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Notes: Background.  Helicobacter pylori is a causative agent of gastritis, and H. pylori infection is thought to be correlated with iron-deficiency anemia (IDA) at puberty. The H. pylori feoB gene product, a high-affinity ferrous iron transporter, plays a central role in iron acquisition and virulence. This study was undertaken to analyze H. pylori feoB status according to clinical data, including antral gastritis with or without IDA.Methods.  Fourteen H. pylori-positive patients aged from 10 to 18 years were categorized into subgroups based on the presence or absence of IDA. Eight patients were diagnosed as having IDA; the other six showed normal hematological findings. Genomic DNA was isolated from H. pylori cultured from each gastric biopsy specimen. Five sets of primers were used for the PCR amplification of the feoB gene. Linking and sequencing of PCR products generated the feoB region, which was 1.93 kb in size. The feoB gene sequences of H. pylori J99 and 26695 were compared with the clinical strains, and the sequences of feoB regions in the IDA (+) and (–) groups were compared.Results.  Sequence analysis of the complete coding region of the feoB gene revealed 16 sites of polymorphism or mutation. Among these, three polymorphisms (E/T254A, I263V, and K511Q) were indigenous to the Korean clinical strains. Although statistically significant differences were observed at four sites (K127T, A273S/P, I438V and I441T) between IDA (+) and (–), the number of specimens was too low to assess the significance of the differences.Conclusion.  The four polymorphisms of the feoB gene observed appear to be related to the clinical phenotype of IDA, but the relation is unclear because of the small number of strains studied. Further studies are required to confirm a correlation between IDA and H. pylori infection.

Notes: Interleukin-18 (IL-18) has multiple important pro-inflammatory effects, including the induction of interferon-γ (IFN-γ) in various diseases. In this study, we investigated the IL-18-producing activities in human pulmonary and pleural tuberculosis (TB) in response to purified protein derivative (PPD) antigen (Ag) from Mycobacterium tuberculosis. The most significant IL-18 production was found in chronic refractory TB (CRTB) patients. However, IFN-γ production in CRTB patients was significantly less than that in healthy tuberculin reactors or in patients with tuberculous pleurisy (TBP). Elevated levels of both IL-18 and IFN-γ were found in pleural fluids from TBP patients. In vitro production of IL-18 was dramatically decreased following an 18 h stimulation with PPD. However, IFN-γ was markedly increased in pleural mononuclear cells from TBP patients after in vitro stimulation with PPD. The mesothelial cell type was the main source of pro-IL-18 in pleural cells from TBP patients, suggesting an important role for these cells in TBP. Taken together, these data indicate that IL-18 is elevated in peripheral blood mononuclear cells from CRTB patients, as well as at the site of TBP, indicating a possible role for IL-18 in both protective immunity and pathologic responses in human TB.

Notes: SUMMARY: Pharmacokinetic studies were performed in 10 stable kidney transplantation patients who received microemulsion formulation (Neoral®) of cyclosporine A (CsA) twice daily. No agents having pharmacokinetic effects on CsA had been used in these patients. The values of various basic pharmacokinetic parameters were similar to those reported in Western literature. The complete area under the blood concentration–time curve (AUC) of CsA for the duration of 12 h (12-h AUC) was determined using the linear trapezoidal rule from seven concentrations at 0, 1, 2, 4, 6, 8, and 12 h after CsA administration. The mean values of 12-h AUC were 4603.63 ± 344.61 ng h/mL. CsA concentrations at 2 h after dosing (not the trough levels) showed the best correlation with the complete AUC (r2 = 0.9322). The abbreviated AUC of CsA was calculated either by stepwise multiple linear regression analysis or by the linear trapezoidal rule from a few sampling time points. Using stepwise multiple linear regression analysis, which was used in calculating abbreviated AUC in all previous studies, the model equation that had the highest correlation and the lowest prediction error with the complete AUC was derived by using CsA concentrations at 2 and 8 h after dosing (12-h AUC = 4.262C2 + 8.390C8− 669.417; r2 = 0.9808, absolute prediction error = 3.97 ± 0.96). Two model equations derived using the linear trapezoidal rule provided the best correlation with the complete AUC: (1) The two time points selected model equation 12-h AUC = 4C2 + 5C8; r2 = 0.9780, absolute prediction error = 6.41 ± 1.22). (2) The three time points selected model equation 12-h AUC = 4C0 + 3C2 + 5C6; r2 = 0.9475, absolute prediction error = 5.00 ± 1.41). When different pharmacokinetic data sets were applied to the model equations derived using regression analysis, the values of coefficients and the constant of the regression equation had changed from the initial equation. Thus, new model equations will emerge every time the new data are applied. In contrast, the values of coefficients in the model equation calculated using trapezoidal rule were unaltered when tested by the new pharmacokinetic data set. Thus, the abbreviated AUC derived using the linear trapezoidal rule would be simpler than and superior to that obtained using stepwise multiple linear regression analysis in prediction of the complete AUC.

Notes: 1. Heavy alcohol consumption has been known as a risk factor for hypertension, although the mechanism by which alcohol intake causes hypertension remains elusive.2. We tested the hypothesis that brief exposure to ethanol augments vascular contractility through the stress response in human chorionic plate arteries.3. Human chorionic plate arteries were mounted in organ baths and exposed to 5% ethanol for 15, 30 or 45 min.4. Brief exposure for 45 min, but not 15 min, not only augmented contractility to KCl and 5-hydroxytryptamine 5 h after the end of exposure, but also increased the expression of heat shock protein (HSP) 70 in the tissues.5. Reverse transcription–polymerase chain reaction showed gradual increases of hsp70 mRNA expression, but not heat shock cognate 70 (hsc70), hsp90α or glucose regulatory protein 78 (grp78) mRNA expression, in an exposure time-dependent manner 3 h after the end of exposure.6. These results indicate that ethanol augments vascular contractility through the stress response.

Notes: Outer membrane proteins (OMPs) of pathogenic bacteria have been used as protective antigens in developing bacterial vaccines. In the present study, we compared the antibody responses to a Pseudomonas aeruginosa OMP vaccine elicited in humans and rabbits by immunization. Immunization with the vaccine induced high titers of serum IgG antibody both in rabbits and humans but reactivities of the induced antibodies with the OMPs were different. The rabbit immune sera recognized most of the OMPs in the vaccine both in immunoblot and immunoprecipitation analyses. In contrast, a great variation in band pattern and intensity was observed among the human immune sera in immunoblot analysis, but not in immunoprecipitation analysis. Denaturation of the OMPs did not affect the binding activity of the rabbit immune sera as determined by ELISA, but substantially reduced those of the human immune sera and anti-OMP IgG purified from a pooled normal human plasma. These data suggest that antibody response to P. aeruginosa OMPs elicited by immunization in humans is mainly directed against discontinuous or conformation-dependent epitopes, which should be taken into account in developing vaccines, especially for OMP-derived synthetic peptides.

Notes: With the introduction of high pressure balloon inflations and antiplatelet therapy, the incidence of stent thrombosis has been markedly reduced, but the incidence of late stent restenosis has not. A new strategy may be local drug delivery, which maintains sustained local concentration and limits systemic complications. To evaluate the efficacy of local nitric oxide (NO) donor delivery on stent thrombosis and complications, local NO donor delivery was performed in stented patients. NO donor (2.0-mg molsidomine) was delivered (1.0 mL/min 〉 10 min) using the Dispatch Catheter after predilation of target lesions in 13 patients (6 angina, 7 myocardial infarction, age 53.1 ± 11.4 years). After local NO donor delivery, Palmaz-Schutz stents were placed using standard methods. Follow-up coronary angiograms were performed 48 hours and 6 months after stenting. None of the patients had hypotensive effects, ischemic symptoms, or ECG changes during and after local NO donor delivery. APTT and CK values were unchanged at 3 and 24 hours after local NO donor delivery and stenting. Follow-up coronary angiograms at 48 hours and 6 months showed all stents patent with TIMI III flow and without intrastent thrombus. No target lesion revascularization and 100% event-free survival were observed during the 6-month clinical follow-up period. Intracoronary stenting may be performed safely and effectively by local NO donor delivery prior to stent implantation.

Notes: Activated caspase-3 is considered an important enzyme in the cell death pathway. To study the specific role of caspase-3 activation in neuronal cells, we generated a stable tetracycline-regulated SK-N-MC neuroblastoma cell line, which expressed a highly efficient self-activating chimeric␣caspase-3, consisting of the caspase-1 prodomain fused to the caspase-3 catalytic domain. Under expression-inducing conditions, we observed a time-dependent increase of processed caspase-3 by immunostaining for the active form of the enzyme, intracellular caspase-3 enzyme activity, as well as poly(ADP-ribose) polymerase (PARP) cleavage. Induced expression of the caspase fusion protein showed predominantly caspase-3 activity without any apoptotic morphological changes. In contrast, staurosporine treatment of the same cells resulted in activation of multiple caspases and profound apoptotic morphology. Our work provides evidence that auto-activation of caspase-3 can be efficiently achieved with a longer prodomain and that neuronal cell apoptosis may require another caspase or activation of multiple caspase enzymes.

Notes: Mixed vegetable juice was acidified to a pH below 4 and subjected to pasteurization regimes between 80 and 100 °C. Hunter colour scale values (L, a and b) of the pasteurized samples were analysed to determine the kinetics of green and total colour degradation. The ‘a’ value was chosen as the physical parameter for green colour while ‘Lab’ represented the total colour. A concept, based on fractional conversion, was used for the determination of kinetic parameters. The results revealed that degradation of green and total colour of the acidified vegetable juice followed first-order kinetics. Temperature dependency of the rate constants for both the green and the total colour could be described by the Arrhenius relationship. The activation energies of green and total colour degradation were 49.19 and 29.00 kJ mol−1, respectively.

Notes: Background  A keloid results from excessive collagen deposition, the cause of which remains elusive. A thorough understanding of the pathophysiology of keloid tissue can help determine the most appropriate treatment strategy.Objectives  To assess the differences in gene expression between keloids and adjacent normal skin in order to define the genes involved in keloid formation.Methods  Three Korean patients with keloids underwent excision of the keloid and adjacent normal skin, which was used as the control. We investigated expression patterns of genes in the keloids and the normal skin using cDNA microarray and in situ hybridization techniques.Results  Nine genes in the keloid tissue were consistently upregulated over the 2·0 ratio compared with the normal control from the cDNA microarray composed of 3063 clones: collagen type I α1 (〈accessionId ref="info:ddbj-embl-genbank/NM_000088"〉NM_000088), DNA segment on chromosome 21 (unique) 2056 expressed sequence (D21S2056E, NNP-1, 〈accessionId ref="info:ddbj-embl-genbank/NM_003683"〉NM_003683), suppressor of Ty 5 homologue (〈accessionId ref="info:ddbj-embl-genbank/NM_003169"〉NM_003169), phosphoglycerate dehydrogenase (〈accessionId ref="info:ddbj-embl-genbank/NM_032692"〉NM_032692), adenosine triphosphate synthase β (〈accessionId ref="info:ddbj-embl-genbank/NM_001686"〉NM_001686), serine (or cysteine) proteinase inhibitor, clade H (heat shock protein 47, 〈accessionId ref="info:ddbj-embl-genbank/NM_001235"〉NM_001235), LIV-1 protein, oestrogen regulated (LIV-1, 〈accessionId ref="info:ddbj-embl-genbank/NM_012319"〉NM_012319), interleukin-11 receptor α (IL11RA, 〈accessionId ref="info:ddbj-embl-genbank/NM_004512"〉NM_004512) and carbonyl reductase 3 (CBR3, 〈accessionId ref="info:ddbj-embl-genbank/NM_001236"〉NM_001236). From the in situ hybridization study, the staining signals in the keloid tissue hybridized with anti sense probes of NNP-1 mRNA were stronger than signals in normal controls. Further, endothelial epithelium, but not the epidermis, expressed the signal equally in both keloid and normal control tissue.Conclusions  We identified nine upregulated genes in keloid tissue using cDNA microarray. Of the nine, the NNP-1 gene was confirmed by topological information using the in situ hybridization technique. We conclude that these nine genes, especially NNP-1, probably contribute either directly or indirectly to keloid formation.