bluuebunny/crossref_metadata_2025_split huggingface dataset:
Efficient Ex Vivo Stimulation ofMycobacterium tuberculosis-Specific T Cells by Genetically DetoxifiedBordetella pertussisAdenylate Cyclase Antigen Toxoids
Authors: Wilkinson Katalin A. , Simsova Marcela , Schölvinck Elisabeth , Sebo Peter , Leclerc Claude , Vordermeier H. Martin , Dickson Stuart J. , Brown Jillian R. , Davidson Robert N. , Pasvol Geoffrey , Levin Michael , Wilkinson Robert J. 
Published: April 2005

ABSTRACTMycobacterium tuberculosisis a significant threat to global health.Mycobacterium bovisBCG vaccine provides only partial protection, and the skin test reagent used to aid diagnosis of both active and latent tuberculosis, purified protein derivative (PPD), lacks specificity and sensitivity. The use of genetically detoxifiedBordetella pertussisadenylate cyclase toxin (CyaA) as a delivery system for two immunodominant proteins ofM. tuberculosisthat are of greater specificity than PPD, early-secreted antigenic target 6-kDa protein (ESAT-6) and culture filtrate protein 10 (CFP-10), was therefore investigated. CyaA toxoids incorporating these antigens were able to restimulate T cells from more than 91% tuberculosis patients and healthy sensitized donors. Delivery of antigen by CyaA decreased by 10-fold the amount of ESAT-6 and CFP-10 required to restimulate T cells, and in low responders, the overall frequency of gamma interferon-producing cells detected by enzyme-linked immunospot assay was increased (P< 0.01 for both antigens). Delivery of ESAT-6 and CFP-10 by CyaA enabled the detection of both CD4+and CD8+T cells: these responses could be blocked by inhibition of major histocompatibility complex class II or class I, respectively. Covalent linkage of antigen to the CyaA vector was required for enhancement to occur, as a mixture of mock CyaA toxoid plus recombinant ESAT-6 did not lead to enhancement. In a simplified whole-blood model to detect tuberculosis infection, the frequency of positive responses to CFP-10 was increased by CyaA delivery, a potentially important attribute that could facilitate the identification of latent infection.
B-Cell and T-Cell Immune Responses to ExperimentalHelicobacter pyloriInfection in Humans
Authors: Nurgalieva Zhannat Z. , Conner Margaret E. , Opekun Antone R. , Zheng Carl Q. , Elliott Susan N. , Ernst Peter B. , Osato Michael , Estes Mary K. , Graham David Y. 
Published: April 2005

ABSTRACTThe acute antibody and T-cell immune response toHelicobacter pyloriinfection in humans has not been studied systematically. Serum fromH. pylori-naive volunteers challenged withH. pyloriand cured after 4 or 12 weeks was tested by enzyme-linked immunosorbent assays for anti-H. pylori-specific immunoglobulin M (IgM) and IgA established using bacterial lysates from homologous (the infecting strain) and heterologousH. pylori. Proteins recognized by IgM antibody were identified by mass spectrometry of immunoreactive bands separated by two-dimensional gel electrophoresis. Mucosal T-cell subsets (CD4, CD8, CD3, and CD30 cells) were assessed by immunohistochemistry. All 18 infected volunteers developedH. pylori-specific IgM responses to both homologous or heterologousH. pyloriantigens.H. pyloriantigens reacted with IgM antibody at 4 weeks postinfection. IgM Western blotting showed immunoreactivity of postinfection serum samples to multipleH. pyloriproteins with molecular weights ranging between 9,000 (9K) to 150K with homologous strains but only a 70K band using heterologous antigens. Two-dimensional electrophoresis demonstrated that production ofH. pylori-specific IgM antibodies was elicited byH. pyloriflagellins A and B, urease B, ABC transporter binding protein, heat shock protein 70 (DnaK), and alkyl hydroperoxide reductase. Mucosal CD3, CD4, and CD8 T-cell numbers increased following infection. IgM antibody responses were detected to a range of homologousH. pyloriantigens 2 to 4 weeks postchallenge. The majority ofH. pyloriproteins were those involved in motility and colonization and may represent targets for vaccine development.
Relapsing Fever Spirochetes Contain Chromosomal Genes with Unique Direct Tandemly Repeated Sequences
Authors: Guyard Cyril , Chester Earl M. , Raffel Sandra J. , Schrumpf Merry E. , Policastro Paul F. , Porcella Stephen F. , Leong John M. , Schwan Tom G. 
Published: April 2005

ABSTRACT

Genome sequencing of the relapsing fever spirochetes
Borrelia hermsii
and
Borrelia turicatae
identified three open reading frames (ORFs) on the chromosomes that contained internal, tandemly repeated amino acid sequences that were absent in the Lyme disease spirochete
Borrelia burgdorferi
. The predicted amino acid sequences of these genes (BH0209, BH0512, and BH0553) have hydrophobic N termini, indicating that these proteins may be secreted.
B. hermsii
transcribed the three ORFs in vitro, and the BH0512- and BH0553-encoded proteins (PBH-512 and PBH-553) were produced in vitro and in experimentally infected mice. PBH-512 and PBH-553 were on the spirochete's outer surface, and antiserum to these proteins reduced the adherence of
B. hermsii
to red blood cells. PCR analyses of 28 isolates of
B. hermsii
and 8 isolates of
B. turicatae
demonstrated polymorphism in each gene correlated with the number of repeats. Serum samples from relapsing fever patients reacted with recombinant PBH-512 and PBH-553, suggesting that these proteins are produced during human infection. These polymorphic proteins may be involved in the pathogenicity of these relapsing fever spirochetes and provide a mechanism for antigenic heterogeneity within their populations.
Mutants of
Mycobacterium tuberculosis
Lacking Three of the Five
rpf
-Like Genes Are Defective for Growth In Vivo and for Resuscitation In Vitro
Authors: Downing Katrina J. , Mischenko Vladimir V. , Shleeva Margarita O. , Young Danielle I. , Young Michael , Kaprelyants Arseny S. , Apt Alexander S. , Mizrahi Valerie 
Published: April 2005

ABSTRACT

Mycobacterium tuberculosis
contains five genes,
rpfA
through
rpfE
, that bear significant homology to the resuscitation-promoting factor (
rpf
) gene of
Micrococcus luteus
, whose product is required to resuscitate the growth of dormant cultures of
M. luteus
and is essential for the growth of this organism. Previous studies have shown that deletion of any one of the five
rpf
-like genes did not affect the growth or survival of
M. tuberculosis
in vitro. In conjunction with the results of whole-genome expression profiling, this finding was indicative of their functional redundancy. In this study, we demonstrate that the single deletion mutants are phenotypically similar to wild-type
M. tuberculosis
H37Rv in vivo. The deletion of individual
rpf
-like genes had no discernible effect on the growth or long-term survival of
M. tuberculosis
in liquid culture, and the ability to resuscitate spontaneously from a nonculturable state in a most probable number assay was also unaffected for the three strains tested (the Δ
rpfB
,
ΔrpfD
, and Δ
rpfE
strains). In contrast, two multiple strains, KDT8 (Δ
rpfA
-mutation
ΔrpfC ΔrpfB
) and KDT9 (Δ
rpfA ΔrpfC ΔrpfD
), which lack three of the five
rpf
-like genes, were significantly yet differentially attenuated in a mouse infection model. These mutants were also unable to resuscitate spontaneously in vitro, demonstrating the importance of the Rpf-like proteins of
M. tuberculosis
in resuscitation from the nonculturable state. These results strongly suggest that the biological functions of the five
rpf
-like genes of
M. tuberculosis
are not wholly redundant and underscore the potential utility of these proteins as targets for therapeutic intervention.
Invasion of Epithelial Cells by Locus of Enterocyte Effacement-Negative EnterohemorrhagicEscherichia coli
Authors: Luck Shelley N. , Bennett-Wood Vicki , Poon Rachael , Robins-Browne Roy M. , Hartland Elizabeth L. 
Published: April 2005

ABSTRACTThe majority of enterohemorrhagicEscherichia coli(EHEC) strains associated with severe disease carry the locus of enterocyte effacement (LEE) pathogenicity island, which encodes the ability to induce attaching and effacing lesions on the host intestinal mucosa. While LEE is essential for colonization of the host in these pathogens, strains of EHEC that do not carry LEE are regularly isolated from patients with severe disease, although little is known about the way these organisms interact with the host epithelium. In this study, we compared the adherence properties of clinical isolates of LEE-negative EHEC with those of LEE-positive EHEC O157:H7. Transmission electron microscopy revealed that LEE-negative EHEC O113:H21 was internalized by Chinese hamster ovary (CHO-K1) epithelial cells and that intracellular bacteria were located within a membrane-bound vacuole. In contrast, EHEC O157:H7 remained extracellular and intimately attached to the epithelial cell surface. Quantitative gentamicin protection assays confirmed that EHEC O113:H21 was invasive and also showed that several other serogroups of LEE-negative EHEC were internalized by CHO-K1 cells. Invasion by EHEC O113:H21 was significantly reduced in the presence of the cytoskeletal inhibitors cytochalasin D and colchicine and the pan-Rho GTPase inhibitor compactin, whereas the tyrosine kinase inhibitor genistein had no significant impact on bacterial invasion. In addition, we found that EHEC O113:H21 was invasive for the human colonic cell lines HCT-8 and Caco-2. Overall these studies suggest that isolates of LEE-negative EHEC may employ a mechanism of host cell invasion to colonize the intestinal mucosa.
Structural and Genetic Diversity of Group BStreptococcusCapsular Polysaccharides
Authors: Cieslewicz Michael J. , Chaffin Donald , Glusman Gustavo , Kasper Dennis , Madan Anup , Rodrigues Stephani , Fahey Jessica , Wessels Michael R. , Rubens Craig E. 
Published: April 2005

ABSTRACTGroup BStreptococcus(GBS) is an important pathogen of neonates, pregnant women, and immunocompromised individuals. GBS isolates associated with human infection produce one of nine antigenically distinct capsular polysaccharides which are thought to play a key role in virulence. A comparison of GBS polysaccharide structures of all nine known GBS serotypes together with the predicted amino acid sequences of the proteins that direct their synthesis suggests that the evolution of serotype-specific capsular polysaccharides has proceeded through en bloc replacement of individual glycosyltransferase genes with DNA sequences that encode enzymes with new linkage specificities. We found striking heterogeneity in amino acid sequences of synthetic enzymes with very similar functions, an observation that supports horizontal gene transfer rather than stepwise mutagenesis as a mechanism for capsule variation. Eight of the nine serotypes appear to be closely related both structurally and genetically, whereas serotype VIII is more distantly related. This similarity in polysaccharide structure strongly suggests that the evolutionary pressure toward antigenic variation exerted by acquired immunity is counterbalanced by a survival advantage conferred by conserved structural motifs of the GBS polysaccharides.
Tumor Necrosis Factor Alpha Plays a Role in the Acceleration of Atherosclerosis byChlamydia pneumoniaein Mice
Authors: Campbell Lee Ann , Nosaka Tadayoshi , Rosenfeld Michael E. , Yaraei Kambiz , Kuo Cho-chou 
Published: April 2005

ABSTRACTThe role of tumor necrosis factor alpha (TNF-α) inChlamydia pneumoniaeatherogenesis was evaluated in TNF-α p55 receptor-deficient C57BL/6J mice fed a high-fat/high-cholesterol diet. No acceleration of atherosclerotic lesion development was observed in infected mice compared to uninfected mice, indicating that TNF-α plays a role in the acceleration of atherosclerosis byC. pneumoniae.
Detection of large deletions in the LDL receptor gene with quantitative PCR methods
Authors: Damgaard Dorte , Nissen Peter H , Jensen Lillian G , Nielsen Gitte G , Stenderup Anette , Larsen Mogens L , Faergeman Ole 
Published: April 2005

Abstract

Background
Familial Hypercholesterolemia (FH) is a common genetic disease and at the molecular level most often due to mutations in the LDL receptor gene. In genetically heterogeneous populations, major structural rearrangements account for about 5% of patients with LDL receptor gene mutations.

Methods
In this study we tested the ability of two different quantitative PCR methods, i.e. Real-Time PCR and Multiplex Ligation-Dependent Probe Amplification (MLPA), to detect deletions in the LDL receptor gene. We also reassessed the contribution of major structural rearrangements to the mutational spectrum of the LDL receptor gene in Denmark.

Results
With both methods it was possible to discriminate between one and two copies of the LDL receptor gene exon 5, but the MLPA method was cheaper, and it was far more accurate and precise than Real-Time PCR. In five of 318 patients with an FH phenotype, MLPA analysis revealed five different deletions in the LDL receptor gene.

Conclusion
The MLPA method was accurate, precise and at the same time effective in screening a large number of FH patients for large deletions in the LDL receptor gene.
Development and preliminary evaluation of the VPS ReplaySuite: a virtual double-headed microscope for pathology
Authors: Johnston Dan J , Costello Sean P , Dervan Peter A , O'Shea Daniel G 
Published: April 2005

Abstract
Background
Advances in computing and telecommunications have resulted in the availability of a range of online tools for use in pathology training and quality assurance. The majority focus on either enabling pathologists to examine and diagnose cases, or providing image archives that serve as reference material. Limited emphasis has been placed on analysing the diagnostic process used by pathologists to reach a diagnosis and using this as a resource for improving diagnostic performance.

Methods
The ReplaySuite is an online pathology software tool that presents archived virtual slide examinations to pathologists in an accessible video-like format, similar to observing examinations with a double-headed microscope. Delivered through a customised web browser, it utilises PHP (Hypertext PreProcessor) to interact with a remote database and retrieve data describing virtual slide examinations, performed using the Virtual Pathology Slide (VPS).
To demonstrate the technology and conduct a preliminary evaluation of pathologists opinions on its potential application in pathology training and quality assurance, 70 pathologists were invited to use the application to review their own and other pathologists examinations of 10 needle-core breast biopsies and complete an electronic survey. 9 pathologists participated, and all subsequently completed an exit survey.

Results
Of those who replayed an examination by another pathologist, 83.3% (5/6) agreed that replays provided an insight into the examining pathologists diagnosis and 33.3% (2/6) reconsidered their own diagnosis for at least one case. Of those who reconsidered their original diagnosis, all re-classified either concordant with group consensus or original glass slide diagnosis. 77.7% (7/9) of all participants, and all 3 participants who replayed more than 10 examinations stated the ReplaySuite to be of some or great benefit in pathology training and quality assurance.

Conclusion
Participants conclude the ReplaySuite to be of some or of great potential benefit to pathology training and quality assurance and consider the ReplaySuite to be beneficial in evaluating the diagnostic trace of an examination. The ReplaySuite removes temporal and spatial issues that surround the use of double-headed microscopes by allowing examinations to be reviewed at different times and in different locations to the original examination. While the evaluation set was limited and potentially subject to bias, the response of participants was favourable. Further work is planned to determine whether use of the ReplaySuite can result in improved diagnostic ability.
Stereophysicochemical variability plots highlight conserved antigenic areas in Flaviviruses
Authors: Schein Catherine H , Zhou Bin , Braun Werner 
Published: April 2005

AbstractBackgroundFlaviviruses, which include Dengue (DV) and West Nile (WN), mutate in response to immune system pressure. Identifying escape mutants, variant progeny that replicate in the presence of neutralizing antibodies, is a common way to identify functionally important residues of viral proteins. However, the mutations typically occur at variable positions on the viral surface that are not essential for viral replication. Methods are needed to determine the true targets of the neutralizing antibodies.ResultsStereophysicochemical variability plots (SVPs), 3-D images of protein structures colored according to variability, as determined by our PCPMer program, were used to visualize residues conserved in theirphysicalchemicalproperties (PCPs) near escape mutant positions. The analysis showed 1) that escape mutations in the flavivirus envelope protein are variable residues by our criteria and 2) two escape mutants found at the same position in many flaviviruses sit above clusters of conserved residues from different regions of the linear sequence. Conservation patterns in T-cell epitopes in the NS3- protease suggest a similar mechanism of immune system evasion.ConclusionThe SVPs add another dimension to structurally defining the binding sites of neutralizing antibodies. They provide a useful aid for determining antigenically important regions and designing vaccines.
Evaluation of the Serodia Treponema pallidum particle agglutination, the Murex Syphilis ICE and the Enzywell TP tests for serodiagnosis of syphilis
Authors: Aktas G , Young H , Moyes A , Badur S 
Published: April 2005

We evaluated the Treponema pallidum haemagglutination assay (TPHA), a treponemal test, with three other treponemal tests, the Serodia T. pallidum particle agglutination assay, the Murex Syphilis ICE IgG + IgM enzyme immunoassay (EIA) and the Enzywell TP IgG + M EIA (a new rapid EIA) for use in conjunction with the rapid plasma reagin test (RPR), a non-treponemal test, for serodiagnosis of syphilis. In all, 124 serum samples were found reactive with RPR and/or TPHA after testing by the routine laboratory protocol. Twenty-three (18.5%) of them were positive only by RPR test and were evaluated as biologically false-positive, 16 were positive only by the TPHA and 84 by both the RPR and TPHA tests; one sample was non-specific (heterophile reaction) in the TPHA. Agreements of the TPHA with the Serodia TPPA, the Murex Syphilis ICE and the Enzywell TP tests were 96.7%, 100% and 99.1%, respectively. We conclude that each one of the tests, the Serodia TPPA, the Murex Syphilis ICE and the Enzywell TP, is an appropriate substitute for screening for serodiagnosis of syphilis.
Book Reviews
Authors: Lev Benjamin 
Published: April 2005

The range of books reviewed is wide, covering theory and applications in operations research, statistics, management science, econometrics, mathematics, computers, and information systems (no software is reviewed). In addition, we include books in other fields that emphasize technical applications. Publishers who wish to have their books reviewed should send them to Professor Benjamin Lev. We list the books received; not all books received can be reviewed because space is limited. Those who would like to review books are urged to send me their names, addresses, and specific areas of expertise. We commission all reviews and do not accept unsolicited book reviews. Readers are encouraged to suggest books that might be reviewed or to ask publishers to send me copies of such books. The authors or editors of the books reviewed in this issue are Benjamin F. Hobbs, Michael H. Rothkopf, Richard P. O’Neill, Hung-Po Chao, David R. Laube, Raymond F. Zammuto, Douglas Long, Charles A. Micchelli, Gianni Di Pillo, and Almerico Murli.
Clinical Performance of a Regenerative Strategy for Intrabony Defects: Scientific Evidence and Clinical Experience
Authors: Cortellini Pierpaolo , Tonetti Maurizio S. 
Published: April 2005

Background: The aim of the present study was to evaluate the clinical performance of a regenerative strategy for the treatment of deep intrabony defects.Methods: This patient cohort study involved 40 patients with one deep interdental intrabony defect each. They were treated with periodontal regeneration using four different regenerative methods (expanded polytetrafluoroethylene [ePTFE] titanium reinforced membranes, bioabsorbable membranes alone, bioabsorbable membranes with a bone replacement graft [combination], or enamel matrix derivative), according to predefined criteria and decision‐making algorithms. Defects were accessed with papilla preservation flaps performed with the aid of an operating microscope and microsurgical instruments. A stringent plaque control regimen was enforced in all the patients during the 1‐year observation period. Outcomes included evaluation of the complete primary closure of the interdental space (CLOSURE), gains in clinical attachment (CAL), and reductions in probing depths (PD).Results: CLOSURE was achieved in all treated defects and was maintained in 90% of cases for the entire healing period. At 1 year the observed CAL gains were 6 ± 1.8 mm on average, corresponding to a resolution of 92.1% ± 12% of the initial intrabony (CAL%) component of the defect. Average PD reduction was 6.1 ± 1.9 mm and was associated with minimal increase in gingival recession (–0.1 ± 0.7 mm). The 12 sites treated with titanium reinforced ePTFE membranes resulted in a 1‐year CAL gain of 6.8 ± 2.2 mm (CAL%: 94.7 ± 13.4); the 11 cases treated with combination therapy showed a 1‐year CAL gain of 5.4 ± 1.7 mm (CAL%: 88.2 ± 9.6); the seven sites treated with bioabsorbable barriers resulted in 5.9 ± 1.2 mm of CAL gain (CAL%: 88.9 ± 11.5); and the 10 sites treated with enamel matrix gained on average 5.9 ± 1.5 mm of CAL (CAL%: 88.9 ± 11.5). No significant differences were observed among the four approaches.Conclusion: The use of an evidence‐based regenerative strategy resulted in clinically relevant amounts of CAL gains, shallow pockets, and minimal gingival recession with the four regenerative approaches. J Periodontol 2005;76:341‐350.
Prospective Assessment of the Use of Enamel Matrix Proteins With Minimally Invasive Surgery
Authors: Harrel Stephen K. , Wilson Thomas G. , Nunn Martha E. 
Published: April 2005

Background: Minimally invasive surgery (MIS) is a surgical technique using very small incisions, indicated for performing regenerative therapy in periodontal defects. Enamel matrix proteins (EMP) have been shown to enhance periodontal regeneration. This study was undertaken to determine the effect of using EMP in combination with an MIS approach.Methods: Patients from two private periodontal practices with chronic periodontitis who, following non‐surgical therapy, had one or more sites with probing depths (PD) of ≥6 mm were included in the study. An MIS approach was utilized for surgical access. Following surgical debridement, EMP was placed into the bony defect. The surgical sites were reevaluated after at least 11 months.Results: Surgical treatment was performed at 160 sites in 16 patients. No significant differences were noted in the results between the two offices and the data were combined. Mean PD reduction (P = 0.002) and attachment level improvements (P = 0.012) were significantly greater than 3 mm with mean post‐surgical PD of 3.17 mm and attachment levels of 4.05 mm, based on subject means. Mean change in recession following surgery was negligible (0.01 mm). All sites were considered to be clinically successful.Conclusion: The combination of MIS and EMP yields significant reductions in probing depths and improvements in attachment levels while producing little or no increase in recession. J Periodontol 2005;76:380‐384.