bluuebunny/crossref_metadata_2025_split huggingface dataset:
In Situ Detection of
Mycobacterium tuberculosis
Transcripts in Human Lung Granulomas Reveals Differential Gene Expression in Necrotic Lesions
Authors: Fenhalls Gael , Stevens Liesel , Moses Lorraine , Bezuidenhout Juanita , Betts Joanna C. , Helden Paul van , Lukey Pauline T. , Duncan Ken 
Published: October 2002

ABSTRACT

We have used RNA-RNA in situ hybridization to detect the expression of several
Mycobacterium tuberculosis
genes in tuberculous granulomas in lung tissue sections from tuberculosis patients. The
M. tuberculosis
genes chosen fall into two classes. Four genes (
icl
,
narX
, and Rv2557 and Rv2558) have been implicated in the persistence of the bacterium in the host, and two genes (
iniB
and
kasA
) are upregulated in response to isoniazid exposure. Both necrotic and nonnecrotic granulomas were identified in all of the patients. Necrotic granulomas were divided into three zones: an outer lymphocyte cuff containing lymphocytes and macrophages, a transition zone consisting of necrotic material interspersed with macrophages, and a central acellular necrotic region. Transcripts of all of the genes studied were found in nonnecrotic granulomas and in the lymphocyte cuff of necrotic granulomas. Mycobacterial gene expression was associated with CD68-positive myeloid cells. Rv2557 and/or its homologue Rv2558,
kasA
, and
iniB
were expressed within the transition zone of necrotic granulomas, whereas
icl
and
narX
transcripts were absent from this area. There was no evidence of transcription of any of the genes examined in the central necrotic region, although mycobacterial DNA was present. The differential expression of genes within granulomas demonstrates that
M. tuberculosis
exists in a variety of metabolic states and may be indicative of the response to different microenvironments. These observations confirm that genes identified in models of persistence or in response to drug treatment in vitro are expressed in the human host.
Modulation of Cytosolic Ca
2+
Concentration in Airway Epithelial Cells by
Pseudomonas aeruginosa
Authors: Jacob Tobias , Lee Rebecca J. , Engel Joanne N. , Machen Terry E. 
Published: October 2002

ABSTRACT

Modulation of cytosolic (intracellular) Ca
2+
concentration (Ca
i
) may be an important host response when airway epithelial cells are exposed to
Pseudomonas aeruginosa
. We measured Ca
i
in Calu-3 cells exposed from the apical or basolateral surface to cytotoxic and noncytotoxic strains of
P. aeruginosa
. Apical addition of either noncytotoxic strains or cytotoxic strains failed to affect Ca
i
over a 3-h time period, nor were changes observed after basolateral addition of noncytotoxic strains. In contrast, basolateral addition of cytotoxic strains caused a slow increase in Ca
i
from 100 nM to 200 to 400 nM. This increase began after 20 to 50 min and persisted for an additional 30 to 75 min, at which time the cells became nonviable.
P. aeruginosa
-induced increases in Ca
i
were blocked by the addition of the Ca channel blocker La
3+
to the basolateral but not to the apical chamber. Likewise, replacing the basolateral but not the apical medium with Ca-free solution prevented
P. aeruginosa
-mediated changes in Ca
i
. With isogenic mutants of PA103, we demonstrated that the type III secretion apparatus, the type III-secreted effector ExoU, and type IV pili were necessary for increased Ca
i
. We propose that translocation of ExoU through the basolateral surface of polarized airway epithelial cells via the type III secretion apparatus leads to release of Ca stored in the endoplasmic reticulum and activation of Ca channels in the basolateral membranes of epithelial cells.
Immunization with C5a Peptidase or Peptidase-Type III Polysaccharide Conjugate Vaccines Enhances Clearance of Group B Streptococci from Lungs of Infected Mice
Authors: Cheng Qi , Debol Steven , Lam Hong , Eby Ron , Edwards Lorri , Matsuka Yury , Olmsted Stephen B. , Cleary P. Patrick 
Published: October 2002

ABSTRACTGroup B streptococci (GBS) are among the most common causes of life-threatening neonatal infections. Vaccine development since the late 1970s has focused on the capsular polysaccharides, but a safe, effective product is still not available. Our quest for a vaccine turned to the streptococcal C5a peptidase (SCPB). This surface protein is antigenically conserved across most if not all serotypes. A murine model was used to assess the impact of SCPB on clearance of GBS from the lungs of intranasally infected animals. Mutational inactivation of SCPB resulted in more-rapid clearance of streptococci from the lung. Immunization with recombinant SCPB alone or SCPB conjugated to type III capsular polysaccharide produced serotype-independent protection, which was evidenced by more-rapid clearance of the serotype VI strain from the lungs. Immunization of mice with tetanus toxoid-type III polysaccharide conjugate did not produce protection, confirming that protection induced by SCPB conjugates was independent of type III polysaccharide antigen. Histological evaluation of lungs from infected mice revealed that pathology in animals immunized with SCPB or SCPB conjugates was significantly less than that in animals immunized with a tetanus toxoid-polysaccharide conjugate. These experiments suggest that inclusion of C5a peptidase in a vaccine will both add another level to and broaden the spectrum of the protection of a polysaccharide vaccine.
Babesia bovisMerozoite Surface Antigen 2 Proteins Are Expressed on the Merozoite and Sporozoite Surface, and Specific Antibodies Inhibit Attachment and Invasion of Erythrocytes
Authors: Mosqueda Juan , McElwain Terry F. , Palmer Guy H. 
Published: October 2002

ABSTRACTTheBabesia bovismerozoite surface antigen 2 (MSA-2) locus encodes four proteins, MSA-2a1, -2a2, -2b, and -2c. With the use of specific antibodies, each MSA-2 protein was shown to be expressed on the surface of live extracellular merozoites and coexpression on single merozoites was confirmed. Individual antisera against MSA-2a, MSA-2b, and MSA-2c significantly inhibited merozoite invasion of bovine erythrocytes. As tick-derived sporozoites also directly invade erythrocytes, expression of each MSA-2 protein on the sporozoite surface was examined and verified. Finally, statistically significant inhibition of sporozoite binding to the erythrocytes was demonstrated by using antisera specific for MSA-2a, MSA-2b, and MSA-2c. These results indicate an important role for MSA-2 proteins in the initial binding and invasion of host erythrocytes and support the hypothesis that sporozoites and merozoites use common surface molecules in erythrocyte invasion.
The N-Terminal Domain of RTX Toxin ApxI of
Actinobacillus pleuropneumoniae
Elicits Protective Immunity in Mice
Authors: Seah J. N. , Frey J. , Kwang J. 
Published: October 2002

ABSTRACT

We expressed three
Actinobacillus pleuropneumoniae
ApxI deletion derivatives to map the domain that could induce protective immunity. Antiserum to ApxI N-terminal covered by residues 40 to 380 was found to neutralize ApxI hemolytic activity but not ApxIII cytotoxicity. When used as a subunit vaccine in mice, this recombinant N-terminal fragment elicited protection against lethal infection with heterologous
A. pleuropneumoniae
serovars.
Role of Type 1 Fimbria- and P Fimbria-Specific Adherence in Colonization of the Neurogenic Human Bladder by
Escherichia coli
Authors: Hull Richard A. , Donovan William H. , Del Terzo Michael , Stewart Colleen , Rogers Margaret , Darouiche Rabih O. 
Published: October 2002

ABSTRACT

Recent clinical studies suggest that the deliberate colonization of the human bladder with a prototypic asymptomatic bacteriuria-associated bacterium,
Escherichia coli
83972, may reduce the frequency of urinary tract infection in individuals with spinal cord injuries. However, the mechanism by which
E. coli
83972 colonizes the bladder is unknown. We examined the role in bladder colonization of the
E. coli
83972 genes
papG
and
fimH
, which respectively encode P and type 1 receptor-specific fimbrial adhesins.
E. coli
83972 and isogenic
papG
Δ and
papG
Δ
fimH
Δ mutants of
E. coli
83972 were compared for their capacities to colonize the neurogenic human bladder. Both strains were capable of stable colonization of the bladder. The results indicated that type 1 class-specific adherence and P class-specific adherence, while implicated as significant colonization factors in experiments that employed various animal model systems, were not required for colonization of the neurogenic bladder in human beings. The implications of these results with regard to the selection of potential vaccine antigens for the prevention of urinary tract infection are discussed.
LOCAL MINIMIZERS FOR THE GINZBURG–LANDAU ENERGY NEAR CRITICAL MAGNETIC FIELD: PART II
Authors: SERFATY SYLVIA 
Published: October 2002

As in Part I, we study local minimizers of the Ginzburg–Landau energy (depending on κ → +∞) for superconductors in a prescribed magnetic field hex. For disc domains, we find and describe stable solutions of the associated equations and show how vortices appear as hex is raised from the first critical field Hc1. We also study the asymptotic limit κ→∞ for hex=Hc1 and prove that the limiting magnetic field in the superconductor satisfies the London equation.
QUEERING JEREMIAH
Authors: MacWilliam Stuart 
Published: October 2002

AbstractSome criticism of queer theory has centred on its alleged ahistoricity in its treatment of historical texts and on its damage to the liberationist efforts of the identity-based feminist and lesbian and gay movements. This paper argues for queer theory's survival as both a valid academic methodological position, and an alternative liberationist approach to issues of gender and sexuality. Queer theory is then applied to the marriage metaphor in Jeremiah chs. 2–3. Recent feminist critiques of the prophetic use of the metaphor postulate a dichotomy between male and female readers' responses to the texts. In reply, this paper argues that there are serious breakdowns in gender divisions within what is a chaotic text; further, that the male reader implied in the text is forced to identify with not the faithful husband but the faithless wife; and finally that critics overlook the positive portrayal of the restored wife in the metaphor's 'third movement'. A queer reading does not wholly rehabilitate the text but suggests tentatively that the metaphor may be read as a subversion rather than a re-inscription of masculinist values.
Role of 4-1BB (CD137) in the functional activation of cord blood CD28−CD8+ T cells
Authors: Kim Young-June , Brutkiewicz Randy R. , Broxmeyer Hal E. 
Published: October 2002

AbstractThe CD28− subset of CD8+ T cells is associated with cytotoxic T lymphocyte (CTL) effector function. We investigated a potential role for 4-1BB, a costimulatory molecule structurally related to members of the tumor necrosis factor (TNF) receptor family, in the generation and functional activation of CD28− CTLs by using human cord blood (CB) cells composed exclusively of naive CD8+ T cells with few or no CD28− CTLs. The 4-1BB was induced preferentially on the CB CD28−CD8+ T cells when CD28 down-regulation was induced by interleukin 15 (IL-15) and IL-12 stimulation. Anti–4-1BB costimulation induced dramatic phenotypic changes in the CD28− CTLs, including restoration of CD28 expression as well as that of memory markers such as CD45RO and CC chemokine receptor 6 (CCR6). Anti–4-1BB costimulation also promoted long-term survival of CD28− CTLs, which were sensitive to activation-induced cell death upon anti-CD3 stimulation. The memory-type CD28+CTLs induced by anti–4-1BB costimulation acquired a greatly enhanced content of granzyme B, a cytolytic mediator, and enhanced cytotoxic activity as compared with CD28− CTLs. Strong cytotoxicity of memory-type CTLs to a 4-1BB ligand–expressing Epstein-Barr virus (EBV)–transformed B-cell line was almost completely abrogated by 4-1BB–Fc, a soluble form of 4-1BB, suggesting involvement of 4-1BB in cytolytic processes. Taken all together, our results suggest that 4-1BB plays a role in the differentiation of effector memory CTLs.
Crystalline-State Photoisomerization of α,β-Unsaturated Thioamide Analyzed by X-rays
Authors: Hosoya Takaaki , Ohhara Takashi , Uekusa Hidehiro , Ohashi Yuji 
Published: October 2002

Abstract
When a crystal of N,N-dibenzyl-1-cyclohexenecarbothioamide was exposed to UV light at 263 K, the cell dimensions were gradually changed with retention of the single crystal form. Since the crystal gradually decomposed after 330 h, the irradiation was stopped and the structure was analyzed by X-rays. The thioamide molecule was converted by 65.0% to the thiolactam molecule. The thioamide and thiolactam molecules were observed as a disordered structure in the crystal. The disordered structure clearly indicates that the hydrogen atom of one of the benzyl groups is extracted by one of the olefin carbon atoms of the cyclohexenyl group and that the benzyl carbon atom makes a bond with another olefin carbon to form a thiolactam ring. The mechanism of the ring formation and the chirality of the produced thiolactam are well explained from the reaction cavity.
Shape behavior of lipid vesicles as the basis of some cellular processes
Authors: Svetina Saša , Žekš Boštjan 
Published: October 2002

AbstractThe basic principles that govern the shape behavior of phospholipid vesicle shapes are discussed. The important membrane parameters of the system are defined by presenting the expressions for the relevant contributions to the system's mechanical energy. In the description of the rather unique shape behavior of lipid vesicles, the emphasis is on providing a qualitative understanding of the dependence of vesicle shape on the parameters of the system. The vesicle shape behavior is then related to biologically important phenomena. Some examples are given of how the results of the shape behavior of lipid vesicles can be applied to the analysis of cellular systems. Red blood cell shape and shape transformations, vesicle fission and fusion processes, and the phenomenon of cellular polarity are considered. It is reasoned that the current biological processes that involve changes of membrane conformation may have their origin in the general shape behavior of closed lamellar membranes. Anat Rec 268:215–225, 2002. © 2002 Wiley‐Liss, Inc.
Motility proteins and the origin of the nucleus
Authors: Dolan Michael F. , Melnitsky Hannah , Margulis Lynn , Kolnicki Robin 
Published: October 2002

AbstractHypotheses on the origin of eukaryotic cells must account for the origin of the microtubular cytoskeletal structures (including the mitotic spindle, undulipodium/cilium (so‐called flagellum) and other structures underlain by the 9(2)+2 microtubular axoneme) in addition to the membrane‐bounded nucleus. Whereas bacteria with membrane‐bounded nucleoids have been described, no precedent for mitotic, cytoskeletal, or axonemal microtubular structures are known in prokaryotes. Molecular phylogenetic analyses indicate that the cells of the earliest‐branching lineages of eukaryotes contain the karyomastigont cytoskeletal system. These protist cells divide via an extranuclear spindle and a persistent nuclear membrane. We suggest that this association between the centriole/kinetosome axoneme (undulipodium) and the nucleus existed from the earliest stage of eukaryotic cell evolution. We interpret the karyomastigont to be a legacy of the symbiosis between thermoacidophilic archaebacteria and motile eubacteria from which the first eukaryote evolved. Mutually inconsistent hypotheses for the origin of the nucleus are reviewed and sequenced proteins of cell motility are discussed because of their potential value in resolving this problem. A correlation of fossil evidence with modern cell and microbiological studies leads us to the karyomastigont theory of the origin of the nucleus. Anat Rec 268:290–301, 2002. © 2002 Wiley‐Liss, Inc.
Determinants of patient satisfaction in chronic illness
Authors: Bidaut‐Russell Michelle , Gabriel Sherine E. , Scott Christopher G. , Zinsmeister Alan R. , Luthra Harvinder S. , Yawn Barbara 
Published: October 2002

AbstractObjectiveTo determine whether primary care provided by generalists versus subspecialists resulted in different levels of patient satisfaction among persons with chronic illness.MethodsA survey containing the Primary Care Provider Questionnaire and the Health Status Questionnaire (HSQ) was mailed to 2 population‐based cohorts of patients with rheumatoid arthritis (RA) or diabetes mellitus (DM). All subjects were at least 35 years old and Rochester, Minnesota residents. Descriptive statistics, Spearman correlation coefficients, and multiple regression models were used to describe and compare the determinants of patient satisfaction.ResultsA total of 86 people (74% female) with RA and 208 people (56% male) with DM responded to the survey. Age range was 41–95 years and median disease duration was 8.7 years (RA) and 13.0 years (DM). Most patients described their health as fair or good. After adjusting for sex differences, RA patients were more likely than DM patients to report having a specialist as their primary care doctor. RA patients, whether reporting seeing a specialist or a generalist, had comparable HSQ physical health, mental health, social functioning, vitality, and bodily pain scores. DM patients seeing a specialist had more bodily pain and poorer physical functioning than those seeing a generalist. Across both chronic illnesses and physician specialties, median scores for patient satisfaction ranged from 17–18 for overall satisfaction (maximum 20); 30–33 for interpersonal skills (maximum 35); 23–26 for technical quality (maximum 30); and 20 for access to care (maximum 25). Multiple linear regression models revealed that 6.8–7.3% of the variation in satisfaction could be explained by HSQ scores, patient demographics, and physician specialty.ConclusionBoth RA and DM patients were highly satisfied with their care, regardless of the specialty of the provider. Physician specialty, patient demographics, and HSQ scores explained only a small proportion in the variation in satisfaction. These findings point to the need for additional research to further elucidate the determinants of patient satisfaction.
Hydrophobicity and helicity of membrane‐interactive peptides containing peptoid residues
Authors: Tang Yan‐Chun , Deber Charles M. 
Published: October 2002

AbstractPeptoid (N‐alkylglycyl) residues in peptides have been studied in a variety of applications, but their behavior in membrane environments has not been systematically investigated. We have synthesized a series of membrane‐interactive peptides of prototypic structure KKAAAXAAAAAXAAWAAXAAAKKKK‐amide, where X corresponds to the peptoid residues Nala (= sarcosine), Nval, Nile, Nleu, Nphe, and Ntrp. Investigation of their relative hydrophobic character by high‐performance liquid chromatography indicated an order of hydrophobicity Ntrp > Nphe > Nleu > Nile > Nval > Nala—largely parallel to the relative scale for these side‐chains in natural amino acids, although all values were significantly more “hydrophilic” than their amino acid correspondents. Conformations of peptoid‐containing peptides measured by circular dichroism spectroscopy were unordered in the presence of SDS micelles but helical for peptides with X = the corresponding amino acids, suggesting a general helix‐breaking tendency for the peptoid residues. However, peptides were able to form helical structures in the solvent n‐butanol, indicating that this conformation is possible if peptides became inserted into micellar phases. The latter notion was confirmed by increasing hydrophobic content of the peptides by embedding peptoid Nala residues in Leu‐rich rather than Ala‐rich sequences, which promoted peptide insertion and helical structure in micelles. The overall results suggest that judicious interspersing of amino acid and peptoid residues in peptide sequences can produce hydrophobic water‐soluble materials with membrane‐partitioning capacity. © 2002 Wiley Periodicals, Inc. Biopolymers 65: 254–262, 2002
Internal dynamics and ionization states of the macrophage migration inhibitory factor: Comparison between wild‐type and mutant forms
Authors: Soares T. A. , Lins R. D. , Straatsma T. P. , Briggs J. M. 
Published: October 2002

AbstractThe macrophage migration inhibitory factor (MIF) is a cytokine that shares a common structural architecture and catalytic strategy with three isomerases: 4‐oxalocrotonate tautomerase, 5‐carboxymethyl‐2‐hydroxymuconate isomerase, and D‐dopachrome tautomerase. A highly conserved N‐terminal proline acts as a base‐acid during the proton transfer reaction catalyzed by these enzymes. Such unusual catalytic strategy appears to be possible only due to the N‐terminal proline pKa shifted to 5.0–6.0 units. Mutations of this residue result in a significant decrease of the catalytic activity of MIF. Two hypotheses have been proposed to explain the catalytic inefficiency of MIF: the lower basicity of primary amines with regard to secondary ones and the increased flexibility resulting from the replacement of a proline by residues like glycine. To investigate that, we have performed molecular dynamics simulations of MIF wild‐type and its mutant P1G, as well as calculated the protonation properties of several mutant forms. It was found that the N‐terminal glycine does not show larger fluctuations compared to proline, but the former residue is more exposed to the solvent throughout the simulations. The apparent pKa of these residues displays very little change (as expected from the structural rigidity of MIF) and is not significantly affected by the surrounding ionizable residues. Instead, the hydrophobic character of the active site seems to be the main factor in determining the pKa of the N‐terminal residue and the catalytic efficiency of MIF. © 2002 Wiley Periodicals, Inc. Biopolymers 65: 313–323, 2002