuberculosis in live cattle and to identify the species or complex of mycobacterium from clinical samples.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.16
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• pp.6955-6960
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• 2014

Background: The cytokinesis-block micronucleus (CBMN) assay is a standard cytogenetic tool employed to evaluate chromosomal damage subsequent to pesticide exposure. Objectives: To evaluate the pooled levels of total micronuclei (MN) and binucleated cells with micronuclei (MNC) in 1000 binucleated lymphocytes among population occupationally exposed to pesticides and further determine the more sensitive biomarker of CBMN. Materials and Methods: A meta-analysis on the pooled levels of MN and MNC in binucleated lymphocytes among occupationally pesticide-exposed populations was conducted using STATA 10.0 software and Review Manager 5.0.24 in this study. Results: We found significant differences in frequencies of MN and MNC in 1000 binucleated lymphocytes between pesticide-exposed groups and controls, and the summary estimates of weighted mean difference were 6.82 [95% confidence interval (95% CI): 4.86-8.78] and 5.08 (95% CI: 2.93-7.23), respectively. However, when we conducted sensitivity analyses further, only the MN remained statistically different, but not the MNC, the summary estimates of weight mean difference were 2.86 (95% CI: 2.51-3.21) and 0.50 (95% CI: -0.16-1.17), respectively. We also observed pesticide-exposed subjects had significantly higher MN frequencies than controls among smokers and nonsmokers, male and female populations, and American, Asian and European countries in stratified analyses. Conclusions: The frequency of MN in peripheral blood lymphocytes might be a more sensitive indicator of early genetic effects than MNC using the CBMN assay for occupationally pesticide-exposed populations.

• Journal of Periodontal and Implant Science
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• v.25 no.2
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• pp.386-396
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• 1995

Interieukin 1${\beta}$ is a potent bone resorptive cytokine which mediates soft tissue destruction through the stimulatidn of prostaglandin production and the induction of collagenase. This constellation of activities suggests a role of IL-1${\beta}$ in the pathogenesis of periodontal disease. The purpose of this study was to evaluate the effects of herbal extracts on production and activity of IL-1${\beta}$. When LPS was added to cultured human blood monocytes, the effects of herbal extracts on the production of IL-1${\beta}$ was evaluate by thymocyte stimulation assay. When rHuIL-1${\beta}$ was added to cultured human gingival fibroblasts, the effects of herbal extracts on production of $PGE_2$ was evaluated by ELISA and when it was added to cultured mouse calvaria, the effects on bone resorption was estimated by .$^{45}Ca$-release bone resorption assay. The herbal extracts that had been used in this study were as follows; Asparagi Radix, Schzandrae Fractus, Zizyphi Fractus and Rhois Galla. The following results were obtained from this study. 1. All these extracts effectively inhibited the production of IL-1${\beta}$ on cultured human blood monocytes. 2. All these extracts effectively inibited the production of $PGE_2$ on cultured human gingival fibroblasts. 3. All these extracts did not effectively inhibit the bone resorption induced by rHulL-1${\beta}$ on cultured mouse calvaria.

• Journal of Periodontal and Implant Science
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• v.40 no.6
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• pp.265-270
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• 2010

Purpose: The aim of this study was to investigate the immunomodulatory effects of canine periodontal ligament stem cells on allogenic and xenogenic immune cells in vitro. Methods: Mixed cell cultures consisting of canine stem cells (periodontal ligament stem cells and bone marrow stem cells) and allogenic canine/xenogenic human peripheral blood mononuclear cells (PBMCs) were established following the addition of phytohemagglutinin. The proliferation of PBMCs was evaluated using the MTS assay. The cell division of PBMCs was analyzed using the CFSE assay. The apoptosis of PBMCs was assessed using the trypan blue uptake method. Results: Periodontal ligament stem cells and bone marrow stem cells inhibited the proliferation of allogenic and xenogenic PBMCs. Both periodontal ligament stem cells and bone marrow stem cells suppressed the cell division of PBMCs despite the existence of a mitogen. No significant differences in the percentages of apoptotic PBMCs were found among the groups. Conclusions: Canine periodontal ligament stem cells have an immunomodulatory effect on allogenic and xenogenic PBMCs. This effect is not a product of apoptosis of PBMCs but is caused by the inhibition of cell division of PBMCs.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.10
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• pp.4329-4333
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• 2015

Background: To investigate the change of frequency and immuno-inhibitory function of myeloid-derived suppressor cells (MDSCs) after treatment of cisplatin (DDP) in A375 human melanoma model. Materials and Methods: BALB/c nude mice were inoculated with A375 cells to establish the human melanoma model and randomly divided into control group given normal saline (NS) and experimental group treated with DDP (5 mg/kg). The percentages of MDSCs in the tumor tissue and peripheral blood after DDP treatment were detected by flow cytometry. The proliferation and interferon-${\gamma}$ (IFN-${\gamma}$) secretion of T cells co-cultured with MDSCs were analyzed through carboxyfluorescein succinimidyl ester (CFSE) labeling assay and enzyme-linked immunospot (ELISPOT) assay, respectively. Results: In A375 human melanoma model, DDP treatment could significantly decrease the percentage of MDSCs in the tumor tissue, but exerted no effect on the level of MDSCs in peripheral blood. Moreover, DDP treatment could attenuate the immuno-inhibitory function of MDSCs. T cells co-cultured with DDP-treated MDSCs could dramatically elevate the proliferation and production of INF-${\gamma}$. Conclusions: DDP can decrease the frequency and attenuate immuno-inhibitory function of MDSCs in A375 melanoma model, suggesting a potential strategy to augment the efficacy of combined immunotherapy.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.9
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• pp.5507-5512
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• 2013

Objective: To investigate the correlation between ERCC1 expression levels in tumor tissue and peripheral blood lymphocytes (PBL) from patients with gastric cancer and assess the relationship between PBL DNA repair rate (DRR) and the efficacy of platinum chemotherapy. Methods: A total of 53 patients with gastric cancer receiving surgery and 20 controls were studied. ERCC1 protein expression in tumour tissue and PBL were determined by immunohistochemical staining. The PBL DRRs of 47 advanced patients and 20 controls were estimated by comet assay. Results: The positive expression rates of ERCC1 were 67. 9%, 56. 6% and 10.0% in tumour tissues, PBLs of gastric cancer patients, and PBLs of the control group. PBL ERCC1 expression correlated with that in tissue (${\chi}^2$=15. 463, p=0.000). Pearson contingency coefficient=0.475). DRRs of cancer patients by tail length (TL) (Z=4. 662, p=0.000) and tail moment (TM) (Z=3. 827, p=0.000) were significantly lower than that of control group. When TL was applied as an indicator, the correlation between DRR and chemotherapy efficacy was significant (Spearman rank correlation r=0.327, p=0.032). Patients with low levels of DRR in PBL presented better short-term efficacy of chemotherapy than those with high levels of DRR. Conclusions: The ERCC1 expression in PBLs may indirectly reflect ERCC1 expression in gastric cancer tissues. Compared with non-cancer populations, patients with gastric cancer may have lower DNA repair capacity. DRR in PBL may predict the short-term efficacy of platinum-based chemotherapy for patients with advanced gastric cancer.

• The Korean Journal of Food And Nutrition
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• v.25 no.2
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• pp.246-252
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• 2012

After a mixed carbohydrate diet, inhibition of ${\alpha}$-amylase and ${\alpha}$-glucosidase involved in the digestion and absorption of carbohydrates can significantly decrease the postprandial increase of blood glucose level. In the course of screening these useful enzyme inhibitors, we selected five kinds of bean, using an in-vitro enzyme inhibition assay method. To evaluate the effect of germination process on the functionality of the bean, we investigated the inhibitory activities of the water extracts of non-germinated bean and germinated bean against ${\alpha}$-amylase and ${\alpha}$-glucosidase, relevant to postprandial hyperglycemia. We also investigated the oxygen radical absorbance capacity(ORAC), total phenolics content, and postprandial blood glucose lowering effect in rats(Sprague-Dawley rat model). Most germinated beans showed significantly higher ${\alpha}$-glucosidase inhibitory activity, compared with non-germinated beans. Among germinated beans, Glycine max had the highest ${\alpha}$-glucosidase inhibitory activity(53.3%). The water extract of germinated Phaseolus vulgaris L. had the highest ${\alpha}$-amylase inhibitory activity(95.1%), followed by Glycine max(58.7%), and Glycine max L. Merr(54.1%). Furthermore, the five germinated beans also showed high antioxidant activities in ORAC assay. Results suggested that the germination process may improve and enhance the anti-hyperglycemia potential and antioxidant activity of the bean.

• Archives of Pharmacal Research
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• v.16 no.4
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• pp.336-338
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• 1993

The immunomodulating activity of Kp, an antitumor protein-polysacchanide preparation from the shake-cultured mycelia of Phellinus linteus, was investigated in ICR mice subcutaneously implanted wit $1\times10^6$ cells of sarcoma 180. The mice were intraperitoneally administered with Kp at a does of 100 mg/kg once daily for five consecutive days starting from 24 hrs after the tumor implantation. Ten days after the last injection, the mice were immunized with $1\times10^7$ or $4\times10^8$ sheep red blood cells (SRBC) and five days later, the antibody-forming immune response were assessed by direct hemolytic plaque assay. To an immunization does of $1\times10^7$ SRBC, the Kp-treated mice elicied a successful humoral immune response despite the turmor-burden and produced $259\times10^3$ plaque-forming cells (PFC)/spleen, while the corresponding tumor-bearing control mice showed virtually no reponse $(2.0\times10^3$ PFC/spleen) (the stimulation index=129.5). However, to an immunization dose of $4\times10^8$ SRBC, both of the control mice and Kp-treated mice showed almost the same level of strong humoral immune response. From these data it is clear that Kp effectively restores the humoral immune response of the turmor-bearing ICR mice.

• Journal of Chest Surgery
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• v.38 no.12 s.257
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• pp.807-814
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• 2005

Background: Cardiopulmonary bypass is an essential process to maintain circulation for saving life during the cardiac surgery, But it is a process in which systemic inflammation was evoked inevitably because of the exposure of blood to foreign surface. The injuries to distal organs during the cardiopulmonary bypass were resulted from systemic inflammation and the disturbances of micro-circulations in the organs. We designed this study to research the effects of leukocyte depletion from pump-oxygenator priming solution on the systemic inflammation, and the micro-circulation of gastric mucosa that is suggested by the gastric mucosal $CO_{2}$ partial pressure and acidity. Material and Method: The dogs were divided into three groups according to the different pump-oxygenator priming solutions; non-hemic crystalloid solution; leukocyte-depleted homologous blood; and non leukocyte-depleted homo-logous blood. Each priming solution group contained five dogs. In all three groups, 2 hours of cardiopulmonary bypass, and 4 consecutive hours of general anesthesia was maintained on the mechanical ventilation. Each dog was evaluated for the gastric mucosal pH, $CO_{2}$ partial Pressure, arterial pH, $CO_{2}$ partial pressure, the exhaled air $CO_{2}$ partial pressure and the level of IL-8 on before the cardiopulmonary bypass, 1 hour after the cardiopulmonary bypass, 2 hours after the cardiopulmonary bypass, 2 hours after the restoration of normal circulation, and 4 hours after the restoration of normal circulation after the cardiopulmonary bypass. The levels of IL-8 were measured with ELISA (enzyme linked immunosorbent assay) technique. Result: 1. There were significant differences of gastric mucosal $CO_{2}$ partial pressure between the leukocyte-depleted homologous blood group and other two groups(vs non leukocyte-depleted homologous blood group; P=0.02, vs non-hemic crystalloid solution group; P=0,01). 2. The gastric mucosal pH of leukocyte-depleted homologous blood group was significantly different from non leukocyte-depleted homologous blood group (p=0.01). 3. The levels of IL-8, which examine the systemic inflammation, showed signi- ficantly better results in leukocyte-depleted homologous blood group and non-hemic crystalloid solution group than non leukocyte-depleted homologous blood group (p=0.01, 0.01). Conclusion: Based upon these results, we concluded that the leukocyte depletion from the pump-oxygenator priming solution has a beneficial effects in reducing systemic inflammation and the preserving of gastric mucosal micro-circulation.

• Parasites, Hosts and Diseases
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• v.47 no.3
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• pp.227-233
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• 2009

Local malaria transmission in the United Arab Emirates (UAE) came to an end in 1997. Nevertheless, UAE has been subjected to substantial importation of malaria cases from abroad, concerning both UAE nationals and immigrants from malarious countries with a total number of 2,119 cases in 2007. To evaluate a new DNA extraction technique using nested PCR, blood samples were collected from 132 individuals who presented to Infectious Diseases Department in Rashid Hospital, Dubai, and Central Department of Malaria Control with fever and persistent headache. Giemsa-stained blood films and ELISA test for malaria antibodies were carried out for detection of Plasmodium infection. Plasmodium infections were identified with the genus-specific primer set and species differentiation using nested PCR. A rapid procedure for diagnosis of malaria infections directly from dried blood spots using for the first time DNA extract from FTA Elute cards was evaluated in contrast to extraction techniques using FTA classic cards and rapid boiling technique. Our new simple technique for DNA extraction using FTA Elute cards was very sensitive giving a sensitivity of 100% compared to 94% using FTA classic cards and 62% in the rapid boiling technique. No complex preparation of blood samples was required prior to the amplification. The production cost of DNA isolation in our PCR assay was much less incomparable to that of other DNA extraction protocols. The nested PCR detected plasmodial infection and could differentiate P. falciparum from P. vivax, and also detected the mixed infection.