bluuebunny/crossref_metadata_2025 huggingface dataset:
Monoclonal antibodies to denatured human ACE (CD 143), broad species specificity, reactivity on paraffin sections, and detection of subtle conformational changes in the C‐terminal domain of ACE
Authors: Balyasnikova I.V. , Metzger R. , Franke F.E. , Danilov S.M.
Published: March 2003

Abstract:Two new mouse monoclonal antibodies (mAbs) were generated to denatured human angiotensin‐converting enzyme (ACE, CD143). The clones 2E2 and 3C5, each of the IgG1 κ chain isotype, detect ACE with high sensitivity, respectively, at 20 ng and 2 ng of protein per lane in Western blotting. They both recognize different epitopes on the C‐domain of ACE located between amino acid residues 740 and 992. In formalin‐fixed and paraffin‐embedded human tissues, immunohistochemistry revealed all known expression sites of ACE, e.g. the epithelial brush borders of proximal kidney tubules, epithelial cells of epididymis, endothelial cells, activated macrophages as well as germ cells during spermatogenesis. In contrast to other mAbs to denatured human ACE, mAbs 2E2 and 3C5 demonstrate cross‐reactivity with a broad spectrum of animal species such as monkey, rat, rabbit, cattle, dog, cat, and guinea pig. In addition, mAb 2E2 recognized mouse ACE in Western blotting and on paraffin sections. Our findings suggest that mAbs 2E2 and 3C5 are useful for identifying even subtle changes in ACE conformation resulting from denaturation. These mAbs are also sensitive tools for the detection of minimal amounts of ACE in biological fluids and tissues using proteomics approaches. Their reactivity in routinely processed tissues of various species may prove useful for correlation of ACE expression in animal models to human diseases.
Biological actions of the epidermal growth factor‐like domain peptides of mouse schwannoma‐derived growth factor and human amphiregulin
Authors: Takenouchi T. , Kadosaka M. , Shin S.Y. , Munekata E.
Published: March 2003

Several members of the epidermal growth factor (EGF) family of growth factors that contain EGF‐like units at their carboxy portion have been isolated and characterized. Schwannoma‐derived growth factor (SDGF) and amphiregulin (AR) are members of this family. SDGF has high sequence homology to AR, and is known to be not only a potent mitogen for astrocytes and fibroblasts but also a neurotrophic factor. We previously confirmed that the synthetic EGF‐like peptides SDGF(38–80) and AR(44–84), corresponding to the EGF‐like domain of mouse SDGF and human AR, respectively, formed similar disulfide bond patterns to that of EGF. In the present study, we further investigated the biological actions of these two EGF‐like peptides on several cultured cell lines. We found that SDGF(38–80) and AR(44–84) have weak mitogenic activity in NIH/3T3 cells and weak binding affinity to the EGF receptor on the surface of A431 cells compared with EGF. However, SDGF(38–80) and EGF induced short neurite outgrowth in PC12 h cells, a subclone of PC12 cells, at 100 nM. In addition, a significant increase in acetylcholinesterase (AChE) activity induced by SDGF(38–80) was observed at a concentration similar range to that of EGF, which is known as a differentiation marker of these cells. The effect of AR(44–84) in PC12 h cells was weaker than those of SDGF(38–80) and EGF, but the AChE activity was significantly increased by the addition of 100 nM AR(44–84), which did not stimulate NIH/3T3 cell growth. These results also suggest that SDGF(38–80) and AR(44–84) may be effective for neuronal differentiation rather than proliferation.
Synthesis of N‐carboxyalkyl and N‐aminoalkyl functionalized dipeptide building units for the assembly of backbone cyclic peptides
Authors: Müller B. , Besser D. , Kleinwächter P. , Arad O. , Reissmann S.
Published: March 2003

Abstract: To improve the assembly of backbone cyclic peptides, N‐functionalized dipeptide building units were synthesized. The corresponding N‐aminoalkyl or N‐carboxyalkyl amino acids were formed by alkylation or reductive alkylation of amino acid benzyl or tert‐butyl esters. In the case of N‐aminoalkyl amino acid derivatives the aldehydes for reductive alkylation were obtained from N,O‐dimethyl hydroxamates of N‐protected amino acids by reduction with LiAlH4. N‐carboxymethyl amino acids were synthesized by alkylation using bromoacetic acid ester and the N‐carboxyethyl amino acids via reductive alkylation using aldehydes derived from formyl Meldrums acid. Removal of the carboxy protecting group leads to free N‐alkyl amino acids of very low solubility in organic solvents, allowing efficient purification by extraction of the crude product. These N‐alkyl amino acids were converted to their tetramethylsilane‐esters by silylation with N,O‐bis‐(trimethylsilyl)acetamide and could thus be used for the coupling with Fmoc‐protected amino acid chlorides or fluorides. To avoid racemization the tert‐butyl esters of N‐alkyl amino acids were coupled with the Fmoc‐amino acid halides in the presence of the weak base collidine. Both theN‐aminoalkyl and N‐carboxyalkyl functionalized dipeptide building units could be obtained in good yield and purity. For peptide assembly on the solid support, the allyl type protection of the branching moiety turned out to be most suitable. The Fmoc‐protected N‐functionalized dipeptide units can be used like any amino acid derivative under the standard conditions for Fmoc‐solid phase synthesis.
Conformational specificity of mini‐αA‐crystallin as a molecular chaperone
Authors: Bhattacharyya J. , Sharma K.K.
Published: March 2003

Abstract: The chaperone activity and biophysical properties of the 19 amino acid peptide DFVIFLDVKHFSPEDLTVK, identified as the functional element in αA‐crystallin and here referred to as mini‐αA‐crystallin, were studied using light scattering and spectroscopic methods after altering its sequence and enantiomerism. The all‐d and all‐l conformers of the peptide do not show marked differences in their chaperone‐like activity against heat‐induced aggregation of alcohol dehydrogenase at 48°C and dithiothreitol‐induced aggregation of insulin. The retro peptide does not show any secondary structure and is also unable to act like a chaperone. Both all‐l and all‐d peptides lose their β‐sheet conformations, hydrophobicity and chaperone‐like activity at temperatures > 50°C. However, upon cooling, a significant portion of those properties was regained, suggesting temperature‐dependent, reversible structural alterations in the peptides under investigation. We propose that both the hydrophobicity and β‐sheet conformation of the functional element of αA‐crystallin are essential for chaperone‐like activity.
Conformation and other biophysical properties of cyclic antimicrobial peptides in aqueous solutions
Authors: Jelokhani‐Niaraki M. , Prenner E.J. , Kay C.M. , McElhaney R.N. , Hodges R.S. , Kondejewski L.H.
Published: March 2003

Abstract:As a step towards understanding the mechanism of the biological activity of cyclic antimicrobial peptides, the biophysical properties and conformations of four membrane‐active cyclic peptide antibiotics, based on gramicidin S (GS), were examined in aqueous environments. These cyclic peptides, GS10 [cyclo(VKLdYP)2], GS12 [cyclo(VKLKdYPKVKLdYP)], GS14 [cyclo(VKLKVdYPLKVKLdYP)] and [d‐Lys]4GS14 [cyclo(VKLdKVdYPLKVKLdYP)] (d‐amino acid residues are denoted bydand are underlined) had different ring sizes of 10, 12 and 14 residues, were different in structure and amphipathicity, and covered a broad spectrum of hemolytic and antimicrobial activities. GS10, GS12 and [d‐Lys]4GS14 were shown to be monomeric in buffer systems with ionic strength biological environments. GS14 was also monomeric at low concentrations, but aggregated at concentrations > 50 µm. The affinity of peptides for self‐assembly and interaction with hydrophobic surfaces was related to their free energy of intermolecular interaction. The effects of variations in salt and organic solvent (trifluoroethanol) concentration and temperature on peptide conformation were also examined. Similar to GS, GS10 proved to have a stable and rather rigid conformation in different environments and over a broad range of temperatures, whereas GS12, GS14 and [d‐Lys]4GS14 had more flexible conformations. Despite its conformational similarity to GS10, GS14 had unique physicochemical properties due to its tendency to aggregate at relatively low concentrations. The biophysical data explain the direct relation between structure, amphipathicity and hydrophobicity of the cyclic peptides and their hemolytic activity. However, this relation with the antimicrobial activity of the peptides is of a more complex nature due to the diversity in membrane structures of microorganisms.
The kinetics of the removal of the N‐methyltrityl (Mtt) group during the synthesis of branched peptides
Authors: Li D. , Elbert D. L.
Published: March 2003

Abstract: The purpose of this short communication is to describe the reaction rate for the removal of the N‐methyltrityl (Mtt) protecting group that is used in solid‐phase peptide synthesis for the production of branched and cyclic peptides. The reaction rate was observed to follow zero‐order kinetics, and we suggest the optimal conditions for the removal of the Mtt group in batchwise synthesis.
Antigenic variation in Porphyromonas gingivalis ribotypes recognized by serum immunoglobulin G of adult periodontitis patients
Authors: Sims T. J. , Ali R. W. , Brockman E. S. , Skaug N. , Page R. C.
Published: March 2003

We obtained clinical isolates of Porphyromonas gingivalis of known ribotype from patients diagnosed with adult periodontitis and used Western blot methodology to evaluate profiles of antigens recognized by IgG in heterologous and homologous patient sera. Our aims were to identify isolates belonging to different serogroups, to learn if serogroup membership is related to ribotype, to assess variation in IgG responses of patients to antigens in homologous and heterologous ribotypes, and to determine the frequency of shared and variable antigens in different biochemical classes recognized across different serogroups and ribotypes. Blots of separation patterns of 28 isolates were developed in sera from patients and bound IgG was quantified by digital image densitometry. The membership of isolates in different serogroups was determined by correlation and hierarchical cluster analysis of isolate whole‐cell IgG binding profiles. Two major isolate clusters, each with two subclusters, were found. Isolates within the same ribotype clustered together in some cases but not others. Homologous isolates ranked high in IgG binding levels relative to those from different patients irrespective of ribotype. Patient subgroups with IgG responses dominant for different ribotypes and serogroups were revealed by correlation analysis. The IgG binding profiles observed for individual protein and proteinase‐resistant antigens across both homologous and heterologous isolates were very dissimilar. Furthermore, the frequency of antigens both shared across all ribotypes and recognized by IgG in patient sera was unexpectedly low. Only two protein antigens (Mr 44 kDa and 27 kDa) were strongly recognized across all ribotypes by different sera. We conclude that the IgG response of patients infected with a particular P. gingivalis serotype or ribotype is directed mainly against antigens that are not shared by other potentially infective clonal types.
The immunogenicity of various peptide antigens inducing cross‐reacting antibodies to a cell surface protein antigen of Streptococcus mutans
Authors: Kato H. , Takeuchi H. , Oishi Y. , Senpuku H. , Shimura N. , Hanada N. , Nisizawa T.
Published: March 2003

A cell surface protein antigen (PAc) of Streptococcus mutans may be involved in the binding of bacteria to the tooth surface, and has long been focused upon as a candidate for a preventive vaccine of dental caries. Previously the peptide PAc (365–377) was shown to raise an antibody in B10.D2 mice which inhibited the binding of salivary components to the PAc molecule. Using this peptide as a unit peptide, two constructs based on multiple antigenic peptides, and several types of tandem repeats of two or three copies were synthesized to estimate the immunogenicity of these peptides. Increase in the immunogenicity was observed for all constructs with the use of an adjuvant compared to the unit peptide alone. However, the tandem repeat constructs generally induced antibody production in the absence of adjuvant, while the multiple antigenic peptide constructs did not induce antibody production under the same condition. Although such a phenomenon may be restricted to this particular peptide sequence, these results may influence the strategy for the design of peptide vaccines.
Physical and genetic map of Streptococcus mutans GS‐5 and localization of five rRNA operons
Authors: Cappiello M. G. , Hantman M. J. , Zuccon F. M. , Peruzzi F. , Amjad M. , Piggot P. J. , Daneo‐Moore L.
Published: March 2003

The physical map of the 2.1 megabase chromosome of Streptococcus mutans GS‐5 has been refined by including all ApaI and SmaI fragments of 5 kbp or greater, and by positioning the fragments generated by the endonuclease I‐CeuI. Sixty‐three new genetic loci have been added to the map, so that it now contains 90 loci. The new loci include those for 35 cloned streptococcal genes of established function and for 23 S. mutans genes of putative function. In addition, five rrn operons were identified and placed on the map of the chromosome. The presence of a SmaI site in each of the rrn operons allowed the direction of transcription of each operon to be deduced. The orientation of the rrn loci indicates that their transcription is directed away from a small region of the chromosome, identifying a possible region for the initiation of chromosome replication.
Expression of β‐defensin genes by human salivary glands
Authors: Bonass W. A. , High A. S. , Owen P. J. , Devine D. A.
Published: March 2003

This study investigated expression of genes encoding human β‐defensins 1 and 2 by human salivary glands. Tissues from surgical biopsies were collected fresh onto ice and stored in liquid nitrogen. Total RNA was extracted using Trizol reagent and human β‐defensin messenger RNA detected by reverse transcriptase polymerase chain reaction amplification. DNA sequencing of amplified fragments, after ligation into pGEM‐T Easy vector and transformation of competent Escherichia coli, confirmed identities of cloned fragments. Human β‐defensin 1 messenger RNA was detected in all 25 samples that generated amplifiable cDNA, as assessed using abl‐specific primers. Three of 13 submandibular gland samples (two normal, one chronically inflamed), and 2 of 2 minor salivary gland samples (one normal, one chronically inflamed) expressed human β‐defensin 2 messenger RNA. All six parotid gland samples studied were negative for human β‐defensin 2 messenger RNA. Thus, human β‐defensin 1 gene expression occurred in all human major and minor salivary glands studied, whereas human β‐defensin 2 expression occurred only in a small number of gland samples.
Adherence of Porphyromonas gingivalis to gingival epithelial cells: modulation of bacterial protein expression
Authors: Agnani G. , Tricot‐Doleux S. , Du L. , Bonnaure‐Mallet M.
Published: March 2003

The protein profiles of Porphyromonas gingivalis (ATCC 33277 and W83) bound to KB gingival epithelial cells were analyzed by SDS‐PAGE and immunoblotting. We found that a 51‐kDa component was formed in bacteria that adhered to the KB cells, whereas 26‐ to 29‐kDa bands were less intensive, in contrast to the protein profile of free bacteria. P. gingivalis ATCC 33277 incubated with protease‐treated KB cells retained the profile of free bacteria. These results demonstrate the specificity of bacterial recognition of eukaryotic membrane components.
Glucose metabolism by Prevotella intermedia and Prevotella nigrescens
Authors: Takahashi N. , Yamada T.
Published: March 2003

Glucose metabolism by Prevotella intermedia and Prevotella nigrescens were investigated. Glucose increased the anaerobic growth of these bacteria and promoted the accumulation of intracellular polysaccharide. The polysaccharide was confirmed to be glycogen‐like glucan by the absorption spectrum of iodine‐polysaccharide complex and the sugar composition. The washed cells consumed glucose anaerobically and converted a part of glucose into the metabolic end‐products acetate, formate and succinate. The rest of glucose was confirmed to be accumulated as intracellular polysaccharide. The cells grown in the presence of glucose produced acetate, formate and succinate without exogenous glucose along with the consumption of intracellular polysaccharide. The metabolism of glucose and intracellular polysaccharide required bicarbonate. Prevotella cells had hexokinase and a set of the usual enzymes of the Embden‐Meyerhof‐Parnas pathway except that phosphofructokinase was pyrophosphate‐dependent. A series of enzymes, including phosphoenolpyruvate carboxylase, phosphoenolpyruvate carboxykinase, malate dehydrogenase, fumarase and fumarate reductase, was found for succinate formation. Another series of enzymes, pyruvate oxidoreductase, pyruvate formate‐lyase, phosphotransacetylase and acetate kinase was found for acetate and formate formation. Glucose 1,6‐bisphosphate‐dependent phosphoglucomutase and fructose 1,6‐bisphosphate‐activated UDP‐glucose pyrophosphorylase were detected for glycogen synthesis, while glycogen phosphorylase was for glycogen degradation. The capacity of intracellular polysaccharide formation in addition to glucose fermentation could be advantageous for survival in the supragingival area as well as in the subgingival area.
Detection of Treponema denticola in endodontic infections by 16S rRNA gene‐directed polymerase chain reaction
Authors: Siqueira J. F. , Rôças I. N. , Favieri A. , Santos K. R. N.
Published: March 2003

A 16S rDNA‐based polymerase chain reaction (PCR) method was used to detect the occurrence of Treponema denticola in root canal infections. Samples were collected from 21 single‐root teeth having carious lesions, necrotic pulps and radiographic evidences of periradicular bone loss. DNA extracted from the samples was amplified using the PCR assay, which yielded specific fragment of T. denticola 16S rDNA. T. denticola was detected in 11 of 21 cases (52.4%), regardless of the presence or absence of symptoms. Since this spirochete was found in a relatively high percentage of the endodontic infections examined and because it is a pathogenic microorganism involved in periodontal diseases, there are reasons to believe that T. denticola can also participate in the pathogenesis of periradicular lesions of endodontic origin.
Subgingival colonization by Porphyromonas gingivalis
Authors: Lamont R. J. , Jenkinson H. F.
Published: March 2003

Porphyromonas gingivalis, a gram‐negative anaerobe, is a major causative agent in the initiation and progression of severe forms of periodontal disease. In order to cause periodontal disease, P. gingivalis must colonize the subgingival region, a process that involves several distinct steps and multiple gene products. The organism must first navigate within the oral fluids in order to reach the hard or soft tissues of the mouth. Retention and growth of bacteria on these surfaces is facilitated by a repertoire of adhesins including fimbriae, hemagglutinins and proteinases. Once established subgingivally, P. gingivalis cells participate in intercellular communication networks with other oral prokaryotic cells and with eukaryotic cells. The establishment of these multiple interactive interfaces can lead to biofilm formation, invasion of root dentin and internalization within gingival epithelial cells. The resulting bacterial and host cellular locations, products and fate contribute to the success of P. gingivalis in colonizing the periodontal region.
Clonal variability among oral Candida albicans assessed by allozyme electrophoresis analysis
Authors: Mata A. L. , Rosa R. T. , Rosa E. A. R. , Gonçalves R. B. , Höfling J. F.
Published: March 2003

A total of 49 Candida albicans strains were isolated from the saliva of 11 healthy children in Piracicaba, Brazil and were analyzed according to their alloenzymatic patterns. Among eight loci assayed, seven were polymorphic and allowed to determine allelic and genotype frequencies, in order to establish the genetic variables for this fungal population. Some children showed just one genetic type, whereas other harbored two or more clones of such yeast, in a multiclonal manner of colonization by C. albicans.
Antifungal activity of histatin‐5 against non‐albicans Candida species
Authors: Nikawa H. , Jin C. , Fukushima H. , Makihira S. , Hamada T.
Published: March 2003

Fungicidal effects of histatin‐5 against 26 oral isolates belonging to 5 non‐albicans Candida species were examined. Fifty μM of histatin‐5 killed more than 95% of Candida tropicalis and Candida guilliermondii isolates and more than 90% of Candida parapsilosis and Candida krusei. However, Candida glabrata was less sensitive to the peptide (mean 62.9%). Our results, taken together, demonstrated that histatin‐5 possessed the fungicidal activity against Candida species other than C. glabrata.
Effects of apigenin and tt‐farnesol on glucosyltransferase activity, biofilm viability and caries development in rats
Authors: Koo H. , Pearson S. K. , Scott‐Anne K. , Abranches J. , Cury J. A. , Rosalen P. L. , Park Y. K. , Marquis R. E. , Bowen W. H.
Published: March 2003

Propolis, a resinous hive product secreted by Apis mellifera bees, has been shown to reduce the incidence of dental caries in rats. Several compounds, mainly polyphenolics, have been identified in propolis. Apigenin and tt‐farnesol demonstrated biological activity against mutans streptococci. We determined here their effects, alone or in combination, on glucosyltransferase activity, biofilm viability, and development of caries in rats. Sprague‐Dawley rats were infected with Streptococcus sobrinus 6715 and treated topically twice daily as follows: (1) tt‐farnesol, (2) apigenin, (3) vehicle control, (4) fluoride, (5) apigenin +tt‐farnesol, and (6) chlorhexidine. Apigenin (1.33 mM) inhibited the activity of glucosyltransferases in solution (90–95%) and on the surface of saliva‐coated hydroxyapatite beads (35–58%); it was devoid of antibacterial activity. tt‐Farnesol (1.33 mM) showed modest antibacterial activity against biofilms and its effects on glucosyltransferases were minimal. The incidence of smooth‐surface caries was significantly reduced by apigenin +tt‐farnesol (60%), fluoride (70%), and chlorhexidine (72%) treatments compared to control (P < 0.05).
Antigen‐presenting cells in human periodontal disease tissues
Authors: Gemmell E. , Carter C. L. , Hart D. N. J. , Drysdale K. E. , Seymour G. J.
Published: March 2003

T cells are present in the inflammatory infiltrates of periodontal disease lesions and require antigen presentation by antigen‐presenting cells (APCs). While it is still not known whether Th1 or Th2 cells predominate in these lesions, it has been reported that different APCs may induce activation of different T‐cell subsets. An immunoperoxidase technique was used to investigate the presence of CD1a+, CMRF‐44+, CMRF‐58+ and CD83+ dendritic cells, CD14+ macrophages or dendritic cell precursors and CD19+ B cells in gingival biopsies from 21 healthy or gingivitis and 25 periodontitis subjects. The samples were divided into three groups according to the size of infiltrate (group 1, small infiltrates; group 2, medium infiltrates; group 3, extensive infiltrates). The presence of numerous CD1a+ Langerhans cells was noted in the epithelium with no differences between the healthy/gingivitis and periodontitis groups. The percentage of CD83+ dendritic cells in the infiltrates was higher than the percentage of CD1a+, CMRF‐44+ or CMRF‐58+ dendritic cells. Endothelial cells positive for CD83 were found predominantly in areas adjacent to infiltrating cells, CD83+ dendritic cells being noted in the region of CD83+ endothelium. The percentage of CD14+ cells in the inflammatory infiltrates was similar to that of CD83+ dendritic cells. B cells were the predominant APC in group 2 and 3 tissues. The percentage of B cells in group 3 periodontitis lesions was increased in comparison with group 1 periodontitis tissues and also in comparison with group 3 healthy/gingivitis sections. Functional studies are required to determine the roles of different APC subpopulations in periodontal disease.
Indoor environment, atopy and the risk of asthma in children in New Zealand
Authors: Wickens Kristin , Pearce Neil , Siebers Robert , Ellis Ian , Patchett Kathryn , Sawyer Gillian , Stone Louise , Tohill Sarah , Kennedy Jo , Slater Tania , Lewis Simon , Fitzharris Penny , Crane Julian
Published: March 2003

The objective of this study was to examine the relationship between the indoor environment, atopy and asthma in 7–9‐year‐old children. Cases and controls were randomly selected from children who participated in the International Study of Asthma and Allergies in Childhood (ISAAC) in Wellington, New Zealand. Cases were children with a previous diagnosis of asthma and current medication use (n = 233) and controls were children with no history of wheezing and no diagnosis of asthma (n = 241). Information was recorded about the indoor environment during the first year of life and currently. Dust was sampled from floors and beds and Der p 1 and Fel d 1 measured using enzyme‐linked immunosorbent assays. Skin‐prick tests were performed with eight common allergens. Sensitization to Dermatophagoides farinae (OR = 3.19; 95% CI 1.74–5.84), Dermatophagoides pteronyssinus (OR = 2.06; 95% CI 1.16–3.65) and cat (OR = 3.89; 95% CI 1.06–14.30) were independently associated with current asthma. The use of a sheepskin in the first year of life (OR = 1.91; 95% CI 1.11–3.33) was also independently associated with current asthma but current Der p 1 levels showed no association with current asthma. Exposures in early life may be more important than current exposures in determining asthma at age 7–9 years. Prospective studies are needed in New Zealand to determine the relative importance of early life exposures to Der p 1 and other risk factors for asthma.
Infants colonized with enterotoxin‐producing staphylococci at 3 months display a decreased frequency of interferon‐γ‐producing CD45RO lymphocytes upon stimulation with staphylococcal enterotoxin A at birth but not at 6 months of age
Authors: Schauer Uwe , Thurau Anja , Seitz Miriam , Nowak Lars , Petri Holger , Leinhaas Claudia , Rieger Christian
Published: March 2003

The aim of the study was to elucidate the relationship between the cytokine response to staphylococcal enterotoxin A (SEA) at birth and subsequent staphylococcal colonization in the first months of life. In a cohort of 45 newborns, cord blood lymphocytes were stimulated with SEA (10 ng/ml) in vitro, re‐stimulated with PMA (phorbol myristate acetate) and ionomycin at day 3 and assessed for CD45RO expression and cytokine generation by flow cytometry. The infants were classified into three groups according to nasal staphylococcal colonization and enterotoxin generation at 3 months: There were 16 infants with either no colonization or non‐enterotoxin‐producing staphylococci, 16 infants with enterotoxins B, C, D and E, and 13 infants colonized with SEA‐producing staphylococci. At birth, the group without subsequent colonization displayed a significantly higher frequency of CD45RO‐positive interferon‐γ‐producing cells (1.7%; range 0.0–9.3%) in comparison to the SEA‐positive group (0.1%; range 0.0–0.4%) and also to the group positive for other enterotoxins (0.50%; range 0.0–2.5%). Comparable but less pronounced results were found for interleukin‐5 but not for interleukins 2 and 4. At 6 months, no differences in cytokine generation were detected between the three groups. The results provide evidence that a non‐specific immunologic immaturity at birth is a risk factor for early bacterial colonization. Furthermore, it is remarkable that this immaturity is similar to that seen in infants destined to be atopic with respect to disequilibrium of interferon‐γ to interleukin‐4 generation. Thus the link between early staphylococcal colonization and subsequent atopy requires further investigation.
Regional variation in the diagnosis of asthma among preschool‐age children
Authors: Csonka Péter , Mertsola Jussi , Kaila Minna , Ashorn Per
Published: March 2003

To study potential regional variation in asthma diagnostics, we retrospectively analyzed the management of 800 acutely wheezing preschool‐age children in two university clinics in Finland. Multivariate modelling indicated that the place of treatment was a strong independent predictor of asthma diagnosis during acute symptoms. Similar regional preferences in diagnostic activity may exist in other countries.
Expression of and responses to CD2 and CD3 in 18‐month‐old children with and without atopic dermatitis
Authors: Jenmalm Maria C. , Aniansson‐zdolsek Helena , Holt Patrick G. , Björkstén Bengt
Published: March 2003

We hypothesize that atopy is associated with a reduced T‐cell function early in life and an imbalance in cytokine production. The purpose of this study was to investigate the expression of and responses to CD2 and CD3 in children who did or did not develop atopic dermatitis early in life. The expression of CD2 and CD3 was analyzed by flow cytometry, and proliferation of CD2 and CD3 was studied by 3H‐thymidine incorporation in phytohaemagglutinin (PHA)‐ and anti‐CD3‐stimulated peripheral blood mononuclear cells (PBMC) of 18‐month‐old children, 25 with and 29 without atopic dermatitis. Exogenous interleukin (IL)‐2 was added to compensate for possible functional differences in accessory cells. Anti‐CD3‐induced secretion of IL‐4, IL‐5, IL‐6, IL‐10, IL‐13, and interferon‐γ (IFN‐γ) was analyzed by enzyme‐linked immunosorbent assay (ELISA). Atopy was associated with a low proportion of CD2+ lymphocytes. Responsiveness to PHA, which activates lymphocytes partly via the sheep erythrocyte receptor, CD2, was reduced in the allergic children. The anti‐CD3‐induced proliferation declined more rapidly with antibody dilution in the allergic than in the non‐allergic children. Atopic dermatitis was associated with high levels of anti‐CD3‐stimulated IL‐5 secretion. The IL‐4/IL‐10 and IL‐4/IFN‐γ ratios were higher in children with elevated total immunoglobulin E (IgE) levels. Skin prick test‐negative children with eczema produced higher levels of IL‐10 than skin prick test‐positive children. In conclusion, atopic children have a reduced T‐cell function. Atopic dermatitis is associated with increased IL‐5 production, while high total IgE levels are associated with high IL‐4/IFN‐γ and IL‐4/IL‐10 ratios.
Expression of CD64 as a potential marker of neonatal sepsis
Authors: Layseca‐Espinosa Esther , Pérez‐González Luis F. , Torres‐Montes Abraham , Baranda Lourdes , De La Fuente Hortensia , Rosenstein Yvonne , González‐Amaro Roberto
Published: March 2003

The aim of this study was to identify a novel immunological indicator useful for the early diagnosis (through a rapid and single determination) of neonatal sepsis (NS). Peripheral blood samples were taken from 63 neonates, who were classified into four groups: proven NS (n = 17); clinical NS (n = 14); disease without infection (n = 17); and healthy newborns (n = 15). Neutrophil expression of CD64, CD43, CD44, CD50, CD62L and Mac‐1, and plasma levels of interleukin (IL)‐1β, IL‐6, tumor necrosis factor‐α (TNF‐α) and soluble L‐selectin (sCD62L), were determined. Expression of CD64 was significantly enhanced in the group with proven sepsis and clinical NS compared to newborns without infection (p < 0.05). Eight newborns with proven or clinical sepsis, but only one with disease without infection, showed an increased percentage of CD64+ cells (diagnostic specificity = 96.8%). No significant differences were found in the expression of the other leucocyte differentiation antigens studied. As previously described, TNF‐α and IL‐6 levels were significantly elevated in newborns with proven or clinical sepsis compared to neonates without infection (p < 0.05). Our results suggest that, through a single determination, the enhanced expression of CD64 is a highly specific indicator of NS, although its diagnostic sensitivity is low (25.8%). In contrast, we found that plasma levels of IL‐1β and sCD62L, as well as the expression of Mac‐1, CD43, CD44, CD50, and CD62L, do not appear to be useful for the diagnosis of NS.
Mycophenolate mofetil in pediatric renal transplantation
Authors: Benfield Mark R. , Symons , Jordan M. , Bynon , Steve , Eckhoff , Devin , Herrin , John , Harmon , William , Kohaut Edward
Published: March 2003

Benfield MR, Symons JM, Bynon S, Echkoff D, Herrin J, Harmon WE, Kohaut EC. Mycophenolate mofetil in pediatric transplantation. Pediatr Transplantation 1999: 3: 33–37. © Munksgaard, 1999 Abstract: The use of mycophenolate mofetil (MMF) in adult renal transplantation has been associated with significantly decreased incidence of acute rejection. However, limited data are available for children after renal transplantation. A total of 67 patients undergoing renal transplantation at the University of Alabama at Birmingham, AL, USA and Children’s Hospital of Boston, MA, USA were enrolled into the Cooperative Clinical Trials in Pediatric Transplantation randomized controlled trial of induction with OKT3® vs. i.v. cyclosporin A (CsA) at the time of transplantation. The first 31 patients entered were begun on azathioprine (AZA), 2 mg/kg on the first post‐operative day. The subsequent 36 patients were begun on MMF, 1000 mg/m2/d. Other maintenance immunosuppression included oral CsA and Prednisone. Biopsy confirmation was obtained for all suspected rejection episodes. Glomerular filtration rate (GFR) was calculated using the Schwartz formula. Data were analyzed using Kaplan–Meier survival curves and compared using log‐rank tests. At the time of analysis, 52 patients (mean age 10.1 ± 5 yr) had completed at least 12 months and 15 others had completed at least 6 months of follow‐up post‐transplantation. Of these, there were 39 male/28 female; 48 white/15 black/4 other; 49 living donor/18 cadaver donor. There were no significant differences in the incidence of rejection episodes, number of rejection episodes, the GFR at 6 and 12 months, allograft, or patient survival between patients receiving MMF vs. AZA. We could demonstrate no significant differences in these outcomes in based on sex, race or induction therapy, leading to the conclusion that pediatric patients treated under a consistent protocol in two institutions have no improvement in short‐term allograft outcome with the addition of MMF therapy.
Ten‐year experience of unrelated bone marrow donor transplants in children with malignant and non‐malignant conditions
Authors: Trigg Michael E. , Rumelhart , Stephen L. , Miller , Jennifer , Goldman , Frederick D. , Holida , Myrl D. , Lee Nita F.
Published: March 2003

Abstract: Children who require a marrow transplant may receive such hematopoietic cells from one of many sources. This study reviews the experience of one center with 58 children who received marrow from unrelated donors over a 10‐year period. These children had a variety of malignant and non‐malignant diseases. During that time period, only three of these children had failed to meet engraftment criteria. All donor marrow specimens were T‐lymphocyte depleted using an antibody/complement methodology. No difference was demonstrated in outcome between donors who were perfectly HLA‐DR–DNA matched versus those who were only partially matched. The increased size of various marrow donor registries has increased the number of potential donors available for these patients. The lack of a requirement for perfect matching means that there is an ever‐increasing number of donors available. No graft‐versus‐host disease (GvHD) or grade III‐IV GvHD was associated with a poorer outcome. Stable, long‐term engraftment with minimal morbidity has been demonstrated in these children as evidenced by stability of survival curves by two years after marrow transplant.
Changing cyclosporin A formulation: An analysis in paediatric cardiac transplant recipients
Authors: Gennery A. R. , O’Sullivan J. J. , Hasan A. , Hamilton J. R. L. , Dark J. H.
Published: March 2003

Abstract: Cyclosporin A is the primary immunosuppressive agent used in cardiac transplantation to maintain chronic immunosuppression. Absorption may be erratic and major side‐effects include nephrotoxicity. A recently introduced formulation (Neoral®) improves absorption in cystic fibrosis heart–lung transplant recipients, but episodes of rejection have been reported on conversion from the oil‐based formulation. A retrospective analysis of 21 paediatric cardiac transplant recipients who had been converted from the oil‐based formulation to the micro‐emulsified formulation was performed. No clinical rejection episodes occurred following conversion. There was a significant reduction in dose following conversion (p < 0.001). The mean trough level was less on the new formulation (p = 0.04), but there was no difference in coefficient of variation or standard deviation. Mean glomerular filtration rate (GFR) was significantly less, but there was no difference in the rate of change of GFR on the new formulation. Erratic absorption and deteriorating renal function remain as significant problems in paediatric cardiac transplant recipients despite conversion to the new formulation.
Can we make some general recommendations regarding immune recovery after hematopoietic reconstitution preceded by ablative therapy?
Authors: Trigg Michael E.
Published: March 2003

Abstract: Children undergoing marrow transplantation have compromised immune systems for variable periods of time after transplant. A number of methods have been utilized to assess recovery of the immune system. Four recommendations are made in regards to the testing that should be done, when the testing should be done, when immunizations should be provided and when antibiotic and other infection prophylaxis can be reduced
Graft‐vs.‐malignancy with allogeneic blood stem cell transplantation: A potential primary treatment modality
Authors: Champlin Richard , Khouri Issa , Giralt Sergio
Published: March 2003

Abstract: The high‐dose chemotherapy and radiation typically used as the preparative regimen for bone marrow transplantation produces considerable morbidity and mortality. An alternative strategy is to utilize a low‐dose, non‐myeloablative, preparative regimen designed not to eradicate the malignancy, but to provide sufficient immunosuppression to achieve engraftment of an allogeneic hematopoietic graft and allow subsequent development of a graft‐vs.‐malignancy effect. We studied this approach in patients who were ineligible for standard myeloablative preparative regimens because of advanced age or comorbidities and demonstrated that purine analog (fludarabine or 2‐CDA) containing non‐myeloablative chemotherapy allows engraftment of HLA‐compatible hematopoietic progenitor cells, and extended remissions were observed in approximately half of chemosensitive patients with recurrent AML or CML. Patients with CLL or lymphoma have been effectively treated using a non‐myeloablative regimen of fludarabine/cyclophosphamide of fludarabine, cytarabine, cisplatin. This chemotherapy is known to be non‐myeloablative and mixed chimerism was anticipated. All patients with engraftment have responded and 67% have achieved complete remission. Maximal responses are slow to develop and occur gradually over a period of several months to one year. Long‐term efficacy must still be determined and controlled trials are necessary comparing this approach with alternative therapies as well as standard transplantation regimens.
T‐cell recovery following marrow transplant: Experience with delayed lymphocyte infusions to accelerate immune recovery or treat infectious problems
Authors: Smith F. O. , Thomson B. G.
Published: March 2003

Abstract: All forms of hematopoietic stem‐cell transplantation are complicated by delayed immune reconstitution, which results in an increased risk of infectious complications and relapse of disease. Donor lymphocyte infusions have been used in an attempt to enhance immune recovery and for the prevention and treatment of specific infections following transplantation. While there is little data to support the use of donor lymphocytes for the enhancement of general immune function post‐transplant, unselected and virus‐specific donor T cells may have efficacy for the prophylaxis and treatment of infections and disease caused by Epstein–Barr virus (EBV) and cytomegalovirus (CMV). While donor lymphocyte infusions may cause significant morbidity and mortality, they are a novel and potentially powerful approach for the treatment of frequently fatal post‐transplant infectious complications.
Post‐transplant epididymitis and orchitis following Listeria monocytogenes septicaemia
Authors: Von Schnakenburg Christian , Hinrichs Bernd , Fuchs Joerg , Kardorff Ruediger
Published: March 2003

Abstract: We report the occurrence of epididymitis and orchitis 1 week after the onset of Listeriosis in an 11‐month‐old boy receiving an orthotopic liver transplantation for biliary atresia. Immunologic implications of Listeria monocytogenes‐induced testicular inflammation are discussed, and the potential role of immunosuppression with tacrolimus is also discussed.
Tailoring immunosuppressive therapy for renal transplant recipients
Authors: Vanrenterghem Yves F. CH.
Published: March 2003

Abstract:During the past decade several new potent immunosuppressive agents with different modes of action and different side‐effect profiles have become available. Nowadays immunosuppression after renal transplantation is no longer one single regimen applicable to all patients. In the selection of the optimal immunosuppressive protocol, individual drug‐related toxicity, recipient‐related risk factors as well as donor organ characteristics have to be taken into account. This article will give an overview of the most recently developed immunosuppressive agents available for clinical use. Their individual mode of action and their different efficacy and safety profile will be described as basis for selection of each of these drugs in an attempt to tailor the optimal therapeutic regimen for the individual patient both in terms of short‐term and long‐term outcome.
Varicella vaccination in pediatric kidney transplant candidates
Authors: Furth Susan L. , Fivush Barbara A.
Published: March 2003

Abstract: Existing studies support the use of varicella vaccine in a two‐dose regimen in patients with renal disease prior to transplantation. Levels of anti‐varicella zoster virus antibody should be monitored on a regular basis after immunization, and where a loss of a previously protective antibody titer occurs, a third booster dose should be considered pretransplant. Further data need to be collected regarding the use of the vaccine in seronegative patients who have already undergone transplantation.
Lectins of members of the Amaryllidaceae are encoded by multigene families which show extensive homology
Authors: Van Damme Els J. M. , Goldstein Irwin J. , Vercammen Guy , Vuylsteke Joan , Peumans Willy J.
Published: March 2003

Screening of complementary DNA (cDNA) libraries constructed from poly(A) rich RNA isolated from young developing ovaries of Narcissus cv. Fortune and Hippeastrum hybr. resulted in the isolation of multiple lectin cDNA clones. Although the lectin clones show a high degree of overall homology within their coding region, they clearly differ from each other at some positions in their nucleotide sequence and deduced amino acid sequence. Moreover, since some differences in the sequence result in different charges along the lectin polypeptides, the different cDNA clones encode lectins with different isoelectric points, which explains the occurrence of multiple lectin isoforms in Narcissus and Hippeastrum at the molecular level. Furthermore, Southern blot analysis of genomic DNA isolated from Narcissus and Hippeastrum yielded numerous restriction fragments hybridizing with lectin cDNA probes leaving no doubt that the Narcissus and Hippeastrum lectins, like the Galanthus nivalis lectin, are encoded by families of closely related lectin genes.
Diurnal and ultradian rhythms in K+ uptake by Trifolium repens under natural light patterns: Evidence for segmentation at different root temperatures
Authors: Macduff J. H. , Dhanoa M. S.
Published: March 2003

The patterns and variability in the diurnal and ultradian fluctuations in net uptake of K+ by 85‐day‐old plants of white clover (Trifolium repens L. cv. Huia) were analysed during 21 days of vegetative growth under natural light conditions, at constant root temperaures of 13 and 25°C. Plants were established in flowing solution culture, and hourly rates of K+ uptake from solutions containing 2.5‐6.5 µM K+ were automatically measured without physical disturbance. Total daily solar radiation ranged between 1.3 and 12.5 MJ m−2 day−1 over the 21 days of measurements. Substantial diurnal changes in rates of K+ uptake, both absolutely and proportionately, were recorded on all but the day of lowest radiation (day 19), during which there was a period of net efflux of K+ from the root system into the flowing nutrient solutions at both temperatures. Typically, uptake rates increased severalfold during the first half of the light period, followed by several hours of relative stability during the afternoon; then a rapid decline throughout the evening and first half of the night, and finally gradual decline during the second half of the night. The data from a sequence of typical days were normalised and indicated that the pattern could be resolved into 4 distinct and linear phases, successively: (1) rapid acceleration, (2) stable uptake, (3) rapid deceleration, and (4) slow deceleration. The full data set (days 1‐21) of hourly K+ uptake rates was Fourier‐transformed and subjected to time‐series analysis to determine whether there were significant ultradian rhythms in K+ uptake. Periodograms indicated a strong 24 h periodicity for uptake at both root temperatures. In addition, there were strong ultradian periodicities of 7 h for uptake at 13°C and 4 h for uptake at 25°C. The results are interpreted in terms of the possible mechanisms responsible for diurnal fluctuations in nutrient uptake.
Regulation of leaf growth of grass by blue light
Authors: Gautier Hélène , Varlet‐Grancher Claude
Published: March 2003

Changes in light quality occur naturally within a canopy when a plant grows from unshaded to shaded conditions, and the reverse occurs after a cut that reduces shading. These changes in light quality could be responsible for the variation in leaf elongation and appearance rates of grasses. The role of blue light in leaf growth was investigated in tall fescue (Festuca arundinacea Schreb.) and perennial ryegrass (Lolium perenne L.). Leaf length was measured daily following a decrease or an increase in blue light to evaluate effects on duration of leaf growth, leaf elongation and the rate of leaf appearance rate. A reduction in blue light increased sheath length by 8 to 14% and lamina length by 6 to 12% for both species. These increases could be reversed by enrichment of blue light. With low blue light treatment, final leaf length was increased due to a greater leaf elongation rate. In tall fescue, but not in perennial ryegrass, this effect was coupled with a greater phyllochron and a longer duration of leaf elongation. Development of successive leaves on a tall fescue tiller were co‐ordinated. A decrease in blue light increased the duration of elongation in the oldest growing leaf and also delayed the appearance of a new leaf, maintaining this co‐ordination. We conclude that final leaf size and phyllochron for tall fescue can be significantly modified by blue light. Perennial ryegrass appeared less responsive, except for displaying longer sheaths and laminae in low blue light, as also occurred for tall fescue. We hypothesize that leaf length could be regulated by the quality of the light reaching the growing region itself.