bluuebunny/crossref_metadata_2025 huggingface dataset:
Major Influence of Oxygen Supply on 13CO2 : 12CO2 Ratio Measurement by Nondispersive Isotope‐Selective Infrared Spectroscopy
Authors: Riecke Björn , Neuhaus Peter , Stockmann Martin
Published: November 2005

Background.  The nondispersive isotope‐selective infrared spectroscopy (NDIRS) is a valid method for the measurement of the 13CO2 : 12CO2 ratio in breath samples. Methodical influences have to be considered to obtain valid results.

Aim.  To evaluate the effect of oxygen supply to patients on the measurement of 13C : 12C ratio in breath samples by NDIRS.

Methods.  Breath samples of 26 healthy volunteers were taken before, immediately after, and 5 minutes after inhalation of 100% oxygen via a continuous positive air pressure (CPAP) mask. Analysis of breath samples was performed by NDIRS.

Results.  Delta ‰ before oxygen inhalation was −25.8 ± 0.2. Immediately after 5 minutes of 100% oxygen inhalation, delta ‰ increased to −14.8 ± 0.5 (delta over baseline [DOB] 11.0 ± 0.4) and after additional 5 minutes of room air inhalation, delta ‰ normalized to −25.6 ± 0.2 (DOB 0.2 ± 0.1).

Conclusions.  Oxygen supply to patients and, therefore, changes in gas composition in breath samples clearly influence 13CO2 measurement by NDIRS. This has to be taken into account in the clinical setting. Thus, oxygen supply during measurement of exhaled 13CO2 by NDIRS has to be avoided or maintained at a strictly constant level.
13C‐Urea Breath Test to Assess Helicobacter pylori Bacterial Load
Authors: Zagari Rocco Maurizio , Pozzato Paolo , Martuzzi Cristina , Fuccio Lorenzo , Martinelli Giuseppe , Roda Enrico , Bazzoli Franco
Published: November 2005

Background.  Some authors have reported, using different protocols, that 13C‐urea breath test (13C‐UBT) is capable of assessing the intragastric Helicobacter pylori bacterial load, whereas others have not confirmed these data. Our aim is to evaluate the correlation between 13C‐UBT values and H. pylori bacterial load.

Materials and Methods.  One hundred ninety‐two patients diagnosed H. pylori‐positive by rapid urease test, histology, and 13C‐UBT were enrolled. H. pylori bacterial load (H. pylori score) and gastritis activity (activity score) were evaluated according to the Updated Sydney System. 13C‐UBT was performed according to the European Standard Protocol. Breath samples were obtained every 10 minutes for 60 minutes in 52 patients and at 30 minutes (T30) in 140 patients and analyzed by mass spectrometry.

Results.  At T30, mean ± SD excess delta 13CO2 excretion was 17.4 ± 1.1, 29.9 ± 2.2, and 48.7 ± 4.8 in patients with H. pylori scores 1, 2, and 3, respectively. This difference was statistically significant: H. pylori score 1 versus 2, p < .005; score 1 versus 3, p < .05; score 2 versus 3, p < .05. A significant positive correlation (G = 0.59) was found between H. pylori score and activity score of chronic gastritis. At T40 and T50 significant correlation between mean excess delta 13CO2 excretion and bacterial load was achieved only in patients with H. pylori scores 1 and 3.

Conclusions.  13C‐UBT European Standard Protocol values correlate with H. pylori bacterial load and the activity of gastritis at T30 breath sampling time.
Asymptomatic Helicobacter pylori Infection Increases Asymmetric Dimethylarginine Levels in Healthy Subjects
Authors: Marra Maurizio , Bonfigli Anna Rita , Bonazzi Patrizia , Galeazzi Renato , Sirolla Cristina , Testa Ivano , Cenerelli Stefano , Boemi Massimo , Testa Roberto
Published: November 2005

Background.  Chronic infections have been demonstrated to be early factors of atherosclerosis and cardiovascular diseases, and their relevance increases when they are caused by agents with extremely broad spectrum of disease outcome such as Helicobacter pylori. The consequent endothelial impairment leads to a reduced bioavailability of nitric oxide. Increasing evidences have pointed out that the endogenous inhibitor of nitric oxide synthase, asymmetric dimethylarginine, defined as a risk factor for cardiovascular disease, may increase in infections and plays an important role impairing the vascular functions of the endothelium. Starting from these findings, we aim to investigate whether H. pylori may affect asymmetric dimethylarginine levels.

Materials and Methods.  The study was carried out on a group of 186 subjects (age 46.2 ± 14.9 years). We evaluated asymmetric dimethylarginine, symmetric dimethylarginine, L‐arginine, presence of H. pylori by 13C‐urea breath test, and the main parameters of glyco and lipo metabolic balance.

Results.  Increased levels of asymmetric dimethylarginine were found in H. pylori‐positive subjects with respect to H. pylori‐negative subjects (0.46 ×/÷ 1.13 versus 0.42 ×/÷ 1.23 mol/l, p < .001, respectively). No differences were detected in L‐arginine levels between the two groups. Multiple regression analysis performed in H. pylori‐positive subjects and H. pylori‐negative subjects showed profound differences in the variables related to asymmetric dimethylarginine (R2 = 66.9%, p < .01 versus 34.3%, p < .01, respectively) and symmetric dimethylarginine (R2 = 39.2%, p < .01 versus 20.6%, p = .09, respectively) levels.

Conclusions.  Our data clearly demonstrate that H. pylori infection increases asymmetric dimethylarginine levels. Moreover, this infection causes a profound metabolic modification that alters the role of the known determinants of asymmetric dimethylarginine levels. We conclude that H. pylori infection must be taken into account as a cause of increased asymmetric dimethylarginine levels and that the eradication of H. pylori may therefore lead to a decrease in asymmetric dimethylarginine levels, which is a further reason for the reduction of the risk for cardiovascular disease in this large portion of population.
Multiple Displacement Amplification of Isolated DNA from Human Gallstones: Molecular Identification of Helicobacter DNA by Means of 16S rDNA‐Based Pyrosequencing Analysis
Authors: Nilsson Isabelle , Shabo Ivan , Svanvik Joar , Monstein Hans‐Jürg
Published: November 2005

Background.  Molecular typing of Helicobacter spp. in clinical biopsy specimens has become increasingly important. By means of nested polymerase chain reaction (PCR) amplification and Southern blot analysis of the PCR amplicons, we have shown that Helicobacter spp. DNA is present in human gallstones. In this study we have investigated the possibility of using multiple displacement amplification (MDA) of isolated gallstone DNA and pyrosequencing analysis for the molecular identification of Helicobacter spp.

Materials and Methods.  DNA isolated from the nucleus of 33 human gallstones and one control strain were used in a MDA assay. Subsequently, pyrosequencing analysis was performed either directly on MDA‐DNA using primers flanking the Helicobacter spp. 16S rDNA variable V3 region or on PCR amplicons derived from broad‐range primers flanking the 16S rDNA variable V3, V4, and V9 regions.

Results.  Pyrosequencing analysis of 16S rDNA derived from MDA‐DNA revealed that Helicobacter spp.‐like DNA was present in 25 of 33 (approximately 76%) gallstones. Using an H. pylori‐specific Southern blot analysis, Helicobacter spp.‐like DNA was present in 20 of 33 [approximately 61%] of the gallstones. Using MDA‐DNA directly in pyrosequencing analysis, Helicobacter spp.‐like DNA was present in 13 of 33 [approximately 39%] gallstones.

Conclusions.  We conclude that multiple displacement amplification combined with pyrosequencing enables a rapid and accurate molecular typing of Helicobacter spp. from small and precious biopsy specimens.
Lower Prevalence of Helicobacter pylori Infection with vacAs1a, cagA‐Positive, and babA2‐Positive Genotype in Erosive Reflux Esophagitis Disease
Authors: Lai Chih‐Ho , Poon Sek‐Kwong , Chen Yun‐Chu , Chang Chi‐Sen , Wang Wen‐Ching
Published: November 2005

Background.  Increased prevalence of esophagitis has been recognized in the West. Helicobacter pylori infection, particularly virulent strains, is proposed as a protective factor against the development of gastroesophageal reflux disease. To evaluate the relationship of reflux esophagitis with virulent H. pylori infection, we studied the prevalence of reflux esophagitis among H. pylori‐infected and ‐uninfected patients and the genotype of isolates in Taiwan.

Methods.  Patients who had routine physical examination were investigated. The severity of esophagitis was evaluated using the Los Angeles grading system. H. pylori status was assessed by histology, rapid urease test, and bacterial culture. Genotyping of vacA, cagA, and babA2 was determined by polymerase chain reaction (PCR). Risk factors for severe esophagitis were evaluated.

Results.  Reflux esophagitis was found in 21.2% of 1622 patients. The prevalence of H. pylori infection was found in 33.0% of 276 patients with reflux esophagitis compared with 67.5% of 378 patients with normal esophagus (p < .001). Esophagitis occurred in a significantly lower rate among H. pylori‐positive patients with peptic ulcer than those without peptic ulcer. cagA, babA2, and vacAs1a were detected in 100% of 143 isolates. Factors that predicted severe esophagitis included age, gender, and hiatus hernia but not H. pylori infection.

Conclusions.  Our study suggests significantly lower incidence of H. pylori infection with the triple‐positive virulent genotype in patients with reflux esophagitis in Taiwan.
A Novel Mechanism of Autolysis in Helicobacter pylori: Possible Involvement of Peptidergic Substances
Authors: Fujita Yukihiro , Yamaguchi Kosaku , Kamegaya Taichi , Sato Hiroko , Semura Kayoko , Mutoh Kenichiro , Kashimoto Takashige , Ohori Hitoshi , Mukai Takao
Published: November 2005

Background.  Helicobacter pylori survival in a hostile acidic environment is known to be caused by its production of urease, which is not released by known secretion pathways. It has been proposed that H. pylori cells undergo spontaneous autolysis during cultivation and that urease becomes surface‐associated only concomitant with bacterial autolysis. The aim of this study was to elucidate mechanisms by which H. pylori cells undergo autolysis during cultivation.

Materials and Methods.  Autolysis of H. pylori KZ109 cells was estimated by measuring the turbidity of the culture, by detection of cytoplasmic protein release into the culture supernatant and by scanning electron microscopic observation of H. pylori cells during cultivation. An autolysis‐inducing factor (AIF) was partially purified from the culture supernatant by a partition method using ethyl acetate.

Results.  Bacterial turbidity of KZ109 cells was drastically decreased after late‐log phase accompanying release of urease and HspB into the extracellular space. Concomitantly, cell lytic activity was detected in the culture supernatant. Scanning electron microscopic observation suggested that partially purified AIF induced cell lysis. It was also shown that the AIF is different from other autolytic enzymes or substances so far reported.

Conclusions.  This study demonstrated the presence of the peptidergic autolytic substances in the culture supernatant of H. pylori KZ109 cells. The results of this study should be useful for further studies aimed at elucidation of the strategy of survival of H. pylori in the gastric environment and elucidation of the mechanisms of pathogenesis induced by H. pylori.
Electrophoretic Deposition of Lead Zirconate Titanate Films on Metal Foils for Embedded Components
Authors: Wu Aiying , Vilarinho P. M. , Kingon A. I.
Published: November 2005

Lead zirconate titanate (PZT) thick films in the thickness range of 5–200 μm on 20 μm copper and 25 μm platinum foils were prepared by electrophoretic deposition (EPD) for application as embedded passive components. The EPD process was conducted in glacial acetic acid medium, and the effects of deposition parameters, such as dc voltage values, processing times, and suspension aging on the film thickness and composition stoichiometry were evaluated. The dependence of the film thickness on the current and aging of the suspension was established and correlated to the chemical modifications occurring in the suspension. Films sintered at 1150°C for 30 min exhibited uniform and dense microstructures with an average grain size of 1.5 μm. A dielectric permittivity of around ∼1330 and loss tangent of 0.05 were measured for films with 8 μm of thickness. The films showed remanent polarization and a maximum polarization value of 24 and 38 μC/cm2, respectively. These properties, comparable with those of bulk PZT ceramics, suggested potential applications of the EPD process for the deposition of ferroelectric thick films on metal foils for embedded component applications.
A novel splice‐site mutation in ECM‐1 gene in a consanguineous family with lipoid proteinosis
Authors: Horev Liran , Potikha Tamara , Ayalon Sharon , Molho‐Pessach Vered , Ingber Arieh , Gany Mohamad Abdel , Edin Basel Sad , Glaser Benjamin , Zlotogorski Abraham
Published: November 2005

Abstract:  Lipoid proteinosis (LP) (OMIM 247100) is a rare, autosomal recessive disorder. Recent studies have shown that LP is the result of reduced expression of the extracellular matrix protein gene (ECM‐1), in which loss‐of‐function mutations have been described. In the present report, we describe a large consanguineous family with LP. We identified a homozygous splice‐site mutation in intron 1 (IVS1 + 1G→C) in three clinically affected patients. This is the first splice‐site mutation reported in LP and is the most 5′ of all ECM‐1 mutations described thus far. It is predicted to result in the removal of the translation initiation site, thus ablating all three known ECM‐1 isoforms (ECM‐1a, ECM‐1b, and ECM‐1c). In addition, we found a novel splicing variant that is not associated with the disease (DQ010946) and results in the generation of a short, prematurely terminating transcript. This case further emphasizes the role of ECM‐1 in LP and highlights the unresolved genotype–phenotype correlation in this disease.
Increased scratching counts depend on a decrease in ability of cutaneous prostaglandin D2 biosynthesis in NC/Nga mice with atopic dermatitis
Authors: Sugimoto M. , Arai I. , Futaki N. , Hashimoto Y. , Honma Y. , Nakaike S.
Published: November 2005

Abstract:  Spontaneous and 2,4,6‐trinitrochlorobenzene (TNCB)‐induced dermatitis models using NC/Nga mice have been recognized as animal models of atopic dermatitis. We reported that scratching behavior leads to dermatitis in a spontaneous dermatitis but not in a TNCB‐induced dermatitis. Prostaglandin D2 (PGD2) suppressed the scratching behavior of NC/Nga mice, suggesting that PGD2 plays a physiological role on inhibiting pruritus. We studied whether there was a difference in skin PG contents between spontaneous and TNCB‐induced dermatitis. Spontaneous dermatitis was induced by cohabitation with NC/Nga mice having severe skin lesions. TNCB‐induced dermatitis was caused by applications of TNCB. PGD2, PGE2, 6keto‐PGF1α, and PGF2α contents in the skin were examined using enzyme‐immunoassay kits. For studying ability to produce skin PGs, PG contents were evaluated after topical treatment of arachidonic acid (AA) or mechanical scratching. In spontaneous dermatitis, PGE2, 6keto‐PGF1α, and PGF2α contents increased with dermatitis, but only PGD2 did not do so. In TNCB‐induced dermatitis, PGD2, PGE2, 6keto‐PGF1α, and PGF2α increased. Determination of skin PG contents after AA treatment or mechanical scratching revealed that skin PGD2 production of conventional group of spontaneous dermatitis was lower than the specific pathogen‐free group. It seemed that ability of skin PGD2 production was attenuated in spontaneous dermatitis. These results suggest that enhancement of scratching behavior in spontaneous dermatitis was caused by the defect of ability to produce PGD2, which plays a physiological role in inhibiting pruritus, resulting in development of dermatitis.
The EP3 receptor stimulates ceramide and diacylglycerol release and inhibits growth of primary keratinocytes
Authors: Konger Raymond L. , Brouxhon Sabine , Partillo Steven , VanBuskirk JoAnne , Pentland Alice P.
Published: November 2005

Abstract:  Primary human keratinocytes (PHKs) are known to express the EP3 subtype of prostaglandin E2 receptor. To better understand the role of EP3 receptors in regulating epidermal function, we characterized their expression, localization, and signaling effects in human skin. Three different splice variants of the EP3 receptor (EP3A1, EP3C, and EP3D) were found to be expressed. Immunohistochemical analysis of human skin demonstrated that EP3 receptors were most prominently expressed in the basal and lower spinous layers of the epidermis. The EP3 receptor agonist sulprostone was then used to examine EP3 receptor‐dependent keratinocyte signaling pathways and functional effects. We observed that sulprostone inhibits keratinocyte growth at doses between 0.02 and 2 nM and induces sn‐1,2‐diacylglycerol (DAG) and ceramide production. Concurrent expression of the cell‐cycle inhibitory protein p21WAF1 also occurred. These data suggest that EP3 receptors produce epidermal growth inhibition through the action of DAG and ceramide second messengers.
Susceptibility to infections in elite athletes: the S‐curve
Authors: Malm C.
Published: November 2005

The susceptibility to upper respiratory tract infections (URTIs) after physical exercise has been described with a J‐shaped curve, suggesting protection from infections with moderate exercise and increased risk for URTI's in elite athletes. Several factors such as time of inoculation, previous infections, pathogen exposure, other stressors than exercise etc. can influence infection outcome. Observed infections in athletes can, therefore, be either the result of increased susceptibility to a novel pathogen, or more severe symptoms of an already established infection. Moreover, the definitions of “strenuous” exercise and “elite” athletes are equivocal, making comparisons between studies difficult.

Because absence of infections is inevitable to become and maintain status as an elite athlete, it is suggested that there is an S‐shaped relationship between exercise load and risk of infections. To become an elite athlete one has to possess state‐of‐the‐art physique, including an immune system able to withstand infections even during severe physiological and psychological stress.
The variable C‐terminal extension of G‐protein‐coupled receptor kinase 6 constitutes an accessorial autoregulatory domain
Authors: Vatter Petra , Stoesser Claudia , Samel Ines , Gierschik Peter , Moepps Barbara
Published: November 2005

G‐protein‐coupled receptor kinases (GRK) are known to phosphorylate agonist‐occupied G‐protein‐coupled receptors. We expressed and functionally characterized mouse GRK6 proteins encoded by four distinct mRNAs generated by alternative RNA splicing from a single gene, mGRK6‐A to mGRK6‐D. Three isoforms, mGRK6‐A to mGRK6‐C differ in their C‐terminal‐most portion, which is known to mediate membrane and/or receptor interaction and regulate the activity of GRK4‐like kinases. One isoform, mGRK6‐D, is identical to the other mGRK6 variants in the N‐terminal region, but carries an incomplete catalytical domain. Mouse GRK6‐D was catalytically inactive and specifically present in the nucleus of transfected cells. Recombinant mouse GRK6‐A to mGRK6‐C were found to be membrane‐associated in cell‐free systems and in transfected COS‐7 cells, suggesting that the very C‐terminus of GRK6‐A, lacking in GRK6‐B and mGRK6‐C and carrying consensus sites for palmitoylation, is not required for membrane interaction. Interestingly, the shortest catalytically active variant, mGRK6‐C, was conspicuously more active in phosphorylating light‐activated rhodopsin than mGRK6‐A and mGRK6‐B, implying that the C‐terminus of the latter two variants may fulfil an autoinhibitory function. Mutation and removal of C‐terminal‐most region of mGRK6‐A by site‐directed mutagenesis revealed that this region contains three autoregulatory elements: two discontinuous inhibitory elements consisting of a single residue, D560, and the sequence between residues S566 and L576, and an intervening stimulatory element. The results suggest that mGRK6‐C may be considered a basic, prototypic representative of the GRK4‐like kinases, which is capable of interacting with both plasma membrane and its receptor substrate, but is resistant to further regulatory modification conferred to the prototype via C‐terminal extension.
Validation of a Monoclonal Antibody-Based Capture Enzyme-Linked Immunosorbent Assay for Detection of
Campylobacter fetus
Authors: Devenish J. , Brooks B. , Perry K. , Milnes D. , Burke T. , McCabe D. , Duff S. , Lutze-Wallace C. L.
Published: November 2005

A monoclonal antibody (MAb)-based antigen capture enzyme-linked immunosorbent assay (ELISA) was compared with the routine culture methodology for the detection of
Campylobacter fetus
subspecies from bovine and ovine field samples inoculated into Clark's transport enrichment medium (TEM). The work was a collaboration between two different diagnostic laboratories, one in Canada and the other in England. In both labs, TEM samples were incubated for 4 days at 35°C and then tested by culture and ELISA. The ELISA consisted of initial screening with MAb M1825 against
C. fetus
subspecies core lipopolysaccharide (LPS). All samples positive on ELISA screening were then retested by ELISA with MAb M1825 and MAbs M1177, M1183, and M1194, which recognize serotype A- and/or serotype B-specific
C. fetus
subspecies LPS epitopes. The Canadian samples consisted of 1,060 preputial washings from 529 bulls, of which 18 were positive by both culture and ELISA and 1,042 were negative by both methods. The English samples consisted of 321 tissue specimens, mostly stomach contents and placentas, from 190 aborted ovine and bovine fetuses. A total of 262 samples were negative by culture and ELISA, 52 samples were positive by culture and ELISA, and 7 samples were culture negative but ELISA positive. The results for all 70 culture-positive isolates were confirmed by conventional biochemical methods as
C. fetus
subsp.
fetus
, with 39 presumptively identified by the ELISA as serotype A and 30 presumptively identified as serotype B and with one sample containing isolates presumptively identified as serotype A and serotype B. A receiver operating characteristic analysis of the combined ELISA data from both countries resulted in an area under the curve of 0.997, with a sensitivity of 100% and a specificity of 99.5% relative to the results of culture. The data confirm that this ELISA method can be used as an excellent test for the screening of field samples in TEM for the presence of
C. fetus
subspecies.
Alterations of Natural Killer Cell and T-Lymphocyte Counts in Adults Infected with Human Immunodeficiency Virus through Blood and Plasma Sold in the Past in China and in Whom Infection Has Progressed Slowly over a Long Period
Authors: Jiang Yongjun , Shang Hong , Zhang Zining , Diao Yingying , Dai Di , Geng Wenqing , Zhang Min , Han Xiaoxu , Wang Yanan , Liu Jing
Published: November 2005

Natural killer (NK) cells, natural killer T (NKT) cells, and T lymphocytes were analyzed by using a flow cytometer in 225 human immunodeficiency virus (HIV)-positive individuals infected through the past sale of blood and plasma without receiving antiretroviral therapy in the People’s Republic of China. According to CD4 T-cell counts these HIV-infected adults were stratified into three groups: long-term slow progressors, HIV-infected subjects, and AIDS patients. NK cell counts in long-term slow progressors were higher compared to HIV infection and AIDS patients (
P
< 0.05) and lower compared to normal controls (
P
< 0.05), whereas NKT cell counts in slow progressors and the HIV infection group were not different from those of normal controls. NK cell counts in HIV-seropositive subjects were positively correlated with CD4 T-cell counts (
P
< 0.05), and NKT cell counts were positively correlated with CD4 T-cell and CD8 T-cell counts (
P
< 0.05). The CD8 T-cell counts were higher in slow progressors compared to those with HIV infection, AIDS patients, and normal controls. These results indicated that HIV infection causes alterations of NK cells and T cells in slow progressors, HIV-infected subjects, and AIDS patient groups, but no difference was found in NKT cell counts and percentages in slow progressors and the HIV-infected group compared to normal controls.
A Strain-Specific Antigen in JapaneseHelicobacter pyloriRecognized in Sera of Japanese Children
Authors: Okuda Masumi , Sugiyama Toshiro , Fukunaga Kenichi , Kondou Masaru , Miyashiro Eikichi , Nakazawa Teruko
Published: November 2005

An enzyme immuno assay (EIA) test based on Japanese strain-derived high-molecular-weight cell-associated proteins (JHM-CAP) was evaluated by comparing with a previously developed EIA test based on a U.S. strain-derived high-molecular-weight cell-associated proteins (HM-CAP). Serum samples of 131 Japanese asymptomatic children (mean age, 5.5 years; range, 0 to 21 years) were tested that include 43 positive and 88 negative children as judged byHelicobacter pyloristool antigen test (HpSA test). Both tests showed comparable and reliable specificities, but the sensitivity of JHM-CAP EIA, at 93.0%, was much higher than that of HM-CAP EIA, at 67.4%. More false-negative results of HM-CAP were obtained in children under 10 years of age. Immunoblot analysis revealed that the JHM-CAP but not the HM-CAP preparation had a 100-kDa antigen recognized by JHM-CAP positive sera. It was concluded that JHM-CAP EIA is highly accurate for the serodiagnosis ofH. pyloriinfection in Japanese young children and that the high sensitivity of JHM-CAP EIA in contrast to HM-CAP EIA is due to the presence of a 100-kDa antigen in Japanese strains that may be recognized by the host immune system at an early stage of infection.
Xylitol Inhibits Inflammatory Cytokine Expression Induced by Lipopolysaccharide fromPorphyromonas gingivalis
Authors: Han Su-Ji , Jeong So-Yeon , Nam Yun-Ju , Yang Kyu-Ho , Lim Hoi-Soon , Chung Jin
Published: November 2005

Porphyromonas gingivalisis one of the suspected periodontopathic bacteria. The lipopolysaccharide (LPS) ofP. gingivalisis a key factor in the development of periodontitis. Inflammatory cytokines play important roles in the gingival tissue destruction that is a characteristic of periodontitis. Macrophages are prominent at chronic inflammatory sites and are considered to contribute to the pathogenesis of periodontitis. Xylitol stands out and is widely believed to possess anticaries properties. However, to date, little is known about the effect of xylitol on periodontitis. The aim of the present study was to determine tumor necrosis factor alpha (TNF-α) and interleukin-1β (IL-1β) expression when RAW 264.7 cells were stimulated withP. gingivalisLPS (hereafter, LPS refers toP. gingivalisLPS unless stated otherwise) and the effect of xylitol on the LPS-induced TNF-α and IL-1β expression. The kinetics of TNF-α and IL-1β levels in culture supernatant after LPS treatment showed peak values at 1 h (TNF-α) and 2 to 4 h (IL-1β), respectively. NF-κB, a transcription factor, was also activated by LPS treatment. These cytokine expressions and NF-κB activation were suppressed by pretreatment with pyrrolidine dithiocarbamate (an inhibitor of NF-κB). Pretreatment with xylitol inhibited LPS-induced TNF-α and IL-1β gene expression and protein synthesis. LPS-induced mobilization of NF-κB was also inhibited by pretreatment with xylitol in a dose-dependent manner. Xylitol also showed inhibitory effect on the growth ofP. gingivalis.Taken together, these findings suggest that xylitol may have good clinical effect not only for caries but also for periodontitis by its inhibitory effect on the LPS-induced inflammatory cytokine expression.
Comparison of Lipopolysaccharide-Binding Functions of CD14 and MD-2
Authors: Koraha Jun , Tsuneyoshi Naoko , Kimoto Masao , Gauchat Jean-Francois , Nakatake Hiroshi , Fukudome Kenji
Published: November 2005

Prior to being recognized by the cell surface Toll-like receptor 4/MD-2 complex, lipopolysaccharide (LPS) in the bacterial outer membrane has to be processed by LPS-binding protein and CD14. CD14 forms a complex with monomeric LPS extracted by LPS-binding protein and transfers LPS to the cell surface signaling complex. In a previous study, we prepared a functional recombinant MD-2 using a bacterial expression system. We expressed the recombinant protein inEscherichia colias a fusion protein with thioredoxin and demonstrated specific binding to LPS. In this study, we prepared recombinant CD14 fusion proteins using the same approach. Specific binding of LPS was demonstrated with a recombinant protein containing 151 amino-terminal residues. The region contained a hydrophilic region and the first three leucine-rich repeats (LRRs). The LRRs appeared to contribute to the binding because removal of the region resulted in a reduction in the binding function. LPS binding to the recombinant MD-2 was resistant to detergents. On the other hand, the binding to CD14 was prevented in the presence of low concentrations of detergents. In the case of human MD-2, the secondary myristoyl chain of LPS added by LpxM was required for the binding. A nonpathogenic penta-acyl LPS mutant lacking the myristoyl chain did not bind to MD-2 but did so normally to CD14. The broader LPS-binding spectrum of CD14 may allow recognition of multiple pathogens, and the lower affinity for LPS binding of CD14 allows transmission of captured materials to MD-2.
Persistent Replication of Human Immunodeficiency Virus Type 1 despite Treatment of Pulmonary Tuberculosis in Dually Infected Subjects
Authors: Kizza Harriet Mayanja , Rodriguez Benigno , Quinones-Mateu Miguel , Mirza Muneer , Aung Htin , Yen-Lieberman Belinda , Starkey Colleen , Horter Libby , Peters Pierre , Baseke Joy , Johnson John L. , Toossi Zahra
Published: November 2005

Tuberculosis (TB) is the most common life-threatening infection in human immunodeficiency virus (HIV)-infected persons and frequently occurs before the onset of severe immunodeficiency. Development of TB is associated with increased HIV type 1 (HIV-1) viral load, a fall in CD4 lymphocyte counts, and increased mortality. The aim of this study was to examine how treatment of pulmonary TB affected HIV-1 activity in HIV-1/TB-coinfected subjects with CD4 cell counts of >100 cells/μl. HIV-1/TB-coinfected subjects were recruited in Kampala, Uganda, and were monitored over time. Based upon a significant (0.5 log10copies/ml) decrease in viral load by the end of treatment, two patient groups could be distinguished. Responders (n= 17) had more rapid resolution of anemia and pulmonary lesions on chest radiography during TB treatment. This group had a significant increase in viral load to levels not different from those at baseline 6 months after completion of TB treatment. HIV-1 viral load in nonresponders (n= 10) with TB treatment increased and at the 6 month follow-up was significantly higher than that at the time of diagnosis of TB. Compared to baseline levels, serum markers of macrophage activation including soluble CD14 decreased significantly by the end of TB treatment in responders but not in nonresponders. These data further define the impact of pulmonary TB on HIV-1 disease. HIV-1 replication during dual HIV-1/TB infection is not amenable to virologic control by treatment of TB alone. Concurrent institution of highly active antiretroviral treatment needs to be evaluated in patients dually infected with pulmonary TB and HIV-1.
Immunological Markers of the R4 Protein of
Streptococcus agalactiae
Authors: Maeland Johan A. , Bevanger Lars , Lyng Randi Valsoe
Published: November 2005

This study focuses on immunological markers of R4, an important
Streptococcus
group B (GBS) protein. The results obtained by using rabbit antisera and purified proteins for antigens in enzyme-linked immunosorbent assay-based experiments provided evidence that R4 possesses two antigenic determinants. One of the determinants is shared with the alpha-like protein 3 (Alp3) of GBS, was named R4/Alp3 common, and was expressed by GBS, which possessed the Alp3-encoding gene
alp3
or the R4-encoding gene
rib
. The other antigenic determinant was detected only in
rib
-positive GBS organisms and was named R4 specific. This determinant probably is an immunological marker unique to the R4 protein. Neither of the antigenic R4 determinants showed serological cross-reactivity with the GBS proteins Cα, Cβ, and R3 or with alpha-like protein 2. Of 60 clinical serotype III GBS strains, 56 (93%) isolates possessed the
rib
gene and 50 (89%) of the
rib
-positive isolates expressed levels of R4 detectable by antibody-based tests, consistent with R4 expression failure or low-level expression in ∼10% of
rib
-positive GBS.
alp3
was not detected in type III GBS but was possessed by six of eight type V strains and six of six type VIII strains. All
alp3
-positive strains were recognized by the R4/Alp3 common antibodies, but none of them were recognized by the R4-specific antibodies. NCTC 9828, a reference strain for R3 and R4, expressed the determinant R4/Alp3 common but not R4 specific. A monoclonal R4 antibody, previously considered to be R4 specific and used in GBS serotyping, targeted R4/Alp3 common and is thus not R4 specific. The results show that failure to discriminate between R4 specific and R4/Alp3 common by antisera designed for GBS serotyping can result in the false identification of Alp3 as R4 or vice versa, whereas anti-R4 antibodies targeting only the determinant R4 specific will detect only R4. Both R4 and Alp3 need further evaluation with respect to the immunobiological function of each distinct antigenic determinant, for instance, with regard to their potential as GBS vaccine components.
Selected RD1 Peptides for Active Tuberculosis Diagnosis: Comparison of a Gamma Interferon Whole-Blood Enzyme-Linked Immunosorbent Assay and an Enzyme-Linked Immunospot Assay
Authors: Goletti Delia , Vincenti Donatella , Carrara Stefania , Butera Ornella , Bizzoni Federica , Bernardini Giuliana , Amicosante Massimo , Girardi Enrico
Published: November 2005

We recently set up a gamma interferon (IFN-γ) enzyme-linked immunospot assay (ELISPOT), using selected early secreted antigenic target 6 (ESAT-6) peptides, that appears specific for active tuberculosis (A-TB). However, ELISPOT is difficult to automate. Thus, the objective of this study was to determine if the same selected peptides may be used in a technique more suitable for routine work in clinical laboratories, such as whole-blood enzyme-linked immunosorbent assay (WBE). For this purpose, 27 patients with A-TB and 41 control patients were enrolled. Our WBE, using the already described selected peptides from ESAT-6 plus three new ones from culture filtrate protein 10, was performed, and data were compared with those obtained by ELISPOT. Using our selected peptides, IFN-γ production, evaluated by both WBE and ELISPOT, was significantly higher in patients with A-TB than in controls (
P
< 0.0001). Statistical analysis showed a good correlation between the results obtained by WBE and ELISPOT (
r
= 0.80,
P
< 0.001). To substantiate our data, we compared our WBE results with those obtained by QuantiFERON-TB Gold, a whole-blood assay based on region of difference 1 (RD1) overlapping peptides approved for TB infection diagnosis. We observed a slightly higher sensitivity with QuantiFERON-TB Gold than with our WBE (89% versus 81%); however, our test provided a better specificity result (90% versus 68%). In conclusion, results obtained by WBE based on selected RD1 peptides significantly correlate with those generated by ELISPOT. Moreover, our assay appears more specific for A-TB diagnosis than QuantiFERON-TB Gold, and thus it may represent a complementary tool for A-TB diagnosis for routine use in clinical laboratories.
Serological Responses in Patients with Severe Acute Respiratory Syndrome Coronavirus Infection and Cross-Reactivity with Human Coronaviruses 229E, OC43, and NL63
Authors: Chan K. H. , Cheng V. C. C. , Woo P. C. Y. , Lau S. K. P. , Poon L. L. M. , Guan Y. , Seto W. H. , Yuen K. Y. , Peiris J. S. M.
Published: November 2005

The serological response profile of severe acute respiratory syndrome (SARS) coronavirus (CoV) infection was defined by neutralization tests and subclass-specific immunofluorescent (IF) tests using serial sera from 20 patients. SARS CoV total immunoglobulin (Ig) (IgG, IgA, and IgM [IgGAM]) was the first antibody to be detectable. There was no difference in time to seroconversion between the patients who survived (
n
= 14) and those who died (
n
= 6). Although SARS CoV IgM was still detectable by IF tests with 8 of 11 patients at 7 months postinfection, the geometric mean titers dropped from 282 at 1 month postinfection to 19 at 7 months (
P
= 0.001). In contrast, neutralizing antibody and SARS CoV IgGAM and IgG antibody titers remained stable over this period. The SARS CoV antibody response was sometimes associated with an increase in preexisting IF IgG antibody titers for human coronaviruses OC43, 229E, and NL63. There was no change in IF IgG titer for virus capsid antigen from the herpesvirus that was used as an unrelated control, Epstein-Barr virus. In contrast, patients who had OC43 infections, and probably also 229E infections, without prior exposure to SARS CoV had increases of antibodies specific for the infecting virus but not for SARS CoV. There is a need for awareness of cross-reactive antibody responses between coronaviruses when interpreting IF serology.
Comparison of Galactomannan Detection, PCR-Enzyme-Linked Immunosorbent Assay, and Real-Time PCR for Diagnosis of Invasive Aspergillosis in a Neutropenic Rat Model and Effect of Caspofungin Acetate
Authors: Scotter Jennifer M. , Chambers Stephen T.
Published: November 2005

The performance of different in vitro diagnostic tests for the diagnosis of invasive aspergillosis (IA) was investigated in a transiently neutropenic rat model. Rats were immunosuppressed with cyclophosphamide and then inoculated intravenously with 1.5 × 10
4
CFU
Aspergillus fumigatus
spores. Animals were then either treated with caspofungin acetate, 1 mg/kg/day for 7 days, or not treated. PCR-enzyme-linked immunosorbent assay (ELISA), real-time PCR, and galactomannan (GM) detection were performed on postmortem blood samples, along with culture of liver, lung, and kidney homogenate. Caspofungin-treated animals showed a decrease in residual tissue burden of  
A. fumigatus
from organ homogenate compared to untreated animals (
P
 < 0.002). PCR-ELISA returned positive results for 11/17 animals treated with antifungal agents and for 10/17 untreated animals. Galactomannan was positive in 8/17 caspofungin-treated animals and 4/17 untreated animals. Real-time PCR was positive in 2/17 treated and 3/17 untreated animals. This study demonstrates that PCR-ELISA is a more sensitive test than either GM detection (
P
= 0.052) or real-time PCR (
P
< 0.01) for diagnosis of IA but that any of the three tests may return false-negative results in cases of histologically proven disease. Galactomannan indices from animals treated with antifungal agents showed a trend (
P
= 0.1) towards higher levels than those of untreated animals, but no effect was observed with PCR-ELISA indices (
P
= 0.29). GM detection, as previously described, may be enhanced by the administration of caspofungin, but PCR-ELISA appears not to be affected in the same way. We conclude that PCR-ELISA is a more sensitive and reliable method for laboratory diagnosis of IA.
Isolation and Characterization of the Promoter and Partial Enhancer Region of the Porcine Inter-α-Trypsin Inhibitor Heavy Chain 4 Gene
Authors: Harraghy Niamh , Mitchell Timothy J.
Published: November 2005

A porcine genomic library was screened for clones containing the promoter of the major acute-phase protein in pigs, inter-α-trypsin heavy chain 4 (ITIH4). Following isolation of the promoter, a functional analysis was performed with Hep3B cells. The promoter was induced by interleukin-6 (IL-6) but not by IL-1β. However, IL-1β was shown to inhibit the IL-6-induced activation of the porcine ITIH4 promoter.
Immunogenicity of Fowlpox Virus Expressing the Avian Influenza Virus H5 Gene (TROVAC AIV-H5) in Cats
Authors: Karaca Kemal , Swayne David E. , Grosenbaugh Deborah , Bublot Michel , Robles Amy , Spackman Erica , Nordgren Robert
Published: November 2005

Vaccination of cats with fowlpox virus expressing the avian influenza (AI) virus H5 hemagglutinin gene (TROVAC AI) resulted in detectable hemagglutination inhibition (HI) antibody responses to the homologous A/Turkey/Ireland/1378/83 (H5N8) (A/tky/Ire/83) AI virus antigen. The HI antibody responses to heterologous A/Chicken/Indonesia/7/03 (H5N1) (A/ck/Indonesia/03) AI virus antigen were also detected in all vaccinated cats, but only after booster vaccinations. The vaccine described in this study and other poxvirus-vectored vaccines may be of value for the prophylaxis of AI virus-associated morbidity and mortality in mammals.
Detection ofBabesia canis rossi,B. canis vogeli, andHepatozoon canisin Dogs in a Village of Eastern Sudan by Using a Screening PCR and Sequencing Methodologies
Authors: Oyamada Maremichi , Davoust Bernard , Boni Mickaël , Dereure Jacques , Bucheton Bruno , Hammad Awad , Itamoto Kazuhito , Okuda Masaru , Inokuma Hisashi
Published: November 2005

BabesiaandHepatozooninfections of dogs in a village of eastern Sudan were analyzed by using a single PCR and sequencing. Among 78 dogs, 5 were infected withBabesia canis rossiand 2 others were infected withB. canis vogeli. Thirty-three dogs were positive forHepatozoon. Hepatozoon caniswas detected by sequence analysis.
MAGNETIC STRUCTURAL EFFECT IN NONEQUILIBRIUM DEFECTIVE SOLIDS
Authors: CHIBIROVA F. KH.
Published: November 2005

Scientific study of the effect of structural memory of nonequilibrium defective solids about the processing in magnetic field (the magnetic structural effect (MSE)) was continued in this paper. The study was aimed to reveal the universal nature of the MSE, which was investigated in several new nonequilibrium defective solids. The results of investigation of the processing in the vortical magnetic field (PVMF) and its effect on the structure of the natural magnetite Fe 3 O 4 and the SnO 2 films were presented. The methods of Mössbauer and X-ray spectroscopy were used. The PVMF reduction of a defectiveness of Fe 3 O 4 structure in the magnetite was detected. The MSE was also observed in the Mössbauer spectra of diamagnetic tin oxide SnO 2 films after the PVMF. One of the possible explanations of the MSE was given in the paper.
MAGNETIC FIELD EFFECT ON ENTROPIC DISORDERING IN SPINEL SPIN GLASS Ga0.8Fe0.2NiCrO4
Authors: SRIVASTAVA J. K. , DORMANN J. L. , RAO S. M. , TREUTMANN W.
Published: November 2005

Mössbauer measurements [T (temperature) = 4.2–298 K (room temperature), H (external magnetic field) = 0, 0.25, 0.5, 2, 4, 12, 40 kOe], carried out in mixed spinel ferrite ( Ga x Fe 1-x)A [NiCr] B O 4 (x = 0.8), show the presence of pinned and entropic disorderings in this spin glass (SG) system, which is in agreement with the earlier studies. Magnetic hysteresis measurements (T = 4.2 K , H = 0-± 60 kOe ) too support this picture. The number of entropic spins in the lattice, N ES , deduced by the Mössbauer effect, is found to increase with decreasing T and decrease, rather sharply, with increasing H. These observations rule out any magnetic cluster formation by entropic spins, reveal their individual paramagnetic nature and confirm their origin in the existence of magnetically frustrated bonds in the lattice. These findings are in agreement with the predictions of the earlier developed Monte Carlo studies and cluster phase transition model.
ON THE RELATION BETWEEN FLUCTUATION AND SCALING-LAW IN GENE EXPRESSION TIME SERIES FROM YEAST TO HUMAN
Authors: NACHER J. C. , OCHIAI T. , AKUTSU T.
Published: November 2005

In this work, the dynamics of fluctuations in gene expression time series is investigated. By using collected data of gene expression from yeast and human organisms, we found that the fluctuations of gene expression level and the average value of gene expression over time are strongly correlated and obey a scaling law. As this feature is found in yeast and human organisms, it suggests that probably this coupling is a common dynamical organizing property of all living systems. To understand these observations, we propose a stochastic model which can explain these collective fluctuations, and predict the scaling exponent. Interestingly, our results indicate that the observed scaling law emerges from the self-similarity symmetry embedded in gene expression fluctuations.
SUPPORTING SOFTWARE UNDERSTANDING WITH AUTOMATED REQUIREMENTS TRACEABILITY
Authors: EGYED ALEXANDER , GRÜNBACHER PAUL
Published: November 2005

Requirements traceability (RT) aims at defining and utilizing relationships between stakeholder requirements and artifacts produced during the software development life-cycle and provides an important means to foster software understanding. Although techniques for generating and validating traceability information are available, RT in practice often suffers from the enormous effort and complexity of creating and maintaining traces. This results in invalid or incomplete trace information which cannot support engineers in real-world problems. In this paper we present a tool-supported approach that requires the designer to specify some trace dependencies but eases trace acquisition by generating others automatically. We illustrate the approach using a video-on-demand system and show how the generated traces can be used in various engineering scenarios to improve software understanding. In a case study using an open source software application we demonstrate that the approach is capable of dealing with large-scale systems and delivers valid results.
A FRAMEWORK FOR COMPARING REQUIREMENTS TRACING EXPERIMENTS
Authors: HAYES JANE HUFFMAN , DEKHTYAR ALEX
Published: November 2005

The building of traceability matrices by those other than the original developers is an arduous, error prone, prolonged, and labor intensive task. Thus, after-the-fact requirements tracing is a process where the right kind of automation can definitely assist an analyst. Recently, a number of researchers have studied the application of various methods, often based on information retrieval after-the-fact tracing. The studies are diverse enough to warrant a means for comparing them easily as well as for determining areas that require further investigation. To that end, we present here an experimental framework for evaluating requirements tracing and traceability studies. Common methods, metrics and measures are described. Recent experimental requirements tracing journal and conference papers are catalogued using the framework. We compare these studies and identify areas for future research. Finally, we provide suggestions on how the field of tracing and traceability research may move to a more mature level.