• 대한한의학회지
• /
• 제22권2호
• /
• pp.109-119
• /
• 2001

Objective : There are many reports that acupuncture has thermoregulatory effects on human and animals. To investigate the effect and mechanism of antipyretic action of acupuncture, we observed the body temperature and cytokine expressions in the hypothalamus of rats. Methods : Lipopolysaccharide (LPS, i.p., 2.5mg/kg) was injected to conscious rats (Sprague-Dawley, male, n=4l) to cause hyperthermia and simple needling (stainless steel, 0.25 mm o.d., 5 mm insertion for 10 sec with no manipulation) was performed bilaterally with the measurement of rectal temperature. Next, we sacrificed rats to remove brain and determined the level of mRNA for interleukin-6 (IL-6), $interleukin-1{\beta}{\;}(IL-1{\beta})$, interleukin-2 (IL-2) and $interferon-{\gamma}{\;}(IFN-{\gamma})$ in the hypothalamus by using reverse transcriptase-polymerase chain reaction (RT-PCR). Resul1s : Needling on forepaw (acupoint HT8) and needling on hindpaw (acupoint BL66 and acupoint LR2) significantly inhibited LPS-induced fever of rats (P<0.01, 10 min after treatment respectively), but same treatment on proximal tail (non-acupoint) did not cause any change on fever. The levels of IL-6 and $IL-1{\beta}$ mRNA in the hypothalamus was significantly enhanced by LPS-injection, while the level of IL-6 and $IL-1{\beta}$ mRNA was markedly reduced after treatment on BL66 (P<0.01). Treatment on forepaw reduced it slightly, but not significantly. Equivalent stimulation on proximal tail did not cause any changes. Conclusions : Our results indicate that acupuncture stimulation on various body parts has differential thermoregulatory effects on LPS-induced fever of rats with site-specificity. And, we suggest that its antipyretic action might be accompanied with the suppression of hypothalamic production of pro-inflammatory cytokine of immune system, IL-6 and $IL-1{\beta}$.

• BMB Reports
• /
• 제41권1호
• /
• pp.41-47
• /
• 2008

Fe65 is characterized as an adaptor precursor (APP) through its PID2 element, as well as with the other members of the APP protein family. With the serum- and glucocorticoid-induced kinase 1 (SGK1) substrate specificity information, we found that the putative site of phosphorylation in Fe65 by SGK1 is present on its $Ser^{566}$ residue in $^{560}CRVRFLSFLA^{569}$(X60469). Thus, we demonstrated that Fe65 and the fluorescein-labeled Fe65 peptide $FITC-^{560}CRVRFLSFLA^{569}$ are phosphorylated in vitro by SGK1. Phosphorylation of the $Ser^{566}$ residue was also demonstrated using a $Ser^{566}$ phospho-specific antibody. The phospho Fe65 was found mainly in the nucleus, while Fe65 S556A mutant was localized primarily to the cytoplasm. Therefore, these data suggest that SGK1 phosphorylates the $Ser^{566}$ residue of Fe65 and that this phosphorylation promotes the migration of Fe65 to the nucleus of the cell.

• 한국방사선학회논문지
• /
• 제6권4호
• /
• pp.275-279
• /
• 2012

16MD CT(n=93) and dual source CT(n=100)를 이용해 촬영된 심장 CT 중 그 결과 협착으로 진단되어 심장영화 촬영을 시행한 환자를 대상으로 심장영화 촬영과의 협착 유무에 대한 비교를 하였다. 그 결과 16MDCT의 경우 가장 높은 민감도, 특이도, 양성예측도, 음성예측도는 LAD(73.5%), RCA(74.5%), LAD(66.7%), LCX(75%), LCX(67.7%)로 나타났고, 16Slice MDCT에서 각 관상동맥 혈관별 정확도는 평균67%로 다소 떨어지는 것으로 나타났고, Somatom Definition dual source CT의 경우 평균 정확도 84%로 16Slice MDCT보다는 17%더 높은 정확도를 나타내었다. 관상동맥영역별(segaments) 구분에서는 16Slice MDCT와 Somatom Definition 모두 근위부(proximal)보다는 원위부로 갈수록 더 높은 정확도를 나타내었다. 하지만 그 결과 값이 10% 오차 범위 내에 있어 명확한 차이라 보기는 힘들 것 이다. 또한 Dual source CT의 경우 역시 관상동맥 영역별 구분에서 큰 차이를 보이지 않았다.

• Bulletin of the Korean Chemical Society
• /
• 제26권3호
• /
• pp.433-439
• /
• 2005

To gain further insight on the relationship between structure and functions of glutathione S-transferase (GST), the three tyrosine 108 mutants, Y108A, Y108F, and Y108W, of human GST P1-1 were expressed in Escherichia coli and purified to electrophoretic homogeneity by affinity chromatography on immobilized GSH. The substitution of Tyr 108 with alanine resulted in significant decrease of the GSH-conjugation activity and the GSH peroxidase activity, but approximately 63% increase of steroid isomerase activity toward ${\Delta}^5$–[androstene 3,17-dione. On the other hand, the substitution of Tyr 108 with phenylalanine resulted in decreases of $k_{cat}\;and\;k_{cat}/K_m{^{EPNP}}$ by 2 orders of magnitude, suggesting that Tyr 108 residue of hGSTP1-1 are considered to be important for the catalysis and the binding of the epoxide substrates. The substitution of Tyr 108 with tryptophan resulted in significant decreases of the specific activities toward EPNP, cumene hydroperoxide and ${\Delta}^5$–ndrostene 3,17-dione, but approximately 2-fold increase on the enzyme-catalyzed addition of GSH to DCNB. We conclude from these results that Tyr 108 in hGST P1-1 plays very different roles depending upon the nature of the electrophilic substrates.

• Archives of Pharmacal Research
• /
• 제24권5호
• /
• pp.455-465
• /
• 2001

The highly potent cytotoxic DNA-DNA cross-linker consists of two cyclopropa[c]pyrrolo[3,4-3]indol-4(5H)-ones insoles [(+)-CPI-I] joined by a bisamido pyrrole (abbreviated to "Pyrrole"). The Pyrrole is a synthetic analog of Bizelesin, which is currently in phase II clinical trials due to its excellent in vivo antitumor activity. The Pyrrole has 10 times more potent cytotoxicity than Bizelesin and mostly form DNA-DNA interstrand cross-links through the N3 of adenines spaced 7 bp apart. The Pyrrole requires a centrally positioned GC base pair for high cross-linking reactivity (i.e., $5^1$-T$AT_2$A*-$3^1$), while Bizelesin prefers purely AT-rich sequences (i.e., $5^1$-T$AT_4$A*-$3^1$, where /(equation omitted) represents the cross-strand adenine alkylation and A* represents an adenine alkylation) (Park et al., 1996). In this study, the high-field $^1$H-NMR and rMD studies are conducted on the 1 1-mer DNA duplex adduct of the Pyrrole where the 5′(equation omitted)TAGTTA*-3′sequence is cross-linked by the drug. A severe structural perturbation is observed in the intervening sequences of cross-linking site, while a normal B-DNA structure is maintained in the region next to the drug-modified adenines. Based upon these observations, we propose that the interplay between the bisamido pyrrole unit of the drug and central C/C base pair (hydrogen-bonding interactions) is involved in the process of cross-linking reaction, and sequence specificity is the outcome of those interactions. This study suggests a mechanism for the sequence specific cross-linking reaction of the Pyrrole, and provides a further insight to develop new DNA sequence selective and distortive cross-linking agents.

• Parasites, Hosts and Diseases
• /
• 제53권1호
• /
• pp.77-83
• /
• 2015

Wuchereria bancrofti, Dirofilaria immitis, and Dirofilaria repens are filarial nematodes transmitted by mosquitoes belonging to Culex, Aedes, and Anopheles genera. Screening by vector dissection is a tiresome technique. We aimed to screen filarial parasites in their vectors by single and multiplex PCR and evaluate the usefulness of multiplex PCR as a rapid xenomonitoring and simultaneous differentiation tool, in area where 3 filarial parasites are coexisting. Female mosquitoes were collected from 7 localities in Assiut Governorate, were microscopically identified and divided into pools according to their species and collection site. Detection of W. bancrofti, D. immitis, and D. repens using single PCR was reached followed by multiplex PCR. Usefulness of multiplex PCR was evaluated by testing mosquito pools to know which genera and species are used by filarial parasites as a vector. An overall estimated rate of infection (ERI) in mosquitoes was 0.6%; the highest was Culex spp. (0.47%). W. bancrofti, D. immitis, and D. repens could be simultaneously and differentially detected in infected vectors by using multiplex PCR. Out of 100 mosquito pools, 8 were positive for W. bancrofti (ERI of 0.33%) and 3 pools each were positive for D. immitis and D. repens (ERI 0.12%). The technique showed 100% sensitivity and 98% specificity. El-Nikhila, El-Matiaa villages, and Sahel Seleem district in Assiut Governorate, Egypt are still endemic foci for filarial parasites. Multiplex PCR offers a reliable procedure for molecular xenomonitoring of filariasis within their respective vectors in endemic areas. Therefore, it is recommended for evaluation of mosquito infection after lymphatic filariasis eradication programs.

• Journal of Periodontal and Implant Science
• /
• 제26권1호
• /
• pp.161-175
• /
• 1996

There were many reports that elevations in the levels of active and latent collagenase in gingival crevicular fluid(GCF) have been correlated positively with periodontal disease activity. To provide a simple diagnostic approach for testing GCF collagenolytic activity, the detection limit of enzyme activity was compared using radiofibril assay(Sodek et.al.1981) and spectrophotometric collagenolytic assay(Nethery et al. 1986). The detection limits of both assay for standard bacterial enzyme were similar and the radiofibril assay showed a little (1/2) lower detection limit for tad pole collagenase. To evaluate the relationship between periodontal tissue destruction and the collagenolytic activity, GCF was collected, and latent and active enzyme activities were measured by a spectrophotometric collagenolytic assay. Twelve subjects showing progressive lesions were selected according to the presence of immediate tissue destruction, frequent abscess formation, and increasing need for tooth extraction, and the absence of underlying systemic disease and previous antibiotic medication history within 6 months. Comparisons were made between sites with either: 1) inflammation with a previous history of progressive loss of periodontal tissue and bone support(2l progressive sites): 2) previous history of bone loss and periodontal destruction but now clinically stable(12 comparably stable sites); or 3) no loss of periodontal tissue and bone support(11 control sites including 5 gingivitis sites and 6 healthy sites). Active collagenase activity was the highest in the progressive sites and decreased in the order of the gingivitis sites, the stable sites, and the healthy sites. The total enzyme activity was $2{\sim}3$ fold higher in the progressive sites and the gingivitis sites, compared to the stable and the healthy sites. The ratio of active to total collagenolytic activity was twice in the progressive sites. Analysis of active collagenase level(5mU) and the ratio of active to total collagenolytic activity(0.8) as a diagnositic test indicates that these measurements have the sensitivity of 0.81 and 0.86, the specificity of 0.70 and 0.65, and the overall agreement of 0.75 and 0.73, respectively. Thus, this method has significant merits as a diagnostic tool to determine wherher the site is in a state of remission or progression.

• Asian Pacific Journal of Cancer Prevention
• /
• 제15권15호
• /
• pp.6081-6086
• /
• 2014

Background: To evaluate the effectiveness of Microlux/DL with and without toluidine blue in screening of potentially malignant and malignant oral lesions. Materials and Methods: In this diagnostic clinical trial clinical examination was carried out by two teams: 1) two oral medicine consultants, and 2) two general dentists. Participants were randomly and blindly allocated for each examining team. A total of 599 tobacco users were assessed through conventional oral examination (COE); the examination was then repeated using Microlux/DL device and toluidine blue. Biopsy of suspicious lesions was performed. Also clinicians opinions regarding the two tools were obtained. Results: The sensitivity and, specificity and positive predictive value (PVP) of Microlux/DL for visualization of suspicious premalignant lesions considering COE as a gold standard (i.e screening device) were 94.3%, 99.6% and 96.2% respectively, while they were 100%, 32.4% and 17.9% when considering biopsy as a gold standard. Moreover, Microlux/DL enhanced detection of the lesion and uncovered new lesions compared to COE, whereas it did not alter the provisional clinical diagnosis, or alter the biopsy site. On the other hand, adding toluidine blue dye did not improve the effectiveness of the Microlux/DL system. Conclusions: The Microlux/DL seems to be a promising adjunctive screening device.

• BMB Reports
• /
• 제36권4호
• /
• pp.409-416
• /
• 2003

The isoform 1 of cyclodextrin glycosyltransferase (CGTase, EC 2.4.1.19) from Paenibacillus sp. A11 was purified by a preparative gel electrophoresis. The importance of histidine, tryptophan, tyrosine, and carboxylic amino acids for isoform 1 activity is suggested by the modification of the isoform 1 with various group-specific reagents. Activity loss, when incubated with diethylpyrocarbonate (DEP), a histidine modifying reagent, could be protected by adding 25 mM methyl-$\beta$-cyclodextrin substrate prior to the modification. Inactivation kinetics of isoform 1 with DEP resulted in second-order rate constants ($k_{inactivation}$) of $29.5\;M^{-1}s^{-1}$. The specificity of the DEP-modified reaction for the histidine residue was shown by the correlation between the loss of isoform activity and the increase in the absorbance at 246 nm of N-carbethoxyhistidine. The number of histidines that were modified by DEP in the absence and presence of a protective substrate was estimated from the increase in the absorbance using a specific extinction coefficient of N-carbethoxyhistidine of $3,200\;M^{-1}cm^{-1}$. It was discovered that methyl-$\beta$-CD protected per mole of isoform 1, two histidine residues from the modification by DEP. To localize essential histidines, the native, the DEP-modified, and the protected forms of isoform 1 were digested by trypsin. The resulting peptides were separated by HPLC. The peptides of interest were those with $R_t$ 11.34 and 40.93 min. The molecular masses of the two peptides were 5,732 and 2,540 daltons, respectively. When the data from the peptide analysis were checked with the sequence of CGTase, then His-140 and His-327 were identified as essential histidines in the active site of isoform 1.

• Journal of Microbiology and Biotechnology
• /
• 제17권4호
• /
• pp.604-610
• /
• 2007

A psychrotrophic strain 7195 showing extracellular lipolytic activity towards tributyrin was isolated from deep-sea sediment of Prydz Bay and identified as a Psychrobacter species. By screening a genomic DNA library of Psychrobacter sp. 7195, an open reading frame of 954 bp coding for a lipase gene, lipA1, was identified, cloned, and sequenced. The deduced LipA1 consisted of 317 amino acids with a molecular mass of 35,210 kDa. It had one consensus motif, G-N-S-M-G (GXSXG), containing the putative active-site serine, which was conserved in other cold-adapted lipolytic enzymes. The recombinant LipA1 was purified by column chromatography with DEAE Sepharose CL-4B, and Sephadex G-75, and preparative polyacrylamide gel electrophoresis, in sequence. The purified enzyme showed highest activity at $30^{\circ}C$, and was unstable at temperatures higher than $30^{\circ}C$, indicating that it was a typical cold-adapted enzyme. The optimal pH for activity was 9.0, and the enzyme was stable between pH 7.0-10.0 after 24h incubation at $4^{\circ}C$. The addition of $Ca^{2+}\;and\;Mg^{2+}$ enhanced the enzyme activity of LipA1, whereas the $Cd^{2+},\;Zn^{2+},\;CO^{2+},\;Fe^{3+},\;Hg^{2+},\;Fe^{2+},\;Rb^{2+}$, and EDTA strongly inhibited the activity. The LipA1 was activated by various detergents, such as Triton X-100, Tween 80, Tween 40, Span 60, Span 40, CHAPS, and SDS, and showed better resistance towards them. Substrate specificity analysis showed that there was a preference for trimyristin and p-nitrophenyl myristate $(C_{14}\;acyl\; groups)$.