bluuebunny/crossref_metadata_2025 huggingface dataset:
Production ofChlamydia pneumoniaeProteins inBacillus subtilisand Their Use in Characterizing Immune Responses in the Experimental Infection Model
Authors: Airaksinen Ulla , Penttilä Tuula , Wahlström Eva , Vuola Jenni M. , Puolakkainen Mirja , Sarvas Matti
Published: May 2003

Due to intracellular growth requirements, large-scale cultures of chlamydiae and purification of its proteins are difficult and laborious. To overcome these problems we produced chlamydial proteins in a heterologous host,Bacillus subtilis, a gram-positive nonpathogenic bacterium. The genes ofChlamydia pneumoniaemajor outer membrane protein (MOMP), the cysteine-rich outer membrane protein (Omp2), and the heat shock protein (Hsp60) were amplified by PCR, and the PCR products were cloned into expression vectors containing a promoter, a ribosome binding site, and a truncated signal sequence of the α-amylase gene fromBacillus amyloliquefaciens. C. pneumoniaegenes were readily expressed inB. subtilisunder the control of the α-amylase promoter. The recombinant proteins MOMP and Hsp60 were purified from the bacterial lysate with the aid of the carboxy-terminal histidine hexamer tag by affinity chromatography. The Omp2 was separated as an insoluble fraction after 8 M urea treatment. The purified proteins were successfully used as immunogens and as antigens in serological assays and in a lymphoproliferation test. The Omp2 and Hsp60 antigens were readily recognized by the antibodies appearing after pulmonary infection following intranasal inoculation ofC. pneumoniaein mice. Also, splenocytes collected from mice immunized with MOMP or Hsp60 proteins proliferated in response to in vitro stimulation with the corresponding proteins.
NF-κB Is Involved in Regulation of CD40 Ligand Expression onMycobacterium bovisBacillus Calmette-Guérin-Activated Human T Cells
Authors: Méndez-Samperio Patricia , Ayala Hilda , Vázquez Abraham
Published: May 2003

Interaction between CD40L (CD154) on activated T cells and its receptor CD40 on antigen-presenting cells has been reported to be important in the resolution of infection by mycobacteria. However, the mechanism(s) by whichMycobacterium bovisbacillus Calmette-Guérin (BCG) up-regulates membrane expression of CD40L molecules is poorly understood. This study was done to investigate the role of the nuclear factor κB (NF-κB) signaling pathway in the regulation of CD40L expression in human CD4+T cells stimulated with BCG. Specific pharmacologic inhibition of the NF-κB pathway revealed that this signaling cascade was required in the regulation of CD40L expression on the surface of BCG-activated CD4+T cells. These results were further supported by the fact that treatment of BCG-activated CD4+T cells with these pharmacological inhibitors significantly down-regulated CD40L mRNA. In this study, inhibitor κBα (IκBα) and IκBβ protein production was not affected by the chemical protease inhibitors and, more importantly, BCG led to the rapid but transient induction of NF-κB activity. Our results also indicated that CD40L expression on BCG-activated CD4+T cells resulted from transcriptional up-regulation of the CD40L gene by a mechanism which is independent of de novo protein synthesis. Interestingly, BCG-induced activation of NF-κB and the increased CD40L cell surface expression were blocked by the protein kinase C (PKC) inhibitors 1-[5-isoquinolinesulfonyl]-2-methylpiperazine and salicylate, both of which block phosphorylation of IκB. Moreover, rottlerin a Ca2+-independent PKC isoform inhibitor, significantly down-regulated CD40L mRNA in BCG-activated CD4+T cells. These data strongly suggest that CD40L expression by BCG-activated CD4+T cells is regulated via the PKC pathway and by NF-κB DNA binding activity.
Humoral Immune Responses to S-Layer-Like Proteins of
Bacteroides forsythus
Authors: Yoneda Masahiro , Hirofuji Takao , Motooka Noriko , Nozoe Koji , Shigenaga Kayoko , Anan Hisashi , Miura Mayumi , Kabashima Hiroaki , Matsumoto Akiko , Maeda Katsumasa
Published: May 2003

Bacteroides forsythus
is one of the important periodontopathic bacteria, and this microorganism is known to have an S-layer outside the outer membrane. The S-layer-like antigens were recently isolated from
B. forsythus
, and they were found to be 270- and 230-kDa proteins in the envelope fraction. In this study, these proteins were confirmed to be specific to
B. forsythus
by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and they were clearly recognized by sera from patients with adult and early-onset periodontitis in Western immmunoblot analysis. We compared the immunoglobulin G (IgG) responses against the purified S-layer-like antigen by enzyme-linked immunosorbent assay. IgG responses against this antigen were low in healthy control subjects, but they were significantly higher in subjects with adult and early-onset periodontitis. Together with the fact that the IgG responses against the crude extract of
B. forsythus
did not rise significantly in patients with periodontitis, S-layer-like proteins are considered to be specific antigens of
B. forsythus
and may play an important role in the progression of periodontitis.
Cervicovaginal Neutralizing Antibodies to Herpes Simplex Virus (HSV) in Women Seropositive for HSV Types 1 and 2
Authors: Mbopi-Kéou Francois-Xavier , Bélec Laurent , Dalessio Julie , Legoff Jérôme , Grésenguet Gérard , Mayaud Philippe , Brown David W. G. , Ashley Morrow Rhoda
Published: May 2003

Antibodies to herpes simplex virus type 1 (HSV-1) and HSV-2 of the immunoglobulin G (IgG) and IgA isotypes were detected in the cervicovaginal secretions (CVS) of 77 HSV-1- and HSV-2-seropositive but clinically asymptomatic African women by type-specific enhanced chemiluminescence Western blotting (ECL-WB). Of the 77 subjects, 34 were HIV negative, shedding HSV-2 DNA in their genital secretions; 20 were HIV positive, shedding HSV-2 DNA; and 23 were HIV negative, not shedding HSV-2 DNA. HSV-specific IgG was detected in CVS of nearly 70% of the women studied. HSV-specific IgA was found in CVS of 50% of the women studied. The distribution of CVS HSV-specific antibodies to each HSV type was highly heterogeneous, with a slight predominance of detectable IgG to HSV-1 (59%) over IgG to HSV-2 (41%), whereas the frequency of detectable IgA to HSV-1 (39%) was similar to that of IgA to HSV-2 (36%). The presence of detectable HSV-specific antibodies was inversely associated with HSV-2 DNA genital asymptomatic shedding but was not affected by HIV seropositivity. In addition, 13 of 77 (17%) CVS samples showed neutralizing activity against HSV-2, as assessed by an HSV-2 in vitro infectivity reduction assay. Neutralizing activity in CVS was associated with the presence of IgG and/or IgA antibodies to HSV-1 and/or to HSV-2 by ECL-WB. Among women whose CVS showed HSV-2-neutralizing activity, the specific activity of HSV-specific neutralizing antibodies was substantially (fivefold) higher in HSV-2 DNA shedders than in nonshedders. In conclusion, HSV-specific antibodies are frequently detected in CVS of asymptomatic African women seropositive for HSV-1 and HSV-2. A subset of these women had functional neutralizing activity against HSV-2 in their CVS. The origin of these antibodies and their role in HSV-2 disease of the female genital tract remain to be determined.
Evaluation of an Immunoglobulin M Capture Enzyme-Linked Immunosorbent Assay for Diagnosis of
Orientia tsutsugamushi
Infection
Authors: Jang Won-Jong , Huh Myung-Suk , Park Kyung-Hee , Choi Myung-Sik , Kim Ik-Sang
Published: May 2003

To differentiate scrub typhus from other acute febrile diseases, a rapid and reliable serological diagnosis is important. We developed an immunoglobulin M (IgM) capture enzyme-linked immunosorbent assay (ELISA) for diagnosis of recent
Orientia tsutsugamushi
infections in humans. The 56-kDa major outer membrane protein of
O. tsutsugamushi
is well known as the most immunodominant antigen in scrub typhus. The test is based on the use of the biotinylated recombinant 56-kDa protein of
O. tsutsugamushi
Boryong, Bor56, which was expressed as a fusion protein with a maltose-binding protein in
Escherichia coli
. In the test, the serum IgM antibodies were captured by anti-human IgM antibodies coated onto a microtiter plate. The captured IgM antibodies were revealed through sequential addition of biotinylated Bor56 antigen and peroxidase-conjugated streptavidin to the plate. The IgM capture ELISA was compared with the immunofluorescence antibody assay (IFA) by testing 176 serum samples from patients with diagnosed cases of rickettsial disease and patients with other acute febrile diseases. Of the 81 IgG IFA-positive samples, 78 tested positive (sensitivity, 96.3%) and all 31 IgM IFA-positive samples tested positive (sensitivity, 100%) by the IgM capture ELISA. The specificity of the IgM capture ELISA was 99%, and 1 of the 95 IFA-negative samples was positive in the assay. These results strongly suggest that IgM capture ELISA using the recombinant Bor56 antigen is a reliable and detailed method for the detection of early
O. tsutsugamushi
infection.
MD-2 Is Necessary for the Toll-Like Receptor 4 Protein To Undergo Glycosylation Essential for Its Translocation to the Cell Surface
Authors: Ohnishi Takahiro , Muroi Masashi , Tanamoto Ken-ichi
Published: May 2003

MD-2 has been reported to be required for the translocation of the Toll-like receptor 4 (TLR4) to the cell surface. However, the mechanism by which MD-2 promotes TLR4 translocation is unknown. We identified the presence of two forms of TLR4 with different molecular masses (approximately 110 and 130 kDa) when TLR4 was expressed together with MD-2. Expressing TLR4 alone produced only the 110-kDa form. Using a membrane-impermeable biotinylation reagent, we found that only the 130-kDa form of TLR4 was expressed on the cell surface. When a cellular extract prepared from cells expressing TLR4 and MD-2 was treated with
N
-glycosidase, the two forms of TLR4 converged into a single band whose size was smaller than the 110-kDa form of TLR4. Mutation of TLR4 at Asn
526
or Asn
575
resulted in the disappearance of the 130-kDa form and prevented TLR4 from being expressed on the cell surface without affecting the ability of TLR4 to associate with MD-2. These results indicate that TLR4 is able to undergo multiple glycosylations without MD-2 but that the specific glycosylation essential for cell surface expression requires the presence of MD-2.
Frequency of Measles Virus-Specific CD4+and CD8+T Cells in Subjects Seronegative or Highly Seropositive for Measles Vaccine
Authors: Ovsyannikova Inna G. , Dhiman Neelam , Jacobson Robert M. , Vierkant Robert A. , Poland Gregory A.
Published: May 2003

The protective effect of measles immunization is due to humoral and cell-mediated immune responses. Little is known about cell-mediated immunity (CMI) to measles vaccine virus, the relative contribution of CD4+and CD8+T cells to variability in such immune responses, and the immunologic longevity of the CMI after measles vaccination in humans. Our study characterizes cellular immune response in subjects seronegative or highly seropositive for measles vaccine immunoglobulin G-specific antibody, aged 15 to 25 years, previously immunized with two doses of measles-mumps-rubella II vaccine. We evaluated the ability of subjects to respond to measles vaccine virus by measuring measles virus-specific T-cell proliferation. We examined the frequencies of measles virus-specific memory Th1 and Th2 cells by an ELISPOT assay. Our results demonstrated that proliferation of T cells in seronegative subjects was significantly lower than that for highly seropositive subjects (P= 0.003). Gamma interferon (IFN-γ) secretion predominated over interleukin 4 (IL-4) secretion in response to measles virus in both groups. The median frequency of measles virus-reactive CD8+T cells secreting IFN-γ was 0.09% in seronegative subjects and 0.43% in highly seropositive subjects (P= 0.04). The median frequency of CD4+T cells secreting IL-4 in response to measles virus was 0.03% in seronegative subjects and 0.09% in highly seropositive subjects (P= 0.005). These data confirm the presence of measles virus-specific cellular immune responses post-measles vaccine immunization in humans. The detection of measles virus-induced IFN-γ and IL-4 production by ELISPOT can be used to identify measles virus-specific low-frequency memory T cells in subjects immunized with measles vaccine. These differences agree in directionality with the observed antibody response phenotype.
Role of Macrophage Migration Inhibitory Factor in Otitis Media with Effusion in Adults
Authors: Kariya Shin , Okano Mitsuhiro , Aoji Katsuya , Kosaka Michiya , Chikumoto Emiko , Hattori Hisashi , Yuen Koji , Nishioka Shinji , Nishioka Keiko , Nishizaki Kazunori
Published: May 2003

Otitis media with effusion (OME) is one of the most common ear diseases. Bacterial endotoxins and several inflammatory cytokines appear to be involved in the pathogenesis of OME in children; however, little is known of the immunological aspects of the onset of OME in adults. We sought to determine the presence of macrophage migration inhibitory factor (MIF) as well as interleukin 1β (IL-1β), tumor necrosis factor alpha (TNF-α), RANTES (regulated upon activation, normal T-cell expressed and presumably secreted), and endotoxin in middle ear effusions (MEEs) from adult patients with OME. In addition, the levels of MIF in MEEs from adults and children were compared. MEE was obtained from 95 adults and 11 children. The levels of MIF, IL-1β, TNF-α, and RANTES were determined by enzyme-linked immunosorbent assay, and the concentrations of endotoxin and total protein were determined by the Endospec assay and bicinchoninic acid assay, respectively. MIF was detected in 97.9% of the MEEs from adults, while endotoxin, IL-1β, TNF-α, and RANTES were detected in 96.8, 12.6, 5.3, and 43.9%, respectively. In addition, the level of MIF was significantly higher than those of endotoxin, IL-1β, and TNF-α. A positive correlation between the levels of MIF and endotoxin was observed. MIF and endotoxin were detected in 81.8 and 72.7%, respectively, of the MEEs from the children. The level of MIF was significantly higher in the children, and conversely that of endotoxin was significantly higher in the adults. These results suggest that the interaction between MIF and endotoxin may promote fluid collection in the middle ear, particularly in adults.
Recombinant Truncated Flagellin of
Burkholderia pseudomallei
as a Molecular Probe for Diagnosis of Melioidosis
Authors: Chen Yao-Shen , Shiuan David , Chen Ssu-Ching , Chye Soi-Moi , Chen Ya-Lei
Published: May 2003

Current serological tests for melioidosis, using impure or uncharacterized cell antigens from
Burkholderia pseudomallei
, have problems in detection sensitivity and specificity. Therefore, we designed and expressed the recombinant flagellin (truncated at both the N- and C-terminal ends), and used the antigen to develop an indirect enzyme-linked immunosorbent assay (ELISA) to diagnose melioidosis. Comparison of the immunoreactivities of the full-length and truncated flagellins reveals that the truncated flagellin performed much better in detection specificity and sensitivity. Only the full-length flagellin was recognized by other bacterial causing septicemia and gave a false-positive result in Western analysis, indicating that the cross-reactive epitopes were located on the more highly conserved N- and C-terminal regions of flagellin. The indirect ELISA using recombinant truncated flagellin as the antigen achieved 93.8% sensitivity and 96.3% specificity and offered a more efficient serodiagnosis of melioidosis.
T- and B-Cell Immune Responses of Patients Who Had Undergone Colectomies to Oral Administration of
Salmonella enterica
Serovar Typhi Ty21a Vaccine
Authors: Kilhamn Jan , Lundin Samuel B. , Brevinge Hans , Svennerholm Ann-Mari , Jertborn Marianne
Published: May 2003

The capacity of an oral live attenuated
Salmonella enterica
serovar Typhi Ty21a vaccine to induce immune responses in patients who had undergone colectomies because of ulcerative colitis was evaluated, and these responses were compared with those of healthy volunteers. Purified CD4
+
and CD8
+
T cells from peripheral blood were stimulated in vitro by using the heat-killed Ty21a vaccine strain, and the proliferation and gamma interferon (IFN-γ) production were measured before and 7 or 8 days after vaccination.
Salmonella
-specific immunoglobulin A (IgA) and IgG antibody responses in serum along with IgA antibody responses in ileostomy fluids from the patients who had undergone colectomies were also evaluated. Three doses of vaccine given 2 days apart failed to induce proliferative T-cell responses in all the six patients who had undergone colectomies, and increases in IFN-γ production were found only among the CD8
+
cells from three of the patients. In contrast, both proliferative responses and increased IFN-γ production were observed among CD4
+
and CD8
+
T cells from 3 and 6 of 10 healthy volunteers, respectively.
Salmonella
-specific IgA and/or IgG antibody responses in serum were observed for five (56%) of nine patients who had undergone colectomies and in 15 (88%) of 17 healthy volunteers. In ileostomy fluids, significant anti-
Salmonella
IgA antibody titer increases were detected in six (67%) of nine patients who had undergone colectomies. The impaired T- and B-cell immune responses found after vaccination in the circulation of patients who have undergone colectomies may be explained by a diminished colonization of the Ty21a vaccine strain due to the lack of a terminal ileum and colon.
Human Papillomavirus-Specific Antibody Status in Oral Fluids Modestly Reflects Serum Status in Human Immunodeficiency Virus-Positive Individuals
Authors: Cameron Jennifer E. , Snowhite Isaac V. , Chaturvedi Anil K. , Hagensee Michael E.
Published: May 2003

Serological assays are valuable tools for studies of the epidemiology of human papillomaviruses (HPVs). The efficacy of a less invasive oral-fluid assay for detection of HPV antibodies was examined. Matched serum, saliva, and oral mucosal transudate (OMT) specimens collected from 150 human immunodeficiency virus-seropositive patients were tested for immunoglobulin G antibodies against HPV-6 and HPV-11 combined (HPV-6/11) and HPV-16 capsids. Antibodies to HPV were detected in both types of oral specimens. Seroprevalence rates were 55% for HPV-6/11 and 37% for HPV-16, whereas oral prevalence rates were significantly lower (for HPV-6/11 in saliva, 31%, and in OMT, 19%; for HPV-16 in saliva, 19%, and in OMT, 17%). HPV antibody detection in OMT more accurately reflected the presence of antibodies in serum than did HPV antibody detection in saliva. More stringent saliva assay cutpoints yielded stronger associations between oropositivity and seropositivity; less stringent OMT cutpoints yielded stronger associations between oropositivity and seropositivity. Although HPV antibodies were detected in oral fluids, further optimization of the assay is necessary before oral-fluid testing can be implemented as a reliable alternative to serum testing for HPV.
Development of an Enzyme-Linked Immunosorbent Assay for Immunoglobulin M Antibodies against Measles Virus
Authors: Roodbari F. , Roustai M. H. , Mostafaie A. , Soleimanjdahi H. , Foroshani R. Sarrami , Sabahi F.
Published: May 2003

Measles is a highly contagious respiratory virus infection, with typical clinical symptoms including maculopapular rash, fever, cough, coryza, and conjunctivitis. Despite implementation of widespread vaccination programs throughout the world, the rates of global morbidity and mortality are still considerable. This study was performed to design a reliable indirect enzyme-linked immunosorbent assay (ELISA) to measure measles-specific immunoglobulin M (IgM). First, human IgM was purified, and then an anti-IgM antibody was produced in rabbits and purified in a multistep process. The rabbit IgG against human IgM was conjugated with peroxidase. Measles virus-infected Vero cells produced viral antigen. One hundred serum samples from infants of 9 to 18 months of age, mostly vaccinated, were evaluated for determining the presence of specific IgM antibodies against measles virus. The samples were also evaluated for neutralizing antibodies against measles virus by a microneutralization test (MNT). By comparing the results of the ELISA with those of MNT, it was demonstrated that ELISA had a sensitivity and specificity of 100 and 92%, respectively. On the other hand, when the results obtained by our ELISA system were compared with those of an imported measles virus IgM ELISA kit (EIAgen; Adaltis Italia SPa, Bologna, Italy), a high level of agreement was shown (
k
= 0.926).
Immunohematological Reference Ranges for Adults from the Central African Republic
Authors: Menard Didier , Mandeng Marie Joelle , Tothy Mesmin Bem , Kelembho Eric Kassa , Gresenguet Gérard , Talarmin Antoine
Published: May 2003

A survey was carried out on 150 healthy adults to establish hematological reference ranges for human immunodeficiency virus (HIV)-negative adults from the Central African Republic (CAR). Immunohematological mean values, medians, and 95th-percentile reference ranges were established. Mean values were as follows: leukocyte (WBC) counts, 5.28 × 10
9
/liter (males) and 5.11 × 10
9
/liter (females); erythrocyte counts, 5.20 × 10
12
/liter (males) and 4.50 × 10
12
/liter (females); hemoglobin, 15.1 g/dl (males) and 12.5 g/dl (females); hematocrit, 45% (males) and 37% (females); lymphocytes, 2,587/μl (males) and 2,466/μl (females); CD4 T cells, 927/μl (males) and 940/μl (females); CD8 T cells, 898/μl (males) and 716/μl (females); and CD4/CD8 T-cell ratio, 1.13 (males) and 1.41 (females). We concluded that (i) the WBC and hemoglobin values of healthy HIV-negative adults from the CAR are lower than the reference values currently used in the CAR and (ii) the absolute CD4 T-cell counts of healthy HIV-negative adults from the CAR are similar to values for Europeans but the absolute CD8 T-cell counts are much higher. Thus, the CD4/CD8 T-cell ratios for healthy adults from the CAR are significantly reduced compared to the ratios for healthy Europeans.
Cross-Reactivity between
Chlamydia trachomatis
Heat Shock Protein 10 and Early Pregnancy Factor
Authors: Betsou Fotini , Borrego Maria José , Guillaume Nicolas , Catry Maria Anjos , Romão Sandra , Machado-Caetano J. A. , Sueur Jean Marie , Mention Jacques , Faille Nicole , Orfila Jeanne
Published: May 2003

Chlamydia trachomatis
heat shock protein 10 (Chsp10) is associated with chronic genital tract infection with
C. trachomatis
. Chsp10 is homologous to human chaperonin 10 (Cpn10) and early pregnancy factor (EPF), a form of human Cpn10 that is specifically secreted at the start of pregnancy. We investigated cross-reactions between serum anti-Chsp10 antibodies and anti-EPF antibodies in pregnant and nonpregnant patients. Pregnancy was found to be associated with the presence of anti-EPF antibodies, which are specifically induced in pregnant women with a history of
C. trachomatis
infection, and with the presence of serum anti-Chsp10 antibodies. We also found that infertility was associated with the presence of anti-Chsp10 and anti-EPF antibodies. The HLA class II haplotype DR8 DQ4 was associated with the presence of anti-Chsp10 antibodies but not of anti-EPF antibodies.
Development of a Recombinant Protein-Based Enzyme-Linked Immunosorbent Assay and Its Applications in Field Surveillance of Rodent Mice for Presence of Immunoglobulin G against
Orientia tsutsugamushi
Authors: Wang Yeau-Ching , Jian Ting-Yu , Tarn Lih-Jeng , Hung Yao-Wen , Chao Hai-Yuan , Ji Da-Der , Liu Hwan-Wun
Published: May 2003

A recombinant protein containing the immunodominant conserved epitope region of the 56-kDa outer membrane protein of the Karp strain of
Orientia tsutsugamushi
was purified to near homogeneity using recombinant DNA techniques. The purified protein was used to immunize rabbits and produced an antibody that could recognize different strains of
O. tsutsugamushi
, as demonstrated both by Western blotting and immunofluorescence assay. An enzyme-linked immunosorbent assay (ELISA) based on this recombinant protein was developed to detect antibody (immunoglobulin G [IgG]) against
O. tsutsugamushi
in mice captured in different districts of Taiwan during 2000 to 2001. A significant difference was found in the antibody seroprevalence rates of
Suncus murinus
mice captured in different districts of Taiwan (χ
2
4, 0.95
= 26.64;
P
< 0.05). Furthermore, a significant difference of IgG seropositivity rates was observed among different kinds of mice (χ
2
5, 0.95
= 93.85;
P
< 0.05). Antibody seropositivity rates were higher in
Bandicota indica
(100%),
Rattus flavipectus
(96.17%), and
Rattus losea
(95.83%) than in
Rattus norvegicus
(86.05%) and
Rattus mindanensis
(83.67%) (χ
2
diff, 5, 0.95
= 12.59,
P
< 0.05). The lowest antibody seropositivity rate (54.4%) was observed in
Suncus murinus
. Antibody seropositivity rates of mice from different districts differed significantly because of the significant difference in antibody seroprevalence rates for
S. murinus
. The results of this study indicated that the recombinant protein ELISA developed in this study could be used to conduct large-scale surveillance of rodent mice for the presence of antibody against
O. tsutsugamushi.
The high seroprevalence rates in rodent mice (except
S. murinus
) suggest that people residing in these districts are at increased risk of developing
O. tsutsugamushi
infection.
Monoclonal Antibodies to
Escherichia coli
-Expressed P46 and P65 Membranous Proteins for Specific Immunodetection of
Mycoplasma hyopneumoniae
in Lungs of Infected Pigs
Authors: Cheikh Saad Bouh K. , Shareck F. , Dea S.
Published: May 2003

The P46 and P65 proteins of
Mycoplasma hyopneumoniae
are two membranous proteins carrying species-specific antigenic determinants. Based on the genomic sequence of the reference strain ATCC 25934, primers were designed for PCR amplification of the genes encoding entire P46 (1,260 bp) and P65 (1,803 bp) and N-terminally truncated P65
c
(1,200 bp). These primers were shown to be specific to
M. hyopneumoniae
since no DNA amplicons could be obtained with other mycoplasma species that commonly colonize the porcine respiratory tract. Both amplified genes were then cloned into the pGEX-4T-1 vector to be expressed in
Escherichia coli
cells as recombinant fusion proteins with glutathione
S
-transferase (GST). Prior to generation of expression constructs, TGA nonsense codons, exceptionally used for tryptophan residues by
M. hyopneumoniae
, had been converted to TGG codons by PCR-directed mutagenesis. Following induction by IPTG (isopropyl-β-
d
-thiogalactopyranoside), both GST-P46 and GST-P65
c
recombinant fusion proteins were recovered by disrupting transformed cells by sonication, purified by affinity chromatography, and then cut with thrombin to release the P46 and P65
c
moieties. The enriched
E. coli
-expressed P46 and P65c proteins were used to immunize female BALB/c mice for the generation of anti-P46 and anti-P65
c
monoclonal antibodies (MAbs). The polypeptide specificities of MAbs obtained was confirmed by Western blotting with cell lysates prepared from the homologous strain. Cross-reactivity study of the anti-P46 and anti-P65
c
MAbs towards two other
M. hyopneumoniae
reference strains (ATCC 25095 and J strains) and Quebec field strains that had been isolated in culture, suggested that the MAbs obtained against both membranous proteins were directed against highly conserved species-specific epitopes. No reactivity to other mycoplasma species tested was demonstrated. Clinical signs and lesions suggestive of enzootic pneumonia were reproduced in specific-pathogen-free pigs that had been inoculated intratracheally with a virulent Quebec field strain (IAF-DM9827) of
M. hyopneumoniae
. Both anti-P46 and anti-P65
c
MAbs permitted effective detection by indirect immunofluorescence and indirect immunoperoxidase assay of
M. hyopneumoniae
in, respectively, frozen and formalin-fixed, paraffin-embedded lung sections from pigs that were killed after the 6- to 7-week observation period.
Effect of Glycosphingolipids Purified fromLeishmania (Leishmania) amazonensisAmastigotes on Human Peripheral Lymphocytes
Authors: Giorgio Selma , Santos Marcia Regina M. , Straus Anita H. , Takahashi Helio K. , Barbiéri Clara Lúcia
Published: May 2003

The effect of purified glycosphingolipids fromLeishmania(Leishmania)amazonensison human lymphoproliferation, on expression of human lymphocyte and monocyte markers (CD3, CD4, CD8, CD14, CD19, and CD45), and on lymphocyte protein kinase C activity was analyzed.
Bactericidal/Permeability-Increasing Protein Is Expressed by Human Dermal Fibroblasts and Upregulated by Interleukin 4
Authors: Reichel Philipp H. , Seemann Christine , Csernok Elena , Schröder Jens-M. , Müller Antje , Gross Wolfgang L. , Schultz Hendrik
Published: May 2003

The bactericidal/permeability-increasing protein (BPI) is an antibiotic- and endotoxin-neutralizing protein of granulocytes and epithelial cells. Constitutive expression of BPI, which increases upon interleukin 4 stimulation, by human dermal fibroblast was demonstrated, suggesting an important role of BPI in gram-negative bacterial clearance and a dampened response to endotoxin in the skin.
One-Step Immunochromatographic Dipstick Tests for Rapid Detection of
Vibrio cholerae
O1 and O139 in Stool Samples
Authors: Nato F. , Boutonnier A. , Rajerison M. , Grosjean P. , Dartevelle S. , Guénolé A. , Bhuiyan N. A. , Sack D. A. , Nair G. B. , Fournier J. M. , Chanteau S.
Published: May 2003

We describe the development and evaluation of a rapid diagnostic test for
Vibrio cholerae
O1 and O139 based on lipopolysaccharide detection using gold particles. The specificity ranged between 84 and 100%. The sensitivity of the dipsticks ranged from 94.2 to 100% when evaluated with stool samples obtained in Madagascar and Bangladesh. The dipstick can provide a simple tool for epidemiological surveys.
Lymphocyte Transformation Test for Medicinal Herbs Yields False-Positive Results for First-Visit Patients
Authors: Mantani Naoki , Kogure Toshiaki , Tamura Jun'ichi , Shimada Yutaka , Terasawa Katsutoshi
Published: May 2003

We performed lymphocyte transformation tests (LTTs) for toki (angelicae radix) and ogon (scutellariae radix) on first-visit patients who had never taken Kampo medicines. LTTs for both herbs were positive in 12 of 14 patients, suggesting that LTTs for these herbs are unreliable for the diagnosis of Kampo medicine-induced liver injury.
Comparison of Complement Fixation and Hemagglutination Inhibition Assays for Detecting Antibody Responses following Influenza Virus Vaccination
Authors: Prince Harry E. , Leber Amy L.
Published: May 2003

Complement fixation (CF) was compared to hemagglutination inhibition (HI) as a method for identifying antibody responses to influenza virus vaccination. CF assays were performed at two different laboratories using paired (pre- and postvaccination) sera from 38 vaccinated laboratory employees; HI assays were performed at a third laboratory. As expected, most vaccinees (31/38 = 82%) responded to at least one of three influenza virus antigens as measured by HI. In contrast, only 21% (8/38) of vaccinees showed a response by CF at laboratory 1, and only 29% (11/38) showed a response by CF at laboratory 2. These findings indicate that due to low sensitivity, CF assays should not be used to assess the antibody response to influenza virus vaccination.
Serological Survey of Toscana Virus Infections in a High-Risk Population in Italy
Authors: Valassina Marcello , Valentini Melissa , Pugliese Agostino , Valensin Pier Egisto , Cusi Maria Grazia
Published: May 2003

Toscana virus is the most important agent responsible for meningitis in central Italy. We report a serosurveillance study, using an immunoenzymatic assay, of 360 serum samples harvested from a high-risk population occupationally exposed to Toscana virus in two regions of Italy, Tuscany and Piedmont. The results indicates a seroprevalence of Toscana virus of 77.2% in the forestry workers, particularly in the Tuscany region. This fact is strictly correlated with the ecological niches specific for the survival of Toscana virus arthropod vector.
Peptidoglycan Induces Mobilization of the Surface Marker for Activation Marker CD66b in Human Neutrophils but Not in Eosinophils
Authors: Mattsson Eva , Persson Terese , Andersson Pia , Rollof Jan , Egesten Arne
Published: May 2003

Peptidoglycan fromStaphylococcus aureusmobilized CD66b in human neutrophils but did not upregulate surface activation markers in eosinophils. In addition, Toll-like receptor 2, implicated in the recognition of peptidoglycan, was detected on the surface of resting neutrophils but not on eosinophils. These findings suggest roles for neutrophils but not eosinophils in innate recognition of peptidoglycan.
A Patient with Crimean-Congo Hemorrhagic Fever Serologically Diagnosed by Recombinant Nucleoprotein-Based Antibody Detection Systems
Authors: Tang Qing , Saijo Masayuki , Zhang Yuzhen , Asiguma Muer , Tianshu Dong , Han Lei , Shimayi Bawudong , Maeda Akihiko , Kurane Ichiro , Morikawa Shigeru
Published: May 2003

We treated a male patient with Crimean-Congo hemorrhagic fever (CCHF). The diagnosis of CCHF was confirmed by reverse transcription-PCR and recombinant nucleoprotein (rNP)-based immunoglobulin G (IgG) and IgM capture enzyme-linked immunosorbent assays of serially collected serum samples. The patient was treated with intravenous ribavirin and recovered with no consequences. The study indicates that rNP-based CCHF virus antibody detection systems are useful for confirming CCHF virus infections. This case also suggests that intravenous ribavirin therapy may be promising for the treatment of CCHF patients.
Anti-Hepatitis A Virus Immunoglobulin M Antibodies in Urine Samples for Rapid Diagnosis of Outbreaks
Authors: de los Angeles Rodríguez Lay Licel , Larralde Díaz Osmany , Martínez Casanueva Raiza , Gutiérrez Moreno Aidonis
Published: May 2003

The main goal of this study was to test the feasibility of using urine for diagnosing hepatitis A virus (HAV) infections. A correlation of 90.78% between the test results of urine and serum samples was obtained. Four outbreaks of hepatitis A were confirmed by testing only urine samples. The levels of anti-HAV immunoglobulin M (IgM) antibodies in urine samples remained stable during 6 months of storage at −70°C but decreased when the samples were stored at 4°C. The results of tests of samples obtained 2 and 6 months after infection suggested that IgM levels decline more rapidly in urine than in serum.
Activation of Coagulation by Administration of Recombinant Factor VIIa Elicits Interleukin 6 (IL-6) and IL-8 Release in Healthy Human Subjects
Authors: de Jonge Evert , Friederich Philip W. , Vlasuk George P. , Rote William E. , Vroom Margaretha B. , Levi Marcel , van der Poll Tom
Published: May 2003

The activation of coagulation has been shown to contribute to proinflammatory responses in animal and in vitro experiments. Here we report that the activation of coagulation in healthy human subjects by the administration of recombinant factor VIIa also elicits a small but significant increase in the concentrations of interleukin 6 (IL-6) and IL-8 in plasma. This increase was absent when the subjects were pretreated with recombinant nematode anticoagulant protein c2, the inhibitor of tissue factor-factor VIIa.
THE KERNEL OF DIRAC OPERATORS ON ${\mathbb S}^3$ AND ℝ3
Authors: ERDŐS LÁSZLÓ , SOLOVEJ JAN PHILIP
Published: May 2003

In this paper we describe an intrinsically geometric way of producing magnetic fields on [Formula: see text] and ℝ3 for which the corresponding Dirac operators have a non-trivial kernel. In many cases we are able to compute the dimension of the kernel. In particular we can give examples where the kernel has any given dimension. This generalizes the examples of Loss and Yau [1].
MICROLOCAL SPECTRUM CONDITION AND HADAMARD FORM FOR VECTOR-VALUED QUANTUM FIELDS IN CURVED SPACETIME
Authors: SAHLMANN HANNO , VERCH RAINER
Published: May 2003

Some years ago, Radzikowski has found a characterization of Hadamard states for scalar quantum fields on a four-dimensional globally hyperbolic spacetime in terms of a specific form of the wavefront set of their two-point functions (termed "wavefront set spectrum condition"), thereby initiating a major progress in the understanding of Hadamard states and the further development of quantum field theory in curved spacetime. In the present work, we extend this important result on the equivalence of the wavefront set spectrum condition with the Hadamard condition from scalar fields to vector fields (sections in a vector bundle) which are subject to a wave-equation and are quantized so as to fulfill the covariant canonical commutation relations, or which obey a Dirac equation and are quantized according to the covariant anti-commutation relations, in any globally hyperbolic spacetime having dimension three or higher.

In proving this result, a gap which is present in the published proof for the scalar field case will be removed. Moreover we determine the short-distance saling limits of Hadamard states for vector-bundle valued fields, finding them to coincide with the corresponding flat-space, massless vacuum states.
ON NON-COMMUTATIVE RUELLE TRANSFER OPERATOR
Authors: MATSUI TAKU
Published: May 2003

In this note, we study a non-commutative analogue of the Ruelle–Perron–Frobenius Transfer operators on UHF algebras.
ON SYMPLECTIC DOUBLE GROUPOIDS AND THE DUALITY OF POISSON GROUPOIDS
Authors: MACKENZIE K. C. H.
Published: May 2003

We prove that the cotangent of a double Lie groupoid S has itself a double groupoid structure with sides the duals of associated Lie algebroids, and double base the dual of the Lie algebroid of the core of S. Using this, we prove a result outlined by Weinstein in 1988, that the side groupoids of a general symplectic double groupoid are Poisson groupoids in duality. Further, we prove that any double Lie groupoid gives rise to a pair of Poisson groupoids (and thus of Lie bialgebroids) in duality. To handle the structures involved effectively we extend to this context the dualities and canonical isomorphisms for tangent and cotangent structures of the author and Ping Xu.
MATCHING MACROSCOPIC PROPERTIES OF BINARY FLUIDS TO THE INTERACTIONS OF DISSIPATIVE PARTICLE DYNAMICS
Authors: DZWINEL WITOLD , YUEN DAVID A.
Published: May 2003

We investigate the role played by conservative forces in dissipative particle dynamics (DPD) simulation of single-component and binary fluids. We employ equations from kinetic theory for matching the coefficients of DPD interparticle force to the macroscopic properties of fluid such as: density, temperature, diffusion coefficient, kinematic viscosity and sound velocity. The sound velocity c is coupled with scaling factor π1 of conservative component of the DPD collision operator. Its value sets up an upper limit on the mass S of a single particle in DPD fluid. The Kirkwood–Alder fluid–solid transition is observed for a sufficiently large S. We emphasize the role of the scaling factor π12 for particles of different types in simulating phase separation in binary fluids. The temporal growth of average domain size R(t) in the phase separation process depends on the value of immiscibility coefficient Δ = π12 - π1. For small immiscibility, R (t) ∝ tβ, where β ≈ 1/2 for R (t) < R H and β ≈ 2/3 for R (t) > R H , R H is the hydrodynamic length. Finally, both phases separate out completely. For larger immiscibility, R(t) increases exponentially at the beginning of simulation, while finally the domain growth process becomes marginal. We also observe the creation of emulsion-like structures. This effect results from an increase of the surface tension on the two-phase interface along with increasing immiscibility.
JACOBI ELLIPTIC FUNCTIONS, NONLINEAR EVOLUTION EQUATIONS AND RECURSION
Authors: HARDY YORICK , STEEB W.-H. , STOOP R.
Published: May 2003

Jacobi elliptic functions play a central role in nonlinear evolution equations. We show how with one recursive call we can calculate the Jacobi elliptic functions sn, cn, and dn. We give an implementation using C++.
THE ATTRACTORS IN SEQUENCE PROCESSING NEURAL NETWORKS
Authors: CHEN YONG , WANG YIN HAI , YANG KONG QING
Published: May 2003

The average length and average relaxation time of attractors in sequence processing neural networks are investigated. The simulation results show that a critical point of α, the loading ratio, is found. Below the turning point, the average length is equal to the number of stored patterns; conversely, the ratio of length and numbers of stored patterns, grow with an exponential dependence exp (Aα). Moreover, we find that the logarithm of average relaxation time is only linearly associated with α and the turning point of coupling degree is located for examining robustness of networks.
EQUILIBRIUM AND QUENCHED ENERGY DISTRIBUTIONS FOR SPIN GLASSES
Authors: BEATON DANIEL A. , DECOSTE CARA , HUNTER D. L. , CONIGLIO ANTONIO
Published: May 2003

In the four-dimensional Edwards–Anderson ±J spin glasses, we observe that the quenched energies distribution from T=2.5 are similar to those observed for the equilibrium states at T=1.0. We show that although these states appear close there are subtle differences which are only seen when we try to quench the system from T=2.5 for a longer number of Monte Carlo steps. The T=2.5 states do not evolve to those seen for the states quenched from T=1.0. We try to identify regions responsible for the subtle differences.
CHOOSEY HOT SAND: REFLECTION OF GRAIN SENSITIVITY ON PATTERN MORPHOLOGY
Authors: ADAMATZKY ANDREW
Published: May 2003

We build and investigate a nonstandard model of pattern formation in a system of discrete entities evolving in discrete space and time. We chose a sandpile paradigm to fit our ideas in the frame of current research. In our model sand is hot because a grain can topple against gradient, i.e., the grain can walk to another node even when a number of grains in its current node is less than a number of neighboring nodes. Sand is choosey because behavior of the grains is not determined by any global parameter or any threshold of a number of neighboring grains (called here a grain sensitivity) but depends on the exact number of grains in the neighboring nodes. Namely, we assume that a grain being at a node x goes to one of the eight neighboring nodes, chosen at random, if there is another grain at the node x or if the number of grains in eight neighboring nodes lies in some set of 2{1,…,8}. These 256 rules of sensitivity are investigated. The classification of the rules if offered, based on the morphology of the patterns generated by each rule. Eight morphological classes are found. Fine structure of every class is investigated and transient phenomena are analyzed. Three kinds of description of class rules by Boolean expressions are offered. Evolution of the classes governed by several one-dimensional parameters is considered.
ENTANGLED QUANTUM STATES AND A C++ IMPLEMENTATION
Authors: STEEB WILLI-HANS , HARDY YORICK
Published: May 2003

Entangled quantum states are an important component of quantum computing techniques such as quantum error-correction, dense coding and quantum teleportation. We determine the requirements for a state in the Hilbert space C4 to be entangled and a solution to the corresponding "factorization" problem if this is not the case. The factorization of nonentangled states is implemented in C++.
SIMPLE AND ACCURATE EXPLICIT BESSEL AND NEUMANN FITTED METHODS FOR THE NUMERICAL SOLUTION OF THE SCHRÖDINGER EQUATION
Authors: SIMOS T. E.
Published: May 2003

Explicit second and fourth algebraic order methods for the numerical solution of the Schrödinger equation are developed in this paper. The new methods have free parameters defined so that the methods are fitted to spherical Bessel and Neumann functions. Based on these new methods we obtained a variable-step algorithm. The results produced based on the numerical solution of the radial Schrödinger equation and the coupled differential equations arising from the Schrödinger equation indicate that this new approach is more efficient than other well known ones.
IDENTIFYING THE BOTTOM LINE AFTER A STOCK MARKET CRASH
Authors: ROEHNER B. M.
Published: May 2003

In this empirical paper we show that in the months following a crash there is a distinct connection between the fall of stock prices and the increase in the range of interest rates for a sample of bonds. This variable, which is often referred to as the interest rate spread variable, can be considered as a statistical measure for the disparity in lenders' opinions about the future; in other words, it provides an operational definition of the uncertainty faced by economic agents. The observation that there is a strong negative correlation between stock prices and the spread variable relies on the examination of eight major crashes in the United States between 1857 and 1987. That relationship which has remained valid for one and a half century in spite of important changes in the organization of financial markets can be of interest in the perspective of Monte Carlo simulations of stock markets.
A GROUP INTERPRETATION OF PARTICLES GENERATED BY ONE-DIMENSIONAL CELLULAR AUTOMATON, WOLFRAM'S RULE 54
Authors: MARTIN BRUNO
Published: May 2003

One-dimensional cellular automata are known to be able to present complex behaviors. In some cases, their evolution may be understood as movings, collisions, or creations of particles. In the case of the special Wolfram's rule 54, Boccara has previously pointed out basic particles. In this paper, we introduce a group which allows the formal study of interactions between these particles. Coming back to the complexity of rule 54 and using the new algebraic classification of Rapaport, we prove that rule 54 is not simple.
PROJECTIVE NOISE CLEANING WITH DYNAMIC NEIGHBORHOOD SELECTION
Authors: KERN A. , STEEB W.-H. , STOOP R.
Published: May 2003

In recent years, several methods of noise cleaning have been devised, of which projective methods have been particularly effective. In our paper, we explain in detail why orthogonal projections are nonoptimal and how the nonorthogonal projections suggested by Grassberger et al., naturally emerge from the SVD method. We show that this approach when combined with a dynamic neighborhood selection yields optimal results of noise cleaning.
DIRECTED BAK–SNEPPEN MODEL FOR FOOD CHAINS
Authors: STAUFFER D. , JAN N.
Published: May 2003

A modification of the Bak–Sneppen model to include simple elements of Darwinian evolution is used to check the survival of prey and predators in long food chains. Mutations, selection, and starvation resulting from depleted prey are incorporated in this model.
COMPUTER SIMULATION OF STRUCTURAL CHANGES IN PHOSPHOLIPID BILAYERS
Authors: SHAHINIAN ARAM A. , POGHOSYAN ARMEN H. , BADALYAN HAMLET G.
Published: May 2003

Using molecular-scale computer simulation, we show that structural changes may take place in phospholipid bilayers in an aqueous solution as we vary the relative concentration of water to phospholipids, the bilayer thickness, the inclination angle of the choline, dipole molecular fragment with respect to the bilayer surface, and the conformation of hydrocarbon chains of phospholipid in the hydrophobic volume of the bilayer. It is shown that in the hydrophobic volume of the bilayer, an interpenetration of the hydrocarbon chains located on opposite sides of the phospholipid bilayer takes place.
MULTIVARIATE TIME SERIES MODELING IN A CONNECTIONIST APPROACH
Authors: KULKARNI D. R. , PARIKH J. C.
Published: May 2003

Multivariate models in the framework of artificial neural network have been constructed for systems where time series data of several variables is known. The models have been tested using computer generated data for the Lorenz and Henon systems. They are found to be robust and give accurate short term predictions. Analysis of the models is able to throw some light on theoretical questions related to multivariate "embedding" and removal of redundancy in the embedding.