bluuebunny/crossref_metadata_2025 huggingface dataset:
SPHERICAL HARMONICS ASSOCIATED TO THE LAPLACE–BESSEL OPERATOR AND GENERALIZED SPHERICAL CONVOLUTIONS
Authors: ALIEV ILHAM A. , RUBIN BORIS
Published: March 2003

A theory of spherical harmonics associated to the Laplace–Bessel differential operator is developed. Natural analogs of the Plancherel theory, the Laplace formula, the Funk–Hecke formula, the product formula, and the addition theorem are obtained. Symmetry properties of the Fourier–Bessel transform, decompositions of smooth functions, and convolution operators are studied.
Projections from the paraventricular nucleus of the thalamus to the rat prefrontal cortex and nucleus accumbens shell: Ultrastructural characteristics and spatial relationships with dopamine afferents
Authors: Pinto Aline , Jankowski Michael , Sesack Susan R.
Published: March 2003

The paraventricular nucleus of the thalamus (PVT) participates in the functional integration of limbic cortical and striatal circuitry. In the rat, the PVT projects to the deep layers of the medial prefrontal cortex (PFC) and to the shell of the nucleus accumbens (NAc). However, the synaptic organization of PVT afferents within these regions remains undescribed. Furthermore, although dopamine (DA) modulates excitatory glutamate transmission in both areas, possible anatomic substrates for specific DA modulation of PVT inputs have not yet been investigated. To address these issues, immunoperoxidase labeling for tyrosine hydroxylase (TH) in DA axons was combined with anterograde tract‐tracing, either by biotinylated dextran amine (BDA) labeled with immunogold–silver or by degeneration after lesions of the PVT. In both regions, and with either tracing method, PVT terminals formed primarily asymmetric axospinous synapses; in the NAc, a proportion of PVT terminals also synapsed onto dendrites. PVT profiles in both regions were often seen in direct apposition to TH‐immunoreactive axons; this association was more evident in the NAc where the DA innervation is denser. Within the PFC, PVT profiles and TH‐labeled axons were occasionally apposed to the same dendrites, but synaptic specializations were not typically seen at these seeming points of convergence. Within the NAc, PVT profiles occasionally made synapses onto spines and distal dendrites that received convergent synapses from TH‐immunoreactive varicosities. These findings represent the first demonstration of postsynaptic convergence between DA and thalamic afferents to a striatal region and are consistent with direct synaptic modulation of PVT transmission by DA in the NAc but not the PFC. J. Comp. Neurol. 459:142–155, 2003. © 2003 Wiley‐Liss, Inc.
A composite statistical procedure for evaluating genotoxicity using cell transformation assay data
Authors: Nishiyama Hiroshi , Omori Takashi , Yoshimura Isao
Published: March 2003

This study proposes a statistical procedure for evaluating data from the BALB/c 3T3 cell transformation assay (CTA). Three steps are applied: (i) a Dunnett type test for identifying clear negativity, (ii) a Margolin procedure for detecting and eliminating downturn data, and (iii) a maximum contrast method for evaluating the dose‐dependent increase of the response. Using Monte Carlo simulation the proposed procedure was compared to three other methods, i.e. a linear regression test, the Simpson–Margolin procedure with linear regression test (trend test), and the Omori procedure. When false positive rates were commonly controlled at a similar level for all methods, the procedure's effectiveness was greatest for dose–response patterns exhibiting non‐linear and down turn parts, being in agreement with a collaborative CTA validation study performed in Japan. Evaluation of CTA data by the proposed procedure satisfied participating researchers. Copyright © 2003 John Wiley & Sons, Ltd.
Toxicokinetic modeling for environmental health problems
Authors: Edler Lutz , Heinzl Harald
Published: March 2003

Toxicokinetic (TK) and physiologically based toxicokinetic (PB‐TK) models enable tissue dosimetry and the definition of the target organ dose after exposure to exogenous toxic compounds. This has qualified PB‐TK models for extrapolation from the experimental animal to the human, from high to low doses, between routes of exposure, between patterns of exposure, and also between robust and susceptible sub‐populations. We show how PB‐TK models are constructed, what assumptions are needed, which mathematical methods are used for model building and by which statistical methods one may assess both model fit and uncertainty of the modeling itself. We will address in particular a new generation of PB‐TK models which include age‐dependent model parameters to model life‐long human exposure with an example from the exposure to dioxins. Finally, we define the role of PB‐TK models for the identification of human health effects after exposure to chemicals in risk assessment. Copyright © 2003 John Wiley & Sons, Ltd.
Core self‐evaluations: a review of the trait and its role in job satisfaction and job performance
Authors: Bono Joyce E. , Judge Timothy A.
Published: March 2003

Over the past five years there has been a growing body of literature that examines the relationships among some of psychology's most studied traits (Neuroticism, self‐esteem, and locus of control). Core self‐evaluation theory posits a conceptual and empirical relationship between these traits and job satisfaction. After briefly reviewing core self‐evaluation theory, we examine the empirical evidence documenting a relationship between these traits and the two central criteria of interest to I/O psychologists—job satisfaction and job performance. We then examine the relationship between core self‐evaluation traits and the Big Five personality traits. We conclude with a discussion of the contributions and limitations of core self‐evaluation research and opportunities for future research. Copyright © 2003 John Wiley & Sons, Ltd.
HLA class I and II typing of the patients with Behçet’s disease in Saudi Arabia
Authors: Yabuki K. , Ohno S. , Mizuki N. , Ando H. , Tabbara K.F. , Goto K. , Nomura E. , Nakamura S. , Ito N. , Ota M. , Katsuyama Y. , Inoko H.
Published: March 2003

Thirteen Saudi Arabian patients with Behçet’s disease (BD) were typed for HLA‐A and ‐B alleles by the conventional serologic typing and for HLA‐DRB1, ‐DQB1 and ‐DPB1 alleles by the polymerase chain reaction‐restriction fragment length polymorphism (PCR‐RFLP) method. As a result, the phenotype frequency of the B51 antigen was significantly increased in the patient group as compared to the ethnically matched control group (76.9% in patients vs. 22.2% in controls), but no significant difference was observed in HLA‐A, ‐DRB1, ‐DQB1 or ‐DPB1 alleles between the patients and controls, as previously observed in Japanese BD. Further, by HLA‐B51 allelic genotyping performed by the polymerase chain reaction‐sequence specific primers (PCR‐SSP) method, all of the B51‐positive patients and controls were found to carry one particular allele, B*5101, except one patient with B*5108.
A new predictive model for insulin‐dependent diabetes mellitus susceptibility based on combinations of molecular HLA‐DRB1 and HLA‐DQB1 pockets
Authors: Djoulah S. , Busson M. , Sasazuki T. , Maillere B. , Yasunaga S. , Kimura A. , Charron D. , Hors J.
Published: March 2003

With a view to establishing an accurate evaluation of the genetic predisposition to insulin‐dependent type I diabetes (IDDM), we have built a model based on the characteristics of the relevant pockets of HLA‐DR and ‐DQ molecules. Three independent populations were investigated. Group I and group II were Caucasoids, while group III was Japanese, including a total of 1,166 IDDM patients and 2,391 healthy controls. We formulate the hypothesis that suceptibility to IDDM is not only explained by the absence of Aspartate 57 (negative charge) from pocket 9 of DQB1 (P9DQ), but also by the presence of an electric charge (+/− vs. neutral), generated by residues 70, 71 and 74 in pockets 4 of DRB1 (P4DR) and DQB1 (P4DQ) molecules. The respective weight of each pocket, was evaluated in a multivariate analysis based on the logistic regression method. The 4 components (2 loci and 2 pockets) were systematically analysed in the computer model. It was clearly shown that the structural characteristics of pockets P9DQ‐P4DR and, to a lesser degree that of P4DQ, account for IDDM predisposition. On applying the model to the whole international series, it appears that the highest risk concerns individuals with P9DQ non‐Asp 57 and both the charged P4 of DRB1 and P4 of DQB1, conferring a 80% prediction of susceptibility. Conversely, P9DQ Asp and neutral P4DR and P4DQ give the lowest risk with a predictive value of 5%. This model of risk susceptibility prediction fits remarkably well with the observed distribution in a worldwide study. It allows a better evaluation of the respective role of HLA‐DR and ‐DQ molecules as a major component of susceptibility to IDDM.
Linkage and association study of the CTLA‐4 region in coeliac disease for Italian and Tunisian populations
Authors: Clot F. , Fulchignoni‐Lataud M.C. , Renoux C. , Percopo S. , Bouguerra F. , Babron M.C. , Djilali‐Saiah I. , Caillat‐Zucman S. , Clerget‐Darpoux F. , Greco L. , Serre J.L.
Published: March 2003

Coeliac disease (CD) is a multifactorial disease for which there is an intensive search for genetic risk factors. Some authors found an association between the CTLA‐4 region and CD. In the present work, we investigate the possible implication of the CTLA‐4 region as a genetic risk factor for CD, through two statistical approaches: the maximum likelihood score (MLS) test in a large Italian sample of affected sib‐pairs using polymorphic genetic markers on chromosome 2, and the transmission disequilibrium test (TDT) in continental Italian and Tunisian families using the CTLA‐4 exon 1 49 A/G polymorphism. None of these approaches provides evidence for linkage or association between the CTLA‐4 region and CD. This might result from a difference in the CTLA‐4 region from population to population, either in its involvement as a risk factor or in the strength of linkage disequilibrium.
Novel associations among HLA‐DQA1 and ‐DQB1 alleles, revealed by high‐resolution sequence‐based typing (SBT)
Authors: Pera C. , Delfino L. , Longo A. , Pistillo M.P. , Ferrara G.B.
Published: March 2003

Althought it is a valuable tool for the identification of HLA alleles, sequence‐based typing (SBT) presents difficulties when used to determine HLA‐DQA1 and ‐DQB1 alleles. Specifically, some HLA‐DQA1 alleles have a three‐base deletion at codon 56 of exon 2 that interferes with the sequencing read. Moreover, the frequently used primers for HLA‐DQB1 may co‐amplify the HLA‐DQB2 pseudogene. To overcome these problems, we amplified DQA1 exon 2 using five group‐specific polymerase chain reactions (PCRs) which allowed separation of deleted from non‐deleted DQA1 alleles. DQB1 exon 2 was amplified using two group‐specific amplifications. To increase typing resolution, we also analyzed DQA1 exons 1, 3 and 4 and DQB1 exon 3 by PCR using sequence‐specific primers (PCR‐SSP) or SBT analysis. Using this method we found some important associations between DQA1 and DQB1 alleles: DQA1*05011 and DQB1*0201, DQA1*0505 and DQB1*03011, DQA1*01021 and DQB1*06, DQA1*01022 and DQB1*0502.
High‐resolution HLA class I typing in the CEPH families: analysis of linkage disequilibrium among HLA loci
Authors: Bugawan T.L. , Klitz W. , Blair A. , Erlich H.A.
Published: March 2003

The HLA region on the short arm of chromosome 6 (6p21.3) contains the most polymorphic coding sequences in the human genome. High‐resolution DNA‐based HLA typing of population samples of the polymorphic class I loci, HLA‐A, ‐B, and ‐C has only recently become feasible. Here, we report molecular HLA typing on family‐based samples of European origin (the CEPH repository), which demonstrated very high polymorphism, with 20 A alleles, 38 B alleles and 19 C alleles in the sample of 248 independent haplotypes. In general, allele frequency distributions are consistently more even (lower observed homozygosity statistic) than expected from a past of selective neutrality, suggesting a history of balancing selection. This was also true for the class II loci, DRB1, DQA1 and DQB1 in these samples, but not for the DPA1 and DPB1 loci, whose allelic frequency distributions were more skewed (higher observed homozygosity statistic) than expected under a neutral model. Although linkage disequilibrium is a prominent feature across the HLA region, only 19% of the eight locus haplotypes were sampled more than once. The relative age of some of the B alleles could be inferred from the pattern of B‐C haplotypic associations. We suggest that the observed patterns of linkage disequilibrium reflect the operation of selection on nearly all HLA alleles.
HLA‐B*15 diversity in the Korean population
Authors: Lee K.W. , Jeon H. , Park J.Y.
Published: March 2003

Acknowledgments:

This research was supported by a grant (KOSEF 981‐0701‐002‐2) from the Basic Research Program of the KOSEF. We thank Dr. C. K. Hurley at Georgetown University for the generous gift of some reference DNAs as well as for a critical review of the manuscript, and Miss J. S. Lee for technical assistance.

Alleles in the HLA‐B*15 group encode molecules belonging to several serologic subgroups, B15 (B62, B63, B75, B76, B77) and B70 (B71, B72), representing many of the most problematic types to assign in routine clinical typing laboratories due to their serologic cross‐reactivity resulting from structural similarity. More than 25% of Koreans express HLA‐B molecules encoded by the HLA‐B*15 alleles. To further characterize HLA‐B*15 in this population, B*15‐specific polymerase chain reaction (PCR) and sequence‐specific oligonucleotide probe (SSOP) hybridization analysis using 39 digoxigenin‐labeled probes were applied to DNA samples obtained from 237 B15/B70 serologically positive unrelated individuals. Nine B*15 alleles were identified. B*1501 was the most frequent allele (64.8%) followed by B*1511 (14.1%), B*1507 (8.6%), and B*1518 (5.5%) comprising more than 90% of B*15‐positive samples. B62 molecules encoded by 4 of the identified alleles (B*1501, B*1507, B*1525, and B*1527) could not be discriminated by serologic reaction patterns. Among the fifteen B15/B70 apparent homozygotes, eight were heterozygotes carrying two different B*15 alleles. Several B*15 alleles exhibited strong associations with specific Cw, DRB1, and A allelic types (e.g., B*1507‐Cw3 (22/22); B*1507‐DRB1*04 (21/22), B*1507‐A24 (17/22)). The data obtained in this study confirmed B*15 diversity in the study population and will be useful in hematopoietic stem cell donor searches as well as in determining the supplementary DNA typing strategy for B15/B70‐positive samples in this population.
Downregulation of the constitutive tapasin expression in human tumor cells of distinct origin and its transcriptional upregulation by cytokines
Authors: Seliger B. , Schreiber K. , Delp K. , Meissner M. , Hammers S. , Reichert T. , Pawlischko K. , Tampé R. , Huber CH.
Published: March 2003

Human tumor cells frequently exhibit abnormalities in the major histocompatibility complex (MHC) class I surface expression which can be due to structural alterations and/or dysregulation of various components of the MHC class I antigen processing machinery, such as HLA class I heavy and light chains, the peptide transporter and the proteasome subunits. Although several cofactors critical for proper MHC class I assembly have been identified, their contribution to the immune escape phenotype of tumor cells has not been analyzed. In order to determine whether tapasin deficits are an integral part of immune escape mechanisms of human tumors, we studied the constitutive and cytokine‐regulated expression pattern of tapasin in malignant cells of distinct histology. Heterogeneous and reduced expression levels of tapasin were found in small‐cell lung carcinoma, pancreatic carcinoma, colon carcinoma, head an neck squamous cell carcinoma and renal cell carcinoma cell lines. Tapasin downregulation was also prominent in surgically removed tumor lesions when compared to normal controls. The impaired tapasin expression is often associated with low MHC class I cell surface expression. In addition, various cytokines, including interferon (IFN)‐α, IFN‐γ, tumor necrosis factor (TNF)‐α and interleukin (IL)‐4, but not granulocyte‐macrophage colony stimulating factor (GM‐CSF), transcriptionally upregulate to a distinct extent and in a time‐dependent manner tapasin expression in tumor cells. Thus, deficient tapasin expression appears to be a frequent event in human tumor cells. Its restoration by cytokines further suggests that impaired tapasin expression in tumors is rather due to dysregulation than to structural alterations.
HLA class I genotyping including HLA‐B*51 allele typing in the Iranian patients with Behçet’s disease
Authors: Mizuki N. , Ota M. , Katsuyama Y. , Yabuki K. , Ando H. , Yoshida M. , Onari K. , Nikbin B. , Davatchi F. , Chams H. , Ghaderi A.A. , Ohno S. , Inoko H.
Published: March 2003

Abstract: It is well known that Behçet’s disease (BD) is strongly associated with human leukocyte antigen (HLA) B51 in many ethnic groups. However, there has been no published report as yet with respect to this association among the Iranian people. Furthermore, since it is now known that the B51 antigen can be encoded by 21 alleles, B*5101‐B*5121, we performed HLA‐B*51 allele typing as well as HLA class I genotyping of 48 Iranian patients with this disease. As a result, the frequency of the B*51 allele was significantly higher (62.1%) in the patient group as compared with the ethnically matched control group (31.8%) (Pc=0.067, R.R.=3.51). In the genotyping of B*51 alleles, 33 out of the 36 B*51‐positive patients possessed B*5101 and the remaining 3 carried B*5108. This study revealed that Iranian patients with BD also had a strong association with HLA‐B51. In addition, this significantly high incidence of HLA‐B*51 was found to be caused by an increase in both the HLA‐B*5101 and HLA‐B*5108 alleles. However, there was no significant difference in the HLA‐B*51 allelic distribution between the patient and control groups.
The HLA Dictionary 2001: a summary of HLA‐A, ‐B, ‐C, ‐DRB1/3/4/5, ‐DQB1 alleles and their association with serologically defined HLA‐A, ‐B, ‐C, ‐DR and ‐DQ antigens
Authors: Schreuder G.M.Th. , Hurley C.K. , Marsh S.G.E. , Lau M. , Maiers M. , Kollman C. , Noreen H.J.
Published: March 2003

Abstract: This report presents the serologic equivalents of 123 HLA‐A, 272 HLA‐B and 155 HLA‐DRB1 alleles. The equivalents cover over 64% of the presently identified HLA‐A, ‐B and ‐DRB1 alleles. The dictionary is an update of the one published in 1999 and also includes equivalents for HLA‐C, DRB3, DRB4, DRB5 and DQB1 alleles. The data summarize information obtained by the WHO Nomenclature Committee for Factors of the HLA System, the International Cell Exchange (UCLA), the National Marrow Donor Program (NMDP) and individual laboratories. In addition, a listing is provided of alleles which are expressed as antigens with serologic reaction patterns that differ from the well‐established HLA specificities. The equivalents provided will be useful in guiding searches for unrelated hematopoietic stem cell donors in which patients and/or potential donors are typed by either serology or DNA‐based methods. These equivalents will also serve typing and matching procedures for organ transplant programs where HLA typings from donors and from recipients on waiting lists represent mixtures of serologic and molecular typings. The tables with HLA equivalents and a questionnaire for submission of serologic reaction patterns for poorly identified allelic products will also be available on the WMDA web page: http://www.worldmarrow.org.
HLA‐B*51 allele analysis by the PCR‐SBT method and a strong association of HLA‐B*5101 with Japanese patients with Behçet’s disease
Authors: Mizuki N. , Ota M. , Katsuyama Y. , Yabuki K. , Ando H. , Shiina T. , Nomura E. , Onari K. , Ohno S. , Inoko H.
Published: March 2003

Abstract: Behçet’s disease (BD) is known to be associated with human leukocyte antigen (HLA) B51 in many different ethnic groups. An increased incidence of HLA‐B51 in the patient group has also been reported in a Japanese population. Recently, the B51 antigen has been identified to comprise 21 alleles, B*5101–B*5121. Further, not only HLA‐B*5101 but also HLA‐B*5108 were found to be relatively increased in the patient groups among Italian and Saudi Arabian populations. Therefore, we performed HLA‐B*51 allele genotyping by the polymerase chain reaction‐sequencing based typing (PCR‐SBT) method in order to investigate whether there is any correlation of one particular B51‐associated allele with Japanese BD. Ninety‐six Japanese patients with BD and 132 healthy Japanese volunteers were enrolled in this study. As a result, the phenotype frequency of the B51 antigen was confirmed to be remarkably increased in the patient group as compared to the ethnically matched control group (59.4% in patients vs. 13.6% in controls; Pc=0.0000000000098, R.R.=9.3). In the B*51 allele genotyping, 56 out of 57 B51‐positive patients were defined as B*5101 and the remaining one was B*5102. In contrast, all of 18 B51‐positive normal controls were B*5101. None of the Japanese patients and healthy controls carried the HLA‐B*5108 allele. This study revealed that B*51 allelic distribution in Japanese was different from those in Italian and Saudi Arabian populations, and that the significantly high incidence of the HLA‐B51 antigen in the Japanese BD patient group was mostly caused by the significant increase of the HLA‐B*5101 allele.
Seven single nucleotide substitutions in human Fcγ receptor IIB gene
Authors: Yasuda K. , Sugita N. , Yamamoto K. , Kobayashi T. , Yoshie H.
Published: March 2003

Abstract: Variation screening for the immunoglobulin G Fc receptor IIB (FcγRIIB) gene was performed with the genomic DNA from 100 healthy Japanese subjects. We identified 3 non‐synonymous and 2 synonymous substitutions and 2 single‐nucleotide polymorphisms in an intron region. These substitutions were found to be located in the ligand‐binding domain and the intron, which might alter the function of FcγRIIb.
Cellular methods used to evaluate the immune response in transplantation
Authors: Reinsmoen N.L.
Published: March 2003

Abstract: Cellular assays have been developed to test for various effector, cytotoxic, and regulatory functions of T cells and have been used throughout the history of clinical transplantation to assess the immune profile of solid organ and marrow recipients. One goal of these cellular studies has been to determine if posttransplant changes in the donor antigen‐specific cellular response could predict good and poor graft outcome, thereby allowing for individualization of immunosuppression. This review outlines the use of established and newly developed cellular assays to assess the dynamic processes of the posttransplant immune response and to provide insights into the mechanisms involved and potential points for intervention.
Non‐mutated tumor‐rejection antigen peptides elicit type‐I allergy in the majority of healthy individuals
Authors: Ohkouchi S. , Yamada A. , Imai N. , Mine T. , Harada K. , Shichijo S. , Maeda Y. , Saijo Y. , Nukiwa T. , Itoh K.
Published: March 2003

Abstract: IgE‐mediated type‐I allergy is generally considered to be a hypersensitivity reaction to foreign antigens, and it is believed that self‐antigens do not evoke this type of allergy. We report here, for the first time, that non‐mutated self‐antigen peptides identified as tumor‐rejection antigen peptides recognized by HLA class I‐restricted and tumor‐specific cytotoxic T lymphocytes (CTLs) elicited a type‐I allergy in the majority of healthy individuals. Peptide‐specific IgE was detectable in sera from certain cases, although the levels did not always correlate with those of type‐I allergy. Repeated vaccinations of nonallergic peptides derived from the same antigens possessing allergic peptides resulted in the suppression of both allergic peptide‐specific IgE responses and type‐I allergy, providing evidence for a new approach to the development of peptide‐based desensitization.
Major histocompatibility complex classI‐restricted antigen processing and presentation
Authors: Van Kaer L.
Published: March 2003

Abstract: Major histocompatibility complex (MHC) class I molecules present antigenic peptides to CD8‐expressing cytotoxic T lymphocytes (CTLs). This antigen recognition system is critically important for immune surveillance against viruses and tumors. Most class I‐binding peptides are generated in the cytosol, as side products from the degradation of misfolded proteins by proteasomes. A subset of the resulting peptides are translocated across the endoplasmic reticulum (ER) membrane by a dedicated peptide transporter, and these peptides are then loaded onto peptide‐receptive class I molecules in the ER. The stable assembly of class I molecules with peptides is controlled by a variety of accessory proteins, including chaperones with general housekeeping functions and factors with dedicated roles in class I assembly. Peptide‐filled class I molecules are then delivered to the cell surface for recognition by CTLs. This highly regulated process permits the host to rapidly counter invading pathogens with strong and sustained CTL responses and, at the same time, avoid misguided attacks. Here, how the class I antigen processing machinery accomplishes this daunting task is reviewed.
Quantitative determination of Fcγ receptor genes by means of fluorescence‐based  real‐time polymerase chain reaction
Authors: Gittinger F.S. , Schindler‐Wuepper L. , Kissel K. , Bux J.
Published: March 2003

Abstract: The granulocyte antigens HNA‐1a, ‐1b and ‐1c reside on the FcγIIIb receptor, which is exclusively expressed on neutrophils. They are involved in autoimmune and alloimmune neutropenias as well as in severe transfusion reactions. Recent family studies show the HNA‐1c antigen to be inheritably linked to HNA‐1a, resulting in individuals carrying three genes. Using sequence‐specific primers, the HNA antigens can be discerned in a polymerase chain reaction (PCR), but not the number of coding FCGR3B genes present in each individual. Therefore a real‐time kinetic PCR method using the LightCycler technique was established to ascertain the amount of FCGR3B genes in each individual. For this purpose, the FCGR3B genes of four HNA‐1a,‐1b,‐1c (+), one HNA‐1b,1c (+) and one HNA‐1b (+) individual were quantified. In addition, two families, in which the mother was FCGR3B deficient, were analyzed. Quantification showed two of the four HNA‐1a,‐1b and‐1c (+) individuals to have three genes and the other two to have only two genes. The HNA‐1b,‐1c (+) individual had also only two genes. As expected, the HNA‐1b donor was typed for two genes. No FCGR3B genes could be detected in the FCGR3B‐deficient mothers of either family; their husbands however, carried two FCGR3B genes. Accordingly, quantitative PCR showed the offspring of both families to have only one FCGR3B gene. Quantitative PCR with the Light Cycler has been revealed to be a fast and reliable method for the determination of FCGR3B genes present. FCGR3B quantification confirms the idea of the HNA‐1c antigen to be inherited not only linked to HNA‐1a, but also to be passed down on its own.
Identification of a new HLA‐B*40 variant, B*4035
Authors: Voorter C.E.M. , Hepkema B.G. , Lems S.P.M. , Van Den Berg‐Loonen E.M.
Published: March 2003

Abstract: In this report, the novel allele B*40351 is presented. The allele was identified in a Caucasian individual by sequence‐based typing. B*4035 is identical to B*4002 in exon 2, but differs in exon 3 at position 463, where it has an A in stead of a C. This results in an amino acid change from arginine to serine at codon 131 of the mature protein. The haplotype carrying the B*4035 was A3 B*4035 Cw2 DR11 DQ3.
Perspective: peptides as mimics of transmembrane segments in proteins
Authors: Deber C.M. , Liu L. ‐P. , Wang C.
Published: March 2003

Abstract: Peptide‐based approaches to protein structure within membranes have proven enormously valuable. When one focusses on the detailed manner through which membrane proteins actually traverse the cell bilayer, a simple observation emerges: helical peptide segments of 20 amino acids each constitute the only tangible connection between the inside and outside of the cell. Thus, a major step towards understanding the key relationships between biological function and membrane protein structure can be taken through characterization, by composition, sequence, chain length, hydrophobicity and conformation, of hydrophobic peptides designed as mimics of transmembrane segments.
N‐terminal myristylation of HBV preS1 domain enhances receptor recognition
Authors: De Falco S. , Ruvo M. , Verdoliva A. , Scarallo A. , Raimondo D. , Raucci A. , Fassina G.
Published: March 2003

Abstract: The N‐terminal portion of the large envelope protein of the human hepatitis B virus (HBV), the preS1 domain, plays a fundamental role in cell attachment and infectivity. Recent investigations have suggested that myristylation of preS1 Gly2 residue is essential for viral infectivity, but the importance of this post‐translational modification on HBV–receptor interaction has not been elucidated completely. In this study we produced, using stepwise solid‐phase chemical synthesis, the entire preS1[1–119] domain (adw2 subtype), and compared its receptor binding activity with the myristylated form, myristyl‐preS1[2–119] in order to define the importance of fatty acid modification. Both synthetic proteins were fully characterized in terms of structural identity using TOF‐MALDI mass spectrometry and analysis of tryptic fragments. Circular dichroism measurements indicated a low content of ordered structure in the preS1 protein, while the propensity of the myristylated derivative to assume a conformationally defined structure was more evident. HBV−receptor binding assays performed with plasma membranes preparations from the hepatocyte carcinoma cell line HepG2 clearly showed that the preS1[1–119] domain recognizes the HBV receptor, and confirmed that binding is occurring through the 21–47 region. The myristylated derivative recognized HBV receptor preparations with higher affinity than the preS1 domain, suggesting that the conformational transitions induced in the preS1 moiety by fatty acid post‐translational modification are important for efficient attachment of viral particles to HBV receptors.
Pseudopeptides containing the 2‐hydrazonoacyl fragment: analogs of the chemotactic agent HCO‐Met‐Leu‐Phe‐OMe
Authors: Torrini I. , Nalli M. , Paglialunga Paradisi M. , Pagani Zecchini G. , Lucente G. , Spisani S.
Published: March 2003

Abstract: In order to further examine the properties of pseudopeptides containing the 2‐hydrazonoacyl fragment, two new series of analogs of the prototypical chemotactic N‐formyl‐tripeptide HCO‐Met‐Leu‐Phe‐OMe were designed and synthesized. The first group contains the new fragment as the N‐terminal residue and is represented by the N‐aryl derivatives p‐Cl‐C6H4‐NH‐N=C(R)‐CO‐Leu‐Phe‐OMe (2 and 3) and by the corresponding N‐aroyl analogs p‐CH3‐C6H4‐CO‐NH‐N=C(R)‐CO‐Leu‐Phe‐OMe (4). The second group contains the new fragment in place of the central Leu residue and is represented by compounds HCO‐Xaa‐NH‐N=C(R)‐CO‐Phe‐OMe (7a and 7b) where Xaa is Nle and Met, respectively. The conformational and biochemical properties of the new products were examined.
Clavaspirin, an antibacterial and haemolytic peptide from Styela clava
Authors: In I.‐H. , Zhao C. , Nguyen T. , Menzel L. , Waring A.J. , Lehrer R.I. , Sherman M. A.
Published: March 2003

Abstract: We cloned the precursor of a novel peptide from a cDNA library prepared from pharyngeal tissues of the tunicate, Styela clava. Its sequence predicted a histidine‐rich, amidated 23‐residue peptide (FLRFigSVIHGIGHLVHHIGVAL‐NH2) that we named clavaspirin. A synthetic clavaspirin was prepared and it was found that it killed Gram‐positive and Gram‐negative bacteria, permeabilized the outer and inner membranes of Escherichia coli, lysed phosphatidylglycerol (POPG) liposomes, and was potently haemolytic towards human and bovine erythrocytes. Each of these activities was performed more effectively at an acidic pH. Circular dichroism measurements of synthetic clavaspirin revealed a largely α‐helical structure and polarized and residue‐specific FTIR spectrometry showed that its association with phospholipid membranes was influenced by pH. Peptides such as clavaspirin may equip tunicate haemocytes to mediate cytotoxicity and participate in antimicrobial defence.
Current status of a mucosal vaccine against dental caries
Authors: Hajishengallis G. , Michalek S. M.
Published: March 2003

The evidence of a specific bacterial cause of dental caries and of the function of the salivary glands as an effector site of the mucosal immune system has provided a scientific basis for the development of a vaccine against this highly prevalent and costly oral disease. Research efforts towards developing an effective and safe caries vaccine have been facilitated by progress in molecular biology, with the cloning and functional characterization of virulence factors from mutans streptococci, the principal causative agent of dental caries, and advancements in mucosal immunology, including the development of sophisticated antigen delivery systems and adjuvants that stimulate the induction of salivary immunoglobulin A antibody responses. Cell‐surface fibrillar proteins, which mediate adherence to the salivary pellicle, and glucosyltransferase enzymes, which synthesize adhesive glucans and allow microbial accumulation, are virulence components of mutans streptococci, and primary candidates for a human caries vaccine. Infants, representing the primary target population for a caries vaccine, become mucosally immunocompetent and secrete salivary immunoglobulin A antibodies during the first weeks after birth, whereas mutans streptococci colonize the tooth surfaces at a discrete time period that extends around 26 months of life. Therefore, immunization when infants are about one year old may establish effective immunity against an ensuing colonization attempts by mutans streptococci. The present review critically evaluates recent progress in this field of dental research and attempts to stress the protective potential as well as limitations of caries immunization.
Effect of inactivation of gtf genes on adherence of Streptococcus downei
Authors: Colby S. M. , McLaughlin R. E. , Ferretti J. J. , Russell R. R. B.
Published: March 2003

The activity of glucosyltransferases (GTF), a group of enzymes that synthesize water‐soluble and ‐insoluble glucans from sucrose, significantly contributes to the cariogenicity of mutans streptococci. Streptococcus downei produces four glucosyltransferases, GTFI, which produces insoluble glucan, and GTFS, GTFT, and GTFU, which synthesize soluble glucans. We have previously reported that inactivation of gtfS results in altered adherence and have now examined its interaction with other enzymes by constructing mutants which were gtfS, gtfS/gtfT, gtfS/gtfI and gtfI. The mutants were tested for their ability to accumulate on wires and on plastic microtiter trays in the presence of sucrose. The gtfS mutant displayed a reduced ability to adhere compared to the wild type but there was no further reduction of adherence in a gtfS/gtfT mutant. In contrast, the gtfS/gtfI double mutant showed a drastic reduction in adherence and when gtfI alone was inactivated, bacteria were unable to adhere to a hard surface. The results confirmed that insoluble glucan is required for strong adherence to a smooth surface but that the amount and structure of this glucan is dependent upon the availability of soluble glucans to act as primer molecules.
Identification of oral mitis group streptococci by arbitrarily primed polymerase chain reaction
Authors: Rudney J. D. , Larson C. J.
Published: March 2003

“Mitis group” streptococci are commensal but may play some role in dental caries, septicemia or endocarditis. Rapid genotypic identification would aid studies of dental plaque ecology, or diagnostic use. AP‐PCR with 58 unpaired arbitrary primers was used to characterize 7 Streptococcus gordonii, 11 Streptococcus sanguis, 2 Streptococcus crista, 5 Streptococcus parasanguis, 18 Streptococcus oralis, and 36 Streptococcus mitis (22 biovar 1 and 14 biovar 2). S. parasanguis 16S rRNA variable region primer RR2 produced species‐specific bands with all S. gordonii and S. sanguis. Human Vβ1 T‐cell receptor primer 434 yielded concordant genotypic identification of all phenotypically defined S. crista and S. parasanguis, 83% of S. oralis, and 74% of S. mitis biovar 1. Amplicon patterns for S. mitis biovar 2 were heterogeneous. Findings suggest that primers RR2 and 434 in succession will allow rapid identification of genotypic groups corresponding closely to mitis group species established by phenotype.
Adherence to and penetration through endothelial cells by oral treponemes
Authors: Peters S. R. , Valdez M. , Riviere G. , Thomas D. D.
Published: March 2003

Oral spirochetes were co‐incubated with monolayers of endothelial cells seeded into multiwell plates or onto filters mounted in plastic chambers. Attachment was assessed using an enzyme‐linked immunosorbent assay and scanning electron microscopy. Invasiveness was determined by monitoring media beneath filters within chambers for spirochetes using darkfield microscopy. Transmission electron microscopy was used to estimate intercellular and intracellular passage of spirochetes through monolayers. All tested treponemes attached to monolayers in a dose‐ and time‐ dependent manner, except Treponema phagedenis. A few treponemes were observed within host cell cytoplasm. Unidentified spirochetes obtained from dental plaque were also invasive. Results indicate that oral spirochetes possess virulence‐associated characteristics shared with pathogenic spirochetes. Further studies should examine the possibility that invasive spirochetes could disseminate from within affected gingiva.
Heterogeneity of Porphyromonas gingivalis strains in the induction of alveolar bone loss in mice
Authors: Baker P. J. , Dixon M. , Evans R. T. , Roopenian D. C.
Published: March 2003

These experiments examine alveolar bone loss in a model in which specific pathogen‐free mice are exposed orally with Porphyromonas gingivalis. Alveolar bone loss was induced as a result of a specific infection with P. gingivalis, rather than other environmental antigens. Infection with live P. gingivalis was required, as significant bone loss did not follow gavage with formalin‐killed P. gingivalis. The virulence of different strains of P. gingivalis was compared. Two laboratory strains of the bacteria (ATCC 53977 and W50) and a mutant strain lacking the 43‐kDa fimbrillin (strain DPG3) induced bone loss. P. gingivalis 381, however, did not induce bone loss. There was a strong immunoglobulin G (IgG) antibody response to infection with each strain but a significant serum IgA response only to strain 381. These studies show that in mice with a background oral microflora bone loss is induced by a specific infection with P. gingivalis and that bacterial strain variation is important in determining whether alveolar bone loss will ensue.
Induction of secretory immunity with bioadhesive poly ( D,L‐lactide‐co‐glycolide) microparticles containing Streptococcus sobrinus glucosyltransferase
Authors: Smith D. J. , Trantolo D. J. , King W. F. , Gusek E. J. , Fackler P. H. , Gresser J. D. , De Souza V. L. , Wise D. L.
Published: March 2003

The effect of mucosal delivery of Streptococcus sobrinus glucosyltransferase (GTF) in bioadhesive poly (
d,l‐lactide‐co‐glycolide) (PLGA) microparticles on induction of salivary IgA and serum IgG antibody responses was measured in Sprague‐Dawley rats. Preparations of GTF/PLGA/gelatin microparticles, or PLGA/gelatin microparticles or GTF in alum, were administered four times at weekly intervals by intranasal or intragastric routes. Two subcutaneous injections of GTF in PLGA/gelatin microparticles or in alum were given to separate groups of rats. Significant elevations in salivary IgA antibody levels to S. sobrinus GTF were observed only in the groups immunized intranasally 28 days after immunizations were begun. Five of six rats given the GTF microparticles intranasally had positive salivary IgA antibody responses to GTF, and the mean salivary IgA antibody level of this group was 30‐fold higher than any other mucosally or systemically immunized group. Salivary IgA responses in the GTF‐microparticle group remained significantly higher than all other mucosally immunized groups for at least 10 weeks after the primary immunization. All rats in this group demonstrated aspects of anamnesis following a more limited secondary course of intranasal administration. Intranasal administration of GTF in microparticles also induced a serum IgG response to GTF in some rats. After secondary intranasal GTF microparticle administration, several rats had sustained serum IgG antibody levels that were within the range of sera from rats subcutaneously injected with GTF in microparticles or in alum. Thus intranasal delivery of GTF‐containing bioadhesive microparticles induced the highest and longest lasting salivary immune response of any mucosal or systemic route or vehicle tested and could be expected to be a useful method for induction of mucosal immunity.
Prostaglandins E2 and I2 downregulate tumor necrosis factor α–induced intercellular adhesion molecule‐1 expression in human oral gingival epithelial cells
Authors: Noguchi K. , Iwasaki K. , Endo H. , Kondo H. , Shitashige M. , Ishikawa I.
Published: March 2003

In the present study, we examined whether prostaglandin (PG) E2 and PGI2 regulated intercellular adhesion molecule‐1 (ICAM‐1) expression in human oral gingival epithelial cells stimulated with tumor necrosis factor α (TNFα). TNFα potently induced ICAM‐1 expression in a dose‐ and time‐dependent fashion. PGE2 and carbacyclin (a stable analogue of PGI2) significantly decreased ICAM‐1 expression in TNFα‐challenged oral gingival epithelial cells. Next, of the four subtypes of PGE2 receptors (EP1, EP2, EP3 and EP4), we examined which subtype(s) mediated inhibition of TNFα‐induced ICAM‐1 expression by PGE2. 11‐deoxy‐PGE2, an EP2/EP4 agonist, significantly suppressed TNFα‐induced ICAM‐1 expression, whereas butaprost, an EP2 agonist, sulprostone, an EP1/EP3 agonist, and ONO‐AP‐324, an EP3 agonist, caused no effect on it. By reverse transcriptase‐polymerase chain reaction, expression of EP4 mRNA was detected in oral gingival epithelial cells. Dibutyryl cAMP, a cAMP analogue, and forskolin, a direct activator of adenylate cyclase, significantly inhibited TNFα‐induced ICAM‐1 expression in oral gingival epithelial cells. From these results, we suggest that PGE2 and PGI2 inhibit TNFα‐elicited ICAM‐1 expression by cAMP‐dependent pathways via EP4 receptors and IP receptors, respectively.
Growth stimulation of Porphyromonas endodontalis by hemoglobin and protoporphyrin IX
Authors: Zerr M. A. , Cox C. D. , Johnson W. T. , Drake D. R.
Published: March 2003

Porphyromonas endodontalis, like other Porphyromonas species, has a complex set of nutritional requirements. In addition to being an obligate anaerobe, the bacterium must be grown in a complex medium consisting of amino acids, reducing agents and heme compounds. P. endodontalis accumulates high concentrations of heme pigments to the extent that colonies appear black on blood agar. This accumulation of heme and the need for these compounds has been characterized as iron requirements by these species. However, in our studies, P. endodontalis demonstrated growth dependence on hemoglobin or protoporphyrin IX but not on free iron. Iron added to other heme compounds actually decreased growth stimulation by porphyrin‐containing compounds. P. endodontalis actively transported free iron, but this process did not appear to be critical for growth. The maximum stimulation of growth by protoporphyrin IX, under conditions of iron deprivation, suggests that P. endodontalis requires the porphyrin moiety as a growth factor.
Altered antigenicity in periodontitis patients and decreased adhesion of Porphyromonas gingivalis by environmental temperature stress
Authors: Amano A. , Premaraj T. , Kuboniwa M. , Nakagawa I. , Shizukuishi S. , Morisaki I. , Hamada S.
Published: March 2003

Periodontopathogenic bacteria survive various environmental changes during the progression of periodontal disease. Alterations in metabolism and protein expression will have to take place to adapt their physiological functions to environmental stress. We examined the effects of an elevation of 2°C in temperature on the adhesive ability and antigenicity of Porphyromonas gingivalis. Elevation of growth temperature of P. gingivalis from 37°C to 39°C remarkably suppressed the expression of surface filamentous structures, such as fimbriae, as well as the adhesive capacities to salivary components and Streptococcus oralis. Sera of severe periodontitis patients revealed a marked increase in serological activity with 39°C cells than with 37°C cells. The alteration of protein profiles of bacterial surface components by temperature elevation was demonstrated by SDS–PAGE, and their Western blot profiles were also different from those of cells grown at 37°C. Although a uniform trend was not found in the altered patterns, sera from severe periodontitis patients detected more antigenic proteins in cells grown at 39°C than 37°C cells. These observations suggest that P. gingivalis downregulates the expression of fimbriae and alters its adhesive capacity and antigenicity by the temperature stress that could occur during the disease progression.
Environmental regulation of recA gene expression in Porphyromonas gingivalis
Authors: Liu Y. , Fletcher H. M.
Published: March 2003

The recA gene product in Porphyromonas gingivalis is involved in DNA repair. Further, disruption of this gene can affect the proteolytic activity and expression of other virulence factors in this organism. Since several known environmental factors can influence virulence gene expression in P. gingivalis, we investigated the influence of these signals on the expression of the recA gene in this organism. A heterodiploid strain of P. gingivalis (designated FLL118) containing a transcriptional fusion of the recA promoter region and the promoterless tetracycline‐resistant gene [tetA(Q)2] and xylosidase/arabinosidase (xa) gene cassette was constructed. The recA promoter activity was assessed by measurement of xylosidase activity in FLL118. The expression remained relatively constant during different growth phases, at different pH levels and in the presence of DNA‐damaging agents. In response to hemin limitation and in the presence of calcium there was a moderate increase in recA promoter activity. Temperature also affected the expression. The highest level of xylosidase activity was observed in cultures at 32°C with a decline of approximately 46% as growth temperature increased to 41°C. Reverse transcriptase polymerase chain reaction analysis revealed that this regulation may be occurring at the transcriptional level. These results suggest that expression of the recA gene in P. gingivalis W83 is responsive to several environmental signals but is not regulated by a DNA damage–inducible SOS‐like regulatory system.
HLA class II alleles and salivary numbers of mutans streptococci and lactobacilli among young adults in Japan
Authors: Ozawa Y. , Chiba J. , Sakamoto S.
Published: March 2003

To investigate the genetic predisposition toward the accumulation of oral microorganisms, the association between the alleles of HLA‐DQA1, ‐DQB1 and ‐DRB1 and the salivary numbers of mutans streptococci and lactobacilli were examined in 102 young adults in Japan. HLA‐DQA1*0102, DQB1*0604, DRB1*0802, and DRB1*1302 were weakly associated with the numbers of lactobacilli in saliva samples (P<0.05). HLA‐DQB1*0601 was associated with the salivary numbers of mutans streptococci (P<0.01). Two HLA class II haplotypes, HLA‐ {DRB1*0802‐DQA1*0301‐DQB1*0302} and HLA‐{DRB1*1302‐ DQA1*0102‐DQB1*0604}, were also weakly associated with the salivary numbers of lactobacilli (P<0.05). These observations suggest that HLA class II alleles may be related to the salivary populations of oral microorganisms, such as mutans streptococci and lactobacilli.