bluuebunny/crossref_metadata_2025 huggingface dataset:
The Reflective Practitioner and the Uses of Rhetoric
Authors: Farmer David John , Patterson Patricia M.
Published: March 2003

This essay invites reflection on four practical uses of rhetoric: as a way to provide persuasive emphasis in administrative communication; as a technique of analysis to improve public administration and expand its limits; as a resource in identifying individual administrative signature; and as a vehicle for reevaluating public administration’s group signature. It reflects on the authors’ experiences and considers possibilities for change toward more conscious and effective group self–representations.
Housing Price Volatility Changes and Their Effects
Authors: Dolde Walter , Tirtiroglu Dogan
Published: March 2003

We examine significant volatility shifts in regional housing price changes, adapting a method of Haugen, Talmor and Torous (1991) independent of predefined sampling blocks. We identify 36 volatility events, most of which are purely regional, but three of which are national. We find significant associations of volatility events and economic conditions, especially national and regional income growth, inflation, and interest rates. During an initial adjustment period after a volatility shift, realized housing returns move opposite to volatility. We find evidence of significant interregional diffusion of volatility increases, but not of decreases. New insights on links between economic conditions and housing volatility and returns should be of value to household investors and mortgage investors.
PARTIAL WAVE ANALYSIS AT LEAR
Authors: Peters Klaus
Published: March 2003

Methods and special techniques for partial wave analysis and dalitzplot analysis in the framework of antiproton nucleon annihilation are presented. The properties of the initial system and the constraints on the final states are discussed as well as many details about the setup and strategy of the analyses. This information was compiled mainly from the experiments Crystal Barrel, Obelix and Asterix but is certainly not complete in many respects.
A STUDY OF THE DYNAMICS OF λsinz
Authors: DOMÍNGUEZ PATRICIA , SIENRA GUILLERMO
Published: March 2003

This paper studies the dynamics of the family λ sin z for some values of λ. First we give a description of the Fatou set for values of λ inside the unit disc. Then for values of λ on the unit circle of parabolic type (λ = exp (i2πθ), θ = p/q, (p, q) = 1), we prove that if q is even, there is one q-cycle of Fatou components, if q is odd, there are two q cycles of Fatou components. Moreover the Fatou components of such cycles are bounded. For λ as above there exists a component Dqtangent to the unit disc at λ of a hyperbolic component.

There are examples for λ such that the Julia set is the whole complex plane. Finally, we discuss the connectedness locus and the existence of buried components for the Julia set.
UNIFORM ASYMPTOTIC EXPANSIONS FOR HYPERGEOMETRIC FUNCTIONS WITH LARGE PARAMETERS II
Authors: OLDE DAALHUIS A. B.
Published: March 2003

In this paper, we obtain an asymptotic expansion for the Gauss hypergeometric function2F1(a + λ, b + 2λ; c; -z), as |λ| → ∞. The expansion holds for fixed values of a, b, and c, and is uniformly valid for z in the domain | ph z| < π.
Obligatory Overt Wh-Movement in a Wh-in-Situ Language
Authors: Simpson Andrew , Bhattacharya Tanmoy
Published: March 2003

Bangla has commonly been assumed to be an SOV wh-in-situ language. Here it is suggested that both of these characterizations are incorrect and that Bangla actually has obligatory overt wh-movement from a basic SVO word order. This is disguised by a conspiracy of factors but revealed in restrictions on wh-scope and certain apparently optional word order possibilities with complement clauses. Adopting a different perspective on the SOV status of Bangla allows for a simple explanation of the patterns observed and raises the possibility that other “wh-in-situ” languages may also have (obligatory) overt wh-movement.
Assessing the joint effects of chlorinated dioxins, some pesticides and polychlorinated biphenyls on thyroid hormone status in Japanese breast‐fed infants
Authors: Yanagawa Takashi , Kobayashi Yosuke , Nagayama Junya
Published: March 2003

Joint effects of dioxin related chemicals (DXNs), hexachlorocyclohexanes (HCHs), DDT, dieldrin, heptachlor‐epoxide (HCE), chlordane and polychlorinated biphenyls (PCB) on the levels of triirodothyronine (T3), thyroxine (T4), thyroid stimulating hormones (TSH) and thyroid binding globulin (TBG) in the peripheral blood of 101 breast‐fed infants are studied. The statistical issue involved is how to estimate the effects based on data from volunteer subjects with possible measurement errors. A chain independent graph is applied for modeling the associations among factors, and dicotomizations of selected factors are performed for estimating the effects. Use of nonparametric methods with careful consideration of over‐adjustment is suggested. It is shown that the estimated odds ratios of DXNs∪DDT, the first principal component of DXNs and DDT, relative to TSH are 3.02 (p‐value=0.03) and 7.15 (p‐value=0.02), respectively, when PCB is not adjusted and adjusted for, respectively. Copyright © 2003 John Wiley & Sons, Ltd.
Important increase of invasive lobular breast cancer incidence in Geneva, Switzerland
Authors: Verkooijen Helena Marieke , Fioretta Gérald , Vlastos Georges , Morabia Alfredo , Schubert Hyma , Sappino André‐Pascal , Pelte Marie‐Françoise , Schäfer Peter , Kurtz John , Bouchardy Christine
Published: March 2003

A recent paper from the United States reported a sharp and unexplained increase in invasive lobular breast cancer incidence since 1977 (Li et al., Cancer 2000;88:2561–9). We investigated if this trend was also present in Geneva, Switzerland, where breast cancer incidence is one of the highest in Europe. We analyzed trends in breast cancer incidence according to histologic subtype, age and stage, to clarify the pattern. Our population‐based study includes all histologically confirmed invasive breast carcinomas (n = 6,247) recorded between 1976 and 1999 at the Geneva Cancer Registry. Breast histology was classified as ductal carcinoma, lobular carcinoma and other. Incidence trends were studied by log‐linear regression analyses. Models including effects of age, period and birth cohorts were used to describe rising incidence trends. The incidence of ductal carcinoma increased 1.2% per year (ptrend < 0.001) from 85.2 to 110.1/100,000. This increase concerned women aged 50–69 years and early‐stage tumors. Lobular cancer incidence increased disproportionately (14.4% per year, ptrend < 0.01) and rose from 2.9 to 20.5/100,000. This increase affected all age categories and both localized and advanced stages. In addition, a strong age‐cohort effect was present (p < 0.05), and women aged 50–59 years born after 1944 experienced the most marked increase. Our study shows a disproportionate increase of lobular breast cancer incidence compared to ductal cancer incidence. Contrary to ductal cancer, trends for lobular cancer are unlikely to be explained by increased use of screening mammography. Other explanations must be researched, in particular the role played by hormone replacement therapy. © 2003 Wiley‐Liss, Inc.
Molecular typing for HLA class I using ARMS‐PCR: Further developments following the 12th International Histocompatibility Workshop
Authors: Tonks S. , Marsh S.G.E. , Bunce M M. , Bodmer J.G.
Published: March 2003

Molecular typing for HLA class I was introduced in the 12th International Histocompatibility Workshop. Following a pilot study using three methods, sequence specific oligotyping (SSO), reverse dot blot and amplification refractory mutation system (ARMS)‐PCR, the ARMS‐PCR method was selected for use. A great advantage of an ARMS‐PCR method is that, unlike the other two methods, it can determine whether sequence motifs are in cis or in trans, as ARMS‐PCR detects two cis located motifs per reaction using forward and reverse sequence specific primers. Resolution was designed to be low to medium level for HLA‐A, ‐B and ‐C alleles. Two hundred and fifty class I kits and 83 HLA‐A2 subtyping kits were distributed. The A2 subtyping kit used a two round nested PCR system to identify all of the A2 alleles known at the time. Typing results on control DNA samples distributed with both the kits showed a very satisfactory performance. Since the 12th Workshop, the kits have been developed with the addition of new primers and primer mixes to increase the resolution of the test.
The origin of Cretan populations as determined by characterization of HLA alleles
Authors: Arnaiz‐Villena A. , Iliakis P. , González‐Hevilla M. , Longás J. , Gómez‐Casado E. , Sfyridaki K. , Trapaga J. , Silvera‐Redondo C. , Matsouka C. , Martínez‐Laso J.
Published: March 2003

The Cretan HLA gene profile has been compared with those of other Mediterranean populations in order to provide additional information regarding the history of their origins. The allele frequencies, genetic distances between populations, relatedness dendrograms and correspondence analyses were calculated. Our results indicate that the Indoeuropean Greeks may be considered as a Mediterranean population of a more recent origin (after 2000 B.C.), while all other studied Mediterraneans (including Cretans) belong to an older substratum which was present in the area since pre‐Neolithic times. A significant Turkish gene flow has not been detected in the Greek or Cretan populations, although Greeks and Turks have two high frequency HLA‐DRB‐DQB haplotypes in common. It is proposed that Imazighen (Caucasoid Berbers living at present in the North African coast and Saharan areas) are the remains of pre‐Neolithic Saharan populations which could emigrate northwards between about 8000–6000 B.C., when desert desiccation began. They also could be part of the stock that gave rise to Sumerians, Cretans and Iberians; this is supported by both linguistic and HLA genetic data.
Immunophenotypic characterization of human bone marrow endosteal cells
Authors: Sillaber C. , Walchshofer S. , Mosberger I. , Gaiger A. , Simonitsch I. , Chott A. , Lechner K. , Valent P.
Published: March 2003

In order to determine the relationship between bone marrow (bm) endosteal cells (EDC) and hemopoietic progenitors, we have analyzed the immunophenotype of EDC using various antibodies (Ab) against mesenchymal antigens. The Ab were applied on paraffin sections of normal bm (iliac crest, n=17; talus, n=1; phalanx, n=1), myeloregenerative bm (after chemotherapy), and hematologic disorders (acute myeloid leukemia (AML), n=8; chronic myeloid leukemia (CML), n=6; myelodysplastic syndromes (MDS), n=14; severe aplastic anemia (SAA), n=4; essential thrombocythemia (ET), n=2; idiopathic (primary) osteomyelo‐fibrosis (IMF), n=1; polycythemia vera (PV), n=1). In normal bm, EDC were found to react with Ab against vimentin, tenascin, α‐smooth muscle actin, osteocalcin, CD51, and CD56, but did not react with Ab against CD3, CD15, CD20, CD34, CD45, CD68, or CD117. An identical phenotype of EDC was found in AML, MDS, SAA, ET, IMF, PV, myeloregenerative bm, and peripheral bones lacking active hemopoiesis (talus, phalanx). In patients with CML, EDC reacted with Ab to CD51, but did not react with Ab to CD56. Based on their unique antigen profile, EDC were enriched from normal bm by enzyme digestion and cell sorting. However, these enriched cells (CD56+, CD45−, CD34−) did not give rise to hemopoietic cells under the culture conditions used, i.e. in the presence of the growth factors IGF‐1, bFGF, SCF, IL‐3, and GM‐CSF. Together, our data do not support the hypothesis that EDC are totipotent mesenchymal progenitors giving rise to hemopoietic cells.
HLA class II polymorphism in a Gabonese Banzabi population
Authors: Migot‐Nabias F. , Fajardy I. , Danze P.M. , Everaere S. , Mayombo J. , Minh T.N. , Renaut A. , Georges A.J.
Published: March 2003

The HLA class II typing of 167 unrelated Gabonese individuals from the Banzabi ethnic group was assessed by polymerase chain reaction‐restriction fragment length polymorphism (PCR‐RFLP). The most frequent alleles at each locus were DRB1*1501–3 (0.31), DQA1*0102 (0.50), DQB1*0602 (0.42) and DPB1*0402 (0.29). The estimation of the haplotype frequencies as well as the observation of the segregation of several haplotypes using additional HLA typing of relatives, revealed that the three‐locus haplotype DRB1*1501–3‐DQA1*0102‐DQB1*0602 was found at the highest frequency (0.31) among these individuals. This haplotype is not typically African and has already been described in Caucasians, but its presence at high frequency is exclusive to populations originating from Central Africa, and can thus be designated as a particular genetic marker of these populations.
Association of HLA class II alleles with different subgroups of diabetes mellitus in Eastern India identify different associations with IDDM and malnutrition‐related diabetes
Authors: Sanjeevi C.b. , Kanungo A. , Shtauvere A. , Samal K.c. , Tripathi B.b.
Published: March 2003

Genetic studies of Malnutrition related diabetes are few. We have analyzed HLA class II gene polymorphism in different types of diabetes mellitus patients from Cuttack in Eastern India. Patients with insulin‐dependent diabetes mellitus (IDDM), non‐insulin‐dependent diabetes mellitus (NIDDM) and malnutrition‐related diabetes mellitus (MRDM), which is subdivided into protein‐deficient diabetes mellitus (PDDM) and fibrocalculous pancreatic diabetes (FCPD), were studied and their associations with autoantibody markers. IDDM and PDDM were associated with DR3 and DQ2 but not DR4 and DQ8. FCPD was positively associated with DQ9 (A*0201‐B*0303). The association of DQ9 with FCPD suggests differences in the genetic background for susceptibility between IDDM and MRDM in the Cuttack population. There is no association seen between HLA‐DR‐DQ and NIDDM patients from Eastern India. Clinical classification of diabetes into IDDM, NIDDM and MRDM does not identify the underlying pathological mechanisms. Presence of autoantibodies to IDDM autoantigens in clinical MRDM and NIDDM identifies the slow‐onset form of IDDM. Due to the absence of autoantibody assays for diagnosis of IDDM in India, slow onset IDDM is not diagnosed and the patients are classified as NIDDM or MRDM. Our study demonstrates that the presence of GAD65 antibody and DR3‐DQ2 positivity in MRDM and NIDDM patients in Eastern India would suggest the presence of slow‐onset IDDM. Our data would indicate alternatively that MRDM can coexist with IDDM in these patients and malnutrition could be one of the reasons for the slower onset in IDDM‐prone individuals.
Characterization of a novel HLA‐A*24 allele containing an HLA‐A*03 sequence motif
Authors: Drábek J. , Bunce M. , Faé I. , Ambruzová Z. , Dunn P. , Ross J. , Fischer G.f.
Published: March 2003

HLA‐A*2418 is described for the first time. It segregates in a Mendelian fashion. Serological analyses indicate that the new allele encodes epitopes from both HLA‐A9 and ‐A3 specificities. Results from nucleotide sequencing analyses of polymerase chain reaction (PCR) amplification products derived from genomic and cDNA are compatible with those findings Note.
N‐linked oligosaccharides can protect target cells from the lysis mediated by NK cells but not by cytotoxic T lymphocytes: role of NKG2‐A
Authors: Sol M.‐A. , Vacaresse N. , Lule J. , Davrinche C. , Gabriel B. , Teissie J. , Ziegler A. , Thomsen M. , Benoist H.
Published: March 2003

We have previously shown that glycophorin A (GPA), inserted by electropulsation into the membrane of K562 cells, protected them from natural killer (NK) cell‐mediated cytotoxicity and the unique N‐linked oligosaccharide of GPA was essential for resistance to occur. The present study demonstrates that the protection level conferred by GPA is similar to the resistance induced by HLA‐Cw3 expressed by transfected K562 cells. A monoclonal antibody against NKG2‐A, an NK inhibitory receptor interacting with HLA class I antigens and belonging to the C‐type lectin receptor, was able to restore the ability of NK cells to lyse K562 cells expressing HLA‐Cw3 at the cell membrane but not electroinserted‐GPA, suggesting that the N‐linked oligosaccharide of GPA cannot be a ligand for NKG2‐A. GPA was then electroinserted into the membrane of two lymphoblastoid B‐cell lines: one was sensitive to NK cell‐mediated lysis, the other was susceptible to cytotoxic CD8+ T‐lymphocyte (CTL)‐mediated cytotoxicity. The electroinserted GPA protected the target cells from NK‐mediated cytotoxicity, whereas it did not modify the cell susceptibility to lysis by CTL. Endoglycosidase F treatment abolished the resistance towards NK cell‐mediated lysis, suggesting that N‐linked glycans could inhibit mechanisms used by NK cells to exert their cytotoxic function in agreement with our previous results.
An assay for peptide binding to HLA‐Cw*0102
Authors: Andersen M.H. , Søndergaard I. , Zeuthen J. , Elliott T. , Haurum J.S.
Published: March 2003

The assembly assay for peptide binding to class I major histocompatibility complex (MHC) molecules is based on the ability of peptides to stabilize MHC class I molecules synthesized by transporter associated with antigen processing (TAP)‐deficient cell. The TAP‐deficient cell line T2 has previously been used in the assembly assay to analyze peptide binding to HLA‐A*0201 and ‐B*5101. In this study, we have extended this technique to assay for peptides binding to endogenous HLA‐Cw*0102 molecules. We have analyzed the peptide binding of 20 peptides with primary anchor motifs for HLA‐Cw*0102. One‐third of the peptides analyzed bound with high affinity, half of the peptides examined did not bind, whereas the remaining peptides displayed intermediate binding activity. Interest in HLA‐C molecules has increased significantly in recent years, since it has been shown that HLA‐C molecules both can present peptides to cytotoxic T lymphocytes (CTL) and in addition are able to inhibit natural killer (NK)‐mediated lysis.
HFE genotypes and haemochromatosis: quantifying the risks of disease
Authors: Lester S. , Bardy P. , McCluskey J.
Published: March 2003

Hereditary haemochromatosis (HH) is an autosomal recessive disease involving mutations in the recently characterised HFE gene linked to HLA‐A in the major histocompatibility complex. The known HFE polymorphisms include the wild‐type allele, a G→A substitution at base 845 (845A) and a C→G substitution at position 187 (187G). Although most cases of HH are accountable by homozygosity of the 845A allele the exact risk of other HFE genotypes, especially those involving the 187G allele has not been determined. We have compiled estimates of disease risk for all known HFE genotypes by re‐analyzing published studies. The data show a hierarchical risk calculated as odds ratio (OR) for each genotype 845A/845A (OR=2101); 845A/187G (OR=24); 187G/187G (OR=9); 845A/Wt (OR=5); 187G/Wt (OR=2). Interestingly, the disease risk of 187G‐genotypes suggests that subtle functional changes in the HFE product can interact with other genetic factors (e.g. trans allele, gender) and environmental factors (e.g. diet) to manifest either as clinical disease, altered iron stores or a normal phenotype. This paradigm is potentially useful in understanding the contribution of HLA alleles to risk of various disorders especially autoimmunity.
Platelet glycoprotein (GP) V polymorphisms in Japanese
Authors: Kunishima S. , Shimizu T. , Kamiya T. , Ozawa K.
Published: March 2003

The glycoprotein (GP) V is a subunit of a platelet receptor for von Willebrand factor, the GPIb/IX/V complex. Recently, polymorphisms in the GPV gene, four involving amino acid changes and five silent, were identified. We developed an allele‐specific restriction method to determine the gene frequencies of the non‐synonymous substitutions in Japanese and Caucasians. The gene frequencies for 341Gly/Arg polymorphism were 0.94 for 341Gly and 0.06 for 341Arg, and 0.95 for 341Gly and 0.05 for 341Arg, in Japanese and Caucasians, respectively. We could not find rare variants for 114Asp/Tyr, 273Met/Ile or 397Leu/Arg. Flow cytometric analysis showed that the 341Arg variant is expressed normally on the platelet membranes, suggesting the possibility of the involvement of the 341Gly/Arg polymorphism as a platelet alloantigen.
LPS upregulates MHC class II I‐A expression in B lymphocytes at transcriptional and at translational levels
Authors: Barrachina M. , Goñalons E. , Celada A.
Published: March 2003

Major histocompatibility complex (MHC) class II molecules are expressed in a limited number of cell types, including B lymphocytes, dendritic cells and macrophages. Lipopolysaccharide (LPS) increases the surface expression of class II molecules in a murine B‐cell line by inducing an increase in I‐A protein and I‐A mRNA levels. LPS does not modify the rate of mRNA degradation; therefore, the increase in mRNA is due to an increase in transcription. In addition, LPS increases the levels of I‐Aα protein, which correlates with an increase in ribosome loading for I‐Aα but not for I‐Aβ mRNA after treatment with LPS. Interestingly, in non‐induced cells, I‐Aα messenger RNA shows a significant peak of free mRNA. Therefore, LPS regulates the expression of MHC class II molecules at translational level in B cells, in addition to the transcriptional control. The actual mechanism implies changes of translation initiation rates, as shown by an increase ribosome loading in polysome gradients.
New HLA class II alleles in the Indonesian population
Authors: Panigoro R. , Greville W.D. , Kennedy A. , Tréjaut J. , Dunckley H.
Published: March 2003

This paper describes two new class II alleles of the major histocompatibility complex (MHC), DRB1*1431 and DRB3*0303, that have been found in the Indonesian population. In addition, the identification of DRB1*0819 is presented as a confirmatory report Note.
Identification of a novel DPB1* allele (DPB1*6901) and the occurrence of HLA haplotypes that extend to DPB1
Authors: Varney M.D. , Tait B.D.
Published: March 2003

Acknowledgments:

The authors gratefully acknowledge the assistance of Alex Gavrilidis (GeneType Pty Ltd) in performing nucleotide sequencing.

The sequence of a novel DPB1 allele, DPB1*6901, observed in a Caucasian bone marrow donor phenotype HLA A2; Cw*0501,*1601; B*4402, *4403; DRB1*0401; *07; DQB1*02; *0301; DPB1*0401; *6901, is described. The sequence is consistent with that previously described for DPB1*0601 with the exception of codon 69. The sequence at this codon is consistent with that previously observed only in the DPB1*1101 and *1501 alleles. It is suggested that DPB1*6901 may have arisen as a result of a recombination event occurring between codons 58 and 64 between DPB1*0601 and DPB1*1101. The sequence of DPB1*1501 from codon 64 is not consistent with DPB1*6901. A linkage disequilibria analysis that examined 212 potential bone marrow recipients in which HLA‐A to DQ haplotypes had been established by family studies showed linkage disequilibrium between HLA‐B, DRB1 and DPB1 in some haplotypes and not others.
HLA‐A*02 allele frequencies and haplotypic associations in Koreans
Authors: Park M.H. , Whang D.H. , Kang S.J. , Han K.S.
Published: March 2003

We have investigated the frequencies of HLA‐A*02 alleles and their haplotypic associations with HLA‐B and ‐DRB1 loci in 439 healthy unrelated Koreans, including 214 parents from 107 families. All of the 227 samples (51.7%) typed as A2 by serology were analyzed for A*02 alleles using polymerase chain reaciton (PCR)‐low ionic strength‐single‐strand conformation polymorphism (LIS‐SSCP) method. A total of six different A*02 alleles were detected (A*02 allele frequency 29.6%): A*0201/9 (16.6%), *0203 (0.5%), *0206 (9.3%), *0207 (3.0%), and one each case of *0210 and *02 undetermined type. Two characteristic haplotypes showing the strongest linkage disequilibrium were A*0203‐B38‐DRB1*1502 and A*0207‐B46‐DRB1*0803. Besides these strong associations, significant two‐locus associations (P<0.001) were observed for A*0201 with B61, DRB1*0901 and DRB1*1401, and for A*0206 with B48 and B61. HLA haplotypes carrying HLA‐A2 showed a variable distribution of A*02 alleles, and all of the eight most common A2‐B‐DR haplotypes occurring at frequencies of ≥1% were variably associated with two different A*02 alleles. These results demonstrate that substantial heterogeneity is present in the distribution of HLA‐A*02 alleles and related haplotypes in Koreans.
DLA‐DQA1 polymorphisms in dogs defined by sequence‐specific oligonucleotide probes (SSOP)
Authors: Kennedy L.J. , Carter S.D. , Barnes A. , Bell S. , Bennett D. , Ollier W.E.R. , Thomson W.
Published: March 2003

Several recent studies have identified DNA sequences for alleles of the DLA‐DQA1 locus in the dog. To date, 10 DQA1 alleles have been reported. No data exists on the frequencies of these alleles within the general dog population, nor is there any indication of whether alleles are breed specific. We have addressed this issue by establishing a molecular‐based sequence‐specific oligonucleotide probing (SSOP) method to identify all published DQA1 alleles and have used these methods to type a large number of dogs. Oligonucleotide probes were designed to detect all the polymorphic sites in exon 2. This allowed assignment at the allele level. Three hundred and thirty dogs were typed for DQA1. All but two of the published DQA1 alleles were identified in these animals. One new allele was identified, and confirmed by DNA cloning and sequencing. This typing method provides a powerful tool for generating data that will be essential for studies investigating the genetic relationships between different breeds.
Sequence‐based typing of HLA class II DQB1
Authors: Dinauer D.M. , Luhm R.A. , Uzgiris A.J. , Eckels D.D. , Hessner M.J.
Published: March 2003

Due to the expanding number of known HLA class II DQB1 alleles, high‐resolution oligotyping is becoming ineffective, therefore a sequence‐based typing (SBT) strategy was developed to provide rapid and definitive typing of HLA‐DQB1. HLA‐DQB1*02, *03, *04, *05, and *06 alleles were individually amplified by polymerase chain reaction (PCR) using exon 2 group‐specific primers. Forward and reverse PCR primers were tailed with M13 universal and M13 reverse sequences, respectively. Subsequent bi‐directional cycle‐sequencing was carried out using Cy5.5‐labeled M13 universal primer and Cy5.0‐labeled M13 reverse primer. Automated sequencing was performed in 30 min using a Visible Genetics, Inc. (VGI) MicroGene ClipperTM Sequencer. Full concordance was observed between this SBT method and oligotyping among 151 individuals.
HLA genes in Mexican Mazatecans, the peopling of the Americas and the uniqueness of Amerindians
Authors: Arnaiz‐Villena A. , Vargas‐Alarcón G. , Granados J. , Gómez‐Casado E. , Longas J. , Gonzalez‐Hevilla M. , Zuñiga J. , Salgado N. , Hernández‐Pacheco G. , Guillen J. , Martinez‐Laso J.
Published: March 2003

Acknowledgments:

This work was supported in part by grants from the Spanish Ministry of Education (PM 57–95 and PM 96–21) and Comunidad de Madrid (06–70–97 and 8.3/14/98). Gilberto Vargas‐Alarcón was supported by the Consejo Nacional de Ciencia y Tecnología and the Fundación Mexicana para la Salud, Mexico.

The HLA allele frequency distribution of the Mexican Mazatecan Indians (Olmec culture) has been studied and compared with those of other First American Natives and worldwide populations (a total of 12,100 chromosomes; 6,050 individuals from 59 different populations). The main conclusions are: 1) An indirect evidence of Olmec and Mayan relatedness is suggested, further supporting the notion that Olmecs may have been the precursors of Mayans; 2) Language and genetics do not completely correlate in microenvironmental studies; and 3) Peopling of the Americas was probably more complex than postulated by Greenberg and others (three peopling waves). Significant genetic input from outside is not noticed in Meso and South American Amerindians according to the phylogenetic analyses; while all world populations (including Africans, Europeans, Asians, Australians, Polynesians, North American Na‐Dene Indians and Eskimos) are genetically related. Meso and South American Amerindians tend to remain isolated in the Neighbor‐Joining, correspondence and plane genetic distance analyses.
Discovery of two novel variants of KIR2DS5 reveals this gene to be a common component of human KIR ‘B’ haplotypes
Authors: Vilches C. , Pando M.J. , Rajalingam R. , Gardiner C.M. , Parham P.
Published: March 2003

Acknowledgments:

This study was supported by grant AI17892 from the U.S. Public Health Service to P.P. C.V. was supported by a fellowship from the “Instituto de Salud Carlos III”, Spain (FIS BAE 98/5105); C.M.G. was supported by a grant from the Leukemia Research Foundation C.V.’s current address: Servicio de Inmunología, H.U. Clínica Puerta de Hierro, San Martín de Porres 4, 28035 Madrid, Spain.

The gene encoding the non‐inhibitory receptor KIR2DS5 has so far been represented by a single cDNA sequence, NKAT9. A previous study by polymerase chain reaction using sequence‐specific primers (PCR‐SSP) failed to detect NKAT9 in genomic DNA of 52 donors, which suggested that KIR2DS5 could be a rare gene. Here, we have characterized two novel variants of KIR2DS5 that differ from NKAT9 by 8 and 10 nucleotide substitutions. The frequency of KIR2DS5 was then re‐assessed by PCR‐SSP using primers specific for conserved sequences of all three known KIR2DS5 variants. We found KIR2DS5 is not a rare gene, but one present in 26% of 34 donors representing the major ethnic groups. Like other non‐inhibitory KIR, the distribution of KIR2DS5 is restricted to the ‘B’ subset of KIR‐gene haplotypes. Transcription of the KIR2DS5 gene was studied by reverse transcriptase (RT)‐PCR in natural killer (NK) cells from one donor and shown to follow the clonal distribution seen for most other KIR genes.
Transracial evidence for the influence of the homologous HLA DR‐DQ haplotype on transmission of HLA DR4 haplotypes to diabetic children
Authors: Park Y.S. , She J.X. , Noble J.A. , Erlich H.A. , Eisenbarth G.S.
Published: March 2003

Abstract: The HLA‐DQB1*0302 allele on DR4 haplotypes is a marker for type 1 diabetes susceptibility and it is an especially high‐risk allele in DR3/4 because of its preferential distribution in Caucasian DR3/4 patients. In Asians, not only DQB1*0302 but also DQB1*0401 on DR4 haplotypes are associated with type 1 diabetes. We investigated whether the contribution of these DQ molecules was also genotype‐dependent in Asians. Although the prevalence of the DR4‐DQB1*0302 haplotype did not differ in patients vs. controls, the DR3/4‐DQB1*0302 genotype had a RR of 12 (P<10−4). Moreover, a significant association of DQB1*0302 with the DR3/4 genotype was found (RR=3, P<10−2). In contrast, the distribution of DQB1*0401 alleles of DR4/X (X: other than 3, 4) is different from that of DR3/4 and DR4/4. Especially a significant association of DQB1*0401 with DR4/X (X: other than 1, 3, 4) was found (RR=3, P<10−3). The frequency of transmission of the DR4‐DQB1*0302 haplotypes to diabetic offspring with DR3 was 80%, while to those without DR3 was 40%. In contrast, the transmission of the DR4‐DQB1*0401 to those with DR3 was 60%, while to those without DR3 was 80%. High‐risk DR4 subtypes were predominant in DR4/X (RR=7, P<10−3), whereas protective DR4 subtypes were observed mainly in the DR3/4 (RR=3, P<0.05). The association with diabetes and transmission to a diabetic offspring of DR4 haplotypes varies depending on the haplotype borne on the homologous chromosome. This might contribute not only to the synergistic effect of DR3/4, but also to the susceptibility influence of HLA DQB1*0401 alleles confined to DR4/X.
Case‐control study with narcoleptic patients and healthy controls who, like the patients, possess both HLA‐DRB1*1501 and ‐DQB1*0602
Authors: Hohjoh H. , Terada N. , Nakayama T. , Kawashima M. , Miyagawa T. , Honda Y. , Tokunaga K.
Published: March 2003

Abstract: In previous studies, we suggested that the tumor necrosis factor (TNF‐α and its receptor 2 (TNFR2) genes could be associated with the susceptibility to human narcolepsy, and that haplotype carrying DRB1*1502 had a negative association with the disorder. To further evaluate these associations, we herein compared narcoleptic patients with healthy individuals who, like the patients, possessed both DRB1*1501 and DQB1*0602. Results agreed with the negative association of DRB1*1502 and positive association of the TNF‐α(−857T) and TNFR2‐196R combination with the disorder. In addition, a significant association of the TNF‐α(−857T) homozygote with the disorder and an increase in a rare haplotype carrying DRB1*1501 and TNF‐α(−857T) in the patients were also observed in the present study.
Resolution of cis‐trans ambiguities between HLA‐DRB1 alleles using single‐strand conformation polymorphisms and sequencing
Authors: Paul P. , Thomas D. , Kawczak P. , Good D. , Cook D.J. , Ball E.J.
Published: March 2003

Abstract: DNA‐based typing of HLA alleles occasionally results in the inability to assign a specific allele because of ambiguity in associating two or more polymorphisms to the same or to alternate homologs (cis/trans ambiguity). Since most individuals are heterozygous at a given HLA locus, the highest level of confidence in definition is obtained when the alleles are tested in isolation. By using single‐strand conformation polymorphisms (SSCP) to separate heterozygous HLA‐DRB1 alleles, followed by sequencing of separated conformers, we have achieved resolution of previously ambiguous assignments without the need for additional probes or primers.
Improved typing procedure for the polymorphic single‐copy RLA‐DQA gene of the rabbit reveals a new allele
Authors: Fain M.A. , Zhao T. , Kindt T.J.
Published: March 2003

Abstract: The DQA gene of the rabbit major histocompatibility complex (MHC, RLA) is highly polymorphic and, in contrast to those reported for other mammalian species, is present as a single copy. These properties allow use of this gene in a method to type the class II locus of RLA by a combination of single‐stranded conformational polymorphism (SSCP) and heteroduplex (HD) analysis. Familial segregation of RLA‐DQA was shown and RLA class II types for rabbits of unknown pedigree were determined using migration patterns of amplified genomic DNA. Typing results were confirmed in experiments where unknown samples were mixed with products from rabbits of RLA types defined by sequence analysis. These analyses detected an RLA‐DQA allele in addition to the five previously described; this new allele is designated RLA‐DQA‐F.
Novel HLA allele, HLA‐B*4013, identified from a potential bone marrow recipient. Putative gene conversion events with donor sequence
Authors: Blanco P.V. , Theiler G. , Marcos C. , Capucchio M. , Lindel N. , Sala L. , Fainboim L. , Satz M.L.
Published: March 2003

Abstract: A new B40 allele was identified in a leukemic Caucasian patient. This allele, designated B*4013, differs in alpha 1 domain from B*4002 at six amino acidic positions: 67, 77, 80, 81, 82 and 83. Most of this substitutions could alter the antigen binding site of the HLA‐B molecule. B*4013 may have originated by gene conversion or reciprocal recombination involving B*4002 as the recipient allele of sequence donated by B*4406. The new allele was serologically typed as a “blank” associated with the Bw4 epitope (Note).
Differences between T‐cell reactivities to major myelin protein‐derived peptides in opticospinal and conventional forms of multiple sclerosis and healthy controls
Authors: Minohara M. , Ochi H. , Matsushita S. , Irie A. , Nishimura Y. , Kira J.
Published: March 2003

Abstract: In Japanese, susceptibility to the conventional form of multiple sclerosis (C‐MS) is associated with the HLA‐DRB1*1501‐DRB5*0101 haplotype while susceptibility to the opticospinal form of MS (OS‐MS) is associated with HLA‐DPA1*0202‐DPB1*0501. To clarify the characteristics of T cells autoreactive to myelin proteins in each MS subtype, we established T‐cell lines reactive to such myelin antigens as myelin basic protein (MBP), proteolipid protein (PLP) and myelin oligodendrocyte glycoprotein (MOG) from 5 of 10 OS‐MS patients, 6 of 11 C‐MS patients and 7 of 13 healthy controls (HCs), and T‐cell epitopes and their restriction molecules were determined. We found that (a) intermolecular epitope spreading was found to be significantly more frequent in MS patients than in HCs (P=0.0128), (b) intramolecular epitope spreading also tended to occur more frequently in MS patients than in HCs (P=0.0584), (c) in OS‐MS, HLA‐DR‐restricted and MOG‐autoreactive T cells were more frequently established as compared with those reactive to MBP or PLP epitopes and (d) in C‐MS, HLA‐DQ‐restricted and PLP‐autoreactive T cells dominated those autoreactive to MBP or MOG epitopes. A DPB1*0501‐restricted MBP‐reactive T‐cell clone from a patient with OS‐MS provided evidence that the first HLA class II anchor amino acid of peptide bound to disease‐susceptible DP5 molecule was distinct from that for the DR2 molecule. Taken together, these differences in specificities of myelin‐autoreactive T cells between C‐MS and OS‐MS as well as the difference in the anchor motif of the binding peptides between each MS subtype‐susceptible HLA class II molecule may contribute to the development of distinct clinical phenotypes.
Identification and sequence characterization of a novel HLA‐A11 serological variant (HLA‐A*1108)
Authors: Palou E. , Santos M. , Gil J. , Campos E. , Nogués N. , Ribera A.
Published: March 2003

Abstract: In this report we describe the identification of a novel HLA‐A*11 allele, HLA‐A*1108, found in two individuals of a Spanish family. This new allele was detected during routine HLA typing by an atypical serological reactivity pattern and by inconclusive patterns obtained in DNA‐based typing methods. The nucleotide sequence of exons 2 and 3 of HLA‐A*1108 was identical to HLA‐A*11011 except for two nucleotide substitutions at codons 152 (GCG→GAG) and 156 (CAG→CGG). These mutations change the non‐charged amino acid alanine, at codon 152, to negative glutamic acid, and also non‐charged glutamine, at codon 156, to positive arginine, which may explain its anomalous serological reactivity.
Identification of HLA‐A*11 variant (A*1107) in the Korean population
Authors: Pyo C.‐W. , Choi H.‐B. , Han H. , Hong Y.‐S. , Kim T.‐G.
Published: March 2003

Abstract: We report here a new HLA‐A*11 allele, A*1107, identified by sequencing based typing in the Korean population. The full‐length sequencing of A*1107 was conducted on cDNA. HLA‐A*1107 differs from HLA‐A*1101 by a single nucleotide at position 399 of codon 109 in exon3 (TTC→TTA), leading to an amino acid change from phenylalanine to leucine. But the serological profile of HLA‐A*1107 did not exhibit the altered HLA‐A11.
Inflammatory bowel disease is associated with a novel promoter polymorphism of natural resistance‐associated macrophage protein 1 (NRAMP1) gene
Authors: Kojima Y. , Kinouchi Y. , Takahashi S. , Negoro K. , Hiwatashi N. , Shimosegawa T.
Published: March 2003

Abstract: Inflammatory bowel diseases (IBD) representing both Crohn's disease and ulcerative colitis are characterized by chronic activation of macrophages. Natural resistance‐associated macrophage protein 1 (NRAMP1) gene regulates macrophage activation of antimicrobial activity and has many pleiotropic effects on macrophage functions. To explore the role of the NRAMP1 gene in IBD susceptibility, we examined the promoter sequence of the NRAMP1 gene whose polymorphisms have been reported to influence the transcriptional activity of the NRAMP1 gene. One novel allele (allele 7) and two previously reported alleles (alleles 2 and 3) have been determined in a Japanese population. We investigated the association of IBD with these three alleles. The allele frequency of allele 7 was significantly higher in patients with Crohn's disease (11.1%) and ulcerative colitis (11.2%) than those in the healthy control group (4.5%) (Pc=0.015, Pc=0.018, respectively). Therefore, our findings suggest that the novel promoter polymorphism of the NRAMP1 gene may influence susceptibility to IBD in the Japanese population.
PRURITUS AND HLA IN HEMODIALYSIS PATIENTS
Authors: Saatçi Ü , Basın E , Özdemir FN , Turan M , Micozkadıoğlu H , Haberal M
Published: March 2003

Pruritus is a common problem among hemodialysis (HD) patients, and the etiopathogenesis of uremic pruritus is still unclear. This study evaluated the relationships between pruritus and human leukocyte antigen (HLA) types in individuals on HD. Seventy‐nine HD patients (32 females and 47 males; mean age: 38.76±14.04 years; age range: 9–70 years) were studied. The HD duration in the group ranged from 9 to 142 months (mean: 52.53±32.48 months). Subjects were assigned to one of two groups, one with pruritus (n=45) and one without pruritus (n=34). Demographic and laboratory parameters were compared in patients with and without pruritus. There were no significant differences between the groups with respect to age, sex, or HD duration. There were also no significant differences in the groups' serum levels of Hb, ferritin, BUN, creatinine, Ca, P, PTH, uric acid, AST, ALT, GGT, total bilirubin and ALP (P>0.05 for all). There were also no differences between the two groups according to the presence of HbsAg and anti‐HCV (P>0.05). The frequency of the HLA‐B35 allele was significantly higher in the pruritus group than in the group without pruritus (P<0.05). The pruritus group also had higher frequencies of HLA‐A3 and ‐DR4, but these differences were not significant (P>0.05). The results of this preliminary study indicate that the HLA‐B35 antigen may contribute to pruritus in HD patients. Further studies are needed to clarify the role that the HLA system plays in pruritus in this patient group.
Elongation of the cytoplasmic domain, due to a point deletion at exon 7, results in an HLA‐C null allele, Cw*0409 N
Authors: Balas A. , Santos S. , Aviles M.J. , García‐Sánchez F. , Lillo R. , Alvarez A. , Villar‐Guimerans L.M. , Vicario J.L.
Published: March 2003

Abstract: The development of molecular techniques for HLA typing has allowed the identification of genes previously assigned as serologic blank alleles. Lack or poor cell surface expression has been found for molecules coded by HLA‐A, ‐B, ‐DRB4, ‐DRB5, and –DPB1 genes. In this report we describe the first HLA‐C gene encoding for a null cell surface molecule. HLA‐Cw*0409 N shows a point deletion at position 1095 within exon 7. This mutation provokes a codon reading shift, generating a new translation stop codon 97 bp downstream to that described in alleles normally expressed. This new stop codon location implies the presence of 32 extra amino acid residues in the cytoplasmic domain. Transfection experiments suggest that elongation of the cytoplasmic domain in Cw*0409 N would be the cause of cell surface expression failure, although Cw*0409 N heavy chain is able to create stable complexes with β2‐microglobulin. HLA‐C fragment length analysis in a small selected group of samples with B44–Cblk haplotypic associations allowed us to identify two additional subjects showing both a serologic silent Cw*04 allele and a point base deletion at the 3′ end of the HLA‐C gene. This finding indicates that the allele frequency of Cw*0409 N within serologic C blank alleles would be appreciable, although basically restricted to the (A23)‐Cw*0409 N‐B*4403‐DR7‐DQ2 haplotype.
Identification of two new HLA‐DRB1 alleles, DRB1*1138 and DRB1*1344
Authors: Kennedy A. , Le T. , Chapman G. , Greville W.D. , Dunckley H.
Published: March 2003

Abstract: Two new HLA‐DRB1 alleles have been identified by sequencing based typing (SBT). HLA‐DRB1*1138 and DRB1*1344 were discovered after following up ambiguous results involving unusual alleles after DRB1 generic typing.
Molecular diversity of the HLA‐A*19 group of alleles in North Indians: Possible oriental influence
Authors: Jaini R. , Naruse T. , Kanga U. , Kikkawa E. , Kaur G. , Inoko H. , Mehra N.K.
Published: March 2003

Abstract: The present study aims to determine the genetic diversity of the HLA‐A19 allelic family in the North Indian and Japanese populations. The HLA‐A*19 group of alleles occurred at similar frequencies in North Indians and Japanese as in Caucasians. All the known serological splits of HLA‐A19 were observed among the North Indians, i.e. A*33 (15.6%), A*32 (8.6%), A*31 (3.5%), A*30 (3%), A*29 (1.2%) and A*74 (0.77%), while only A*30 (0.7%), A*31 (17.6%) and A*33 (11.7%) were observed in the Japanese. High resolution analysis indicated that the A*29, A*30, A*31 and A*32 alleles were represented by only single subtypes among the North Indians while the HLA‐A*33 group comprised two alleles, A*3301 (4.3%) and A*3303 (43.7%). All 15 of the HLA‐A*33 positive samples from the Tamil population of South India were found to be A*3303. One novel subtype of A*33, A*3306 was also observed in the North Indian sample. Conversely, only one subtype each of A*30, A*31 and A*33 was encountered in the Japanese population, of which A*3101 and A*3303 were the most frequent (58.5% and 39%, respectively, among the HLA‐A*19 group of alleles). All other subtypes of A19 were not found in the Japanese in the present study. The study suggests a significant amount of genetic admixture in the North Indian gene pool from other racial groups, with profound oriental influence.
Analysis of the T‐cell receptor β‐chain variable‐region (Vβ) repertoire in chronic human Chagas' disease
Authors: Fernández‐Mestre M.T. , Jaraquemada D. , Bruno R.E. , Caro J. , Layrisse Z.
Published: March 2003

Abstract: Previous studies have shown that infection with the protozoan Trypanosoma cruzi (Chagas' disease) is associated with genetic components [human leukocyte antigen (HLA) genes and T‐cell receptor (TCR) genes]. We studied the TCR Vβ repertoire of peripheral blood lymphocytes of 23 unrelated serologically positive subjects using reverse transcriptase‐polymerase chain reaction (RT‐PCR). The patients, previously tested for HLA genotypes, were clinically classified as asymptomatic, arrhythmic and cardiopathic patients. Statistical analysis showed the significant increment of the Vβ7 family in chagasics with arrhythmia compared with asymptomatic and cardiopathic patients, indicating that the frequency of this family is variable in different clinical forms of the disease and possibly that these T cells might be a marker of the progression of Chagas' disease. Based on the calculation of a Δ score the order of variability in the TCR repertoire was: patients with heart failure > asymptomatic > arrhythmic patients. The major histocompatibility complex (MHC) of the individual may influence the use of particular V genes in T‐cell response to foreign antigens. We found a significant increase of the Vβ7 family in arrhythmic patients who were DRB1*01 DQB1*0501 DPB1*0401, a marker associated with susceptibility to cardiac damage in Chagas' disease. If confirmed by further studies in a larger cohort, a possible association between the TCR Vβ repertoire and the MHC haplotype of chagasic patients could be postulated.