• Tuberculosis and Respiratory Diseases
• /
• 제41권5호
• /
• pp.489-493
• /
• 1994

연구배경: NAD glycohydrolase(NADase)는 세포 표면에 위치하며 glycosylphatidylinositol(GPI)로 세포표면에 부착되어 있으며 bacterial PI-specific phospholipase C(PI-PLC)에 의해 분리된다. 최근 결핵환자에서 NADase가 증가한다는 보고가 있었으나 결핵균자체가 NADase가 높기 때문에 절대적인 NADase치가 증가했는 지는 확실치 않다. 따라서 저자들은 정상 대조군과 결핵환자의 적혈구에서 순수한 NADase activity를 측정하였다. 방법: 19명의 정상 건강 대조군과 16명의 과거 결핵의 진단 및 치료를 받지 않은 결핵환자를 대상으로하여 NADase activity를 측정하였고 결핵환자는 결핵치료 3개월후 재 검사하였다. NADase activity는 [carbonyl-$^3H$] nicotinamide 동위원소를 사용하여 측정하였다. 결과: 건강 대조군에서 $2031{\pm}824.0pmol/min/10^6$ erythrocytes, 결핵 감염군에서 $3339{\pm}1568.0$, 그리고 isoniazide와 rifampin으로 치료한 3개월 후 $2238.6{\pm}1013.1$을 얻어 결핵 감염시 NADase activity가 유의한 차이를 가지고 증가하며(p<0.05), 치료전에 비하여 치료후 유의있게 감소하였다. 결론: 결핵감염시 NADase activity가 올라가고 결핵 치료시 NADase activity가 정상화 되어 NADase activity가 결핵감염의 진단 및 치료에 대한 새로운 지표가 될수 있을 것으로 사료된다.

• Animal cells and systems
• /
• 제8권2호
• /
• pp.125-133
• /
• 2004

We have extended our previous work that cross-linking CD4 molecules using specific MAb induced antigen nonspecific, MHC unrestricted killing of virally infected target cells by CD$4^+$We have extended our previous work that cross-linking CD$4^+$ molecules using specific MAb induced antigen nonspecific, MHC unrestricted killing of virally infected target cells by CD$4^+$ T cells. The killing activity of antibody activated CD$4^+$T cells was completely blocked by herbimycin A, a protein tyrosine kinase (PTK) inhibitor, but not by bisindolylamaleimide, a protein kinase C (PKC) inhibitor. Herbimycin A treated human or bovine peripheral blood CD$4^+$T cells lacked PTK activity and failed to kill virally infected target cells even after cross-linking of CD4 molecules. The CD$4^+$cross-linking failed to induce effector cell proliferation or the transcription of TNF${\beta}$ Upregulation of TNF${\beta}$ was induced by incubating the antibody activated effector cells with BHV-1 infected D17 target cells for 10 h. Anti-TNF${\beta}$ antibody partially abolished (13-44%) the direct effector cell-mediated antiviral cytotoxicity. However, this antibody neutralized 70 to 100% of antiviral activity of effector and target cell culture supernatants against BHV-1 infected D17 cells. The inhibition level of the antiviral activity by the antibody was dependent on the effector and target cell ratio. These results support the hypothesis that increased p$56^ICK enzyme activity in effector cells transduces a signal critical for effector cell recognition of viral glycoproteins expressed on the target cells. Following target cell recognition, lytic cytokines known to participate in target cell killing were produced. A better understanding of the killing activity displayed by CD$4^+$T lymphocytes following surface receptor cross-linking will provide insight into the mechanisms of cytotoxic activity directed toward virally-infected cells.T cells. The killing activity of antibody activated CD$4^+$T cells was completely blocked by herbimycin A, a protein tyrosine kinase (PTK) inhibitor, but not by bisindolylamaleimide, a protein kinase C (PKC) inhibitor. Herbimycin A treated human or bovine peripheral blood CD4T cells lacked PTK activity and failed to kill virally infected target cells even after cross-linking of CD4molecules. The CD4 cross-linking failed to induce effector cell proliferation or the transcription of TNF$\beta$. Upregulation of TNF$\beta$ was induced by incubating the antibody activated effector cells with BHV-1 infected D17 target cells for 10 h. Anti-TNF$\beta$ antibody partially abolished (13-44%) the direct effector cell-mediated antiviral cytotoxicity. However, this antibody neutralized 70 to 100% of antiviral activity of effector and target cell culture supernatants against BHV-1 infected D17 cells. The inhibition level of the antiviral activity by the antibody was dependent on the effector and target cell ratio. These results support the hypothesis that increased $56^ICK enzyme activity in effector cells transduces a signal critical for effector cell recognition of viral glycoproteins expressed on the target cells. Following target cell recognition, lytic cytokines known to participate in target cell killing were produced. A better understanding of the killing activity displayed by CD$4^+$T lymphocytes following surface receptor cross-linking will provide insight into the mechanisms of cytotoxic activity directed toward virally-infected cells.

• 한국수소및신에너지학회논문집
• /
• 제19권2호
• /
• pp.124-131
• /
• 2008

Crude cytoplasmic fraction of phototrophic purple sulfur bacterium, Thiocapsa roseopersicina NCIB 8347, were initially prepared and purified by sonication, ultracentrifugation, ammonium sulfate fractionation and heat-treatment and it has been previously reported. Using various applications of chromatography far the purification of membrane-bound and soluble hydrogenases from heat-treated enzyme fraction were studied at present report. When the heat-treated enzyme preparation was applied to the anion column chromatography using Q-sepharose, Fraction I and II, which were extracted with the KCl 0-0.5 M gradient, showed the specific evolution hydrogenase activity 3.86 and 2.27 U/mg-protein respectively. Specific hydrogenase activitys of Fraction I and II were further increased to 4.35 and 7.46 U/mg-protein for Fraction I and to 2.49 and 4.41 U/mg-protein fur Fraction II respectively, when hydrophobic interaction column, Phenyl superose, and anion exchange column, Mono-Q, were applied. Size exclusion chromatography using superdex 200 concentrated the hydrogenase Fraction I and II to 9.19 and 7.84 U/mg-protein respectively at the final step of purification.

• Journal of Microbiology and Biotechnology
• /
• 제14권3호
• /
• pp.520-524
• /
• 2004

A protein that exhibited a high similarity to a major serine transporter of Escherichia coli, SdaC, was found in Haemophilus injluenzae Rd. Also, $Na^+$-stimulated serine transport activity was detected in the cells. The sdaC of H. injluenzae was cloned and the properties of the transporter were investigated. The activity of serine transport was stimulated by $Na^+$. Uptake of $Na^+$ elicited by L-serine influx into cells was also observed, which supports the idea that L-serine is transported by a mechanism of $Na^+$serine symport. No uptake of $H^+$ elicited by L-serine influx was detected. This result was not consistent with that obtained with the homologous protein, SdaC of E. coli, which uses $H^+$as a coupling cation. The serine transport via the SdaC of H. influenzae was not inhibited by other amino acids such as threonine or D-serine like the SdaC of E. coli. Thus, the SdaC of H. influenzae is a $Na^+$-dependent L-serine specific symporter and an unusual natural mutant. The $K_m$ and the $V_{max}$, value for the serine transport in the SdaC of H. influenzae were $7.6\mu$M and 22.9 nmol/min/mg protein, respectively.

• 한국미생물·생명공학회지
• /
• 제25권2호
• /
• pp.159-165
• /
• 1997

We collected 3,237 plant samples, mainly leaves of various trees, from many provinces in Korea and a total of 1,925 Bacillus thuringiensis isolates were obtained and characterized. The isolates were characterized in terms of crystal morphology, PAGE pattern of the toxin proteins, plasmids pattern, biochemical characteristics, and bioassay. The microscopic observation showed that 49.1% of the isolates have bipyramidal shape crystals, 7.1% of spherical shape crystals, 1.4% of rhomboidal shape crystals, and others have small or amorphous inclusions. The insecticidal activities of the spore-crystal mixtures of isolates were tested against Plutella xylostella, Bombyx mori, Culex pipiens, and Agelastica coerulea. Bioassay showed that 51.3% of the isolates were shown to be active; lepidopteran-specific (44.8%), dipteran-specific(4.9%) and coleopteran-specific (1.6%). The remainder(48.8%) did not show any activity against the insects we tested. Interestingly though, some of these non-active isolates were shown to have bipyramidal crystals. By serotyping 22 isolates of our collection, we found that there are various kind of subspecies such as aizawai, amagiens, canadensis, darmstadiensis, galleriae, finitimus, kurstaki, morrisoni and neoleonensis, and three isolates have been classified into a new serotype, H49, and one of them, the type strain, named subsp. muju. From this study it was found that phylloplane is a good source for the isolation of Bacillius thuringiensis, and Bacillus thuringiensis is distributed widely in Korea.

• 대한의생명과학회지
• /
• 제15권4호
• /
• pp.287-293
• /
• 2009

Phospholipase D (PLD) is an enzyme hydrolyzing phosphatidylcholine to phosphatidic acid (PA) and choline. We investigated the involvement of PLD1 in the uptake of norepinephrine (NE) in PC12 cells, pheochromocytoma cells. NE uptake was specific in PC12 cells because nomifensine, a specific blocker of NE transporter, blocked NE uptake. Inhibition of PLD function in PC12 cells by the treatment of butanol suppressed the NE uptake. In contrast, overexpression of PLD1 in PC12 cells increased NE uptake efficiently. These results suggest that PLD activity is involved in NE uptake. We explored the action mechanism of PLD in NE uptake. PA phosphatase inhibitor, propranolol, blocks the formation of PKC activator diacylglycerol from PA. Propranolol treatment to PC12 cells blocked dramatically the uptake of NE. Specific PKC inhibitors, GF109203X and Ro31-8220, blocked NE uptake. Taken together, we suggest for the first time that PLD1 activity is involved in NE uptake via the activation of PKC.

• Biomolecules & Therapeutics
• /
• 제1권1호
• /
• pp.121-124
• /
• 1993

To elucidate the effect of antipsychotics and antidepressants on phosphoinositide(Pl) second massenger system, we studied the dose-dependent inhibition of the phosphoinositide-specific phospholipase C(PLC) isozymes, ${\beta}_1,\;{\gamma}_1$ and${\delta}_1,$ by fluphenazine and haloperidol as antipsychotics, and amitriptyline, maprotiline and mianserin as antidepressants. All the antipsychotics and antidepressants tested showed inhibition on at least one of the PLC isozymes with $IC_{50}$ at the concentration between 25 and $250 {\mu}M.$ Maprotiline, mianserin and amitriptyline inhibited 80 to 90% of the activities of all three PLC isozymes at the concentration of $250{\mu}M,$ while haloperidol and fluphenazine inhibited PLC ${\beta}_1$ and${\gamma}_1$ But baclofen didn't inhibit any PLC isozyme. These results suggested that PLC isozymes are inhibited by antipsychotics and antidepessants even though the concentration is high, and these drugs may affect PI signal transduction system by direct inhibition of PLC isozymes.

• Journal of Microbiology
• /
• 제43권3호
• /
• pp.244-250
• /
• 2005

The effect of cytochrome $c_{550}$ encoded by cccA in Bacillus subtilis during the event of sporulation was investigated. The sporulation of cccA-overexpressing mutant was significantly accelerated, while disruptant strain showed delayed sporulation in spite of the same growth rate. Activity of sporulation stage-0-specific enzyme, extracellular $\alpha-amylase$ of mutant strains was similar to that of the control strain, but cccA-overexpressing mutant exhibited higher activity of stage-II-specific alkaline phosphatase and stage-III-specific glucose dehydrogenase when compared to deletion mutant and control strain. Northern blot analysis also revealed that cccA-overexpressing mutant showed high level of spo0A transcripts, while the disruptant rarely expressed spo0A. These results suggested that although cytochrome $c_{550}$ is dispensable for growth and sporulation, expression of cccA may play an important role for initiation of sporulation through regulation of spo0A expression.

• 한국발생생물학회지:발생과생식
• /
• 제17권1호
• /
• pp.1-8
• /
• 2013

Osteoprotegerin (OPG) is a secreted glycoprotein that regulates bone resorption by inhibiting differentiation and activation of osteoclast, thereby potentially useful for the treatment of many bone diseases associated with increased bone loss. In this study, we designed a novel cDNA expression cassette by modifying the potent and mammary gland-specific goat ${\beta}$-casein/hGH hybrid gene construct and examined human OPG (hOPG) cDNA expression in transgenic mice. Six transgenic mice all successfully expressed hOPG in their milk at the level of 0.06-2,000 ${\mu}g/ml$. An estimated molecular weight of the milk hOPG was 55 kDa in SDS-PAGE, which is the same as a naturally glycosylated monomer. This hOPG expression was highly specific to the mammary glands of transgenic mice. hOPG mRNA was not detected in any organs analyzed except mammary gland. Functional integrity of milk hOPG was evaluated by TRAP (tartrate-resistant acid phosphatase) activity assay in bone marrow cell cultures. OPG ligand (OPG-L) treatment increased TRAP activity by two fold but it was completely abolished by co-treatment with transgenic milk containing hOPG. Taken together, our novel cDNA expression cassette could direct an efficient expression of biologically active hOPG, a potential candidate pharmaceutical for bone diseases, only in the mammary gland of transgenic mice.

• 한국환경보건학회지
• /
• 제30권3호
• /
• pp.283-292
• /
• 2004

Valid evaluation of community reaction to road-traffic noise exposure is important for the policy making and quality of life. The purpose of this study is to evaluate the relationship between community reaction and nighttime road-traffic noise. The study measured nighttime roadside noise caused by high traffic which is greater than LAeq(22:00-07:00) 65dB(A) and lower than 65dB(A) at Chonju city, from March to April, 2003. Three hundred sixty seven subjects, aged from 20 to 65, were selected from those who are residing close to the major roads. They were asked to answer the questions regarding noise source, stress(PWI-SF), annoyance, disturbance of specific activity, disturbance of sleep, somatic symptom, and four suggested confounding factors. The relationship between annoyance, somatic symptom and noise exposure was not significant. Adjusted odds ratio(95% C.I.) for disturbance of communication　 and disturbance of attention and rest　 were 1.59(1.03-2.71), 1.64(1.06-2.81), respectively. On the other hand sleep disturbance　 was indicated as 1.34(0.77-2.32). The results suggested that federal policy-making about road-traffic noise should consider community reaction evaluated by various perspective including annoyance, disturbance of specific activity, disturbance of sleep, and confounding factors.