bluuebunny/crossref_metadata_2025 huggingface dataset:
Interleukin-15 Enhances Cytotoxicity, Receptor Expression, and Expansion of Neonatal Natural Killer Cells in Long-Term Culture
Authors: Choi Sunwoong S. , Chhabra Vaninder S. , Nguyen Quoc H. , Ank Bonnie J. , Stiehm E. Richard , Roberts Robert L.
Published: September 2004

Newborn infants have a higher susceptibility to various pathogens due to developmental defects in their host defense system, including deficient natural killer (NK) cell function. In this study, the effects of interleukin-15 (IL-15) on neonatal NK cells was examined for up to 12 weeks in culture. The cytotoxicity of fresh neonatal mononuclear cells (MNC) as assayed by K562 cell killing is initially much less than that of adult MNC but increases more than eightfold after 2 weeks of culture with IL-15 to a level equivalent to that of adult cells. This high level of cytotoxicity was maintained for up to 12 weeks. In antibody-dependent cellular cytotoxicity (ADCC) assays using CEM cells coated with human immunodeficiency virus gp120 antigen, IL-15 greatly increased ADCC lysis by MNC from cord blood. IL-15 increased expression of the CD16+CD56+NK markers of cord MNC fivefold after 5 weeks of incubation. Cultures of neonatal MNC with IL-15 for up to 10 weeks resulted in a unique population of CD3−CD8+CD56+cells (more than 60%), which are not present in fresh cord MNC. These results show that IL-15 can stimulate neonatal NK cells and sustain their function for several weeks, which has implications for the clinical use of IL-15.
Different Methods of Identifying New Antigenic Epitopes of Human Papillomavirus Type 16 E6 and E7 Proteins
Authors: Nakagawa Mayumi , Kim Kevin H. , Moscicki Anna-Barbara
Published: September 2004

Human papillomavirus (HPV) infection is the most common cause of sexually transmitted viral infection and is the main cause of cervical cancer. Identification of HPV T-cell epitopes would be instrumental not only in our understanding of the protective immune response but also in the development of vaccines and immunotherapies. In contrast to viruses which cause systemic infection, identification of HPV epitopes is technically challenging because HPV causes a localized mucosal infection and the frequency of pathogen-specific T lymphocytes in peripheral blood is expected to be low. Here we describe three new antigenic epitopes (E7 7-15 [TLHEYMLDL], E6 52-61 [FAFRDLCIVY], and E7 79-87 [LEDLLMGTL]) of HPV 16 E6 and E7 proteins which have oncogenic activities. E7 7-15 was identified among peptides previously shown to bind to human leukocyte antigen (HLA)-A2.1 molecule, but it was found likely to be restricted by the HLA-B48 molecule. E6 52-61 (likely to be restricted by HLA-B57) and E7 79-87 (likely to be restricted by HLA-B60) were detected, based on the magnitude of the T-cell immune responses, in another individual. In particular, T-cell clones specific for the E6 52-61 epitope were isolated effectively by magnetically selecting them based on gamma interferon secretion. This is an efficient method of identifying new epitopes of antigens for which the number of specific T lymphocytes in the circulation is expected to be small, and it should be widely applicable in identifying new T-cell epitopes.
Prospective Study of Serological Conversion as a Risk Factor for Development of Leprosy among Household Contacts
Authors: Douglas J. T. , Cellona R. V. , Fajardo T. T. , Abalos R. M. , Balagon M. V. F. , Klatser P. R.
Published: September 2004

Although the prevalence of leprosy has declined over the years, there is no evidence that incidence rates are falling. A method of early detection of those people prone to develop the most infectious form of leprosy would contribute to breaking the chain of transmission. Prophylactic treatment of serologically identified high-risk contacts of incident patients should be an operationally feasible approach for routine control programs. In addition, classification of high-risk household contacts will allow control program resources to be more focused. In this prospective study, we examined the ability of serology used for the detection of antibodies to phenolic glycolipid I of
Mycobacterium leprae
to identify those household contacts of multibacillary leprosy patients who had the highest risk of developing leprosy. After the start of multidrug therapy for the index case, a new case of leprosy developed in one in seven of the 178 households studied. In households where new cases appeared, the seropositivity rates were significantly higher (
P
< 0.001) than those in households without new cases. Seropositive household contacts had a significantly higher risk of developing leprosy (relative hazard adjusted for age and sex [aRH], 7.2), notably multibacillary leprosy (aRH = 24), than seronegative contacts.
Increased Production of Proinflammatory Cytokines following Infection with Porcine Reproductive and Respiratory Syndrome Virus andMycoplasma hyopneumoniae
Authors: Thanawongnuwech Roongroje , Thacker Brad , Halbur Patrick , Thacker Eileen L.
Published: September 2004

Induction of the proinflammatory cytokines interleukin-1 (IL-1) (α and β), IL-6, IL-8, IL-10, IL-12, and tumor necrosis factor alpha (TNF-α) in pulmonary alveolar macrophages (PAMs) was assessed following experimental infection with porcine reproductive and respiratory syndrome virus (PRRSV) and/orMycoplasma hyopneumoniaeby using in vivo and in vitro models. The in vivo model consisted of pigs infected with PRRSV and/orM. hyopneumoniaeand necropsied at 10, 28, or 42 days postinfection. Pigs infected with both pathogens had a greater percentage of macroscopic lung lesions, increased clinical disease, and slower viral clearance than pigs infected with either pathogen alone. The pigs infected with both PRRSV andM. hyopneumoniaehad significantly increased levels of mRNA for many proinflammatory cytokines in PAMs collected by bronchoalveolar lavage (BAL) at all necropsy dates compared to those in uninfected control pigs. Increased levels of IL-1β, IL-8, IL-10, and TNF-α proteins in BAL fluid, as measured by enzyme-linked immunosorbent assay, confirmed the increased cytokine induction induced by the pathogens. An in vitro model consisted ofM. hyopneumoniae-inoculated tracheal ring explants cultured with PRRSV-infected PAMs. PAMs were harvested at 6 or 15 h postinfection with either or both pathogens. The in vitro study detected increased IL-10 and IL-12 mRNA levels in PAMs infected with PRRSV at all time periods. In addition, IL-10 protein levels were significantly elevated in the culture supernatants in the presence ofM. hyopneumoniae-inoculated tracheal ring explants. The increased production of proinflammatory cytokines in vivo and in vitro associated with concurrentM. hyopneumoniaeand PRRSV infection may play a role in the increased rates of pneumonia associated with PRRSV infection. The increased levels of IL-10 may be a possible mechanism that PRRSV andM. hyopneumoniaeuse to exacerbate the severity and duration of pneumonia induced by PRRSV and modulate the respiratory immune response.
Comparison of Oral Fluid Collectors for Use in a Rapid Point-of-Care Diagnostic Device
Authors: Holm-Hansen Carol , Tong Gary , Davis Cheryl , Abrams William R. , Malamud Daniel
Published: September 2004

Orally based diagnostic testing is emerging as an alternative, noninvasive method for analyzing a variety of analytes. These analytes include pathogens, antibodies, drugs, and nucleic acids. In the present study we developed a protocol for evaluation of collectors that could be used in orally based, point-of-care diagnostics. A performance comparison was carried out with a number of commercially available collectors, and their ability to deliver fluid, proteins, bacteria, and nucleic acid from pathogens compatible with PCR was assessed. The collectors were all capable of picking up and delivering test materials, albeit at various levels.
Differential Immune Responses to Primary Measles-Mumps-Rubella Vaccination in Israeli Children
Authors: Rager-Zisman Bracha , Bazarsky Elina , Skibin Agneta , Tam Guy , Chamney Shlomo , Belmaker Ilana , Shai Iris , Kordysh Ella , Griffin Diane E.
Published: September 2004

Measles remains an important cause of morbidity and mortality worldwide, primarily due to problems associated with delivery of the live attenuated vaccine to susceptible populations. In some developed countries, there is concern about the effects of immunization on the immune system. In this study, we analyzed the responses of 12-month-old Bedouin and Jewish children living in Israel to routine measles-mumps-rubella (MMR) vaccination. Seroconversion to measles was 99% in Bedouin and 79% in Jewish children (
P
< 0.01), and that to mumps and rubella was 92 to 100% in both groups. Measles neutralizing antibody titers were higher in Bedouin (333 ± 39 mIU/ml) than Jewish (122 ± 60 mIU/ml) children (
P
< 0.002). Immunoglobulin G levels were higher in Bedouin than Jewish children (
P
= 0.007) and increased after vaccination (
P
= 0.0009). Leukocyte (
P
< 0.02) and lymphocyte (
P
= 0.04) counts were higher and CD4 lymphocyte percentages were lower (
P
< 0.001) in Bedouin than Jewish children before and after vaccination. Leukocyte counts and natural killer cell numbers did not change after vaccination, but lytic activity increased in Bedouin children (
P
< 0.005). Spontaneous proliferation of cultured peripheral blood mononuclear cells increased with vaccination, but there were no changes in the proliferative responses to phytohemagglutinin or tetanus toxoid. In summary, no adverse effects of MMR vaccination on immune function were detected. However, there were differences in underlying immunologic parameters and in response to the measles component of the vaccine between Bedouin and Jewish children. It is not known whether genetic differences or environmental exposure accounts for these differences.
Mass Value Assignment of Total and Subclass Immunoglobulin G in a Human Standard Anthrax Reference Serum
Authors: Semenova V. A. , Steward-Clark E. , Stamey K. L. , Taylor T. H. , Schmidt D. S. , Martin S. K. , Marano N. , Quinn C. P.
Published: September 2004

An anti-Anthrax Vaccine Adsorbed (anti-AVA) standard human reference serum pool, AVR414, has been prepared, and the total and protective antigen (PA)-specific immunoglobulin G (IgG) were quantified. AVR414 was prepared by plasmapheresis of healthy adults who had received a minimum of four subcutaneous injections of AVA. Mass values (in milligrams per milliliter) for total IgG and IgG subclasses 1 to 4 were determined by radial immunodiffusion. Anti-PA-specific IgG assignment (in micrograms per milliliter) was done by consensus of two complementary approaches: homologous enzyme-linked immunosorbent assay (ELISA) with affinity-purified anti-PA IgG as a calibrator and summation of mean PA-specific IgG subclass concentrations determined by IgG subclass-specific ELISA using the United States National Reference Preparation for Human Serum Proteins as a standard. The total IgG concentration assigned to AVR414 reference serum was 8.33 mg/ml. IgG subclass concentrations were the following: for IgG1, 4.48 mg/ml; for IgG2, 3.35 mg/ml; for IgG3, 0.37 mg/ml; and for IgG4, 0.30 mg/ml. The assigned mass value for total anti-PA-specific IgG was 141.2 μg/ml. Anti-PA-specific IgG subclass concentrations were the following: for IgG1, 79.6 μg/ml; for IgG2, 35.3 μg/ml; for IgG3, 3.2 μg/ml; and for IgG4, 25.3 μg/ml. Human reference serum pool AVR414 will have direct application in the standardization of anthrax serological assays, in reagent qualification, and as a standard for quantification of PA-specific IgG in humans who have been vaccinated with or otherwise exposed to
Bacillus anthracis
PA.
Comparison of Western Immunoblotting and the C6 Lyme Antibody Test for Laboratory Detection of Lyme Disease
Authors: Mogilyansky Elena , Loa Chien Chang , Adelson Martin E. , Mordechai Eli , Tilton Richard C.
Published: September 2004

Three commercial Lyme disease Western immunoblotting (WB) kits and the C6
Borrelia burgdorferi
(Lyme) enzyme-linked immunosorbent assay (ELISA) kit were compared using two commercially available performance panels from the Centers for Disease Control and Prevention (CDC) and Boston Biomedica (BBI). Combined, the panels consisted of 52 characterized specimens. Immunoglobulin G (IgG) sensitivity was similar for the three WB products. The BBI and Marblot WBs were more specific for IgG antibodies, while the Virablot was the most sensitive for IgM antibody. The BBI WB was 100% specific for IgM, while Marblot was 97% and Virablot was 77% specific for IgM. The C6 ELISA was found to be 100% sensitive. Four false-positive C6 results were identified in patients that had clinically and microbiologically confirmed Lyme disease but were not detected by the CDC reference methods. No one WB product showed overall superiority. The C6 ELISA shows promise as the first ELISA for Lyme disease that would not require a supplemental test such as a WB.
Low CD4
+
T-Lymphocyte Values in Human Immunodeficiency Virus-Negative Adults in Botswana
Authors: Bussmann Hermann , Wester C. William , Masupu Kereng V. , Peter Trevor , Gaolekwe Sarah M. , Kim Soyeon , Reich Ann Marie , Ahn Sam , Wu Ying , Thior Ibou , Essex Max , Marlink Richard
Published: September 2004

CD4
+
-lymphocyte counts (LCs) play a crucial role in the management and monitoring of HIV infection. Variability in CD4
+
LCs has been reported to occur as a result of measurement techniques and/or biological variations. We report on the CD4
+
LCs of healthy human immunodeficiency virus (HIV)-seronegative adults in Botswana. Samples were obtained from HIV-seronegative blood donors. The median CD4
+
LC was 726 cells/mm
3
(for females, 782 cells/mm
3
; for males, 698 cells/mm
3
). The median CD8
+
LC was 488 cells/mm
3
(for females, 494 cells/mm
3
; for males, 485 cells/mm
3
). The median CD4
+
-to-CD8
+
ratio was 1.57 (for females, 1.66; for males, 1.51). Our findings of low CD4
+
LCs among HIV-negative adults in Botswana are significant and have important implications for the management of HIV disease in the population of this sub-Saharan African country.
Leukotriene B4Receptor (BLT-1) Modulates Neutrophil Influx into the Peritoneum but Not the Lung and Liver during Surgically Induced Bacterial Peritonitis in Mice
Authors: Scott Melanie J. , Cheadle William G. , Hoth J. Jason , Peyton James C. , Subbarao Krishnaprasad , Shao Wen-Hai , Haribabu Bodduluri
Published: September 2004

Leukotriene B4(LTB4) is a rapidly synthesized, early neutrophil chemoattractant that signals via its cell surface receptor, BLT-1, to attract and activate neutrophils during peritonitis. BLT-1-deficient (BLT-1−/−) mice were used to determine the effects of LTB4on neutrophil migration and activation, bacterial levels, and survival after cecal ligation and puncture (CLP). Male BLT-1−/−or wild-type (WT) BALB/c mice underwent CLP. Tissues were harvested for determination of levels of bacteria, myeloperoxidase (MPO), LTB4, macrophage inflammatory protein 2 (MIP-2), and neutrophil (polymorphonuclear leukocyte [PMN]) numbers at 4 and 18 h after CLP. PMN activation was determined by an assessment of phagocytosis ability and CD11b expression. Survival was also determined. BLT-1−/−mice had decreased numbers of PMNs in the peritoneum at both 4 and 18 h after CLP but increased numbers of PMNs in the blood at 18 h compared with WT mice. Liver and lung MPO levels were significantly higher in BLT-1−/−mice at both 4 and 18 h after CLP, with increased bacterial levels in the blood, the liver, and peritoneal fluid at 4 h. Bacterial levels remained higher in peritoneal fluid at 18 h, but blood and liver bacterial levels at 18 h were not different from levels at 4 h. PMN phagocytosis and CD11b levels were decreased in BLT-1−/−mice. LTB4levels were similar between the groups before and after CLP, but MIP-2 levels were decreased both locally and systemically in BLT-1−/−mice. Survival was significantly improved in BLT-1−/−mice (71%) compared with WT mice (14%) at 48 h post-CLP. Thus, LTB4modulates neutrophil migration into the mouse peritoneum, but not the lung or liver, after CLP. Despite higher bacterial and PMN levels at remote sites, there was increased survival in BLT-1−/−mice compared to WT mice. Decreased PMN activation may result in less remote organ dysfunction and improved survival.
Epitope-Specific Antibody Levels Demonstrate Recognition of New Epitopes and Changes in Titer but Not Affinity during Treatment of Tuberculosis
Authors: Bothamley Graham H.
Published: September 2004

Antibody levels rise during treatment of tuberculosis. This study examined when this rise occurred, whether there was recognition of new antigen binding sites (epitopes) on the same or different antigens, and how long specific antibody persisted. Forty patients with smear-positive pulmonary tuberculosis provided serum before and during treatment. Antibody levels were measured using a monoclonal antibody competition assay to epitopes restricted to theMycobacterium tuberculosiscomplex and an enzyme-linked immunosorbent assay for lipoarabinomannan. Significant increases in antibody levels were apparent after 7 days of treatment. Five samples (12.5%) had positive titers to all epitopes at the start of treatment, and this increased to 23 (58%) during treatment. Antibody to epitopes with the poorest sensitivity (the TB23 epitope of the 19-kDa antigen and the TB78 epitope of hsp65) showed the greatest increases after treatment. Antibody to these two epitopes was also absent in some patients with relapsed tuberculosis until after treatment. Antibody titers showed a biphasic response, with a fall at 2 to 3 months of treatment. Sera from two patients showed changes in the affinity of epitope-specific antibody during treatment, whereas the majority did not. Those infected with isoniazid-resistant strains ofM. tuberculosisshowed a late rise in antibody. Antibody to the TB68 epitope of the 16-kDa α-crystallin homolog was short-lived, but it recurred with bacteriological relapse during treatment. Positive antibody titers persisted for at least 3 to 18 months after treatment. Diagnostic tests for tuberculosis should be evaluated using only pretreatment sera. Delayed antigenic recognition could be due to active suppression and/or failure to engage internal antigens ofM. tuberculosis.
Analysis of Possible Factors Affecting the Specificity of the Gamma Interferon Test in Tuberculosis-Free Cattle Herds
Authors: Cagiola M. , Feliziani F. , Severi G. , Pasquali P. , Rutili D.
Published: September 2004

Bovine tuberculosis (TB) is still a zoonotic problem in the world. Despite the fact that eradication programs for bovine TB are being implemented in many countries, it remains a public health problem. These programs are mainly based on a single intradermal tuberculin test using bovine tuberculin purified protein derivative (PPD), isolation, and slaughtering of infected animals. The aim of this study was to assess the specificity of the gamma interferon (IFN-γ) test in TB-free cattle herds, by using not only Australian tuberculins but also tuberculins produced at our institute, and to correlate the response with the type of production (beef cattle, dairy cattle, and a dual-purpose breed), the housing system, and the age of the animals. We studied 800 animals selected from 20 TB- and paratuberculosis-free herds. The animals were tested in parallel, after stimulation with Australian tuberculins and tuberculins produced at our institute, by using the skin test and two IFN-γ assays. The results of this trial showed that the specificity of the IFN-γ test is higher than that of the skin test (96.8%) and ranges from 97.3% (using only Australian tuberculins) to 98.6% (using tuberculins produced at our institute). We found that different categories of cattle could influence the specificity of the skin test but that these differences tended to be reduced in the IFN-γ assay, especially when Italian PPDs were used.
Response to Superantigen Stimulation in Peripheral Blood Mononuclear Cells from Children Perinatally Infected with Human Immunodeficiency Virus and Receiving Highly Active Antiretroviral Therapy
Authors: McCloskey Thomas W. , Haridas Viraga , Pontrelli Lucy , Pahwa Savita
Published: September 2004

Our understanding of the pathogenesis of perinatal human immunodeficiency virus (HIV) infection is still evolving. We sought to characterize the response to the bacterial superantigenStaphylococcusenterotoxin B (SEB) of lymphocytes from HIV-infected children receiving treatment with highly active antiretroviral therapy (HAART). Using the flow cytometric methodology, we quantified apoptosis, proliferation, cytokine production, and activation antigen upregulation in CD4 and CD8 T lymphocytes following in vitro stimulation of peripheral blood mononuclear cells (PBMCs) with SEB. The levels of proliferation, CD4 interleukin-2 (IL-2) production, CD8 gamma interferon (IFN-γ) production, and upregulation of CD69 expression by cells from HIV-infected children were indistinguishable from those by cells from controls. However, stimulation with SEB dramatically decreased the ratio of resting apoptotic cells to cycling apoptotic cells in the controls but not in the patients. In addition, unstimulated spontaneous apoptosis of CD4 T cells remained greater in the patients than in the controls. The percentages of IL-2-positive CD8 T cells and IFN-γ-positive CD4 T cells following SEB stimulation were significantly lower in the patients than in the controls. Our multiparameter approach was able to demonstrate differences in lymphocyte superantigen responsiveness in HIV-infected children receiving HAART in comparison to that in uninfected controls, notably, an apoptotic versus a proliferative response to stimulation.
CXCR2 Blockade InfluencesAnaplasma phagocytophilumPropagation but Not Histopathology in the Mouse Model of Human Granulocytic Anaplasmosis
Authors: Scorpio Diana G. , Akkoyunlu Mustafa , Fikrig Erol , Dumler J. Stephen
Published: September 2004

Anaplasma phagocytophilumis an obligate intracellular bacterium that infects neutrophils and causes human granulocytic anaplasmosis. Infection induces neutrophil secretion of interleukin-8 or murine homologs and perpetuates infection by recruiting susceptible neutrophils. We hypothesized that antibody blockade of CXCR2 would decreaseA. phagocytophilumtissue load by interrupting neutrophil recruitment but would not influence murine hepatic pathology. C3H-scidmice were treated with CXCR2 antiserum or control prior to or on day 14 after infection. Quantitative PCR and immunohistochemistry forA. phagocytophilumwere performed and severity of liver histopathology was ranked. Control mice had more infected cells in tissues than the anti-CXCR2-treated group. The histopathological rank was not different between treated and control animals. Infected cells of control mice clustered in tissue more than in treated mice. The results support the hypothesis of bacterial propagation through chemokine induction and confirm that tissue injury is unrelated toA. phagocytophilumtissue load.
Membrane-Anchored CD14 Is Important for Induction of Interleukin-8 by Lipopolysaccharide and Peptidoglycan in Uroepithelial Cells
Authors: Shimizu Toshiaki , Yokota Shin-ichi , Takahashi Satoshi , Kunishima Yasuharu , Takeyama Koh , Masumori Naoya , Takahashi Atsushi , Matsukawa Masanori , Itoh Naoki , Tsukamoto Taiji , Fujii Nobuhiro
Published: September 2004

We investigated the induction of interleukin-8 (IL-8) by bacterial lipopolysaccharide (LPS) and peptidoglycan (PGN) in the bladder cancer cell lines T24, 5637, UM-UC-3, and HT1197. T24 and 5637 cells strongly induced IL-8 after stimulation with LPS or PGN in a dose- and time-dependent manner, whereas UM-UC-3 and HT1197 cells did so very weakly. The expression of CD14 at the mRNA, total cellular protein, and cell surface protein levels differed among these cell lines, but the expression levels of Toll-like receptors 2 and 4 (TLR2 and TLR4) were not significantly different. The CD14 expression levels were found to correlate with the inducibility of IL-8 by LPS or PGN. Treatment of T24 and 5637 cells with phosphatidylinositol-specific phospholipase C to eliminate CD14 from the cell surface dramatically suppressed the induction of IL-8. On the other hand, UM-UC-3 cells transfected with CD14 cDNA expressed membrane-anchored CD14 and showed more efficent induction of IL-8 by LPS stimulation than untransfected controls. These results suggest that the presence of the membrane-anchored, but not the soluble, form of CD14 is a strong factor in IL-8 induction in bladder epithelial cells in response to bacterial components. The presence of the membrane-anchored form of CD14 may thus be a determinant for the inflammatory response of uroepithelial cells.
Antigen-Specific Gene Expression Profiles of Peripheral Blood Mononuclear Cells Do Not Reflect Those of T-Lymphocyte Subsets
Authors: McLaren Paul J. , Mayne Michael , Rosser Stuart , Moffatt Teri , Becker Kevin G. , Plummer Francis A. , Fowke Keith R.
Published: September 2004

Advances in microarray technology have allowed for the monitoring of thousands of genes simultaneously. This technology is of particular interest to immunologists studying infectious diseases, because it provides tremendous potential for investigating host-pathogen interactions at the level of immune gene expression. To date, many studies have focused either on cell lines, where the physiological relevance is questionable, or on mixed cell populations, where the contributions of individual subpopulations are unknown. In the present study, we perform an intrasubject comparison of antigen-stimulated immune gene expression profiles between a mixed population of peripheral blood mononuclear cells (PBMC) and the two predominant cell types found in PBMC, CD4
+
and CD8
+
T lymphocytes. We show that the microarray profiles of CD4
+
and CD8
+
T lymphocytes differ from each other as well as from that of the mixed cell population. The independence of the gene expression profiles of different cell types is demonstrated with a ubiquitous antigen (
Candida albicans
) as well as with a disease-specific antigen (human immunodeficiency virus p24). This study has important implications for microarray studies of host immunity and underscores the importance of profiling the expression of specific cell types.
Age-Specific Immune Response to HspA in
Helicobacter pylori
-Positive Persons in Mexico
Authors: Eamranond Peter P. , Torres Javier , Muñoz Onofre , Pérez-Pérez Guillermo I.
Published: September 2004

The immune response to heat shock protein A (HspA) in
Helicobacter pylori
-positive adults increases with age in developed countries. This response has not been studied with children or in developing countries (G. I. Pérez-Pérez, J. M. Thiberge, A. Labigne, and M. J. Blaser, J. Infect. Dis. 174:1046-1050, 1996). As determined by using a specific enzyme-linked immunosorbent assay, HspA seropositivity among 592 individuals in Mexico was <10% in children and increased to >40% in adults.
Is the Perceived Association between
Chlamydia pneumoniae
and Vascular Diseases Biased by Methodology?
Authors: Maraha Boulos , Berg Hans , Kerver Marjolein , Kranendonk Steef , Hamming Jaap , Kluytmans Jan , Peeters Marcel , van der Zee Anneke
Published: September 2004

Inter- and intralaboratory inconsistencies in detection rates of
Chlamydia pneumoniae
in vascular specimens have been demonstrated. In this study, 66 vascular tissue specimens from 66 patients with vascular disease were tested by three PCR assays: a 16S PCR-based reverse line blot (RLB) assay, a single-step PCR, and a nested PCR. Also, we explored the impacts of different DNA polymerase enzymes on the results based on gel electrophoresis and hybridization. The PCR results by gel electrophoresis in the single-step PCR depended on which DNA polymerase was used. All samples were negative with AmpliTaq Gold DNA polymerase, and 54.5% (36 of 66) were positive with the conventional
Taq
DNA polymerase. All samples were negative after hybridization with a
C. pneumoniae
-specific probe. In the nested PCR, all specimens were negative by gel electrophoresis and after hybridization. The RLB assay failed to detect
C. pneumoniae
in any specimen; however, 20 specimens were
Chlamydia
sp. positive. The sequence analysis of six of these samples demonstrated
Chlamydia
-like organisms. RLB detected
Chlamydia
sp. DNA in water and in the elution buffer after passage of the Qiagen columns (11 of 40). This study identified factors that may influence the detection of
C. pneumoniae
DNA in vascular tissues and consequently bias the perception of a link between
C. pneumoniae
and vascular diseases. The following are strongly recommended: to use DNA polymerases that have to be activated, to decontaminate with dUTP-uracil-DNA glycosylase, to hybridize with specific probes, to include sufficient controls, and to use molecular grade water.
Evaluation ofTrichinella spiralisLarva Group 1 Antigens for Serodiagnosis of Human Trichinellosis
Authors: Escalante Marcela , Romarís Fernanda , Rodríguez Mercedes , Rodríguez Esperanza , Leiro José , Gárate María T. , Ubeira Florencio M.
Published: September 2004

To identifyTrichinellaantigens suitable for high-specificity and high-sensitivity serodiagnosis of human trichinellosis, we evaluated assays using four antigens: (i) crude first-stage larval extract (CLE), (ii) O-deglycosylated CLE, (iii) tyvelose-bearing antigens (Trichinella spiralislarva group 1 [TSL-1] antigens) purified by US4 affinity chromatography and coupled directly to enzyme-linked immunosorbent assay (ELISA) plates (pTSL-1 antigens), and (iv) TSL-1 antigens immobilized on ELISA plates with the monoclonal antibody (MAb) US4 (cTSL-1 antigens). Assays using these antigens were compared by analysis of sera from healthy individuals (n= 224) (group 1), individuals with noninfectious intestinal pathologies (n= 114) (group 2), individuals with other parasitic infections (n= 107) (group 3), and individuals with confirmed trichinellosis (n= 42) (group 4). Our results indicate that capture ELISA using cTSL-1 antigens is the most effective method for serodiagnosis of human trichinellosis; this was the only method showing 100% specificity and 100% sensitivity at the patent stage of the infection, and it was also the most sensitive for sera obtained prior to patency in indirect immunofluorescence (IIF). Indirect ELISA with pTSL-1 antigens was also 100% specific but was slightly less sensitive, particularly with sera obtained before IIF patency. Inhibition ELISA with MAb US4 indicated (i) that inTrichinella-infected patients the immune response to TSL-1 antigens is directed mostly against tyvelose-containing epitopes (mean of 84.2% of total anti-TSL-1 immunoglobulin G1 [IgG1] antibody response [range, 51.3 to 97.6%]) and (ii) that in most individuals a large proportion of anti-CLE IgG1 antibodies (mean, 49.5%; range, 7.3 to 92.6%) are directed against tyvelose epitopes.
A Predominant and Virulent
Legionella pneumophila
Serogroup 1 Strain Detected in Isolates from Patients and Water in Queensland, Australia, by an Amplified Fragment Length Polymorphism Protocol and Virulence Gene-Based PCR Assays
Authors: Huang Bixing , Heron Brett A. , Gray Bruce R. , Eglezos Sofroni , Bates John R. , Savill John
Published: September 2004

In epidemiological investigations of community legionellosis outbreaks, knowledge of the prevalence, distribution, and clinical significance (virulence) of environmental
Legionella
isolates is crucial for interpretation of the molecular subtyping results. To obtain such information for
Legionella pneumophila
serogroup 1 isolates, we used the standardized amplified fragment length polymorphism (AFLP) protocol of the European Working Group on Legionella Infection to subtype
L. pneumophila
SG1 isolates obtained from patients and water sources in Queensland, Australia. An AFLP genotype, termed AF1, was predominant in isolates from both patients (40.5%) and water (49.0%). The second most common AFLP genotype found in water isolates was AF16 (36.5%), but this genotype was not identified in the patient isolates. When virulence gene-based PCR assays for
lvh
and
rtxA
genes were applied to the isolates from patients and water, nearly all (65 of 66) AF1 strains had both virulence genes,
lvh
and
rtxA
. In contrast, neither the
lvh
nor the
rtxA
gene was found in the AF16 strains, except for one isolate with the
rtxA
gene. It appears that this may explain the failure to find this genotype in the isolates from patients even though it may be common in the environment. In view of the evidence that the AF1 genotype is the most common genotype among strains found in patients and water sources in this region, any suggested epidemiological link derived from comparing the AF1 genotype from patient isolates with the AF1 genotype from environmental isolates must be interpreted and acted on with caution. The use of virulence gene-based PCR assays applied to environmental samples may be helpful in determining the infection potential of the isolates involved.
Phenotypic and Molecular Characteristics of
Streptococcus agalactiae
Isolates Recovered from Milk of Dairy Cows in Brazil
Authors: Duarte Rafael S. , Miranda Otávio P. , Bellei Bruna C. , Brito Maria Aparecida V. P. , Teixeira Lúcia M.
Published: September 2004

Information on the characteristics of
Streptococcus agalactiae
obtained from bovine sources in Brazil is still very limited. The aim of this study was to assess the phenotypic and genotypic diversity among
S. agalactiae
isolates from milk of dairy cows presenting clinical or subclinical mastitis in the southeast region of Brazil. Phenotypic characterization was based on physiological and serological tests. Antimicrobial susceptibility tests were carried out by the disk method. Genetic diversity was evaluated by using random amplified polymorphic DNA-PCR (RAPD-PCR) (by using the primer 1254) and pulsed-field gel electrophoresis (PFGE) (by using SmaI as the restriction enzyme) and by PCRs for detection of genes associated with resistance to erythromycin and tetracycline as well as PCRs for detection of genes coding for cell surface-associated proteins. According to the results of physiologic tests, 45 (52.9%) isolates showed beta-hemolysis and 44 (51.7%) were susceptible to bacitracin. Fourteen different biotypes were detected. The two most frequent biotypes comprised strains that were non-beta-hemolytic; fermented galactose, lactose, and salicin; produced protease; and were negative for DNase production. Serotype III was predominant (66 isolates [77.6%]), followed by serotypes II, Ia, Ib, and VI. Resistance to tetracycline and erythromycin was found in 38 (44.7%) and 9 (10.5%) isolates, respectively, with
tet
(O) (31.7%) and
erm
(B) (100%) being the most frequently occurring resistance genes. Three genes coding for surface proteins,
bca
,
lmb
, and
scpB
, were detected in 55 (64.7%), 7 (8.2%), and 43 (50.5%) isolates, respectively. In most cases, isolates from animals in the same herd presented closely related genetic profiles (determined by either RAPD-PCR or PFGE), which were distinct from those of isolates from different herds.
Whole-Blood Counting Immunoassay as a Short-Turnaround Test for Detection of Hepatitis B Surface Antigen, Anti-Hepatitis C Virus Antibodies, and Anti-Treponema pallidumAntibodies
Authors: Kudo Toyoichiro , Kido Aiko , Nishiyama Yukiko , Koganeya Hiroshi , Okuda Takako , Nabeshima Motoshige , Iinuma Yoshitsugu , Ichiyama Satoshi
Published: September 2004

Whole-blood samples were used for a counting immunoassay (CIA) with the aim of developing a short- turnaround test. After optimization of the CIA, hepatitis B surface antigen (HBsAg), anti-hepatitis C virus antibodies (anti-HCV), and anti-Treponema pallidumantibodies (anti-TP) were detected as efficiently as by an enzyme immunoassay (EIA) with serum samples. The correlations between whole-blood CIA and serum EIA were 99.8, 97.1, and 99.4% for HBsAg, anti-HCV, and anti-TP, respectively. Whole-blood CIA may be of value when rapid screening of many samples is required.
Uses of
Staphylococcus aureus
GeneChips in Genotyping and Genetic Composition Analysis
Authors: Dunman P. M. , Mounts W. , McAleese F. , Immermann F. , Macapagal D. , Marsilio E. , McDougal L. , Tenover F. C. , Bradford P. A. , Petersen P. J. , Projan S. J. , Murphy E.
Published: September 2004

Understanding the relatedness of strains within a bacterial species is essential for monitoring reservoirs of antimicrobial resistance and for epidemiological studies. Pulsed-field gel electrophoresis (PFGE), ribotyping, and multilocus sequence typing are commonly used for this purpose. However, these techniques are either nonquantitative or provide only a limited estimation of strain relatedness. Moreover, they cannot extensively define the genes that constitute an organism. In the present study, 21 oxacillin-resistant
Staphylococcus aureus
(ORSA) isolates, representing eight major ORSA lineages, and each of the seven strains for which the complete genomic sequence is publicly available were genotyped using a novel GeneChip-based approach. Strains were also subjected to PFGE and ribotyping analysis. GeneChip results provided a higher level of discrimination among isolates than either ribotyping or PFGE, although strain clustering was similar among the three techniques. In addition, GeneChip signal intensity cutoff values were empirically determined to provide extensive data on the genetic composition of each isolate analyzed. Using this technology it was shown that strains could be examined for each element represented on the GeneChip, including virulence factors, antimicrobial resistance determinants, and
agr
type. These results were validated by PCR, growth on selective media, and detailed in silico analysis of each of the sequenced genomes. Collectively, this work demonstrates that GeneChips provide extensive genotyping information for
S. aureus
strains and may play a major role in epidemiological studies in the future where correlating genes with particular disease phenotypes is critical.
Potential Screening Assay for Undetectable Viruses on the Basis of Their Capacity To Induce Alpha Interferon Production
Authors: Schmidt Barbara , Ashlock Brittany , Neipel Frank , Levy Jay A.
Published: September 2004

High interferon production by plasmacytoid dendritic cells (PDC) was unexpectedly noted after their coculture with CD4
+
cells from a healthy donor whose cells subsequently showed human herpesvirus type 6 and 7 infections. This release of interferon was not observed with uninfected normal CD4
+
cells. Induction of PDC interferon production could help screen for covert virus infections.
Multiplex PCR Method for Identifying Recombinant Vaccine-Related Polioviruses
Authors: Kilpatrick David R. , Ching Karen , Iber Jane , Campagnoli Ray , Freeman Christopher J. , Mishrik Nada , Liu Hong-Mei , Pallansch Mark A. , Kew Olen M.
Published: September 2004

The recent discovery of recombinant circulating vaccine-derived poliovirus (recombinant cVDPV) has highlighted the need for enhanced global poliovirus surveillance to assure timely detection of any future cVDPV outbreaks. Six pairs of Sabin strain-specific recombinant primers were designed to permit rapid screening for VDPV recombinants by PCR.
Detection and Identification of
Actinobacillus pleuropneumoniae
Serotypes 1, 2, and 8 by Multiplex PCR
Authors: Schuchert Jennifer A. , Inzana T. J. , Angen Ø. , Jessing S.
Published: September 2004

Multiplex PCR assays were developed to identify
Actinobacillus pleuropneumoniae
serotypes 1, 2, and 8. Primers designed for the conserved capsular polysaccharide (CP) export region amplified a 489-bp DNA fragment from all serotypes. Primers specific to the CP biosynthesis regions of serotypes 1, 2, and 8 amplified fragments of 1.6 kb, 1.7 kb, and 970 bp from only their respective serotypes.
Edible ectomycorrhizal mushrooms: challenges and achievements
Authors: Yun Wang , Hall Ian R
Published: September 2004

There are about 2500 recorded species of edible mushrooms. The most expensive and sought after of these mushrooms belong to the mycorrhizal group and include Tuber melanosporum Vitt. (Périgord black truffle), Tuber magnatum Pico & Vitt. (Italian white truffle), Tricholoma matsutake (Ito & Imai) Sing. (matsutake), Boletus edulis Bull: Fr. sensu lato (porcini) Cantharellus cibarius Fr.: Fr. (chanterelle), and Amanita caesarea (Scop.: Fr.) Pers: Schw. (Caesar's mushroom). The total market for these is measured in billions of US dollars. Over the past 100 years, harvests of many mycorrhizal mushrooms have declined dramatically, which has prompted interest in the development of methods for their cultivation. So far only a few species of truffles have been produced in commercial quantities, although methods have been developed that may see the cultivation of species such as Cantharellus cibarius, Lyophyllum shimeji (Kawam.) Hongo, and Lactarius deliciosus (L.: Fr.) Gray. Despite this, many of the most expensive mycorrhizal mushrooms, including Tuber magnatum Pico & Vitt. and Tricholoma matsutake, have defied cultivation. Our paper will attempt to highlight possible reasons why mycorrhizal mushrooms have proven to be so difficult to grow and how we might better manage mycorrhizal mushroom forests to sustain natural production.Key words: edible, ectomycorrhiza, mushroom, truffle.
Dissipated strain energy method for determining preconsolidation pressure
Authors: Wang L B , Frost J D
Published: September 2004

The dissipated strain energy method (DSEM), a new method for determining the preconsolidation pressure, is presented in this paper. Compared with the energy method, the DSEM uses dissipated strain energy and the slope of the unloadingreloading cycle (in the strain energy  effective consolidation stress space) for the plot to minimize the sample disturbance effects and eliminate the effect of elastic deformation. Dissipated strain energy, in terms of micro mechanics, is directly related to the irreversible process of consolidation and can be supported by theories dealing with consolidation and compaction. The use of the unloadingreloading slope to simulate the elastic reloading for the recompression stage can minimize sample disturbance effects. Examples presented indicate that the proposed new method is less operator dependent than most of the existing methods.Key words: dissipated strain energy, preconsolidation pressure, consolidation, energy method, dissipated strain energy method.
Variation of paternal contribution in a seed orchard of Cryptomeria japonica determined using microsatellite markers
Authors: Moriguchi Yoshinari , Taira Hideaki , Tani Naoki , Tsumura Yoshihiko
Published: September 2004

We investigated gene flow and pollen contamination in a seed orchard of Cryptomeria japonica D. Don using six microsatellite markers. The quality of a seed crop is determined by many factors, including the rate of pollen contamination from outside sources, degree of self-fertilization, and equality of the clones as pollen donors. The surveyed seed orchard consisted of 62 clones and a total of 360 ramets. The seeds from 12 mother trees in the seed orchard were investigated using six highly polymorphic loci with high multipaternity exclusion probability (0.999). The estimated average pollen contamination rate was high at 47.78% (±4.12%), and the clonal self-fertilization rate was 2.22% (±0.94%) when we assumed that null alleles were present. The high pollen contamination rate could dramatically reduce the quality of the seed crop. The contamination rate differed from tree to tree, but the differences were not related to the location of each individual. The levels of contribution as pollen donors differed significantly amongst clones in the seed orchard. Differences in flowering phenology and relative pollen fecundity may be responsible for the differences in paternal contributions. The effective distance of pollen dispersal in this C. japonica seed orchard seemed to be greater than previously estimated.
FREE FIELD CONSTRUCTIONS FOR THE ELLIPTIC ALGEBRA $\rscr{A}_{q,p}(\widehat{sl}_2)$ AND BAXTER'S EIGHT-VERTEX MODEL
Authors: SHIRAISHI J.
Published: September 2004

Three examples of free field constructions for the vertex operators of the elliptic quantum group [Formula: see text] are obtained. Two of these ( for p1/2=±q3/2, p1/2=-q2) are based on representation theories of the deformed Virasoro algebra, which correspond to the level 4 and level 2 Z-algebra of Lepowsky and Wilson. The third one (p1/2=q3) is constructed over a tensor product of a bosonic and a fermionic Fock spaces. The algebraic structure at (p1/2=q3), however, is not related to the deformed Virasoro algebra. Using these free field constructions, an integral formula for the correlation functions of Baxter's eight-vertex model is obtained. This formula shows different structure compared with the one obtained by Lashkevich and Pugai.
SOFTWARE WAREHOUSE: ITS DESIGN, MANAGEMENT AND APPLICATION
Authors: DAI HONGHUA , DAI WEI , LI GANG
Published: September 2004

To have an effective and efficient mechanism to store, manage and utilize software sources is essential to the automation of software engineering. The paper presents an innovative approach in managing software resources using software warehouse where software assets are systematically accumulated, deposited, retrieved, packaged, managed and utilized, driven by data-mining and OLAP technologies. The results lead to streamlined high efficient software development process and enhance the productivity in response to modern challenges of the design and development of software applications.
IDENTIFYING SOFTWARE COMPONENT ASSOCIATION WITH GENETIC ALGORITHM
Authors: YAN XIAOWEI , ZHANG CHENGQI , ZHANG SHICHAO
Published: September 2004

Identifying software component association is useful for component management and component retrieval. In this paper we design an evolutionary strategy to understand software structure better and identify software component association, by using genetic algorithm. Our mining strategy is effective for global search, especially when the searched space is so large that it is hardly possible to use deterministic search method.
APPLICATION OF CLUSTERING TECHNIQUES TO SOFTWARE COMPONENT ARCHITECTURE DESIGN
Authors: LO SHU-CHUAN , CHANG JIA-HUI
Published: September 2004

This paper describes the application of two variants of clustering techniques to a matrix of requirements (use cases) vs. design units (classes), with the aim of augmenting the design of software component architectures. One algorithm used a modified COMO matrix-clustering algorithm, while the other is average-linkage agglomerative clustering based on similarity among classes as a measure of distance. Our results provide a shortcut for software engineers with limited experience in Component-Based Software Development (CBSD) who are constructing component architectures. This paper also evaluates the correlations between these two clustered results. We solicited feedback from experienced software developers who confirmed that the inferred hierarchies conformed to their conceptualizations of the software structure.
DATA MINING FOR VALIDATION IN SOFTWARE ENGINEERING: AN EXAMPLE
Authors: KAJKO-MATTSSON MIRA , CHAPIN NED
Published: September 2004

Consider two independently done software engineering studies that used different approaches to cover some of the same subject area, such as software maintenance. Although done differently and for different purposes, to what extent can each study serve as a validation of the other? Within the scope of the subject area overlap, data mining can be applied to provide a quantitative assessment. This paper reports on the data mining that attempted to cross validate two independently done and published software engineering studies of software maintenance, one on a corrective maintenance maturity model, and the other on an objective classification of software maintenance activities. The data mining established that each of the two independently done studies effectively and very strongly validates the other.