bluuebunny/crossref_metadata_2025 huggingface dataset:
Both the HLA‐DPB1 and ‐DRB1 alleles correlate with risk for multiple sclerosis in Japanese: clinical phenotypes and gender as important factors
Authors: Fukazawa T. , Yamasaki K. , Ito H. , Kikuchi S. , Minohara M. , Horiuchi I. , Tsukishima E. , Sasaki H. , Hamada T. , Nishimura Y. , Tashiro K. , Kira J.
Published: March 2003

The purpose of this study was to clarify the association of HLA‐DRB1 and ‐DPB1 alleles with multiple sclerosis (MS) in Japanese, to determine whether optico‐spinal MS (OS‐MS) and conventional MS are immunogenetically distinct, and to verify the role of gender difference in HLA associations of MS. We studied HLA‐DRB1 and ‐DPB1 polymorphisms in 166 Japanese patients with MS. Forty‐seven patients were classified as having the optico‐spinal MS (OS‐MS) and 119 as having conventional MS. A lack of DPB1*0301 and a higher frequency of DPB1*0501 compared with controls (corrected P<0.0074; odds ratio=9.48) were found in OS‐MS. By contrast, we found for the first time an association of DPB1*0301 with conventional MS in Japanese (corrected P=0.0444; odds ratio=3.28). Logistic analysis, adjusted for sex and age, revealed independent associations of DPB1*0301 (P=0.0004, adjusted odds ratio (aOR)=4.70), DPB1*0501 (P=0.0081, aOR=2.50) and DRB1*1501 (P=0.0252, aOR=2.21) with conventional MS. However, the frequencies of DRB1*1501 and DPB1*0501 in male patients with conventional MS were equal to those in male controls while the DPB1*0301 frequency was increased in both male and female patients. We did not find any association of these HLA alleles with disease course and severity. In conclusion, OS‐MS is a DPB1*0501‐associated distinct subtype of MS, and DPB1*0301 is the most strongly associated allele with conventional MS in Japanese. In addition, gender plays an important role in HLA association with MS.
HLA genes in Arabic‐speaking Moroccans: close relatedness to Berbers and Iberians
Authors: Gómez‐Casado E. , Del Moral P. , Martínez‐Laso J. , García‐Gómez A. , Allende L. , Silvera‐Redondo C. , Longas J. , González‐Hevilla M. , Kandil M. , Zamora J. , Arnaiz‐Villena A.
Published: March 2003

The gene profile of Arabic‐speaking Moroccans has been compared with those of other Mediterranean populations in order to provide additional information about the history of their origins. Our HLA data suggest that most Moroccans are of a Berber (Imazighen) origin and that Arabs who invaded North Africa and Spain in the 7th century A.D. did not substantially contributed to the gene pool; however, they imposed their advanced culture and their religion. Present‐day Egyptians are also related to Moroccan Berbers and this supports an ancient Saharan origin for part of the present‐day Mediterraneans, particularly for the Arabic‐speaking ones (also Algerians) and also for the older substratum of Mediterranean people.
IMGT/HLA Database – a sequence database for the human major histocompatibility complex
Authors: Robinson J. , Malik A. , Parham P. , Bodmer J.G. , Marsh S.G.E.
Published: March 2003

The IMGT/HLA Database is a specialist database for sequences of the human major histocompatibility (MHC) system. It includes all the HLA sequences officially recognised and named by the WHO Nomenclature Committee for Factors of the HLA System. The database provides users with online tools and facilities for the retrieval and analysis of these sequences. These include allele reports, alignment tools and a detailed database of all source cells. The online IMGT/HLA submission tool allows the submission of both new and confirmatory allele sequences directly to the WHO Nomenclature Committee for Factors of the HLA System. The latest version (release 1.4.1, November 1999) contains 1,015 HLA alleles from over 2,270 component sequences derived from the EMBL/GenBank/DDBJ databases. From its release in December 1998 until December 1999 the IMGT/HLA website received approximately 100,000 hits. The database currently focuses on the human major histocompatibility complex but will be used as a model system to provide specialist databases for the MHC sequences of other species.
Evolution of Pacific/Asian populations inferred from HLA class II allele frequency distributions
Authors: Mack S.J. , Bugawan T.L. , Moonsamy P.V. , Erlich J.A. , Trachtenberg E.A. , Paik Y.K. , Begovich A.B. , Saha N. , Beck H.P. , Stoneking M. , Erlich H.A.
Published: March 2003

The allele frequency distributions for the HLA class II loci, DRB1, DQB1 and DPB1, in eight Pacific/Asian populations: Hawaiian, Samoan, Malay, Papua New Guinea (PNG) Highlands, and two Indonesian and PNG Lowland groups, were determined using high‐resolution polymerase chain reaction/sequence‐specific oligonucleotide probe (PCR/SSOP) typing methods. The allele frequency distributions for the HLA‐DRB1 locus were determined for a third Indonesian population as well as for an additional Filipino population. DRB1 alleles in the DR2 serogroup (or allelic lineage) are very common in this region; in some populations, more than 50% of the alleles belong to this serogroup. The DRB1*1502 allele is frequent in nine of the ten populations studied, reaching a frequency of 0.48 in one Indonesian population and among Filipinos. Extensive DR‐DQ haplotype diversity was detected in these populations. Seven different DR2‐DQB1 haplotypes were observed in the Indonesian and PNG Lowland populations, eight in the PNG Highlands and ten in Malays and Filipinos. The DRB1*0410 allele, commonly observed in Australia, is observed in the PNG Highlands at a low frequency (f=0.03) and is absent in the other populations. Two additional DRB1 alleles commonly observed in Australia, DRB1*0405 and *1407, are also observed in the PNG Highlands at high frequencies (f=0.132 and 0.126), while they are rare in the PNG Lowlands (f=0.039 and 0.013). These alleles are generally rare or absent in the other populations. The DPB1*0501 allele, common in Chinese and Japanese populations, is most frequent in the Samoan, Hawaiian, Indonesian, and Malay populations, and the *0401 allele is the most frequent DPB1 allele in the PNG Lowlands. Both of these alleles have the same very high frequency (f=0.34) in the PNG Highlands. Analyses of homozygosity (the Ewens‐Watterson F statistic) in these and other populations indicate that, while most allele frequency distributions are consistent with balancing selection, values of F for the Indonesian and Javan populations may reflect positive directional selection. Phylogenetic trees constructed using the allele frequencies at the DRB1 locus of the populations reported here, as well as those for additional Pacific, Asian, and Australian populations, indicate that the PNG Highland population is more closely related to Australian populations than to PNG Lowland populations, while the PNG Lowlands are more closely related to other Melanesian populations.
Cytomegalovirus‐infected neuroblastoma cells exhibit augmented invasiveness mediated by β1α5 integrin (VLA‐5)
Authors: Scholz M. , Blaheta R.A. , Wittig B. , Cinatl J. , Vogel J.‐U. , Doerr H.W. , Cinatl J.
Published: March 2003

Previously, experimental in vivo results showed that the productively and persistently human cytomegalovirus (HCMV)‐infected neuroblastoma cell line UKF‐NB‐4AD169 exhibits a more malignant phenotype than the non‐infected variant UKF‐NB‐4. To prove the assumption that enhanced malignancy may be due to enhanced invasive potential of the infected cells we studied interactions of both lines with monolayers of cultured endothelial cells. UKF‐NB‐4AD169 cells adhered to and transmigrated through endothelial monolayer to a significantly higher extent compared with UKF‐NB‐4. Furthermore, the adhesion of UKF‐NB‐4AD169 but not of UKF‐NB‐4 resulted in focal disruption of the monolayer integrity which facilitates tumor cell transmigration. Blocking antibodies directed against the β1 integrin chain as well as β1α5 on the tumor cells specifically inhibited adhesion in a concentration‐dependent manner. When UKF‐NB‐4 were pretreated with a β1 integrin activating antibody, focal disruption of the endothelial integrity also occurred. These findings lead us to suggest that HCMV infection activates β1α5 in the host neuroblastoma cell which in turn enables these cells to tightly adhere to endothelial cells. In the presence of the protease inhibitor phenantroline, β1α5‐mediated adhesion was not impaired whereas UKF‐NB‐4AD169‐mediated endothelial monolayer permeabilization was dose dependently inhibited. We conclude that human cytomegalovirus infection contributes to augmented neuroblastoma invasiveness via adhesion of activated β1α5 and subsequent matrix digestion by proteases.
CDw108 expression during T‐cell development
Authors: Mine T. , Harada K. , Matsumoto T. , Yamana H. , Shirouzu K. , Itoh K. , Yamada A.
Published: March 2003

We recently reported a gene encoding the human CDw108, a glycosylphosphatidylinositol (GPI)‐anchored membrane glycoprotein that is preferentially expressed on activated T lymphocytes and erythrocytes. The present study investigated the expression of CDw108 on various tissues and cells, particularly on T cells during development. The murine CDw108 cDNA was cloned initially, and it was highly homologous to the human CDw108 (88.0% or 89.3% similarity at the nucleotide or amino acid level, respectively) or identical to the murine semaphorin K1/Sema7A. The CDw108 mRNA was demonstrated in a few tissues including thymus and brain with the highest expression coming on day 7 in whole embryo followed by relatively consistent expression during development. Cell‐surface expression of the CDw108 during T‐cell development was further examined by flow cytometry in the human umbilical cord blood and thymus. It was preferentially expressed on a CD34+ stem cell population of umbilical cord blood, and CD3dull CD34+/‐ CD117 (c‐kit)+ CD4bright CD8bright cells in the thymus that are involved in the stage of positive selection. These results suggest the contribution of CDw108 in T‐cell development, especially in the stage of positive selection in the thymus (
Note).
Significant association of the tumor necrosis factor receptor 2 (TNFR2) gene with human narcolepsy
Authors: Hohjoh H. , Terada N. , Kawashima M. , Honda Y. , Tokunaga K.
Published: March 2003

Acknowledgments:

This work was supported by Grants‐in‐Aid from the Ministry of Education, Science, Sports and Culture of Japan and by the grant provided by the Uehara Memorial Foundation.

We report on the association study of the tumor necrosis factor receptor 2 (TNFR2) gene with human narcolepsy. A single‐nucleotide polymorphism in TNFR2, which is involved in an amino acid substitution [methionine(M)/arginine(R)] at position 196, was investigated in 149 Japanese narcoleptic patients and 204 healthy individuals as controls. Results reveal that the frequency of the TNFR2–196R allele significantly increased in the patients as compared with that in the controls (P=0.029), suggesting that TNFR2 is likely associated with the susceptibility to narcolepsy. In addition, the analyses of the relationship of TNFR2 and TNF‐α with the susceptibility to narcolepsy indicate the possibility that an additive effect on the susceptibility to the disorder lies between TNFR2–196R and TNF‐α(–857T) alleles.
Allele frequencies and haplotypic associations defined by allelic DNA typing at HLA class I and class II loci in the Japanese population
Authors: Saito S. , Ota S. , Yamada E. , Inoko H. , Ota M.
Published: March 2003

HLA class I and class II allelic genotypes were determined in 371 unrelated individuals and 309 members of 81 families inhabiting the central Japan area. A total of 20 HLA‐A alleles, 16 HLA‐Cw alleles, 38 HLA‐B alleles, 27 HLA‐DRB1 alleles, 15 HLA‐DQB1 alleles and 12 HLA‐DPB1 alleles were detected. By the two‐, three‐, four‐, five‐ and six‐locus allelic association analyses extracted from the HLA‐A to ‐DPB1 locus, 26 HLA‐Cw‐B haplotypes, 25 HLA‐DRB1‐DQB1 haplotypes, 42 HLA‐Cw‐B‐DRB1 haplotypes, 37 HLA‐Cw‐B‐DRB1‐DQB1 haplotypes, 29 HLA‐A‐Cw‐B‐DRB1‐DQB1 haplotypes and 21 HLA‐A‐Cw‐B‐DRB1‐DQB1‐DPB1 haplotypes with the frequencies of higher than 0.005 were recognized. Among 19 HLA‐B alleles with the high allele frequencies (above 0.007), 9 HLA‐B alleles, B*0702, B*1301, B*3701, B*3901, B*4006, B*4403, B*5201, B*5901 and B*6701 were found to be tightly associated with single HLA‐Cw alleles. Most of HLA‐DRB1 alleles showed strong associations with single HLA‐DQB1 alleles, but DRB1*0802 and DRB1*1401 were associated with two different DQB1 alleles. Extended haplotypes carrying infrequent class I alleles with the allele frequencies of lower than 0.007 were defined by family studies. Gene frequencies and haplotypic associations within the entire HLA classical loci elucidated at the high resolution (four‐digital) allelic level will provide useful information on anthropology, marrow donor registry, legal medicine and disease‐association studies.
Single nucleotide polymorphisms in Intron 2 of the human interleukin‐1 receptor antagonist (IL‐1Ra) gene: further definition of the IL‐1β and IL‐1Ra polymorphisms in North American Caucasians and Taiwanese Chinese
Authors: Tseng L.‐H. , Chen P.‐J. , Lin M.‐T. , Shau W.‐Y. , Chaung S.‐M. , Martin P.J. , Hansen J.A.
Published: March 2003

Abstract: Previous studies have suggested that a variable number tandem repeat (VNTR) polymorphism in the second intron of the interleukin‐1 receptor antagonist (IL‐1Ra) gene and the single nucleotide polymorphisms at positions −511 and +3954 of the IL‐1β gene might be associated with increased risks of chronic inflammatory diseases, autoimmune diseases and gastric cancer. In the present study, IL‐1β and IL‐1Ra genotypes were analyzed among Asians in Taiwan and Caucasians in North America. We identified a novel polymorphism with 3 nucleotide substitutions in the IL‐1Ra VNTR 2‐repeat allele. One of the substitutions corresponds with the fourth 3′ end nucleotide of the reverse primer that is often used for analysis of the IL‐1Ra‐associated VNTR locus. Mismatching between this primer and the 2‐repeat allele can cause misleading amplification results when stringent conditions are used for annealing. The estimated haplotype frequencies of the variant IL‐1 genes were significantly different between Taiwanese and Caucasians. The frequency of the pro‐inflammatory IL‐1Ra 2‐repeat allele was significantly lower in Taiwanese than in Caucasians. In contrast, the frequencies of the pro‐inflammatory IL‐1β–511T allele and +3954C allele were significantly higher among Taiwanese compared with Caucasians.
Lack of association between NRAMP1 gene polymorphisms and Trypanosoma cruzi infection
Authors: Calzada J.E. , Nieto A. , López‐Nevot M.A. , Martín J.
Published: March 2003

Abstract: Genetic analysis in mice and humans have established the key role of the human natural resistance‐associated macrophage protein 1 (NRAMP1) in resistance to intracellular infections. In the present study we investigated whether four NRAMP1 polymorphisms (5′(GT)n, −236 C→T, D543N, and 3′UTR deletion) were important in determining the susceptibility to Trypanosoma cruzi infections as well as in the development of chagasic cardiac disease. Genotyping for these variants was assessed in 83 seropositive (asymptomatic, n=51, cardiomyopathic, n=32) and 85 seronegative individuals from a Peruvian population where T. cruzi is endemic. No statistically significant differences either between patients and controls or between asymptomatic and cardiomyopathic individuals were observed with respect to NRAMP1 variants. Our data suggest that the NRAMP1 genetic polymorphism analysed do not play a major role in the pathogenesis of T. cruzi infection in this Peruvian sample.
HLA‐DQ alleles in patients with celiac disease in Turkey
Authors: Tüysüz B. , Dursun A. , Kutlu T. , Sökücü S. , Çine N. , Süoğlu Ö. , Erkan T. , Erginel‐Ünaltuna N. , Tümay G.
Published: March 2003

Abstract: The prevalence of the HLA‐DQA1 and DQB1 alleles in 55 Turkish children with celiac disease and 50 control subjects was investigated by using an allele‐specific DNA‐based polymerase chain reaction‐sequence‐specific primer (PCR‐SSP) method. The frequency of the DQA1*0501 and DQB1*02 alleles was higher in celiac patients than in controls. The DQA1B1 (*0501; *02) haplotype was present in 46 (83.6%) patients and only in 12 (24%) controls. The remaining 9 celiac patients which were negative for DQA1B1 (*0501;*02) carried the DQA1B1 (*03;*0302) haplotype. We found an excess homozygosity of the DQB1*02 allele and the DQA1B1 (*0501;*02) haplotype in the patients. No statistically significant correlation was found between the homozygosity of this haplotype or the DQB1*02 allele and an earlier onset of the disease.
Nomenclature for factors of the dog major histocompatibility system (DLA), 2000: Second report of the ISAG DLA Nomenclature Committee
Authors: Kennedy L.J. , Angles J.M. , Barnes A. , Carter S.D. , Francino O. , Gerlach J.A. , Happ G.M. , Ollier W.E.R. , Thomson W. , Wagner J.L.
Published: March 2003

Abstract: The ISAG DLA Nomenclature Committee met during the “Comparative Evolution of the Mammalian MHC” meeting in Manchester, England on 10th September 2000. The main points discussed were the naming of class I genes and alleles, and the inclusion of alleles from other canidae.
Comparative analysis of the association of HLA‐B*51 suballeles with Behçet’s disease in patients of German and Turkish origin
Authors: Kötter I. , Günaydin I. , Stübiger N. , Yazici H. , Fresko I. , Zouboulis C.C. , Adler Y. , Steiert I. , Kurz B. , Wernet D. , Braun B. , Müller C.A.
Published: March 2003

Abstract: The distribution of the different HLA‐B*51 suballeles among patients with Behçet’s disease (BD) of German (n=33) and Turkish (n=92) origin in comparison to their presence in the respective ethnically matched healthy control groups (German: n=325, Turkish: n=93) was studied. HLA‐B*51x was significantly increased in both patient groups in comparison to the controls (Germans: 58% vs. 12%, OR 9.76, P<0.001; Turkish: 75% vs. 25%, OR 9.13, P<0.001). Molecular subtyping of B*51x revealed HLA‐B*51011 and B*5108 as the predominant suballeles in both patient groups and controls although with a slightly increased frequency of HLA‐B*5108 in the diseased individuals. HLA‐B*5105 was the only further HLA‐B*51x subtype detected in one Turkish patient heterozygous also for HLA‐B*5101. HLA‐B*5107 although present in a Turkish as well as German control was absent in the patient groups. There was also a tendency towards a higher degree of homozygosity for HLA‐B*51x in both patient groups versus the matched controls (Germans: 10% in patients vs. 2,5% in controls; Turkish: 27% in patients vs. 13% in controls). Our study further supports previous hypothesis of an association of BD with B51 suballeles which share amino‐acid residues at positions 63 and 67 as well as at positions 77–83 for specific peptide binding and natural killer (NK)‐cell interactions. This applies to HLA‐B*5101 and B*5108, but not to HLA‐B*5107 different at position 67, which could be negatively associated with BD.
HLA class II polymorphism in Aka Pygmies and Bantu Congolese and a reassessment of HLA‐DRB1 African diversity
Authors: Renquin J. , Sanchez‐Mazas A. , Halle L. , Rivalland S. , Jaeger G. , Mbayo K. , Bianchi F. , Kaplan C.
Published: March 2003

Abstract: HLA‐DRB1, ‐DQB1 and ‐DPB1 polymorphisms were investigated in two African populations, the Basse Lobaye Aka Pygmies of the Central African Republic, and a Bantu‐speaking group from the Democratic Republic of Congo Kinshasa. Allelic and haplotypic frequency distributions reveal marked differences between the two populations in spite of their geographical proximity: the Aka exhibit high frequencies for several alleles, especially at the DPB1 locus (0.695 for DPB1*0402), probably due to rapid genetic drift, while the Bantu distributions are more even. Genetic distances computed from DRB1 allelic frequencies among 21 populations from North and sub‐Saharan Africa were applied to a multidimensional scaling analysis. African populations genetic structure is significantly shaped by linguistic differentiation, as confirmed by an analysis of molecular variance. However, selective neutrality tests indicate that many African populations exhibit an excess of heterozygotes for DRB1, which is likely to explain the genetic similarity observed between some North African and Bantu populations. Overall, this study shows that natural selection must be taken into account when interpreting the patterns of HLA diversity, but that this effect is probably minor in relation to the stochastic events of human population differentiations.
West Mediterranean islands (Corsica, Balearic islands, Sardinia) and the Basque population: contribution of HLA class I molecular markers to their evolutionary history
Authors: Grimaldi M.‐C. , Crouau‐Roy B. , Amoros J.‐P. , Cambon‐Thomsen A. , Carcassi C. , Orru S. , Viader C. , Contu L.
Published: March 2003

Abstract: The genetic structure of Balearic islands (Corsica and Sardinia), situated on the same trans‐Mediterranean maritime routes and having very similar histories, were compared and their position among the neighbouring Caucasian populations was inferred. For this purpose, three HLA loci (HLA‐A, ‐B and ‐Cw) were typed at the DNA level in these populations and the allelic and haplotypic frequencies were estimated. Because previous studies have shown common genetic features in the Sardinians and Basques, HLA‐Cw molecular typing was also performed in a sample of French Basques in order to establish the haplotypic structure of this population for a more accurate comparison with the three others. By its allelic composition, the Corsican population has an intermediate position between the two other islander populations. Its close relationship with the Sardinian population, however, was clearly revealed by the phylogenetic analysis which also suggests a proximity with eastern Mediterranean peoples, whereas the Balearic islands are more narrowly related to Spain and western Europe. Peculiarities were observed in the distributions of some common haplotypes in the populations of the islands that confirm the results of the phylogenetic analysis and could be related to their history. Noteworthy is the presence of the HLA‐A30‐Cw*0501‐B18 haplotype at frequencies ≈2% in Corsica and the Balearic islands, yet the estimated frequencies of this haplotype are much lower than in the Sardinian and Basque populations.
Novel HLA‐B alleles formed by an inter‐locus recombination with HLA‐C, HLA‐B*0713 and B*6702
Authors: Steiner N.K. , Baldassarre L. , Koester R. , Ng J. , Hartzman R.J. , Hurley C.K.
Published: March 2003

Abstract: This paper describes two novel HLA‐B locus alleles. B*0713 appears to have resulted from an interlocus recombinational event which inserted a sequence from an HLA‐C allele into B*0702. This event altered a significant portion of the alpha‐1 domain alpha helix. B*6702 shares much of its exon 2 sequence with B*0713 but is identical in exon 3 to several HLA‐B locus alleles including B*67012.
Two new HLA‐B alleles, B*4422 and B*4704, identified in a study of families with autoimmunity
Authors: Grams S.E. , Moonsamy P.V. , Mano C. , Oksenberg J.R. , Begovich A.B.
Published: March 2003

Abstract: HLA‐B typing of approximately 1 262 individuals from a study of 372 simplex families with multiple sclerosis has led to the identification of two new alleles (HLA‐B*4422 and HLA‐B*4704). Sequencing confirmed that both of these new alleles represent novel combinations of previously described sequence motifs, reinforcing the notion that recombination and/or gene conversion‐like events play an important role in generating HLA allelic diversity. The identification of these new alleles brings the total number of HLA‐B alleles to 465.
Sequence of the HLA‐B*1573 allele
Authors: Morante M.P. , Vidal S. , Mosquera A.M. , Querol S. ,   J. García, Rodríguez‐Sánchez J.L.
Published: March 2003

Abstract: We report here the sequence of an HLA B*15 allele, B*1573. Initially, a cord blood unit from the Umbilical Cord Bank of Barcelona was typed by sequence‐specific oligonucleotide hybridization and revealed an unusual hybridization pattern. After the cloning, the sequence‐based typing assigned two different alleles: B*07021, and a second allele identical to B*1558 in exons 2 and 3, except for a single nucleotide substitution in exon 3, which changed codon 140 from Phe to Leu (ΤΤΧ→ΤΤΑ).
POLYMORPHISMS OF ANGIOTENSIN‐CONVERTING ENZYME, ANGIOTENSINOGEN, ANGIOTENSIN II TYPE 1 AND TYPE 2 RECEPTORS AND ENDOTHELIAL NITRIC OXIDE SYNTHASE IN TURKISH RENAL TRANSPLANT RECIPIENTS: A PRELIMINARY STUDY
Authors: Ataç FB , Akçay A , Özdemir Fn , Verdi H , Terzioğlu M , Çolak T , Emiroğlu R , Haberal M
Published: March 2003

Many researchers are now focusing on ways in which the renin‐angiotensin system (RAS) and RAS gene polymorphism affect progressive renal damage, hypertension, cardiac complications, and renal allograft function. In this study, we investigated the frequency of polymorphism in the genes for angiotensin‐converting enzyme (ACE), angiotensinogen (AGT), angiotensin II type 1 and 2 receptors (AT1 and AT2), and endothelial nitric oxide synthase (ec‐NOS) in Turkish renal transplant recipients. Angiotensin‐converting enzyme insertion/deletion (I/D) polymorphism was assessed in 125 patients. M235T polymorphism of the angiotensinogen gene, 1166 A/C polymorphism of the AT1 receptor gene, and 3123C/A polymorphism of the AT2 receptor gene, and intron 4 VNTR polymorphism in the ec‐NOS gene were assessed in 40 of the patients. The latter list was also investigated in 50 healthy controls from the general Turkish population. DD/ID/II genotypes of the ACE gene were detected in 56%, 34%, and 10% of the kidney recipient group, respectively. In the 40 recipients who were tested for the other polymorphisms, the corresponding figures for the AGT TT/TM/MM genotypes were 0%, 82.5%, and 17.5%, respectively; for the AT1 CC/CA/AA genotypes were 0%, 2.5%, and 97.5%, respectively; for the AT2 AA/CA/CC genotypes were 0%, 0%, and 100%, respectively; and for the ec‐NOS intron 4 aa/ab/bb genotypes were 0%, 12%, and 88%, respectively. The results revealed that Turkish renal transplant recipients exhibit a wide‐range of RAS gene and ec‐NOS polymorphisms.
New analogues of bradykinin containing a conformationally restricted dipeptide fragment in their molecules
Authors: Derdowska I. , Prahl A. , Lammek B. , Neubert K. , Hartrodt B. , Kania A. , Dobrowolski D. , Malhem S. , Trzeciak H.I. , Wierzbad T.
Published: March 2003

Abstract: The present paper describes the synthesis and some pharmacological properties of two new bradykinin analogues containing the ethylene‐bridged dipeptide Phe‐Phe in their molecules. In a further two peptides this modification was combined with acylation of the N‐terminus with 1‐adamantaneacetic acid. Finally, we synthesized four analogues by removing the Ser6 residue from the four peptides mentioned above. The activity of the new analogues was assayed on isolated rat uterus (RUT) and in rat blood pressure tests (BPT). The results clearly indicate that the proposed modification, alone or in combination with other changes, resulted in either a drop in antiuterotonic activity or even in conversion to an agonism. Although this tendency is not so distinct in blood pressure assays, the antagonistic potency of the new analogues is also diminished. Nevertheless, it was demonstrated that the d‐amino acid in position 7 which, until recently, was considered necessary for antagonism, may be replaced, together with the amino acid occupying position 8, by a suitable, sterically restricted L,L‐dipeptide unit.
Enzymatic synthesis of chromogenic substrates for Glu,Asp‐specific proteinases
Authors: Milgotina E.I. , Shcheglov A.S. , Lapa G.B. , Chestukhina G.G. , Voyushina T.L.
Published: March 2003

Abstract  Glu,Asp‐specific endopeptidases represent a new subfamily of chymotrypsin‐like proteolytic enzymes. These enzymes prefer Glu or Asp residues in the P1 position of the substrates. p‐Nitroanilides of N‐acylated di‐, tri‐ and tetrapeptides with C‐terminal glutamic or aspartic acid residues have been obtained. Acyl peptide p‐nitroanilides were synthesized via acylation of glutamic or aspartic acid p‐nitroanilides using methyl esters of the respective N‐acylated peptides, generally with good yields. The reactions were performed in organic solvents using subtilisin 72 sorbed on silica as a catalyst. The kinetic parameters for the hydrolysis of these p‐nitroanilides with proteinases from Bacillus intermedius and Bacillus licheniformis were determined.
11‐Demethylcyclosporins exhibit a single conformation in methanol and dimethylsulfoxide
Authors: Sedmera P. , Jegorov A. , Buchta M. , Cvak L.
Published: March 2003

Abstract:NMR studies showed that 11‐demethylcyclosporin A (cyclosporin E) and 11‐demethylcyclosporin B exist as single species both in polar and nonpolar solvents. They adopt the same conformation that was found in the solid state.
Molecular dynamics simulation of the antimicrobial salivary peptide histatin‐5 in water and in trifluoroethanol: a microscopic description of the water destructuring effect
Authors: Iovino M. , Falconi M. , Marcellini A. , Desideri A.
Published: March 2003

Abstract: The results of 520 ps molecular dynamics simulation of histatin‐5, a small peptide present in human saliva and possessing antimicrobial activity, dissolved in water and in 2,2,2‐trifluoroethanol, are reported. The simulations indicate that histatin‐5 is destabilized in water and begins to unfold after 250 ps, while in organic solvent it maintains a regular secondary structure throughout the trajectory. Analysis of the peptide−solvent hydrogen bonds indicates that 2,2,2‐trifluoroethanol is a poorer proton acceptor than water. The fluorine atom of the alcohol is almost never engaged in a hydrogen bond and the organic solvent interacts mainly with the peptide through its hydroxyl group. For some residues analysis of the solvent residence time indicated longer values for 2,2,2‐trifluoroethanol than for water. The most striking difference is related to the number of times the solvent enters and leaves the first coordination shell of the peptide. This value was more than one order of magnitude higher for water than for the alcohol, suggesting that this may be the main cause of α‐helix destabilization perpetrated by water.
Novel cyclic peptide inhibits intercellular adhesion molecule‐1‐mediated cell aggregation
Authors: Shannon J.P. , Silva M.V. , Brown D.C. , Larson R.S.
Published: March 2003

Abstract: Leukocyte adherence mediated by intercellular adhesion molecule‐1 (ICAM‐1) binding to leukocyte function‐associated antigen (LFA‐1) is required for proper inflammatory and immune function. Inhibition of ICAM‐1\LFA‐1 binding using monoclonal antibodies (mAb) has been shown to be efficacious at inhibiting lymphoma metastasis as well as leukocyte emigration into tissue in a number of inflammatory diseases such as ischemia−reperfusion injury, septic shock and rheumatoid arthritis. In this report, we describe the development and characterization of a small peptide antagonist of ICAM‐1‐dependent cell aggregation. By using repeated selection of a cyclic nonapeptide phage display library on purified ICAM‐1, we identified phage that were competitively eluted with anti‐ICAM‐1 mAb. The peptide sequences were determined by nucleotide sequencing, and the peptide sequence (C*LLRMRSIC*) (IP01) that occurred most frequently was chosen for further study. Phage expressing this peptide sequence specifically bound ICAM‐1 over a range of 5 × 106 to 1 × 108 phage/µL. A cyclic IP01 peptide, linear IP01 peptide, a cyclic nonapeptide with a scrambled IP01 sequence, and a random, cyclic nonapeptide were synthesized. The cyclic and linear IP01 peptides were able to inhibit ICAM‐1‐mediated cell aggregation at a concentration of 1 mm, whereas the random and scrambled peptide sequences did not alter aggregation. Cyclic IP01 had a half‐maximal inhibitory concentration of ≈ 970 µm. Cyclic IP01 did not inhibit cellular aggregation that was dependent on ICAM‐2 or ICAM‐3. Alanine substitutions in the cyclic IP01 identified at least four amino acids necessary for inhibition of ICAM‐1 dependent cell aggregation; leucine 2, leucine 3, methionine 5, and arginine 6. Finally, we showed that cyclic IP01 can inhibit firm adhesion of neutrophils to endothelium, a critical event in inflammatory diseases, in an assay that recapitulates physiologic flow conditions. Homology of IP01 with the primary amino acid sequences of the α or β subunit of LFA‐1 was not identified. Thus, we identified a unique molecule that inhibits ICAM‐1 dependent cell adhesion, but is not related to the primary sequence of the ICAM‐1 ligand LFA‐1. Due to the small size and ability to block cell−cell adhesion, IP01 may serve as a useful tool for study of ICAM‐1 and LFA‐1 biology as well as for the development of small molecule therapeutics.
Synthesis of enantiopure non‐natural α‐amino acids usingtert‐butyl (2S)‐2‐[bis‐(tert‐butoxycarbonyl)amino]‐5‐oxopentanoate as key‐intermediate:the first synthesis of(S)‐2‐amino‐oleic acid
Authors: Constantinou‐Kokotou V. , Magrioti V. , Markidis T. , Kokotos G.
Published: March 2003

Abstract:A general method for the synthesis of enantiopure non‐natural α‐amino acids is described. The key intermediatetert‐butyl (2S)‐2‐[bis(tert‐butoxycarbonyl)amino]‐5‐oxopentanoate was obtained froml‐glutamic acid after suitable protection and selective reduction of the γ‐methyl ester group by DIBALH. Wittig reaction of this chiral aldehyde with various ylides led to a variety of δ,ε‐unsaturated α‐amino acids. This methodology was applied to the synthesis of (S)‐2‐amino‐oleic acid.
Synthesis of novel protected Nα(ω‐thioalkyl) amino acid building units and their incorporation in backbone cyclic disulfide and thioetheric bridged peptides
Authors: Gazal S. , Glukhov E. , Gilon C. , Gellerman G.
Published: March 2003

Abstract: General methods for the preparation of protected Nα(ω‐thioalkyl) amino acids building units for backbone cyclization using reductive alkylation and on‐resin preparation are described. The synthesis of non‐Gly Fmoc‐protected S‐functionalized N‐alkylated amino acids is based on the reaction of readily prepared protected ω‐thio aldehyde with the appropriate amino acid. Preparation of Fmoc‐protected S‐functionalized N‐alkylated Gly building units was carried out using two methods: reaction of glyoxylic acid with Acm‐thioalkylamine and an on‐resin reaction of bromoacetyl resin with Trt‐thioalkylamines. Three model peptides were prepared using these building units. The GlyS2 building unit was incorporated into a backbone cyclic analog of somatostatin that contains a disulfide bridge. Formation of the disulfide bridge was performed by on‐resin oxidation using I2 or Tl(CF3COO–)3. Both methods resulted in the desired product in a high degree of purity in the crude. The AspS3 building unit was also successfully incorporated into a model peptide. In addition, the in situ generation of sulfur containing Gly building units was demonstrated on a Substance P backbone cyclic analog containing a thioether bridge.
Disulfide pairing and secondary structure of ASP1, an olfactory‐binding protein from honeybee (Apis mellifera L)
Authors: Briand L. , Nespoulous C. , Huet J.‐C. , Pernollet J.‐C.
Published: March 2003

Abstract: In insects, the transport of airborne, hydrophobic odorants and pheromones through the sensillum lymph is accomplished by olfactory‐binding proteins (OBPs). We report the structural characterization of a honeybee OBP called ASP1 found in workers and drones, previously observed to bind queen pheromone components. A novel method based on ion‐spray mass spectrometry analysis of cyanylation‐induced cleavage products of partially reduced protein with Tris(2‐carboxyethyl)phosphine was needed to determine the recombinant ASP1 disulfide bond pairing. It was observed to be Cys(I)‐Cys(III), Cys(II)‐Cys(V), Cys(IV)‐Cys(VI), similar to those already described for other OBPs from honeybee and Bombyx mori suggesting that this pattern occurs commonly throughout the diverse family of insect OBPs. Circular dichroism revealed that ASP1 is an all‐alpha protein in accordance with NMR preliminary data, but unlike lipocalin‐like vertebrate OBPs.
Synthesis and analysis of potent, more lipophilic derivatives of the bradykinin B2 receptor antagonist peptide Hoe 140
Authors: Kennedy K.J. , Orwig K.S. , Dix T.A. , Christopher J. , Jaffa A.A. , Dix T.A.
Published: March 2003

Abstract:  Bradykinin (BK) is an endogenous peptide that has been implicated in several pathological conditions, hence antagonists of its activity have therapeutic potential. The decapeptide Hoe 140 is currently one of the best BK antagonists, but interest remains in finding even more potent compounds. A library of Hoe 140 derivatives was synthesized that incorporated non‐natural analogs of the cationic, naturally occurring amino acids arginine (Arg) and lysine (Lys). The modified amino acids were designed to form enhanced ionic interactions due to an increase in local hydrophobicity, which promotes desolvation of the cation in water. The potencies of the resulting peptides were determined by competitive binding assays in human A431 cells expressing the BK B2 receptor. Two of the peptides synthesized were equipotent to Hoe 140 (IC50s 2.99 and 3.36 nm) and the most potent was demonstrated as a functional antagonist in vitro by blocking BK‐mediated phosphorylation of mitogen‐activated protein (MAP) kinases. The new derivatives are more hydrophobic than Hoe 140 and thus may exhibit changes in pharmacokinetic properties when evaluated in vivo.
Synthesis of cyclic peptides from unprotected precursors using removable Nα‐(1‐(4‐methoxyphenyl)‐2‐mercaptoethyl) auxiliary
Authors: Cardona V.M.F. , Hartley O. , Botti P.
Published: March 2003

Abstract: A new method to cyclize unprotected peptides is presented. The method involves the use of a 1‐phenyl‐2‐mercaptoethyl derivative on the N‐terminal glycine. This template acts as an auxiliary thiol‐containing group in order to drive cyclization with a counterpart thioester moiety on the same molecule. Subsequent facile removal of the derivative generates products with only native peptide structure. The successful, high‐yield cyclization of the peptide GSPYSSDTTPA is described.
Enhanced interleukin‐8 production in THP‐1 human monocytic cells by lipopolysaccharide from oral microorganisms and granulocyte‐macrophage colony‐stimulating factor
Authors: Baqui A. A. M. A. , Meiller T. F. , Falkler W. A.
Published: March 2003

Granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) has been used to assist in bone marrow recovery during cancer chemotherapy. Interleukin‐8 (IL‐8) plays an important role in macrophage mediated inflammatory processes including exacerbation of periodontal diseases, one of the most common complications in GM‐CSF receiving cancer patients. The effect of GM‐CSF supplementation on IL‐8 production was investigated in a human monocyte cell line THP‐1, stimulated with lipopolysaccharide extracted from two oral microorganisms, Porphyromonas gingivalis and Fusobacterium nucleatum. Resting THP‐1 cells were treated with lipopolysaccharide (1 μg/ml) of P. gingivalis or F. nucleatum and/or GM‐CSF (50 IU/ml) for varying time periods. The production of IL‐8 in THP‐1 cells was measured by a solid‐phase enzyme‐linked immunosorbent assay (ELISA). A very low level of the cytokine IL‐8 was produced constitutive in THP‐1 cells. Starting from 8 h of treatment and afterwards GM‐CSF alone significantly increased IL‐8 production in THP‐1 cells. Lipopolysaccharide (1 μg/ml) extracts from either F. nucleatum or P. gingivalis amplified IL‐8 production 500–800 times in comparison to resting THP‐1 cells. When lipopolysaccharide of F. nucleatum or P. gingivalis was supplemented with 50 IU/ml of GM‐CSF, there was a statistically significant enhanced production of IL‐8 by THP‐1 cells after 1 day to 7 days of treatment as compared with lipopolysaccharide treatment alone. GM‐CSF (50 IU/ml) also significantly increased IL‐8 production from 2–7 days of treatment of THP‐1 cells when supplemented with a positive control, phorbol‐12‐myristate‐13 acetate (PMA), as compared to PMA treatment alone. These investigations using the in vitro THP‐1 human monocyte cell model indicate that there may be an increase in the response on a cellular level to oral endotoxin following GM‐CSF therapy as evidenced by enhanced production of the tissue‐reactive inflammatory cytokine, IL‐8.
Proteolysis and utilization of albumin by enrichment cultures of subgingival microbiota
Authors: Wei G. X. , Van Der Hoeven J. S. , Smalley J. W. , Mikx F. H. M. , Fan M. W.
Published: March 2003

Subgingival dental plaque consists mainly of microorganisms that derive their energy from amino acid fermentation. Their nutrient requirements are met by the subgingival proteolytic system, which includes proteases from microorganism and inflammatory cells, and substrate proteins from sulcus exudate, including albumin. To determine the selective effect of individual proteins on microbiota, we used albumin as the main substrate for growth. Eight subgingval plaque samples from untreated periodontal pockets of patients with adult periodontitis were inoculated in peptone yeast medium with bovine albumin (9 g/l). After three subculture steps, cell yields of the enrichment cultures at the medium with 0, 1.25, 2.5, 5, 10, and 20 g/l albumin were determined. Proteolytic activity (U/absorbance at 550 nm) of the enrichment cultures and different isolates derived from the cultures was estimated by the degradation of resorufin‐labeled casein. It was observed that the yield of the mixed culture was albumin limited, and the proteolytic activities of the cultures in albumin broth were higher than in control (peptone broth). Among the isolates from the enrichment cultures, Peptostreptococcus micros, Prevotella melaninogenica, Prevotella buccae and Prevotella bivia demonstrated proteolysis. The frequent occurrence of Streptococcus gordonii and Streptococcus anginosus in the albumin cultures is explained by their ability to utilize arginine as an energy source for growth. Albumin in the medium was partly degraded by pure cultures but completely consumed in enrichment cultures, indicating synergy of bacterial proteinases. It is concluded that the subgingival microbiota possesses proteolytic activity and may use albumin as a substrate for their growth. Enrichment cultures on albumin may serve as a relatively simple in vitro model to evaluate the effects of proteinase inhibitors.
Candida albicans triggers interleukin‐6 and interleukin‐8 responses by oral fibroblasts in vitro
Authors: Dongari‐Bagtzoglou A. , Wen K. , Lamster I. B.
Published: March 2003

Oral candidiasis is the most frequent opportunistic infection associated with an immunocompromised host. Production of proinflammatory cytokines, such as interleukin‐6 (IL‐6) and IL‐8, by host cells in response to Candida albicans can be expected to have a major impact in the activation of immune effector cells against the invading microorganism. Using a human cell–C. albicans coculture model system, we determined that this microorganism can trigger secretion of these potent chemoattractant and proinflammatory cytokines by oral mucosal fibroblasts. This response varied depending on the infecting strain and required fungal viability, germination of yeast into hyphae and mannose‐mediated direct contact between the host cell and Candida. The secretion of proinflammatory cytokines by oral mucosal fibroblasts in response to C. albicans suggests that these cells have the potential to enhance the host defense against this organism in vivo. This may have important implications in controlling fungal overgrowth in the oral cavity.
Heterogeneity of the prtC gene of Porphyromonas gingivalis
Authors: Wittstock M. , Schmidt H. , Flemmig T. F. , Karch H.
Published: March 2003

In this study, the nucleotide sequences of the prtC genes of six clinical Porphyromonas gingivalis isolates obtained from patients with periodontitis and from reference strain 53977 were determined. All analyzed genes were heterogeneous in their nucleotide composition and differed in up to 13 nucleotides. Moreover, substantial differences were found in comparison to prtC of reference strain 53977. The prtC genes of 45 Porphyromonas gingivalis isolates were amplified by polymerase chain reaction (PCR) and the PCR products were also digested with restriction endonucleases Tsp509I, NlaIII and DraII (PCR–restriction fragment‐length polymorphism). Nine different restriction pattern combinations were observed, with four being most frequent (28.9%, 26.7%, 17.8% and 11.1%). The data presented here demonstrates that prtC genes are heterogeneous in their nucleotide sequence and therefore may be used as a target for molecular epidemiological studies. The observed heterogeneity of prtC genes may be a result of microevolution processes.
Porphyromonas gingivalis induction of mediator and cytokine secretion by human gingival fibroblasts
Authors: Steffen M. J. , Holt S. C. , Ebersole J. L.
Published: March 2003

We hypothesized that bacterial viability and strain characteristics of Porphyromonas gingivalis could affect the induction of pro‐inflammatory mediator secretion by human gingival fibroblast cultures. Both killed and viable P. gingivalis elicited production of prostaglandin E2, interleukin‐1β (IL‐1β), IL‐6 and IL‐8, although killed P. gingivalis induced generally higher levels, particularly IL‐6 and IL‐8, compared with the viable bacteria. P. gingivalis strains, which exhibited wild‐type levels of trypsin‐like protease activity, stimulated human gingival fibroblasts to secrete increased levels of prostaglandin E2 and IL‐1β, although minimal levels of IL‐6 and IL‐8 were noted in supernatants from the gingival fibroblast cells. P. gingivalis strains BEI and NG4B19, which have either decreased or undetectable levels of trypsin‐like protease, respectively, induced significantly greater IL‐6 and IL‐8 levels in gingival fibroblast cultures compared with the other strains. The ability of antibody to P. gingivalis to alter human gingival fibroblast production of pro‐inflammatory mediators was tested using nonhuman primate antisera. Both immune and nonimmune sera altered the P. gingivalis–generated pattern of mediators from the gingival fibroblasts. We conclude that: (i) viable and killed P. gingivalis were capable of inducing various pro‐inflammatory cytokines from human gingival fibroblasts; (ii) strain differences in cytokine induction were noted, and the expression of a trypsin‐like protease activity was related to decreased extracellular levels of IL‐6 and IL‐8; and (iii) the presence of serum, particularly with specific antibody to P. gingivalis, significantly altered human gingival fibroblast cytokine production compared with P. gingivalis alone.
Identification and antimicrobial susceptibility of enterococci isolated from the root canal
Authors: Dahlén G. , Samuelsson W. , Molander A. , Reit C.
Published: March 2003

Enterococci are occurring in opportunistic infections involving the oral cavity. This study has identified enterococcal species in 29 endodontic infections undergoing treatment with Ca (OH)2 dressings. The in vitro antimicrobial susceptibility of 29 isolated enterococcal strains was determined. Enterococcus faecalis was speciated for 26 isolates and Enterococcus faecium for three isolates. In vitro antimicrobial susceptibility testing revealed enterococcal isolates resistant to benzylpenicillin, ampicillin, clindamycin, metronidazole and tetracycline but sensitive to erythromycin and vancomycin. Due to low sensitivity to antimicrobial agents, enterococci may be selected in root canals undergoing standard endodontic treatment and significantly contribute to endodontic treatment failures.