• Title/Summary/Keyword: Blood Assay

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Multiple-biometric Attributes of Biomarkers and Bioindicators for Evaluations of Aquatic Environment in an Urban Stream Ecosystem and the Multimetric Eco-Model (도심하천 생태계의 수환경 평가를 위한 생지표 바이오마커 및 바이오인디케이터 메트릭 속성 및 다변수 생태 모형)

  • Kang, Han-Il;Kang, Nami;An, Kwang-Guk
    • Journal of Environmental Impact Assessment
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    • v.22 no.6
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    • pp.591-607
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    • 2013
  • The objectives of the study were to evaluate the aquatic environment of an urban stream using various ecological parameters of biological biomarkers, physical habitat quality and chemical water quality and to develop a "Multimetric Eco-Model" ($M_m$-E Model) for the ecosystem evaluations. For the applications of the $M_m$-E model, three zones including the control zone ($C_Z$) of headwaters, transition zone ($T_Z$) of mid-stream and the impacted zone ($I_Z$) of downstream were designated and analyzed the seasonal variations of the model values. The biomarkers of DNA, based on the comet assay approach of single-cell gel electrophoresis (SCGE), were analyzed using the blood samples of Zacco platypus as a target species, and the parameters were used tail moment, tail DNA(%) and tail length (${\mu}m$) in the bioassay. The damages of DNA were evident in the impacted zone, but not in the control zone. The condition factor ($C_F$) as key indicators of the population evaluation indicator was analyzed along with the weight-length relation and individual abnormality. The four metrics of Qualitative Habitat Evaluation Index (QHEI) were added for the evaluations of physical habitat. In addition, the parameters of chemical water quality were used as eutrophic indicators of nitrogen (N) and phosphorus (P), chemical oxygen demand (COD) and conductivity. Overall, our results suggested that attributes of biomarkers and bioindicators in the impacted zone ($I_Z$) had sensitive response largely to the chemical stress (eutrophic indicators) and also partially to physical habitat quality, compared to the those in the control zone.

Hematological Change of Korean Native Goats Experimentally Infected with Bovine Leukemia Virus (Bovine Leukemia Virus에 실험감염된 한국재래산양의 혈액상의 변동)

  • 이필돈;김종호;전무형
    • Korean Journal of Veterinary Service
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    • v.18 no.1
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    • pp.1-21
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    • 1995
  • To elucidate pathogenesis of bovine leukemia virus(BLV) in Korean native goats, the goats experimentally infected with BLV were studied especially for the aspects of infectivity and hematological changes. The experimental goats were examined for 27 months by agar-gel immunodiffusion(AGID) test and syncytium formation assay. During this period, changes of total leucocyte, absolute Iymphocyte and atypical Iymphocyte were examined, and the distribution of surface immunoglobulin ( sIg ) -bearing cells and rosette forming cell (RFC) in the peripheral Iymphocyte were also investigated. By indirect immunofluorescence (IFA) and complement dependent antibody cytotoxicity (CDAC) assay using monoclonal antibody(Mab) against bovine leukosis tumor-associated anti-gen(BL-TAA), changes of BL-TAA positive Iymphocyte in peripheral blood were measured. The results obtained through the experiment were summarized as follows. 1. Antibody titers were measured by AGID using gP51 and P24 antigens. The animals were serologically converted at 2 months post-inoculation(pi) in gP51 antigen, whereas sero-converted at 4 months pi in P24 antigen. In comparison with antibody titers for gP51, P24 antigen showed lower titers throughout the trial period. 2. The peripheral lymphocytes from all of the infected goats, as co-cultivated with F8l cells manifested syncytial formation at 4 months pi. 3. On counting total leucocyte, Iymphocyte and atypical Iymphocyte, two out of four infected goats showed normal distribution, while No 2 of the remaining two revealed temporal and No 3, Persistant increasing number of the cells. 4. The optimal condition of rosette formation of the peripheral Iymphocyte of normal Korean native goats was shown in the sheep erythrocyte treated with 0.1M AET for 30 nun at $37^{\circ}C$. When the Iymphocytes were treated in nylon wool column, the number of sIg-bear-ing cell were increased in the nylon wool adherent cells, but RFC was increased in the non-adherent cells. Of the infected goats, No 2 and No 3 showed significantly increasing number of sIg-bearing cells at 18 months pi. 5. The Iymphocytes of No 2 and No 3 goats reacted positively in IFA using Mab against BL-TAA at 12 months pi and 18 months pi, respectively. In CDAC test, all of four infected goats revealed positive reaction at 24 months pi. The higher positive rates were observed in No 2 and No 3 as compared with the remainders.

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Diallyl Sulfides (DAS) and Diallyl Disulfides (DADS) Exhibit a Suppressive Effect on the Proliferation and Migration of Vascular Smooth Muscle

  • Kim, Min-Ju;Kwak, Jung-Hyun;Baek, Seung-Han;Yeo, Hyun-Yang;Song, Ju-Hyun;Cho, Bong-Jun;Kim, Oh-Yoen
    • Preventive Nutrition and Food Science
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    • v.15 no.2
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    • pp.137-142
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    • 2010
  • Previous studies report that organo-sulfur compounds derived from garlic inhibited smooth muscle cell (SMC) proliferation and induced apoptosis of cancer cells. Recently, lipid-soluble compounds such as diallyl sulfides (DAS) and diallyl disulfides (DADS) have been reported to more effectively suppress tumor cell proliferation. However, there were few studies on the suppressive effects of lipid-soluble garlic sulfur compounds on the proliferation and migration of vascular smooth muscle cells (VSMC). Therefore, this study investigated the effect of DAS and DADS on VSMC proliferation/migration induced by oleic acid (OA), a principal fatty acid in circulating triglyceride of blood stream. Assays performed include a tetrazole (MTT) assay, a wound healing assay and a Western blots. VSMC proliferations were enhanced by OA in a dose-dependent manner at concentrations of $10{\sim}50\;{\mu}M$ and inhibited by DAS and DADS compared to non-treated control. OA-induced proliferations were also attenuated by DAS and DADS. OA-induced cell migrations were 2.5 times higher than non-treated control, and they were significantly attenuated by DAS (32% at $150\;{\mu}M$ and 50% at $200\;{\mu}M$) and DADS (40% at $150\;{\mu}M$ and 46% at $200\;{\mu}M$). OA-induced cell migration was also attenuated by PD98059 (ERK inhibitor), SB203580 (P38 inhibitor) and particularly by LY204002 (PI3K inhibitor) and SP600125 (JNK2 inhibitor). Additionally, Western blot assays showed that OA-induced JNK1/2-phosphorylation was down-regulated after treatment with DAS and DADS. In conclusion, the findings of our study support the idea that DAS and DADS may have a suppressive effect on the proliferation and migration of OA-induced VSMC and that this effect may be partly associated with PI3K and JNK2 pathways.

Cytokine Expressions with Lectins from Allomyrina dichotoma and Bombyx mori (장수풍뎅이(Allomyrina dichotoma)와 누에(Bombyx mori) 렉틴의 사이토카인 생성 양상 비교)

  • Kim, Se-Jin;Chung, See-Ryun;Jeune, Kyung-Hee
    • Korean Journal of Pharmacognosy
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    • v.36 no.2 s.141
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    • pp.129-135
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    • 2005
  • Lectins from Allomyrina dichotoma (ADL) and Bombyx mori (BML) were partially purified by physiological saline extraction, ammonium sulfate fractionation, anion exchange column chromatography on DEAE Sephadex A-50 and gel filtration column chromatography on Sephadex G-200. An assay for cytokine expression was carried out by using reverse transcription polymerase chain reaction(RT-PCR). mRNA isolated from PBMC(human peripheral blood mononuclear cells) were stimulated with ADL(O.D.=0.2) and BML(O.D.=0.1) for various times(1,4,8,24,48 and 72 h) and various cytokine mRNA assessed by RT-PCR were shown as follows: The patterns of bands for IL-1 mRNA of BML were very similar with those from ADL and these bands were decreased along the increasing reaction times after showing a strong band at 1 h. However mRNA expressions for IL-2, IL-6, $IFN{\gamma}$ and $TNF{\alpha}$ showed different patterns between ADL and BML. With the effect of ADL, the expression of IL-2 and IL-6 mRNA were continuously detected until 72 h with the strongest band of IL-2 mRNA at 24 h. The strong bands of $IFN{\gamma}$ mRNA were observed from 4 to 8 h but the strongest one of $TNF{\alpha}$ was just observed at 1 h. Meanwhile with BML, the bands for IL-2 and $IFN{\gamma}$ were increased along the increasing reaction times until 72 h. The strongest bands were showed from 4 to 8 h with IL-6 and at 8 h with $TNF[\alpha}$. To verify quantitatively ELISA was used for assay of protein secretions of the cytokine gene with IL-2 and $IFN{\gamma}$ expressed markedly different in RT-PCR. The highest cytokine secretion for IL-2 was demonstrated at 48 h. The production of $IFN{\gamma}$ was markedly increased at 24 h and secreted highest at 72 h. These result suggest that ADL and BML, as inducers of cytokines, can elicit detectable cytokine mRNA from PBMC within the first few hours of stimulation and maintain the production of cytokines for a few days by the methods of RT-PCR and ELISA.

The Effects of Melatonin and Ge-132 on Acute Hematopoietic Syndrome following Radiation Exposure (방사선피폭 후 급성조혈계증후군에 대한 Melatonin과 Ge-132의 효과)

  • Jang, Seong-Soon
    • Journal of Radiation Protection and Research
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    • v.29 no.4
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    • pp.237-242
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    • 2004
  • The radioprotective effects of Melatonin and Ge-132 on acute hematopoietic injury was investigated in mice exposed to an acute whole-body radiation dose of 8 Gy. Melatonin was administered intraperitoneally 1 hour before irradiation at a dose of 200 mg/kg, and Ge-132 was administered orally from days 5 to 20 after irradiation at a dose 130 - 150 mg/kg/d. The radioprotective effects were evaluated for spleen using TUNEL assay, and in peripheral blood by counting lymphocyte & WBC. The 4 experimental groups (irradiation-only, melatonin pretreatment, Ge-132 posttreatment, and melatonin pretreatment plus Ge-132 posttreatment) were observed for survival analysis up to 30 days following irradiation. The apoptotic index (47.8% vs 45.9%, p=0.385), and the number of lymphocytes ($97/{\mu}{\ell}\;vs\;101/{\mu}{\ell}$, p=0.898) were not significantly different between the irradiation-only and the melatonin pretreatment group, But the number of WBCs ($147/{\mu}{\ell}\;vs\;306/{\mu}{\ell}$, p=0.010) was higher in the melatonin pretreatment group. The irradiation-only, melatonin, Ge-132, and melatonin plus Ge-132 treatments resulted in survival rate at 30 days of 21.4%, 100%, 35.7%, and 85.7%, respectively. The melatonin pretreatment group in survival analysis between groups was showed significantly higher survival than the irradiation-only(p=0.000), or Ge-132 posttreatment group(p=0.0003). These results indicate that the melatonin may have a potential as an effective radioprotector on acute hematopoietic syndrome following radiation exposure.

Effect of different drying methods on anti-oxidation and anti-thrombosis activities of Salicornia europaea (함초의 건조방법에 따른 항산화 및 항혈전 활성의 변화)

  • Sung, Hwa-Jung;Kim, Deuk Hoi;Sohn, Ho-Yong
    • Food Science and Preservation
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    • v.24 no.5
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    • pp.658-665
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    • 2017
  • The current study was performed to investigate the changes of bio-activities of Salicornia europaea (SE) depending on the different dry methods. The ethanol extracts were prepared from the freeze-drying SE (FD), hot-air drying SE (HD), and shade drying SE (SD). Their anti-oxidant and anti-thrombosis activities were compared. The yields of ethanol extraction in FD, HD, and SD were 14.4, 13.2 and 11.9%, respectively. The highest contents of total polyphenol (4.6 mg/g) and total sugar (23.4 mg/g) were shown in FD, whereas, the highest content of reducing sugar (14.6 mg/g) was observed in HD. In anti-oxidation activity assay, the FD and HD showed similar radical scavenging activities and reducing power. However, in SD, nitrite scavenging activity and reducing power were severely decreased. In anti-coagulation activity assay, the thrombin time (TT), prothrombin time (PT) and activated partial thromboplastin time (aPTT) of FD, HD and SD did not show significant changes at 5 mg/mL concentration. But the HD at 7 mg/mL had strong inhibitions against thrombin and blood coagulation factors. The platelet aggregation and hemolysis activities were not affected by dry methods. Our results suggest that both FD and HD are effective to maintain the functional ingredient of SE, and HD is economic and efficient dry process for production of functional food.

Experimental infection of piglets with a field isolate of Aujeszky's disease virus in Korea: Pathogenecity, excretion, distribution and immunogenicity of virus (국내분리 Aujeszky's disease virus의 실험적 감염 자돈에 대한 바이러스학적 연구)

  • Park, Jeong-woo;Jun, Moo-hyung;An, Soo-hwan
    • Korean Journal of Veterinary Research
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    • v.30 no.2
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    • pp.177-186
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    • 1990
  • To investigate the etiology, pathogenicity and virological properties of NYJ-1-87 strain of Aujeszky's disease virus (ADV) that was isolated from the diseased piglet in Korea, the virus at $10^{6.0}TCID_{50}/0.1ml$ was inoculated intranasally and subcutaneously into 30 to 35 days-old piglets. Results obtained through the experiments were summarized as follows. 1. Ten of the infected piglets were clinically observed for 15 days. On the 2nd day post-inoculation(pi), the signs of pyrexia, anorexia and convulsion were noted. On the 4th to 7th days pi, nervous signs of incoordination and intermittent spasm were shown in the most of piglets, and one out of 5 piglets infected intranasally was died with severe nervous signs at the 7th day pi. The signs became relieved on the 8th day pi and all of remainder were completely recovered on the 13th to 14th days pi. 2. In hematological study, prominent decrease in the number of total leukocyte and lymphocyte was shown in the ADV-infected piglets on the 6th day pi. On the 8th day pi, the cell numbers were slightly increased and returned to normal level on the 10th day pi. 3. Viral excretion of the ADV-inoculated piglets was examined by swabbing of nasal and oral cavities, and rectal feces. During the periods of the 3rd to 11th days pi, the virus was excreted intermittently from nasal and oral cavities, and rectal feces. The nasal excretions were shown the highest virus concentration of $10^{5.2}TCID_{50}/0.1ml$ at the 5th day pi. 4. Recovery of the inoculated virus from various organs of the piglets that were died or experimentally slaughtered was attempted, and the virus was isolated from the tissues of brain and tonsil by the cultured cell-inoculation method. The highest recovery rate was noted in the tonsil. By indirect immunofluorescence antibody assay using ADV-monoclonal antibody, the viral antigens were detected in tissues of spleen and liver as well as brain and tonsil on the 7th to 9th days pi. The virus was not isolated from blood and the tissues of lung and kidney throughout the experiments. 5. Titers of virus neutralizing antibody in the piglets experimentally infected with ADV became increased after the 6th to 9th days pi in both of intranasal and subcutaneous inoculation showing the highest titers of 64 to 128 on the 29th day pi. When the antibody levels were measured by radial immunodiffusion enzyme assay, the reactive diameter was enlarged to be positive after the 4th to 6th days pi in both of intranasal and subcutaneous inoculation showing the largest diameter of 13 to 14mm on the 29th day pi.

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Effect of Glycyrrhizae Radix on the Immune Responses(II) - Immuno-regulatory Action of Glycyrrhizin and Glycyrrhetinic Acid - (감초가 면역반응에 미치는 영향(II) - Glycyrrhizin 및 Glycyrrhetinic acid의 면역조절작용 -)

  • 한종현;오찬호;은재순
    • YAKHAK HOEJI
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    • v.35 no.3
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    • pp.174-181
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    • 1991
  • These experiments were conducted to investigate the effects of glycyrrhizin(GL) and glycyrrhetinic acid(GA) on histamine synthesis, lymphocyte blastogenesis in C57BL/6J mice splenocytes, IL-1 production, $Ca^{2+}$ uptake by macrophage-like P388D$_{1}$ cells and plaque forming cell assay against SRBC. Histamine contents, lymphocyte blastogenesis, IL-1 activity, $Ca^{2+}$ uptake and plaque forming cell were determined by enzyme isotope method, [sup 3/H]-thymidine incorporation, C3H/HeJ mouse thymocytes proliferation, the addition of 5 $\mu$Ci/ml $^{45}$Ca$^{2+}$ to P388D$_{1}$, cell suspension and assay to sheep red blood cell, respectively. Cytotoxicity, which was expressed as 50% mortality, was occurred by the addition of GL(10$^{-3}$M) and GA(10$^{-4}$M). Histamine production in mouse spleen cell culture was significantly increased by the addition of 0.25 $\mu\textrm{g}$/ml of Con A, after 48 hour incubation. Con A dependent T-lymphocyte proliferation was also enhanced by the addition of 0.25 .mu.g/ml of Con A. The effects of GL on histamine contents and T-lymphocyte proliferation were significantly decreased at high dose (10$^{-5}$M), while IL-1 activity was remarkably suppressed by 10$^{-8}$~10$^{-4}$M of GL. $Ca^{2+}$ uptake was not changed, but antibody production was increased by GL(10 mg/kg). GA inhibited histamine contents at 10$^{-9}$~10$^{-7}$ and depressed Con A (0.25 $\mu\textrm{g}$/ml) dependent T-lymphocyte proliferation at 10$^{-7}$~10$^{-5}$M of GA, but increased suboptimal dose (Con A 0.1 $\mu\textrm{g}$/ml) at 10$^{-9}$~10$^{-7}$M of GA. IL-1 activity was suppressed by 10$^{-8}$~10$^{-4}$M of GA and $Ca^{2+}$ uptake was enhanced by 10$^{-9}$~10$^{-6}$ of GA, but antibody production was not changed by GA. From the above results, it is suggested that GL and GA have immuno-regulatory action. GL decreased cell-mediated immune response, and increased humoral immune response at high dose. On the other hand, low dose of GA enhanced cell-mediated immune response, while high doses of GA decreased humoral immune reaction.

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Comparative Study on Human Risk by Ionizing Radiation and Pesticide as Biological Information about Environmental Disaster (환경재해에 관한 생물정보로서의 이온화 방사선과 살충제의 인체 위해성 비교 연구)

  • Kim, Jin-Kyu;Hyun, Soung-Hee
    • Journal of Radiation Protection and Research
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    • v.26 no.4
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    • pp.385-392
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    • 2001
  • Environmental risk factors such as ionizing radiations, heavy metals, and pesticides can cause environmental disasters when they exist in excess. The increases in use of ionizing radiation and agricultural pesticide are somewhat related to the possibility of the disaster. The risk of radiation and pesticide was evaluated by means of the single cell gel electrophoresis (SCGE) assay on the human blood lymphocytes. The lymphocytes were irradiated with $0{\sim}2.0Gy$ of $^{60}Co$ gamma ray. Another groups of lymphocytes were exposed to various concentrations of parathion. Significantly increased tail moment, which was a marker of DNA strand breaks in SCGE assay, showed a clear dose- or concentration-response relationship. Parathion of a recommended concentration for agricultural use ($1mg {\ell}^{-1}$ ) has a strong cytotoxic effect on lymphocytes, which is equivalent to damage induced by 0.1 Gy of ${\gamma}$-ray. Furthermore, $2mg{\ell}^{-1}$ of parathion can give rise to DNA damage equivalent to that induced by 0.25 Gy at which the radiation-induced damage can start to develop into clinical symptoms. The comparative results of this study can provide an experimental basis and biological information for the prevention of environmental disaster.

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Association Study between Genetic Polymorphisms of CYP2C19 Gene and Essential Hypertension in Koreans (한국인에서 CYP2C19 유전자 다형성과 본태성 고혈압 간의 연관성 연구)

  • Park, Ah-Ram;Shin, Eun-Soon;Son, Nak-Hoon;Jang, Yang-Soo;Shin, Dong-Jik
    • Journal of Life Science
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    • v.20 no.5
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    • pp.799-804
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    • 2010
  • In humans, CYP2C19, a member of the cytochrome P450 subfamily, metabolizes arachidonic acid to produce epoxyicosanoid acids, which are involved in vascular tone and regulation of blood pressure (BP). Recent findings suggest that CYP2C19 gene polymorphisms might be considered as a novel candidate gene for cardiovascular disease. We thus focused on the Korean population to explore the association of two polymorphisms ($CYP2C19^*2$ and $^*3$) in this gene and essential hypertension (EH). A total of 1,241 participants (537 hypertensive subjects and 704 healthy controls) were recruited from the Yonsei Cardiovascular Genome Center in Korea. The CYP2C19 polymorphisms were genotyped using the $SNaPShot^{TM}$ assay. The allele and genotype frequencies of $CYP2C19^*3$ showed significant difference between hypertensives and normotensives (P=0.019 and P=0.023, respectively). Logistic regression analysis indicated that the $CYP2C19^*3$ A allele carriers were significantly associated with EH (OR, 0.723; 95% CI, 0.538-0.972, P=0.032) under a dominant model. In addition, CYP2C19 G-A haplotype ($2C19^*2\;G-^*3$ A combination) was found to significantly reduce EH risk (OR, 0.714, P=0.015). We believe this provides evidence that $CYP2C19^*3$ polymorphism may contribute to a protective effect in the development of EH.