bluuebunny/crossref_metadata_2025 huggingface dataset:
Recognition of ADP‐ribosylation factor 4‐like by HLA‐A2‐restricted and tumor‐reactive cytotoxic T lymphocytes from patients with brain tumors
Authors: Nonaka Y. , Tsuda N. , Shichijo S. , Ito M. , Maeda Y. , Harada M. , Kamura T. , Shigemori M. , Itoh K.
Published: March 2003

Abstract: Although specific immunotherapy is one candidate treatment of brain tumor, the molecular basis of T‐cell‐mediated recognition of brain tumors has not yet been elucidated. In this study, we tried to identify brain tumor antigens using HLA‐A2‐restricted and tumor‐reactive cytotoxic T lymphocytes (CTLs). As an HLA‐A2‐restricted OK‐CTL line contained CTLs capable of responding to HLA‐A2+ malignant glioma cells, this cell line was used for identification of brain tumor antigens. After screening a cDNA library from brain tumor cells, this CTL line was found to produce interferon (IFN)‐γ when cultured with COS‐7 cells, which were cotransfected with both a cDNA clone (clone 1) and HLA‐A0207 cDNA. Data base searches indicated that the clone 1 was 98% identical to that of the human ADP‐ribosylation factor 4‐like (ARF4L). Two peptides, ARF4L 15–24 and ARF4L 69–77, possessed the ability to induce HLA‐A2‐restricted and tumor‐reactive CTLs from peripheral blood mononuclear cells of patients with brain tumors. Although ARF4L seemed to be ubiquitously expressed at the mRNA level, ARF4L‐reactive CTLs failed to exhibit cytotoxicity against normal lymphoid blasts. These results indicate that these two ARF4L peptides could be targets for immunotherapy of HLA‐A2+ patients with brain tumors.
The CD1 family and T cell recognition of lipid antigens
Authors: Dutronc Y. , Porcelli S.A.
Published: March 2003

Abstract: For many years it was thought that T lymphocytes recognized only peptide antigens presented by MHC class I or class II molecules. Recently, it has become clear that a wide variety of lipids and glycolipids are also targets of the T cell response. This novel form of cell‐mediated immune recognition is mediated by a family of lipid binding and presenting molecules known as CD1. The CD1 proteins represent a small to moderate sized family of β2‐microglobulin‐associated transmembrane proteins that are distantly related to MHC class I and class II molecules. They are conserved in most or all mammals, and control the development and function of T cell populations that participate in innate and adaptive immune responses through the recognition of self and foreign lipid antigens. Here we review the current state of our understanding of the structure and function of CD1 proteins, and the role of CD1‐restricted T cell responses in the immune system.
Analysis of microsatellite polymorphism around the HLA‐B locus in Iranian patients with Behçet's disease
Authors: Mizuki N. , Yabuki K , Ota M. , Katsuyama Y. , Ando H. , Nomura E. , Funakoshi K. , Davatchi F. , Chams H. , Nikbin B. , Ghaderi A.A. , Ohno S. , Inoko H.
Published: March 2003

Abstract: We have previously suggested that in a Japanese population the susceptible locus for Behçet's disease (BD) is HLA‐B51 itself. To confirm this finding in another population, we performed HLA class I typing using the PCR‐SSP method and analyzed eight polymorphic markers distributed within 1100 kb around the HLA‐B gene using automated sequencer and subsequent automated fragment detection by fluorescent‐based technology with the DNA samples of 84 Iranian patients with BD and 87 healthy ethnically matched controls. As a result, three microsatellite alleles (MICA‐A6, MIB‐348, C1‐4‐1‐217) and HLA‐B51 were found to be strongly associated with BD. Of these alleles HLA‐B51 is the most strongly associated allele. There were no alleles that were increased in allele frequency at any microsatellite loci centromeric of MICA or telomeric of HLA‐B51. Therefore, HLA‐B51 was confirmed to be by far the most strongly associated gene with BD in an Iranian population.
Genetic epidemiology of rheumatoid arthritis
Authors: Harney S , Wordsworth B.P.
Published: March 2003

Abstract: Building on the spectacular success of molecular genetics in defining the biological basis of many rare single gene disorders over the past decade, epidemiologists have turned their attention to unravelling the complex genetic mysteries of common disorders, such as rheumatoid arthritis (RA). As a prelude to any such endeavour it is obviously important to establish that there is a significant genetic component to the disease. The classical approaches of twin and other family recurrence risk studies, coupled with prevalence studies in different ethnic and migrant populations, have been used to estimate the environmental and genetic contributions to RA. However, developing a consensus on these estimates has proved difficult, thereby providing an early warning to the unwary investigator that the road to gene discovery in RA is likely to be a rough ride.
Enhanced cleavage of diaminopimelate‐containing isopeptides by leucine aminopeptidase and matrix metalloproteinases in tumors: application to bioadhesive peptides
Authors: Yamazaki Y. , Savva M. , Mokotoff M. , Yamazaki Y. , Oka S. , Kleinman H.K.
Published: March 2003

We prepared (2S,6S)‐Z‐Dpm(Z)(OMe) (4) by protease‐mediated hydrolysis of (R,R/S,S)‐Z‐Dpm(Z)(OMe)‐OMe (3), converted it to (2S,6S)‐Dpm(Z)(OMe) (6) via PCl5 to an NCA intermediate and hydrolysis, protected the amino group with Boc to give (2S,6S)‐Boc‐Dpm(Z)(OMe) (7), which upon ammonolysis of the Me ester afforded (2S,6S)‐Boc‐Dpm(Z)(NH2) (8). Hydrogenolysis of 8 and protection with Fmoc gave (2S,6S)‐Boc‐Dpm(Fmoc)(NH2) (10). Using 10 and SPPS, we prepared three Dpm‐containing peptides and their corresponding Lys peptides. Enzymatic studies with mLAP and cLAP showed that the Leu moiety in Ac‐Gly‐(2S,6S)‐Dpm(Leu)(NH2)‐Ala (14) was hydrolyzed 68‐fold and> 1000‐fold more rapidly, respectively, than that in Ac‐Gly‐Lys(Leu)‐Ala (12). The enhanced rate of Leu formation from 14 compared to 12 was also observed with homogenates of mouse C3 sarcomas. This homogenate also hydrolyzed Ac‐Gly‐(2S,6S)‐Dpm(Ac‐Gly‐Pro‐Gln‐Gly‐Leu)(NH2)‐Ala (16) to Ac‐Gly‐(2S,6S)‐Dpm(NH2)‐Ala (13), Leu and Ac‐Gly‐Pro‐Gln‐Gly (17). This implies the side chain is cleaved first by endopeptidases, such as matrix metalloproteinases (MMPs), and then the remaining Leu is cleaved by LAP‐like exopeptidases. The rate of liberation of 17 from 16 and the corresponding Lys isopeptide, Ac‐Gly‐Lys(Ac‐Gly‐Pro‐Gln‐Gly‐Leu)‐Ala (15), was not significantly different. The rate of formation of 13 was faster from 16 than Ac‐Gly‐Lys‐Ala (11) was from 15. Thus, the entire isopeptide side chain can be removed by the cooperative action of LAP‐like and MMP‐like peptidases present in tumor tissue, which occurs faster in the Dpm peptide 16 than in the Lys peptide 15. The rate of formation of 13 from 16 by lung, liver, and intestine homogenates (from the same C3 tumor‐bearing mice) was comparable to or higher than from the tumor homogenates, but the rate by blood was only 4% the value of the tumor homogenates. Analogs of a bioadhesive fragment from the laminin α1 chain were prepared by replacing the essential Lys with Dpm(NH2) (20) and Dpm(Leu)(NH2) (21). Both Dpm‐containing peptides were active, although considerably weaker than the corresponding Lys peptides 18 and 19, in a cell attachment assay with human fibrosarcoma HT‐1080 cells.
Solid‐phase precipitation and extraction, a new separation process applied to the isolation of synthetic peptides
Authors: Bonner A.G. , Udell L.M. , Creasey W.A. , Duly S.R. , Laursen R.A.
Published: March 2003

Abstract: A new method for separation and purification is described. The process, referred to as solid‐phase precipitation and extraction (SPPE), was developed and applied to postcleavage isolation of synthetic peptides. The technique uses normal approaches of chromatography and solid‐phase extraction sorbents with a precipitation or drying procedure so that the sorbent becomes a support matrix for thin‐film deposition of the compounds of interest. This procedure causes precipitated compounds of interest to be trapped on the large surface area or in the pores of the matrix so that by‐products and impurities can be removed by strong wash solvents. In application to solid‐phase peptide synthesis chemistry, by‐products from the cleavage and deprotection are selectively extracted from the crude sample mixture under mild conditions. In comparison to the ether precipitation method used in peptide chemistry, the SPPE process provides isolated products that are 14–17% (w/w) higher purity.
Somatostatin analogs
Authors: Janecka A. , Zubrzycka M. , Janecki T.
Published: March 2003

Abstract:  Somatostatin is a hypothalamic peptide hormone that inhibits the secretion of growth hormone, glucagon, insulin, gastrin and secretin, and also plays a role in neural transmission. Because of its wide range of possible clinical applications hundreds of somatostatin analogs have been synthesized and bioassayed to date. This review gives a historical perspective, summarizing approximately 30 years of research on somatostatin. The main focus is on the structure−activity relationships and conformational studies of the last generation of somatostatin agonists and their selectivity for five somatostatin receptor subtypes. Achievements in the synthesis of nonpeptide somatostatin analogs, as well as the first somatostatin antagonists, are also discussed. Finally, the use of a cyclic somatostatin scaffold to design ligands for other G‐protein‐coupled receptors, such as opioid and melanocortin receptors, is mentioned.
Conformational comparison of µ‐selective endomorphin‐2 with its C‐terminal free acid in DMSO solution, by 1H NMR spectroscopy and molecular modeling calculation
Authors: In Y. , Minoura K. , Ohishi H. , Ishida T. , Minakata H. , Kamigauchi M. , Sugiura M.
Published: March 2003

Abstract: In order to make clear the structural role of the C‐terminal amide group of endomorphin‐2 (EM2, H‐Tyr‐Pro‐Phe‐Phe‐NH2), an endogenous µ‐receptor ligand, in the biological function, the solution conformations of endomorphin‐2 and its C‐terminal free acid (EM2OH, H‐Tyr‐Pro‐Phe‐Phe‐OH), studied using two‐dimensional 1H NMR measurements and molecular modeling calculations, were compared. Both peptides were in equilibrium between the cis and trans isomers around the Tyr‐Pro ω bond in a population ratio of ≈ 1 : 2. The lack of significant temperature and concentration dependence of NH protons suggested that the NMR spectra reflected the conformational features of the respective molecules themselves. Fifty possible 3D structures for the each isomer were generated by the dynamical simulated annealing method under the proton−proton distance constraints derived from the ROE cross‐peaks. These energy‐minimized conformers, which were all in the φ torsion angles estimated from JNHCαH coupling constants within ± 30°, were then classified in groups one or two according to the folding backbone structures. All trans and cis EM2 conformers adopt an open conformation in which their extended backbone structures are twisted at the Pro2–Phe3 moiety. In contrast, the trans and cis conformers of EM2OH show conformational variation between the ‘bow’‐shaped extended and folded backbone structures, although the cis conformers of its zwitterionic form are refined into the folded structure of the close disposition of C‐ and N‐terminal groups. These results indicate clearly that the substitution of carboxyl group for C‐terminal amide group makes the peptide flexible. The conformational requirement for µ‐receptor activation has been discussed based on the active form proposed for endomorphin‐1 and by comparing conformational features of EM2 and EM2OH.
Synthesis, X‐ray crystallographic structures of thio substituted N‐acetyl N′‐methylamide alanine and evaluation of sp2 sulfur parameters of the CFF91 force field
Authors: Tran T.T. , Burgess A.W. , Treutlein H. , Perich J.
Published: March 2003

Abstract  Acetyl thioalanine N‐methyl (Ac‐Alat‐NHMe) and thioacetyl alanine N‐methyl (Act‐Ala‐NHMe) were synthesized, crystallized and their X‐ray diffraction structures determined for the first time. Both molecules adopted β‐sheet conformations and showed similar hydrogen bonding patterns with one molecular surface forming two oxo hydrogen bonds and the other forming two thio hydrogen bonds. The crystal structure data for the two thioamides provided a validation of the thioamide parameters for the newly derived CFF91 force field because the observed crystal (φ, ψ) angles were situated in the global minimum regions of the theoretical (φ, ψ) map predicted using the parameters. In addition, the parameters were further validated because conformational energy minimization of the crystal structure produced low deviations in unit cell dimensions, bond lengths, bond angles and torsional angles, and a 120‐ps molecular dynamics simulation also gave a low deviation for the most probable N‐H…S=C bond distance.
Engineering the prion protein using chemical synthesis
Authors: Ball H.L. , King D.S. , Cohen F.E. , Prusiner S.B. , Baldwin M.A.
Published: March 2003

Abstract: In recent years, the technology of solid‐phase peptide synthesis (SPPS) has improved to the extent that chemical synthesis of small proteins may be a viable complementary strategy to recombinant expression. We have prepared several modified and wild‐type prion protein (PrP) polypeptides, of up to 112 residues, that demonstrate the flexibility of a chemical approach to protein synthesis. The principal event in prion disease is the conformational change of the normal, α‐helical cellular protein (PrPC) into a β‐sheet‐rich pathogenic isoform (PrPSc). The ability to form PrPSc in transgenic mice is retained by a 106 residue ‘mini‐prion’ (PrP106), with the deletions 23–88 and 141–176. Synthetic PrP106 (sPrP106) and a His‐tagged analog (sPrP106HT) have been prepared successfully using a highly optimized Fmoc chemical methodology involving DCC/HOBt activation and an efficient capping procedure with N‐(2‐chlorobenzyloxycarbonyloxy) succinimide. A single reversed‐phase purification step gave homogeneous protein, in excellent yield. With respect to its conformational and aggregational properties and its response to proteinase digestion, sPrP106 was indistinguishable from its recombinant analog (rPrP106). Certain sequences that proved to be more difficult to synthesize using the Fmoc approach, such as bovine (Bo) PrP(90−200), were successfully prepared using a combination of the highly activated coupling reagent HATU and t‐Boc chemistry. To mimic the glycosylphosphatidyl inositol (GPI) anchor and target sPrP to cholesterol‐rich domains on the cell surface, where the conversion of PrPC is believed to occur, a lipophilic group or biotin, was added to an orthogonally side‐chain‐protected Lys residue at the C‐terminus of sPrP sequences. These groups enabled sPrP to be immobilized on either the cell surface or a streptavidin‐coated ELISA plate, respectively, in an orientation analogous to that of membrane‐bound, GPI‐anchored PrPC. The chemical manipulation of such biologically relevant forms of PrP by the introduction of point mutations or groups that mimic post‐translational modifications should enhance our understanding of the processes that cause prion diseases and may lead to the chemical synthesis of an infectious agent.
Adherence of Peptostreptococcus micros morphotypes to epithelial cells in vitro
Authors: Kremer B. H. A. , Herscheid A. J. , Papaioannou W. , Quirynen M. , Van Steenbergen T. J. M.
Published: March 2003

Peptostreptococcus micros, which is associated with oral and non‐oral mixed anaerobic infections, occurs in three colony morphotypes, the smooth type, the rough type and the smooth variant of the rough type. These types differ in surface structures; the rough type expresses large fibrillar surface appendages, which are absent on the surface of both the smooth and the smooth variant of the rough type. To determine the role of these surface structures in adherence we characterized the adherence of the three morphotypes of P. micros to epithelial cells in vitro. Although all three types adhered well to epithelial cells, adhering numbers of the rough type were significantly lower than those of the smooth and the smooth variant of the rough type. Protease treatment increased the adherence of the rough type to the level of the two other types. The adherence of all three types was reduced more than 85% by treatment with 10 mM sodium periodate. Furthermore, the adherence was pH independent and could not be blocked by incubation with antisera to the bacteria. In addition, we determined the capacity to invade epithelial cells by P. micros. In an acridine orange assay such invasion could not be detected. Our results suggest that the adherence of P. micros to epithelial cells is mediated by periodate‐sensitive extracellular polysaccharides and that the protruding fibril‐like protein surface structures of the rough type have an obstructive effect on the adherence.
Oxygen metabolism by Treponema denticola
Authors: Caldwell C. E. , Marquis R. E.
Published: March 2003

Treponema denticola strains ATCC 35405 and ASLM were found to have moderately active oxygen metabolism and consumed some 0.46 μmol O2/h/mg cell protein in anaerobic growth medium or about ten times this amount in aerobic medium. There appeared to be no differences between the two strains in their oxidative metabolism. The spirochetes showed significant endogenous O2 utilization, which was stimulated only slightly by added glucose or arginine, moderately by glycine, but markedly by casamino acids or brain‐heart infusion broth. O2 metabolism by intact cells was insensitive to cyanide and so did not appear to involve cyanide‐sensitive cytochrome oxidases. Moreover, difference spectra of cell extracts and membranes did not reveal heme profiles. However, the spirochetes did have very active reduced nicotinamide adenine dinucleotide (NADH) oxidase(s) and also contained the protective enzymes NADH peroxidase and superoxide dismutase. Both the oxidase(s) and the peroxidase had rather broad substrate specificities. Either NADH or reduced nicotinamide adenine dinucleotide phosphate could serve as reductant, and the enzymes were active with a variety of oxidants. Enzyme activity in fresh cell extracts was only somewhat stimulated by added flavins, but after frozen storage, the activity became much more activated by flavin adenine nucleotide, and to a lesser extent, by flavin mononucleotide. The enzymes were insensitive to fluoride, which inhibits heme‐based but not flavin‐based oxidases at low pH values. Clearly, these anaerobic spirochetes have significant oxygen metabolism, even at the low levels of O2 measured in periodontal pockets and contain enzymes that offer at least moderate protection against damage by reactive oxygen species.
Isolation and chemical analysis of a lipopolysaccharide from the outer membrane of the oral anaerobic spirochete Treponema pectinovorum
Authors: Walker S. G. , Xu X. , Altman E. , Davis K. J. , Ebersole J. L. , Holt S. C.
Published: March 2003

Isolation of a putative lipopolysaccharide from the surface of the oral treponeme, Treponema pectinovorum, revealed it to contain larger amounts of 3‐deoxy‐
d‐manno‐octulosonic acid compared with other oral Treponema species. This molecule was isolated from the outer membrane of T. pectinovorum and had chemical characteristics of a putative lipopolysaccharide. The yield of lipopolysaccharide was between 0.6% and to 1.1% of the bacterial dry weight. The purified molecule was resistant to the action of proteinases and consisted of both sugars and lipids. 3‐Deoxy‐
d‐manno‐octulosonic acid and hexoses accounted for 6.1–8.7% and 17.6–20.2%, respectively of the dry weight. Carbohydrate compositional analysis revealed the presence of glucose, galactose, 2‐acetamido‐2‐deoxy‐glucose, rhamnose and 6‐deoxy‐talose in the molar ratio of 1.00: 0.96: 0.19: 0.88: 0.98, respectively. No heptose was detected. The fatty acid analysis determined the presence of straight chain, C13:00, C14:00, C15:00 and C17:00 acids, as well as branched chain, C13:00, C14:00 and two species of C15:00, acids. Electrophoretic analysis indicated that the lipopolysaccharide was present as two major species.
Impaired secretory immunity in dystrophic epidermolysis bullosa
Authors: Sweet S. P. , Ballsdon A. E. , Harris J. C. , Roberts G. J. , Challacombe S. J.
Published: March 2003

Dystrophic epidermolysis bullosa is a congenital disorder characterized by blistering of the skin and oral mucosa. This study investigated the hypothesis that children with dystrophic epidermolysis bullosa have impaired oral secretory immunity. Immunoglobulin A (IgA), secretory IgA and IgG concentrations, and IgA and secretory IgA antibody levels to Candida albicans, Lactobacillus casei and Streptococcus mutans were measured in whole saliva from 22 children with dystrophic epidermolysis bullosa and 22 matched controls. Salivary total IgA and total IgG concentrations were significantly raised in dystrophic epidermolysis bullosa due to serum leakage from oral blistering, but the converse was seen with secretory IgA. This suggestion of a mucosal immune defect was supported by decreased secretory IgA antibody responses to all three microorganisms tested. This apparent defect in secretory immunity in dystrophic epidermolysis bullosa may be due to mucosal involvement and damage resulting in impaired antigen sampling in mucosal associated lymphoid tissue or to impaired transport of secretory IgA across the salivary gland mucosa.
Use of the Actinobacillus actinomycetemcomitans leukotoxin promoter to drive expression of the green fluorescent protein in an oral pathogen
Authors: Lippmann J. E. , Froeliger E. H. , Fives‐Taylor P. M.
Published: March 2003

The gene for the green fluorescent protein, gfp, was cloned, under the control of the Actinobacillus actinomycetemcomitans leukotoxin (ltx) promoter, in the A. actinomycetemcomitans shuttle vector, pSU20. A. actinomycetemcomitans containing the ltx‐gfp construct emitted bright green fluorescence in the standard invasion assay using epifluorescence microscopy. These data demonstrate that the green fluorescent protein will be a useful tool for the live analysis of A. actinomycetemcomitans interactions with host cells, and that the ltx promoter can be used to drive the expression of non–A. actinomycetemcomitans genes.
Production of inflammatory cytokines by human gingival fibroblasts stimulated by cell‐surface preparations of Porphyromonas gingivalis
Authors: Imatani T. , Kato T. , Okuda K.
Published: March 2003

Porphyromonas gingivalis is a gram‐negative rod associated with the progression of human periodontal disease. Inflammatory cytokines are believed to be the major pathological mediators in periodontal diseases. We therefore investigated the productions of interleukin‐1β (IL‐1β), interleukin‐6 (IL‐6), interleukin‐8 (IL‐8), and tumor necrosis factor‐α (TNF‐α) in human gingival fibroblasts treated with lipopolysaccharide, polysaccharide and outer‐membrane proteins from P. gingivalis ATCC 53977. Outer‐membrane protein from P. gingivalis enhanced the production of IL‐6 and IL‐8 from the cells of periodontium in vitro as well as lipopolysaccharide did. The IL‐8 production activity of polysaccharide from P. gingivalis was higher than that of other cell‐surface components. The levels of IL‐6 and IL‐8 released from the P. gingivalis lipopolysaccharide‐treated human gingival fibroblasts were lower than those of the same cells treated with lipopolysaccharides from Actinobacillus actinomycetemcomitans or Escherichia coli. Rabbit antisera against either outer‐membrane protein or lipopolysaccharide inhibited the IL‐6 and IL‐8 production derived from human gingival fibroblasts stimulated sonicated supernatants from P. gingivalis. The present study suggests that, in addition to lipopolysaccharide, outer‐membrane protein and polysaccharide of P. gingivalis are also pathological mediators in periodontal diseases.
Oligoclonal accumulations of T‐cell clones in gingivitis and periodontitis lesions
Authors: Itoh H. , Ohsawa Y. , Yoshie H. , Yamazaki K.
Published: March 2003

Gingivitis and periodontitis have distinct clinical and immunopathological characteristics. We have previously demonstrated that T cells infiltrating periodontitis lesions recognize a restricted repertoire of antigens or antigenic epitopes. However, the clonality of T cells in the gingivitis lesion is not known. Therefore, we carried out a clonal analysis of T cells infiltrating gingivitis lesions using combined reverse transcription–polymerase chain reaction and single‐strand conformation polymorphism (SSCP) analysis. As with periodontitis lesions, SSCP analysis demonstrated the emergence of a number of distinct bands suggesting clonal accumulation in the gingivitis lesion. Although the mean number of distinct bands in gingival tissue was significantly higher than that in peripheral blood mononuclear cells, numerical analysis clearly demonstrated that there was no difference in the total number of bands in gingival tissue specimens between the different disease types. Although there were slight variations in the number of distinct bands in each Vβ family, there was no significant difference between gingivitis lesions and periodontitis lesions. These results demonstrate that antigen‐specific T‐cell responses also take place in gingivitis lesions. It remains to be determined, however, what role these antigen‐specific T cells play and what antigens the T cells recognize in the pathogenesis of periodontal disease.
Denaturing gradient gel electrophoresis analysis to study bacterial community structure in pockets of periodontitis patients
Authors: Zijnge V. , Harmsen H. J. M. , Kleinfelder J. W. , Van Der Rest M. E. , Degener J. E. , Welling G. W.
Published: March 2003

Bacteria are involved in the onset and progression of periodontitis. A promising molecular technique, denaturing gradient gel electrophoresis (DGGE), to study microbial population dynamics in the subgingival pocket is presented. Twenty‐three samples were taken from the subgingival pockets of nine patients and six healthy family members. From four periodontitis patients, 12 samples were evaluated before, 1 day after and 3 months after treatment. Part of the 16S rRNA gene of all bacteria was amplified by PCR and separated by DGGE, creating banding patterns representative of the community structure. Shifts in composition and diversity of the microbial population could be determined semiquantitatively, and this showed that treatment resulted in a decrease in the diversity of the population. After 3 months a microbial population 33–47% different from the population before treatment had re‐established. Intense bands representing Exiguobacterium aurantiacum were present in 13 out of 25 samples, indicating that this species may play a role in periodontal disease.
Comparison between serial skin‐prick tests and specific serum immunoglobulin E to mite allergens
Authors: Schuetze Gerd , Gravesande Karin Storm van’s , Sparhold Susanne , Frischer Thomas , Kuehr Joachim
Published: March 2003

Sensitization to dust mite allergens can be determined by means of a skin‐prick test (SPT) or by measurement of specific IgE antibodies in serum (sIgE). In our study, concordance of the results of both methods was analyzed on the basis of reproducible SPT results. Three consecutive SPTs were performed on 138 school children (age 6–8 years) at one‐year intervals. SIgE was determined at the end of a two‐year observation period. Seven common inhalant allergens (Dpt, Df, birch pollens, hazel pollens, grass pollens and cat and dog dander) were analyzed. The majority of subjects with positive SPT reactions to the respective allergen also showed sIgE (Dpt: 82/86; Df: 53/53; cat dander: 31/32; dog dander: 6/9; birch pollens: 29/31; hazel pollens: 22/22; grass pollens: 37/37). A significant correlation between the SPT [weal diameter (P1) or allergen/histamine ratio (P2)] and sIgE was found for Dpt (P1 = 0.004/P2 = 0.016), birch pollens (P1 = 0.002/P2 = 0.0001) and grass pollens (P1 = 0.0005/P2 = 0.0001). There was also a significant correlation between sIgE to Dpt and to either Der p 1 (p = 0.0001) or Der p 2 (p = 0.0001), as well as between sIgE of both major allergens (p = 0.0001). In the analysis of co‐sensitization of Dpt and Df, most subjects sensitized to Dpt were also sensitized to Df (57/91). Children with sIgE to Dpt (n = 87) usually showed sIgE to Df (n = 83). In this study, SPT and sIgE results are concordant and appear equivalent when using reproducible SPTs. Therefore, in the case of a positive Dpt result, additional testing for sensitization to Df can be regarded as redundant when Dpt and Df are the major contributors to the allergen content of house dust.
Effect of pre‐immunization by killed Mycobacterium bovis and vaccae on immunoglobulin E response in ovalbumin‐sensitized newborn mice
Authors: Tükenmez Faruk , Bahçeciler Nerin Nadir , Barlan Işıl Berat , Başaran Müjdat Mustafa
Published: March 2003

A recently advanced hypothesis suggests that decreased exposure to T‐helper (Th) 1‐inducing agents cause Th2‐biased differentiation in response to concomitant allergens. We therefore examined the effect of pre‐immunization with killed Mycobacterium bovis and killed M. vaccae which are known to be very potent inducers of Th1 immune response, on serum IgE response in ovalbumin (OVA)‐sensitized newborn mice. Eighty‐four newborn Balb/c mice were divided into four groups and were immunized intraperitoneally 24 h after birth with 50 µl of 5 × 104 colony‐forming units (c.f.u.) of killed M. bovis in group I (M. bovis group, n = 19), with 25 µl of 2.5 × 108 c.f.u. of killed M. vaccae plus 25 µl of 5 × 104 c.f.u. of killed M. bovis in group II (M. vaccae+ M. bovis group, n = 28) and with 50 µl of only phosphate‐buffered saline (PBS) in group III (no mycobacterial immunization, n = 18). No injection was applied to mice in group IV (control group, n = 19). Starting from 8 weeks of age, all mice except the control group were sensitized with 0.5 ml of 20 mg/ml OVA administered intraperitoneally 7 times every other day. Thirty days after the final injection, all animals except those in the control group were challenged with an aerosol of 2 mg/ml OVA. Forty‐eight hours later, blood was collected from all mice for determination of serum IgE levels. A statistically significant difference was observed in the serum total IgE levels between groups III and IV (p = 0.0099), indicating that the mice were successfully sensitized with OVA. Serum total IgE values of the female mice in M. bovis group were found to be significantly lower than group III (p = 0.009), while no difference was observed in males. Serum total IgE levels of the M. vaccae+M. bovis group were found to be significantly lower than group III both in male and female mice (p < 0.0001 and p = 0.0001, respectively), showing even lower values in females in comparison with controls (p = 0.0092). Pre‐immunization in the newborn period with killed M. bovis alone or in addition to M. vaccae may potentially be helpful in down‐regulating an IgE response.
Atopic diseases in Sami and Norse schoolchildren living in northern Norway
Authors: Selnes A. , Bolle R. , Holt J. , Lund E.
Published: March 2003

Among children in the western world, atopic diseases are a major cause of morbidity. However, several prevalence studies have indicated that the frequency of these diseases displays both geographic and ethnic variations. In 1995, we conducted a questionnaire‐based, cross‐sectional survey in northern Norway. Atopic diseases among 8676 schoolchildren, aged 7–13 years, including 491 children with Sami ethnicity, were studied. The role of ethnicity (Sami/white Caucasian) was determined by comparing the reported atopic disease rate in each of the respective groups. In the areas under investigation (the cumulative incidence, the point prevalence of asthma and allergic rhinoconjunctivitis and the cumulative incidence of atopic dermatitis), the Sami children scored higher than the white Caucasian Norwegian children. The relative risks (RR) in Sami children were: current asthma RR = 2.01 [95% confidence interval (CI) 1.48–2.73]; current allergic rhinoconjunctivitis RR = 1.51 (95% CI 1.14–1.99); lifetime atopic dermatitis RR = 1.39 (95% CI 1.18–1.63). We thus conclude that there is an association between Sami ethnicity and asthma and allergy among schoolchildren in northern Norway.
Diagnosis of food allergy in Finland: Survey of pediatric practices
Authors: Kaila M. , Vanto T. , Valovirta E. , Koivikko A. , Juntunen‐Backman K.
Published: March 2003

Food‐related symptoms are common in the first years of life, and food allergy should be diagnosed using an elimination challenge test. We surveyed Finnish hospital‐based pediatricians using a self‐completion questionnaire to ascertain the current clinical practice: 24 of the 25 pediatricians (representing 24 of 25 hospitals) so approached gave evaluable responses. Food allergies were diagnosed using a clinical elimination challenge test in patients with suspected allergy to cow's milk or cereals (wheat, rye, barley, oats). Of the 24 departments, four reported that they performed challenge in all patients before diagnosis was confirmed, and 14 performed challenge in most patients before diagnosis was confirmed. The duration of the challenge varied from 0.5 to 7 days (median 4 days). A 1‐week challenge was used in eight hospitals. The double‐blind placebo‐controlled challenge was used in seven of the hospitals, and in none routinely. Altogether, 16 of the respondents agreed that there is a need to establish clinical guidelines for the diagnosis of food allergy. In conclusion, despite a long tradition of medical education on the subject of food allergy, practices vary for its diagnosis. There is therefore a requirement for appropriate clinical guidelines.
Asthma in children: Prevalence, treatment, and sensitization
Authors: Hesselmar Bill , Åberg Nils , Åberg Birgitta , Eriksson Bo
Published: March 2003

This study compares the prevalence of asthma and sensitization in children from two Swedish regions with different climates: Göteborg on the southwest coast and Kiruna in the northern inland, north of the Arctic Circle. The 412 children of a population‐based sample, 203 in Göteborg and 209 in Kiruna, were investigated at age 7–8 and 12–13 years. Questionnaire reports and interviews were obtained from all children at 7–8 years of age, and 192 children were skin‐prick tested for common aeroallergens in Göteborg and 205 in Kiruna. At the follow‐up, 5 years later, almost all the children were re‐investigated. The prevalence of asthma, wheeze, and sensitization had increased with increasing age during the follow‐up period. The questionnaire reports revealed that the prevalence of asthma was 8.5% at 12–13 years of age. All children who in the questionnaire reported current asthma, were using asthma medication. The interviews indicated that the prevalence of a clinically significant asthma might be even higher, reaching ≈ 12%. Asthma and wheeze were as common in Göteborg as in Kiruna despite large differences in prevalence of sensitization. Sensitization, and especially sensitization to animals, was far more common in Kiruna than in Göteborg. This study shows that asthma and wheeze are increasingly prevalent even in school age children and that sensitization does not necessarily reflect the prevalence of asthma in a population.
Immunoglobulin G antibodies of children exposed to microorganisms in a water‐damaged school
Authors: Savilahti Risto , Uitti Jukka , Laippala Pekka , Husman Tuula , Reiman Marjut
Published: March 2003

The aim of this study was to determine whether exposure to fungi (molds and yeasts) among children attending a water‐damaged school was reflected by the children's immunoglobulin G (IgG) response to microorganisms typical of water damage and whether the presence of these IgG antibodies was associated with respiratory symptoms and morbidity. The relationships between positive IgG antibodies and atopy, described as elevated allergen‐specific immunoglobulin E (IgE) antibodies, were also examined. The study population consisted of a randomly selected group of exposed children attending a water‐damaged school and a group of unexposed children of the same age. Serum samples for analyses of IgG and IgE antibodies were drawn from the children. The respiratory morbidity, the number of positive IgG antibodies to nine microorganisms indicating water damage, and IgE sensitization to common environmental allergens (Phadiatop®) were studied. The mean number of positive IgG findings was significantly higher among the exposed children. The number of positive IgG antibodies did not correlate with respiratory illnesses or symptoms at the individual level even though the exposed children who had positive IgG antibodies to four or more microorganisms in the total group comparison tended to have higher respiratory morbidity. In the exposed group, a negative correlation was found between the number of positive IgG antibodies and the total value of allergen‐specific IgE antibodies. As among adults exposed to microorganisms at work, IgG antibodies in children seem to be a relevant indicator of exposure to microorganisms in a water‐damaged school on the group level.
Skin‐prick test findings in atopic asthmatic children: A follow‐up study from childhood to puberty
Authors: Kaleyias Joseph , Papaioannou Dimitris , Manoussakis Manolis , Syrigou Ekaterini , Tapratzi Polixeni , Saxoni‐Papageorgiou Photini
Published: March 2003

In a prospective cohort study we investigated the course of allergic sensitization from childhood to puberty in a group of children with atopic asthma. An attempt was made to correlate the findings with the persistence of asthma. A total of 150 children with atopic asthma established at 7 years of age were evaluated when 8–10 years of age. A battery of skin‐prick tests (SPTs) to common environmental allergens, a detailed clinical history for asthma severity classification, and spirometric analyses, were performed. In 127 of these children a re‐evaluation was performed at puberty. A variety of statistical methods were used to analyze the results regarding changes in skin test reactivity to individual aeroallergens and atopic index (degree of atopy), as well as to determine any correlation between these changes and the persistence of asthma in puberty. A wide spectrum of modification in skin reactivity to common environmental allergens was observed, including the complete loss of sensitization to some allergens or the development of a new one to others. Specifically, 34% of asthmatic children sensitive to Dermatophagoides pteronyssinus and 52.7% sensitive to cat lost their sensitivity in puberty, while only 7.5% and 11.1%, respectively, became sensitized (p = 0.03 and p = 0.001, respectively). In contrast, regarding pollen sensitivity, 30.2% and 24% of asthmatic children became sensitive in puberty to olive pollen and grasses mix, respectively, and only 11.7% and 12.5%, respectively, lost their sensitivity to these allergens (p = 0.04). No correlation was shown between the skin test reactivity changes to individual allergens and the persistence of asthma, but a significant correlation was found between atopic index to indoor allergens in childhood and the persistence of asthma at puberty (p = 0.04). Interestingly, multi‐sensitivity to allergens (≥ 4 allergens) in childhood was also found to correlate with the persistence of asthma at puberty [p = 0.05, odds ratio (OR) = 2.65, 95% confidence interval (CI) 1.2–7.2]. Our findings indicate that significant modification of skin reactivity to common environmental allergens in atopic children with asthma in puberty can occur. However, no association between these changes and the persistence of asthma could be demonstrated, although children with indoor allergic sensitization and multi‐reactivity were found to have a higher probability of maintaining their asthma in puberty.
Human malignant osteopetrosis: Pathophysiology, management and the role of bone marrow transplantation
Authors: Fasth Anders , Porras Oscar
Published: March 2003

Abstract: steopetrosis is a heterogeneous group of diseases characterized by lack of osteoclast function. Osteopetrosis is found spontaneously in most mammalian species and many transgenic animals have been created, but so far no animal model has been found that genetically corresponds to human malignant autosomal recessive osteopetrosis. The only curative treatment for malignant osteopetrosis is bone marrow transplantation. A review of the literature and preliminary data from IBMTR shows that infants transplanted with marrow from an HLA‐identical sibling or unrelated volunteer donor have an actuarian five‐year survival with a functioning graft of 50–70%, while those transplanted with a T‐cell‐depleted mismatched marrow have a very poor survival of only about 10%.
Long‐term follow‐up in hematopoieticstem‐cell transplant patients
Authors: Fisher Vicki L.
Published: March 2003

Abstract: Hematopoietic stem‐cell transplantation (HSCT) has increasingly become an accepted treatment for many childhood diseases and disorders. Potential HSCT recipients can be children with hematological malignancies or solid tumors, as well as congenital and acquired disorders. In the past decade, the use of HSCT in the treatment of pediatric disorders has grown exponentially while advances in supportive care have improved survival rate, contributing to a rapidly growing population of transplant survivors. Although numerous similarities can be found between pediatric and adult long‐term HSCT survivors, this article provides a brief overview of the pediatric patients, emphasizing the aspects of surveillance and late effects. Understanding the long‐term complications that can occur after HSCT is important in determining the appropriate evaluations and medical treatment for the patient involved. The goal of this article is to assist caregivers in providing optimal care for long‐term survivors of HSCT. The initial section of this work comprises the three major causes of late effects in HSCT. It will then encompass a system review of the different potential complications that are seen with HSCT.
Hepatic veno‐occlusive disease
Authors: Ribaud Patricia , Gluckman Eliane
Published: March 2003

Abstract:
Cryptosporidial infections after solid organ transplantation in children
Authors: Gerber D. A. , Green M. , Jaffe R. , Greenberg D. , Mazariegos G. , Reyes J.
Published: March 2003

Abstract: The diagnosis and treatment of moderate‐to‐severe diarrhea in solid organ transplant recipients is often a challenge because of the variety of infectious and non‐infectious causes. The morbidity associated with this clinical condition is of particular significance in the pediatric population where malnutrition may lead to poor growth and development. Rarely, Cryptosporidium has been identified as the cause of clinically significant diarrhea in pediatric solid organ transplant patients. A retrospective review identified cases of cryptosporidiosis among the 1160 non‐renal, abdominal organ transplant recipients cared for at the Children’s Hospital of Pittsburgh between 1981 and June 1998. Four cases of clinically significant diarrhea were identified in three liver transplant recipients and one small bowel transplant recipient. Endoscopy and biopsy with histologic confirmation diagnosed three cases; ova and parasitic examination of stool specimens identified the fourth case. Therapy varied among the patients depending on when they had been diagnosed as, over the years, different and newer agents have been indicated for the treatment of cryptosporidiosis. All four patients resolved their infections. Hence, endoscopy and biopsy is recommended for pediatric transplant patients who present with chronic diarrhea of unknown etiology. The patients who may be at a higher risk for cryptosporidial infections include those with an increased immunosuppressive state (i.e. pre‐existing immunodeficiency, malignancy, re‐transplantation, and those receiving higher doses of immunosuppressive therapy). While cryptosporidiosis is a non‐lethal complication, it allows the clinician to gain further insight into the degree of immunosuppression of their patient.
Lipoprotein abnormalities are highly prevalent in pediatric heart transplant recipients
Authors: Chin Clifford , Rosenthal David , Bernstein Daniel
Published: March 2003

Abstract: The role of hyperlipidemia in graft coronary artery disease (GCAD) is controversial although hyper‐triglyceridemia is an independent risk factor. Recent studies show that 3‐hydroxy‐3‐methyl‐glutaryl‐coenzyme A (HMG‐CoA) inhibitors decrease the incidence of GCAD in adults. The incidence of GCAD in pediatric patients is lower than in adults; it is not clear whether age‐related differences in lipid metabolism account for some of this protection. This study was performed to: characterize the lipoprotein profile in children after heart transplantation; demonstrate that total cholesterol (TC) is a poor marker for underlying lipoprotein abnormalities; and to compare lipid abnormalities in patients who had been converted from cyclosporin A (CsA) to tacrolimus. Seventy‐one determinations of fasting lipoprotein profiles were performed in a cohort of 28 children. Each child had at least two determinations on separate occasions. TC, low‐density lipoprotein (LDL), and serum triglyceride (TG) levels were categorized as abnormal if greater than the 75th percentile for age and gender. A high‐density lipoprotein (HDL) level less than the 25th percentile was considered abnormal. Immunosuppression included CsA or tacrolimus, azathioprine, and prednisone. We found that 90% of the patients studied had abnormalities of either TG or HDL. In contrast, LDL tended to be normal when adjusted for age and gender. TC was a poor indicator of any underlying abnormality in TG, LDL, or HDL. In patients converted to tacrolimus, no significant differences were found in the levels of TG, LDL or HDL compared with each patient’s respective values while receiving CsA. Hence, lipoprotein abnormalities among pediatric heart transplant recipients are highly prevalent. TC is a poor screening tool in the evaluation for lipid abnormalities. Lipoprotein profiles remain statistically unchanged after conversion from CsA to tacrolimus.
Allograft rejection in pediatric liver transplantation: Comparison between cadaveric and living related donors
Authors: Toyoki Yoshikazu , Renz John F. , Mudge Christine , Ascher Nancy L. , Roberts John P. , Rosenthal Philip
Published: March 2003

Abstract: The aim of this study was to determine whether living related liver transplantation has an immunological advantage compared with cadaveric liver transplants in children. The records of 100 pediatric primary liver transplant recipients performed between January 1992 and December 1998 at the University of California, San Francisco Medical Center, were reviewed retrospectively. Ten children who died or required a second graft within the first 14 post‐operative days were excluded from this study group. Two children with combined kidney‐liver transplants were also excluded. As a result, the study group included 51 children in the cadaveric liver transplantation (CLT) group and 37 children in the living‐related liver transplantation (LRLT) group. Until 1995, primary immunosuppression consisted of cyclosporin A, azathioprine and steroids. Since 1995, primary immunosuppression consisted of cyclosporin A, mycophenolate mofetil and steroids. Actuarial graft survival rates at 1, 2 and 5 yrs were 90%, 80.9% and 80.9% in the CLT group vs. 94.6%, 91.6% and 78.5% in the LRLT group, respectively (NS). Rejection was diagnosed in 40 of 51 cadaveric first grafts (78.4%) and 25 of 37 living‐related primary grafts (67.6%). Rejection episodes were diagnosed greater than 1 yr post‐transplantation in 11 of 51 cadaveric first grafts (21.6%) and none of 37 living‐related primary grafts (0%) (p < 0.05). LRLT succeeded in reducing the immunosuppressive therapy compared with CLT at 24 months after transplantation (p < 0.05). The overall incidence of rejection and graft survival rate were comparable in CLT and LRLT; however, rejection episodes in LRLT recipients diagnosed greater than 1 yr post‐transplant were significantly fewer than CLT recipients. LRLT have a partial immunological advantage compared with CLT.