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  • 1
    ISSN: 1432-1211
    Keywords: Key words NOD mouse ; GAD65 ; I-Ag7 ; Peptide ; Vaccine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Insulin-dependent diabetes mellitus (IDDM) develops in nonobese diabetic (NOD) mice through the destruction of the B cells in pancreatic Langerhans islets by islet autoantigen-specific T cells. The islet autoantigen glutamic acid decarboxylase 65 (GAD65) is thought to be a major target autoantigen in IDDM. In the present report, we established GAD65-specific T-cell clones using overlapping peptides that cover the amino acid sequences of mouse GAD65. T-cell epitopes of GAD65 were characterized by proliferation and binding assays using various analogue peptides and wild-type or mutant I-Ag7 transfectants. The efficacy of the peptide vaccine in IDDM was determined by administering T-cell epitope peptides to NOD mice and evaluating the histopathology of their insulitis. We obtained two types of T-cell clone, one specific for peptide p316–335 and another specific for p531–545 of GAD65. The p531–545 site has already been identified, but we report the p316–335 site for the first time. T-cell clones recognized those peptides in the wild-type I-Ag7 but not in the mutant I-Ag7 in which the serine at position 57 of the β-chain was replaced by an aspartic acid. Both the p316–335 and p531–545 peptides bound weakly to I-Ag7. Some peptides with amino acid substitutions had antagonistic activity, and administration of a large amount of wild-type peptide reduced the severity of insulitis in NOD mice. Our results suggest that peptide vaccine therapy may be useful in autoimmune diseases, including IDDM.
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  • 2
    ISSN: 1432-1211
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-1211
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The ampibian Xenopus laevis is the most primative vertebrate in which the major histocompatibility complex (MHC) has been defined at the biochemical, functional, and molecular genetic levels. We previously described the isolation and characterization of cDNA clones encoding X. laevis MHC class II β chains. In the present study, genomic clones encoding class II β chains were isolated from X. laevis homozygous for the MHC f haplotype. Three class II β chain genes, designed Xela-DAB, Xela-DBB, and Xela-DCB, were identified. Seqeunce analysis of these genes showed that Xela-DBB and Xela-DCB corresponding to the previously characterized cDNA clones F3 and F8, respectively, whereas Xela-DAB encodes a third, hitherti unidentified class II β chain of the MHC f haplotype. As a representative of X. laevis class II β chain genes, the Xela-DAB gene underwent detailed structural analysis. In addition,the nucleotide sequence of Xela-DAB f cDNA clones was determined. The Xela-DAB gene is made up of a least six exons, with an exon-intron organization similar to that of a typical mammalian class II β chain gene. The 5′-flanking region of the Xela-DAB gene contains transcriptional control elements known as X1, X2, and Y, but lacks typical TATA or CCAAT boxes. A notable feature of the X. laevis class II β chain genes is that sizes of the introns are larger than those of their mammalian counterparts. As assessed by northern blot analysis, the three class II β chain genes had similar expression patterns, with the highest level of transcription detected in the intestine. Identification of the Xela-DAB,-DBB, and -DCB genes is consistent with our previous observations, which suggested that the MHC of the tettraploid frog X. laevis is diploidized at the genomic level and contains three class II β chain genes per haplotype that cross-hybridize to one another under reduced stringency conditions.
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  • 4
    ISSN: 1432-1211
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The peptide motif of the HLA-DR53 (DRB4*0101) molecule, which is associated with autoimmune diseases including Vogt-Koyanagi-Harada’s syndrome, was determined by peptide binding assay using human L plastin p581 – 595 peptide and its substituted analogues. L plastin p581 – 595 peptide is one of the naturally processed peptides bound to HLA-DR9/DR53 (DRB1*0901/DRB4*0101) molecules. The binding affinity of each peptide to the HLA-DR53 molecule was measured by fluorescence intensity of biotinylated peptides to L cell transfectants expressing HLA-DR53 molecules, followed by treatment with avidin-fluorescence. Binding of biotinylated peptides to HLA-DR53 molecules was not inhibited by all single-alanine-substituted nonbiotinylated peptides, indicating that the replaced position was important for binding to the HLA-DR53 moleule. The inhibitory motif is considered to be an HLA-DR53-specific binding motif, composed of a positively charged residue (K) at position 1, a hydrophobic residue (I) at position 4, positively charged residue (R or K) at position 8 or 9, and another hydrophobic residue (I) at position 10. This predicted motif is different from the binding motifs of other HLA-DR molecules.
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  • 5
    ISSN: 1432-1211
    Keywords: Key words HLA ; Peptide ; VKH disease ; Tyrosinase ; T-cell response
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Human T-cell-mediated autoimmune diseases are often genetically linked to particular alleles of HLA class II genes. Vogt-Koyanagi-Harada’s (VKH) disease, which is regarded as an autoimmune disorder in multiple organs containing melanocytes, has been found to be associated with HLA-DR4 (DRB1*0405) and HLA-DR53 (DRB4*0101). Tyrosinase is a melanoma antigen (Ag) expressed by normal melanocytes as well as melanoma cells against which responses by autologous T cells have been detected. We established a T-cell line from the peripheral blood of a patient with VKH disease which responded to synthetic peptides corresponding to tyrosinase. The T-cell line was generated which recognized the tyrosinase p188 – 208 peptide when presented by the HLA-DR4 (DRB1*0405) molecule on the surface of HLA class II-expressing L-cell transfectants. The minimal antigenic peptide which induced T-cell responses was an 11-amino-acid sequence and located at tyrosinase p193 – 203 (E-I-W-R-D-I-D-F-A-H-E). This peptide contained the DRB1*0405-binding peptide motif (hydrophobic residues (Y, F, W) at position 1 as an anchor residue, and negatively charged residues (D, E) at position 9), which corresponded to the W at p195 and the D at p203. These observations demonstrate that tyrosinase peptides are immunogenic, and may be a candidate for an autoantigen in VKH disease, suggesting that probing the T-cell responses against synthetic peptides is a productive approach for identifying the autoantigenic peptides associated with autoimmune diseases including VKH disease.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1432-1211
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    s.l. ; Stafa-Zurich, Switzerland
    Materials science forum Vol. 561-565 (Oct. 2007), p. 2009-2012 
    ISSN: 1662-9752
    Source: Scientific.Net: Materials Science & Technology / Trans Tech Publications Archiv 1984-2008
    Topics: Mechanical Engineering, Materials Science, Production Engineering, Mining and Metallurgy, Traffic Engineering, Precision Mechanics
    Notes: Structures and morphologies of Ge precipitates in an Al-Ge alloy were characterized by acombination of transmission electron microscopy and three-dimensional electron tomography.Faceting of the precipitates was clearly seen using transmission electron microscopy and varietiesof precipitate morphologies were identified by three-dimensional electron tomography
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1440-1797
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: SUMMARY: 22-oxa-calcitriol (OCT), a vitamin D analogue, suppresses parathyroid hormone (PTH) secretion and has less calcaemic activity than 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] in vivo. In this study, we evaluated the effect of OCT on PTH secretion in vitro using human hyperplastic parathyroid tissue obtained during surgery for advanced renal hyperparathyroidism and normal bovine parathyroid glands to compare the effects of 1,25(OH)2D3. 22-oxa-calcitriol suppressed PTH secretion by nodular hyperplastic parathyroid tissue and normal bovine tissue in a dose dependent manner, the same as 1,25(OH)2D3. We showed the additive effect of extracellular calcium level on suppression of PTH secretion by OCT and 1,25(OH)2D3. These results suggest that OCT suppresses PTH secretion, the same as 1,25(OH)2D3 not only in normal parathyroid cells but also on hyperplastic parathyroid cells.
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  • 9
    ISSN: 1077-3118
    Source: AIP Digital Archive
    Topics: Physics
    Notes: Characteristics of Schottky barriers formed on homoepitaxial diamond film have been studied. Current–voltage characteristics of Al contacts on both the as-grown film and the oxidized film show rectification. On the other hand, ohmic property is observed on Au/as-grown film while Au/oxidized film shows rectification. These results imply that the mechanism of the barrier formation on the as-grown diamond is drastically changed by oxidation. The difference of electrical properties between the as-grown film and the oxidized film is also observed from capacitance–voltage characteristics. This result suggests that additional acceptors which are not related to boron, exist in the as-grown film and disappear after oxidation. © 1995 American Institute of Physics.
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 1432-2323
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Résumé Malgré des progrès thérapeutiques récents, l-hyperparathyroïdie secondaire en rapport avec l'insuffisance rénale nécessite une parathyroïdectomie. L'hyperparathyroïdie récidivante est un des problèmes les plus importants. Dans cette étude, l'incidence de la récidive de l'hyperparathyroïdie a été évaluée par rapport à des données d'anatomie pathologique. L'analyse par cytométrie de flux de l'AND a été réalisée pour évaluer le pouvoir de prolifération du tissu parathroïde. Deux cent quarante huit patients ayant eu une parathyroïdectomie entre 1973 et 1991 ont été étudiés. La fréquence de la récidive de l'hyperparathyroïdie après parathyroïdectomie subtotale était de 4/19 (21.1%), celle de récidives du greffon après exérèse du tissu résiduel et autotransplantation à l'avant-bras était de 2/4 (50%). La fréquence de récidive du greffon après parathyroïdectomie totale et autotransplantation à l'avant-bras a été de 16/212 (7.5%). La fréquence de récidive était plus élevée (p〈0.01) lorsque le tissu greffé était de siège d'une hyperplasie nodulaire (17/68; 25%) que lorsque le greffon etait le siège d'une hyperplasie diffuse (1/105; 〈1%). Les 58 échantillons étudiés an cytométrie de flux ont montré une diploïdie d'ADN. Le nombre de cellules dispersées ayant une disposition non diploïde était plus important (p〈0.01) en cas d'hyperplasie nodulaire avant l'autotransplantation et en cas d'exérèse de tissu parathyroïde pour récidive qu'en cas d'hyperplasie diffuse. Ces données cliniques et les résultats d'analyse cytométrique indiquent clairement que le tissu parathyroïde hyperplasique a une puissance de croissance élevée et, pour éviter les récidives du greffon, il ne faut pas autotransplanter de tissu hyperplasique.
    Abstract: Resumen A pesar de recientes avances terapéuticos, el hiperpatiroidismo secundario a falla renal severa hace necesaria la paratiroidectomía, pero el hiperparatiroidismo recurrente constituye uno de los problemas más significativos. En el presente estudio se analizó la incidencia de hiperpatiroidismo recurrente en relación con los hallazgos histopatológicos. Se practicó análisis de DNA por citometría de imagen con el fin de estimar el potencial de proliferación del tejido paratiroideo. El estudio comprendió 248 pacientes sometidos a paratiroidectomía entre 1973 y 1991. La frecuencia hiperparatiroidismo recurrente después de paratiroidectomía subtotal fue 4/19 (21.1%), y la tasa de recurrencia dependiente del injerto, después de la resección paratiroidea con autoinjerto en el antebrazo, fue de 2/4 (50%). La frecuencia de recurrencia dependiente del injerto después de paratiroidectomía total con autoinjerto en el antebrazo fue de 16/212 (7.5%). La frecuencia de recurrencia apareció significativamente mayor (p〈0.01) cuando se autotrasplantó tejido hiperplásico nodular (17/68; 25%) que cuando se autotrasplantó tejido hiperplásico difuso (1/105; 〈1%). La totalidad de los 58 especímenes sometidos a análisis de DNA por citometría de imagen exhibió un patrón nuclear diploide. Sin embargo, el número de células dispersas que exhibieron valores citométricos de DNA nuclear por fuera del pico principal del histograma diploide fue significativamente mayor (p〈0.01) en el tejido hiperplásico nodular antes de ser autoinjertado y en el tejido paratiroideo resecado en el curso de la reoperación realizada por recurrencia, que en el tejido hiperplásico difuso. Estos hallazgos clínicos y los resultados del análisis de DNA indican claramente que el tejido paratiroideo hiperplásico nodular posee un mayor grado de potencial de crecimiento, y se llega a la conclusión que para prevenir la recurrencia dependiente del injerto no se debe autoinjertar el tipo nodular de tejido hiperplásico.
    Notes: Abstract In spite of recent therapeutic advances, severe overt secondary hyperparathyroidism due to chronic renal failure necessitates parathyroidectomy and recurrent hyperparathyroidism is one of the most significant problems in this patient population. In the present study, the incidence of recurrent hyperparathyroidism was evaluated in relation to the histopathological features. Image cytometric DNA analysis was performed to estimate the proliferative potential of parathyroid tissue. The study comprised 248 patients who underwent parathyroidectomy from 1973 to 1991. The frequency of recurrent hyperparathyroidism after subtotal parathyroidectomy was 4 (21.1%) of 19 patients, the rate of graft-dependent recurrence after removal of residual parathyroid tissue with forearm autograft was 2 (50%) of 4 patients. The frequency of graft-dependent recurrence after total parathyroidectomy with forearm autograft was 16 (7.5%) of 212 patients. The frequency of recurrence was significantly higher (p〈0.01) when nodular hyperplastic parathyroid tissue was autografted (17 of 68 patients, 25%) than when diffuse-hyperplastic tissue was grafted (1 of 105 patients, 〈1%). All 58 specimens subjected to image cytometric DNA analysis showed a diploid nuclear pattern cytometrically. However, the relative number of scattered cells that displayed cytometric nuclear DNA values outside the main diploid histogram peak was significantly greater (p〈0.01) in nodular hyperplastic tissue before being autografted and in parathyroid tissue removed at re-operation performed for recurrence than in diffuse hyperplastic tissue. These clinical findings and results of DNA analysis clearly indicated that nodular hyperplastic parathyroid tissue has a higher growth potential, and it is concluded that to prevent graft-dependent recurrence, the nodular type of hyperplastic tissue should not be autografted.
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