bluuebunny/crossref_metadata_2025 huggingface dataset:
Sampling of periodontal pathogens by paper points: evaluation of basic parameters
Authors: Hartroth B. , Seyfahrt I. , Conrads G.
Published: March 2003

Paper points are widely established for the collection of subgingival plaque or other oral samples to analyze the microbiota, especially the presence of periodontal pathogenic bacteria such as Actinobacillus actinomycetemcomitans or Porphyromonas gingivalis. In contrast to the high frequency of usage of paper points in oral sampling, very few data are available about the parameters influencing the sampling process. Therefore, in four different in vitro experiments (6–9 repeats), we inoculated paper points with standardized suspensions (2×109 colony‐forming units/ml) of A. actinomycetemcomitans and P. gingivalis to test the influence of the origin (kind) of paper point (“manufacturer”), size (according to the International Organization for Standardization ISO 25–80), sampling time (5 to 60 s), and elution time (5 to 60 s). Sampled bacteria were detected and (semi‐) quantified using 16S rRNA/DNA‐directed oligonucleotide probes. The bacterial load was categorized and calculations performed with index values ranging between 0 (below the detection limit of 102 to 103 colony‐forming units) and 9 (>106 colony‐forming units). We found statistically significant differences in the efficiency of bacterial sampling between the 5 paper point manufacturers tested, expressed in a mean bacterial index between 4.4 and 7.8. ISO 45 Paper points were statistically proven to work most efficiently. According to our results, a sampling time of 60 s seems to be optimum. However, shorter times between 5 and 30 s did not significantly reduce the sampling efficiency. We found an interval of 20 s best to elute bacteria from the paper points by vortexing. The evaluation of basic parameters for subgingival plaque sampling by paper points might help to optimize microbiologically based diagnostics in periodontal diseases.
Protective and destructive immune reactions in apical periodontitis
Authors: Márton I. J. , Kiss C.
Published: March 2003

Apical periodontitis is initiated primarily by the mixed microflora of infected root canals. Continuous flow of bacteria and their products through the apical foramen induces influx, activation and coordinated interaction of immune‐inflammatory cells within the periapical area. Successful mobilization of host defense mechanisms prevents abundant extraradicular bacterial invasion. However, anti‐infective effector mechanisms are not restricted to killing the invading microorganisms but also destroy normal tissue components and induce bone absorption, resulting ultimately in the loss of the affected teeth. Moreover, autocrine and paracrine loops of stimulation may lead to the perpetuation of the local inflammatory lesion and may also alter the function of remote tissues and organs. This review attempts to summarize current knowledge about the pathogenic mechanism of apical periodontitis, focusing on the formation of a special granulation tissue that effectively fights bacteria originated from the infected pulp chamber and, by exerting this protective function, also contributes to harmful local and distant events. The dynamic equilibrium between defensive and destructive mechanisms may provide a pathobiological basis for better understanding of clinical signs and symptoms of various forms of apical periodontitis lesions and influence treatment strategy and practice.
Salivary immunoglobulin A antibodies to gp41 in human immunodeficiency virus–seropositive patients: lack of correlation with disease progression
Authors: Toro M. J. , Escalona L. , Chaiyarit P. , Warner N. A. , Gregory R. L.
Published: March 2003

Mucous membranes are the main route of transmission of human immunodeficiency virus (HIV). Interestingly, some viral inhibitory activities have been found in saliva. The purpose of this study was to determine the level of salivary immunoglobulin A (IgA) antibodies to gp41 in HIV+ patients at various disease stages to identify whether gp41was able to induce vigorous humoral responses. Unstimulated saliva samples were obtained from three groups of subjects (n=37): group A (HIV−), group B (HIV+, CD4+ <200/mm3), and group C (HIV+, CD4+ >200/mm3). IgA antibody levels to purified gp41 were determined by enzyme‐linked immunosorbent assay (ELISA). Western blot analyses were performed using HIV+ saliva to confirm IgA reactivity to gp41. ELISA demonstrated that HIV+ subjects had higher IgA antibody to gp41 than HIV− individuals. No significant differences were noted between HIV+, CD4+ <200/mm3 and CD4+ >200/mm3 subjects. High (81.25%) IgA reactivity to gp41 was demonstrated by Western blotting of saliva from all HIV+ individuals. In conclusion, gp41 responses are important in the HIV disease process, as indicated by the high IgA levels and gp41 reactivity in saliva of HIV+ patients.
Interaction of Porphyromonas gingivalis with KB cells: comparison of different clinical isolates
Authors: Eick S. , Rödel J. , Einax J. W. , Pfister W.
Published: March 2003

The ability of different Porphyromonas gingivalis strains (15 clinical isolates and ATCC 33277) to attach to and invade KB cells, in relation to other properties such as release of interleukin (IL)‐6 and IL‐8, cytotoxicity, proteolytic activity and types of fimbriae genes present, was examined. A hierarchical cluster analysis based on adherence and internalization resulted in four groups. Eight of the 15 clinical isolates belonged to a cluster group whose adherence and internalization were about 10% those of the ATCC strain. A negative correlation between lysine‐specific protease activity and adherence was found. In all cases the released concentrations of IL‐6 and IL‐8 were very low. Only one strain was found to be cytotoxic to KB cells. Principal components analysis demonstrated correlations between adherence, internalization and autoaggregation. Most strains had fimA type I and II, type I being associated with elastase‐like activity. The ability of P. gingivalis to invade epithelial cells may be a key factor for maintaining periodontal disease.
Cloning and expression of a Porphyromonas gingivalis gene for protoporphyrinogen oxidase by complementation of a hemG mutant of Escherichia coli
Authors: Kusaba A. , Ansai T. , Akifusa S. , Nakahigashi K. , Taketani S. , Inokuchi H. , Takehara T.
Published: March 2003

Porphyromonas gingivalis, a bacterium implicated in periodontal pathogenesis, has a growth requirement for iron protoporphyrin IX. By complementation with a P. gingivalis 381 chromosomal DNA library, we were able to isolate a clone that enhanced the poor growth of a hemG mutant of Escherichia coli. The DNA sequence analysis of this clone revealed three open reading frames (ORFs). ORF3 encoded a protein of 466 amino acids with a calculated molecular weight of 51 695 Da. The deduced amino acid sequence of the ORF3 gene had significant similarity to sequences of protoporphyrinogen oxidase (PPO) from Myxococcus xanthus (30% identical residues). When the ORF3 gene was overexpressed in E. coli, the extract had much higher PPO activity than a control extract, and this activity was inhibited by acifluorfen, a specific inhibitor of PPO. Thus, ORF3 was named PgHemG. Furthermore, several porphyrin‐related genes, including hemD, hemN and hemH, were identified in the data bases on the websites available on‐line. We postulated that a porphyrin biosynthetic pathway to heme from preuroporphyrin may be conserved in P. gingivalis.
Cytokines in the oral mucosa of mice infected with Candida albicans
Authors: Farah C. S. , Gotjamanos T. , Seymour G. J. , Ashman R. B.
Published: March 2003

Oropharyngeal candidiasis is associated with defects in cell‐mediated immunity, and is commonly seen in immunocompromised patients. We have previously shown that T‐cell‐deficient BALB/c nude (nu/nu) mice are extremely susceptible to oropharyngeal candidiasis, and that recovery from a chronic infection is dependent on CD4 T lymphocytes. In this study we describe the local tissue cytokine profile in lymphocyte‐reconstituted immunodeficient mice and their euthymic counterparts. Mice were infected orally with 108 cells of the yeast Candida albicans, and oral tissues sampled on days 0, 4, 8, and 14. Nude mice were reconstituted with 3 × 107 naïve lymphocytes following oral inoculation. Interleukin (IL)‐6, interferon (IFN)‐γ and tumour necrosis factor (TNF)‐α were identified in the oral tissues of infected euthymic mice recovering from oral infection, as well as naïve controls. TNF‐α was identified in nude oral tissue on days 4 and 8, but only after lymphocyte reconstitution. No IL‐2, IL‐4 or IL‐10 was detected in either euthymic or athymic mice at any time‐point throughout the experiment. This study confirms the functional activity of T lymphocytes in reconstituted nude mice, and suggests that TNF‐α may be an important mediator in the recovery from oropharyngeal candidiasis.
Study on the Prevention of Allergy in Children in Europe (SPACE): Allergic sensitization in children at 1 year of age in a controlled trial of allergen avoidance from birth
Authors: Halmerbauer Gerhard , Gartner Christian , Schierl Michael , Arshad Hassan , Dean Tara , Koller Dieter Y. , Karmaus Wilfried , Kuehr Joachim , Forster Johannes , Urbanek Radvan , Frischer Thomas ,   the SPACE collaborative study team
Published: March 2003

Several studies have demonstrated that early intervention may modulate the natural course of atopic disease. Our objective was to prevent sensitization to house‐dust mite and food allergens, as well as the development of atopic symptoms during infancy, by the combination of an educational package and the use of mite allergen‐impermeable mattress encasings. A multicentre European, population‐based, randomized, controlled study of children at increased atopic risk [Study on the Prevention of Allergy in Children in Europe (SPACE)] was performed in five countries (Austria, Germany, Greece, the UK, and Lithuania), and included three cohorts – schoolchildren, toddlers, and newborns. We report on the newborn cohort. A total of 696 newborns were included from Austria, the UK, and Germany. Inclusion criteria were: a positive history of parental allergy; and a positive skin‐prick test or specific immunoglobulin E (IgE) (IgE ≥ 1.43 kU/L) against at least one out of a panel of common aeroallergens in one or both parents. At 1 year of age, the overall sensitization rate against the tested allergens [dust‐mite allergens: Dermatophagoides pteronyssinus and Dermatophagoides farinae (Der p and Der f)] and food allergens (egg, milk) in the prophylactic group was 6.21% vs. 10.67% in the control group. The prevalence of sensitization against Der p was 1.86% in the prophylactic group vs. 5% in the control group. In conclusion, we were able to demonstrate, in a group of newborns at risk for atopic diseases, that the sensitization rate to a panel of aero‐ and food allergens could be effectively decreased through the use of impermeable mattress encasings and the implementation of easy‐to‐perform preventive measures.
The National Asthma Campaign Manchester Asthma and Allergy Study
Authors: Custovic Adnan , Simpson Bridget M. , Murray Clare S. , Lowe Lesley , Woodcock Ashley ,   on behalf of the NAC Manchester Asthma and Allergy Study Group
Published: March 2003

The NACManchester Asthma and Allergy Study is a prospective study of the development of asthma and allergies in childhood. The subjects (995 children at age 3 years) were recruited in utero by screening parents in the antenatal clinic using skin prick testing and a questionnaire regarding allergic diseases. Children were assigned to risk groups according to parental atopic status (high risk, both parents atopic; medium risk, one parent atopic; low risk, neither parent atopic). A subgroup of those at high risk (with no pets in the home) was randomized to stringent environmental control (allergen impermeable covers for the parental and infant bed, hot washing of bedding weekly, HEPA vacuum cleaner, hard floor for the nursery), and the remainder followed a normal regime. The children have been followed prospectively. The environmental influences are very clearly defined. Measurements of environmental exposures include levels of house dust mite; cat and dog allergens during pregnancy and early life; pet ownership and exposure; childcare arrangements; number of siblings; vaccination uptake; thorough dietary questionnaire; and endotoxin exposure. Further unique objective outcome in the cohort is the assessment of lung function in preschool children using specific airways resistance, which at age 3 years clearly reflects both genetic and environmental influences.
Prediction of asthma in children at 6 and 12 years of age: Perth infant asthma follow‐up study
Authors: Le Souëf Peter
Published: March 2003

Longitudinal studies, starting in infancy, provide an opportunity to assess risk factors prior to long‐term exposure to environmental factors. The status of the infant soon after birth is determined by genetic background and factors related to in utero influences. As a result of recent advances in molecular biology, the genetic component can now be evaluated.
Perinatal risk factors for recurrent wheeze in early life
Authors: Lødrup Carlsen Karin C. , Carlsen Kai‐håkon , Nafstad Per , Bakketeig Leiv
Published: March 2003

The possible value of tidal flow volume (TFV) loops measured at birth in relation to the risk of developing recurrent or persistent bronchial obstruction within two years of life was assessed. TFV loops were measured at a mean age of 2.7 days in 802 neonates enrolled in the ‘Environment and Childhood Asthma’ (ECA) study in Oslo. Of these, 77 children developed recurrent or persistent bronchial obstruction (cases) and were included in a nested case–control study within the ECA study; 88 controls (the child born closest in time to the case), with no history of bronchial obstruction in the first two years of life, were also included. Information on socio‐economic factors, parental atopic diseases and parental smoking habits during the pregnancy was collected from a questionnaire completed by the parents in the maternity ward, and cord blood IgE (CB‐IgE) was determined as part of routine sampling in the delivery ward. Mean tPTEF/tE (time to reach peak flow to total expiratory time) was slightly lower in cases (0.31; 95% CI 0.28–0.34) than in controls (0.33; 0.31–0.35) (difference not significant), whereas geometric mean CB‐IgE was significantly higher among cases (0.39; 0.30–0.52) than controls (0.27; 0.23–0.33). No significant differences between cases and controls were found for respiratory rate, peak tidal expiratory flow or expiratory volume. However, the odds ratio for developing recurrent or persistent bronchial obstruction was 3.5 (1.1–11.6) if tPTEF/tE was < 0.20 and 4.1 (1.1–14.5) with maternal daily smoking during the pregnancy, after adjusting for age, weight, sex, CB‐IgE, parental atopy, maternal education and family income. The TFV parameter tPTEF/tE < 0.20 measured within the first week of life as well as maternal daily smoking during pregnancy are significant, independent risk factors for developing recurrent or persistent bronchial obstruction within the first two years of life.
Identification of immunogenic maize proteins in a casein hydrolysate formula
Authors: Frisner Henrik , Rosendal Annette , Barkholt Vibeke
Published: March 2003

Cow's milk‐based formulas used for infants with cow's milk allergy are based on hydrolyzed proteins. The formulas that are successful in preventing allergic responses are extensively hydrolyzed. Nevertheless, reactions to such formulas are occasionally reported, and protein material of higher molecular weight than expected has been detected by binding immunoglobulin E (IgE) from patients’ sera. This paper presents the identification of high‐molecular‐weight material in the extensively hydrolyzed casein formula, Nutramigen. The material was concentrated by simple centrifugation. The proteins in the pellet were separated by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE), and protein‐containing bands were analyzed by protein sequencing after electroblotting. The proteins were identified as maize zeins, which are water‐insoluble proteins of apparent Mr 20 000 and 23 000, presumably originating from the maize starch in Nutramigen. Rabbits immunized with this formula developed antibodies against zeins but not against milk proteins. The maize zeins are probably identical to the recently reported components in Nutramigen (1), detected by binding of IgE from milk allergic patients, but not correlated to clinical allergic reactivity. The clinical relevance of maize proteins in Nutramigen remains to be established.
Low levels of interferon‐γ in nasal fluid accompany raised levels of T‐helper 2 cytokines in children with ongoing allergic rhinitis
Authors: Benson Mikael , Strannegård Inga‐lisa , Wennergren Göran , Strannegård Örjan
Published: March 2003

The T‐helper 2 (Th2) cytokines interleukin‐(IL‐) 4, IL‐5, IL‐6, IL‐10 and the Th1 cytokine IFN‐γ and their associations with eosinophils, eosinophilic cationic protein (ECP) and immunoglobulin (Ig) E were studied in nasal lavage fluid from 60 school children with allergic seasonal rhinitis and 36 nonatopic healthy controls, before and during the pollen season. Eosinophil differential counts and IgE increased significantly in the patients during the pollen season. The eosinophil differential counts, ECP and IgE were all significantly higher during the season than in specimens simultaneously obtained from the nonatopic controls. Before season, the levels of ECP and IgE, but not eosinophils, were significantly higher in the patients than in the controls. During the season the nasal lavage fluid levels of IFN‐γ were significantly lower and the IL‐4/IFN‐γ quotients significantly higher in the allergic than in the control children. In the allergic children, but not in the controls, the nasal fluid levels of the Th2 cytokines IL‐4, IL‐5 and IL‐10 increased during the season, and together with IL‐6, were correlated with the differential counts of eosinophils, and with the levels of ECP and IgE. These findings are compatible with the hypothesis that a deficient release of the Th1 cytokine IFN‐γ plays an important role in the pathogenesis of allergic inflammation. Regardless of whether the defective IFN‐γ secretion is primary or a consequence of suppression by other cytokines, it will in the atopic subjects enhance the release of Th2 cytokines, which in turn will facilitate the development of allergic inflammation.
Hyperimmunoglobulin‐E syndrome with recurrent infection: A review of current opinion and treatment
Authors: Erlewyn‐Lajeunesse Michel D. S.
Published: March 2003

Hyperimmunoglobulin E (hyper‐IgE) syndrome with recurrent infection is a rare idiopathic primary immunodeficiency. It consists of a severe dermatitis with recurrent abscess formation, respiratory tract infections and very high titres of serum immunoglobulin E (IgE). Hyper‐IgE syndrome is also associated with skeletal abnormalities. Variability of presentation makes it easy to confuse the diagnosis with that of severe atopy or other rare immunodeficiencies. The aim of this article is to review the literature in order to consider the clinical findings, etiology and treatment of this syndrome.
Airway nitric oxide in infants with acute wheezy bronchitis
Authors: Ratjen Felix , Kavuk Ilker , Gärtig Sebastian , Wiesemann Hans‐Georg , Grasemann Hartmut
Published: March 2003

Concentrations of nitric oxide (NO) in exhaled air are increased in children and adults with asthma, and NO measurements are used as a non‐invasive marker to monitor airway inflammation in these patients. To define the role of NO in infants with acute wheezy bronchitis, we measured nasal and end‐tidal NO concentrations in 17 infants with acute virus‐associated wheezy bronchitis, in 22 term infants without respiratory disease, and in nine premature infants. Nasal NO measurements were performed with an olive placed in the infant's nose; end‐tidal NO concentrations were assessed during tidal breathing through a snugly fitting face mask. Both end‐tidal NO concentrations and nasal NO concentrations were reduced in infants with acute wheezy bronchitis. There were no differences in NO concentrations between term infants and premature infants. Measurements by both techniques were highly reproducible, as assessed by repeated measurements three times daily on three consecutive days in eight premature infants. Reduced airway NO concentrations in infants with virus‐associated acute wheezy bronchitis are in contrast to findings in adults where both upper and lower airway NO levels are increased in patients with asthma. Whether this reflects a different inflammatory reaction to upper airway infections in acutely wheezy infants or pathophysiologic differences in airway response remains to be determined.
Critical evaluation of prognostic factors in childhood asthma
Authors: Van Bever H. P. , Desager K. N. , Hagendorens M.
Published: March 2003

Current knowledge of the natural history of asthma is improving through the establishment of a more precise definition of asthma linked with information from a number of large‐scale longitudinal studies. Risk factors for the development of childhood asthma are now more clearly understood. They include gender, atopic status, genetic and familial factors, respiratory infections, and outdoor and indoor pollution (1). In the present review two types of asthma and their prognosis will be discussed:

Asthma in preschool children and its risk factors for evolution towards persistent childhood asthma.

Asthma in older children and its risk factors for evolution towards adult asthma.
Exposure to pets and atopic dermatitis during the first two years of life. A cohort study
Authors: Zirngibl Angelika , Franke Kaethe , Gehring Ulrike , Von Berg Andrea , Berdel Dietrich , Bauer Carl Peter , Reinhardt Dietrich , Wichmann H.‐Erich , Heinrich Joachim ,   for the GINI study group
Published: March 2003

The aim of this study was to assess the association between keeping pets in early childhood and the occurrence of atopic dermatitis in an ongoing birth cohort followed up to the age of 2 years. We analyzed data of 4578 children in the intervention and observation part of an ongoing cohort study. The children were recruited at birth in the two study regions Wesel and Munich between January 1996 and June 1998. Information on atopic diseases and pet ownership was obtained by questionnaire at the child's first and second birthday. The logistic regression model showed a negative association between ‘keeping any pet’ and in particular ‘keeping dogs’ in the 1st year of life and the development of atopic dermatitis in the 1st and the 2nd years of life. The protective effects remained statistically significant after adjusting for several possible confounding variables (1st yearany pet OR 0.71, 95% CI [0.55;0.92], 1st yeardog OR 0.62, 95% CI [0.39;0.98], 2nd yearany pet OR 0.74, 95% CI [0.57;0.97], 2nd yeardog OR 0.63, 95% CI [0.40;0.98]). Ownership of small furred pets (hamster, rabbit and guinea pig) also showed a borderline protective effect for the 1st year. We assume an association between keeping pets and undefined environmental factor(s) that contribute protectively to the development of atopic dermatitis in early life, presumably by effects on the maturation of the immune system.
High‐dose therapy with stem cell rescue for pediatric solid tumors: Rationale and results
Authors: Chen Allen R.
Published: March 2003

Abstract: Metastatic and recurrent pediatric solid tumors usually respond to chemotherapy but are likely to recur. Because of steep dose–response relationships, HDT requiring hematopoietic rescue may improve outcome. This strategy has recently been shown to be effective for metastatic neuroblastoma. Metastatic Ewing’s sarcoma appears to be a closely analogous situation, and promising phase II studies suggest that a definitive trial of efficacy would be appropriate. Phase I or II trials remain appropriate and are needed to define further the efficacy of HDT for most other poor prognosis pediatric solid tumors.
Use of calcium‐channel blockers in pediatric renal transplant recipients
Authors: Silverstein Douglas M. , Palmer JoAnn , Baluarte H. Jorge , Brass Caroline , Conley Susan B. , Polinsky Martin S.
Published: March 2003

Abstract: Hypertension (HTN) is a significant problem in pediatric renal transplant (TP) recipients, predisposing the individuals to the development of cardiovascular disease and graft dysfunction. Calcium channel blockers (CCB) are considered excellent agents to treat post‐TP HTN. We compared the efficacy and adverse effects of the two most commonly prescribed CCBs in our pediatric renal TP population: nifedipine (ProcardiaR, or P) and amlodipine (NorvascR, or N). All patients (n = 24) had been started on a CCB for systolic (SBP) and/or diastolic BP (DBP) > 95%. There were no other changes in adjunctive antihypertensive medications or doses during the cross‐over period. Post‐TP, pretreatment (pretx) SBP was 137.6 ± 10.9 mmHg. The post‐treatment SBP were (in mmHg): 128.5 ± 11.9 (all patients, n = 24) (p = 0.009 vs. pretx); 126.4 ± 10.0 (P alone, n = 15) (p = 0.007 vs. pretx); 132.8 ± 14.4 (P + other antihypertensive(s), n = 9) (p = 0.331, NS vs. pretx). The post‐TP, pretreatment DBP was 88.2 ± 11.1 mmHg. The post‐treatment DBP were (in mmHg): 78.5 ± 6.9 (all patients, n = 24) (p = 0.03 vs. pretx); 77.2 ± 7.4 (P alone, n = 15) (p = 0.008 vs. pretx); 80.7 ± 6.1(P + other antihypertensive(s), n = 9) (p = 0.063, NS vs. pretx). P and N were equally effective in reducing SBP (p = 0.843, NS) and DBP (p = 0.612, NS). Cyclosporin A (CyA) dose (p = 0.81) and trough levels (p = 0.19) were similar in P‐ and N‐treated patients. Calculated GFR was virtually identical in P‐ and N‐treated patients (p = 0.89). Patients (or parents of) reported a higher incidence of various side‐effects while receiving P, including headache, flushing, dizziness and leg cramps. Furthermore, 22/24 (91.7%) reported some degree of gingival hyperplasia during treatment with P, and all these patients reported a stabilization or reduction of hypertrophy after the switch from P to N. We conclude that CCBs (N) are efficacious drugs for the purpose of BP control and renal protection in pediatric renal TP recipients.
The management of Epstein–Barr virus associated post‐transplant lymphoproliferative disorders in pediatric solid‐organ transplant recipients
Authors: Green Michael , Michaels Marian G. , Webber Steven A. , Rowe David , Reyes Jorge
Published: March 2003

Abstract: Despite a growing understanding of the pathogenesis and spectrum of Epstein–Barr virus (EBV) and EBV‐associated post‐transplant lymphoproliferative disease (PTLD) in organ transplant recipients, the optimal management of this complication remains controversial. The absence of comparative data evaluating potential therapeutic strategies explains the lack of uniformly accepted guidelines for the management of PTLD. The purpose of this review is to provide an overview of potential therapies and offer a set of guidelines for the management of EBV‐associated PTLD in children.
Unusual evolution of an Epstein–Barr virus‐associated leiomyosarcoma occurring after liver transplantation
Authors: Brichard Bénédicte , Smets Françoise , Sokal Etienne , Clapuyt Philippe , Vermylen Christiane , Cornu Guy , Rahier Jacques , Otte Jean Bernard
Published: March 2003

Abstract: We report the case of a child who developed, 2 yr after orthotopic liver transplantation (OLTx) for biliary atresia, a multi‐focal hepatic tumor with lymphonodular metastases, identified as an Epstein–Barr virus (EBV)‐associated leiomyosarcoma. Chemotherapy was given without tumor response. Subsequently, slow growth of the tumor was observed. Immunosuppression was tapered and stopped 9 yr after transplantation. At the present time, 12 yr after the discovery of the first hepatic lesions, the patient is alive and completely symptom‐free, the abdominal masses are stable, and liver function tests are completely normal. Smooth muscle tumors are increasingly recognized in children with various immunodeficiencies occurring after organ transplantation. This unusual evolution of a clinically aggressive tumor into a stable disease after restoration of immunity confirms that the immune status of the patient is a crucial factor.
Reversal of protein‐losing enteropathy in a child with Fontan circulation is correlated with central venous pressure after heart transplantation
Authors: Holmgren D. , Berggren H. , Wåhlander H. , Hallberg M. , Myrdal U.
Published: March 2003

Abstract: We report on the reversal of protein‐losing enteropathy (PLE) after heart transplantation (HTx) in a 10‐yr‐old boy with Fontan circulation, previously treated unsuccessfully with heparin for several months. The protein loss continued immediately after the Tx. During the following month, however, a gradual decrease in protein loss was observed, which correlated with a decrease in the inferior vena cava (IVC) pressure. The patient is doing well with a normal serum albumin level and a normal IVC pressure, 2 yr after Tx.
Ureterocystoplasty (bladder augmentation) with a solitary kidney
Authors: Kurzrock Eric A. ,   Lavjay Butani, V. Perez Richard
Published: March 2003

Abstract: Children who develop end‐stage renal disease (ESRD) as a result of obstructive uropathies require evaluation and treatment of associated bladder dysfunction to ensure a good outcome following renal transplantation. Bladder dynamics can often be optimized medically, although surgical intervention is occasionally necessary. For those patients who require bladder augmentation, the use of a dilated native ureter (ureterocystoplasty) is preferred to the more commonly used intestine or stomach (enterocystoplasty), which carry a higher risk of complications. Unfortunately, most patients do not have a suitable anatomy for ureterocystoplasty and, by necessity, intestine or stomach has to be utilized. Herein, we describe the successful application of ureterocystoplasty in the presence of ESRD and a solitary kidney prior to renal transplantation. We believe that owing to the many advantages of native urothelium, every effort should be made to use ureter and avoid the use of intestine.
Application of MARS artificial liver support as bridging therapy before split liver retransplantation in a 15‐month‐old child
Authors: Hommann M. , Kasakow L. B. , Geoghegan J. , Kornberg A. , Schotte U. , Gruhn B. , Fuchs D. , Hermann J. , Zintl F. , Scheele J.
Published: March 2003

Abstract: Molecular Adsorbent Recirculating System (MARS) is a blood‐filtering system designed to provide biological artificial liver support. We describe its use in a small child to illustrate its effectiveness and practicality in this age group. A 15‐month‐old male underwent split liver transplantation for acute liver failure following bone marrow transplantation. After development of graft dysfunction we instituted MARS‐dialysis. MARS therapy led to a dramatic fall in serum bilirubin and transaminases. Liver synthetic function was not affected. This was accompanied by a stabilization of the patients clinical condition until repeat split liver transplantation was performed 2 weeks after the first graft. MARS‐dialysis is practical in the small child. In this case, it did not provide definitive treatment but was an excellent bridging therapy before retransplantation.
Neurodevelopmental outcome in high‐risk patients after renal transplantation in early childhood
Authors: Qvist Erik , Pihko Helena , Fagerudd Pia , Valanne Leena , Lamminranta Sirkka , Karikoski Jukka , Sainio Kimmo , Rönnholm Kai , Jalanko Hannu , Holmberg Christer
Published: March 2003

Abstract:Patient and graft survival rates of pediatric renal transplant recipients are currently excellent, but there are few reports regarding the long‐term neurodevelopmental outcome after renal transplantation (Tx) in early childhood. Children with renal failure from infancy would be expected to have a less favorable developmental prognosis. We report the neurodevelopmental outcome in 33 school‐age children transplanted between 1987 and 1995 when < 5 yr of age. We prospectively performed a neurological examination, magnetic resonance imaging (MRI) of the brain, electroencephalograms (EEGs), audiometry, and neuropsychological tests (NEPSY), and measured cognitive performance (WISC‐R); we related these results to school performance and to retrospective risk factors prior to Tx. Twenty‐six (79%) children attended normal school and 76% had normal motor performance. Six of the seven children attending a special school had brain infarcts on MRI. The EEG was abnormal in 11 (35%), and five (15%) received anti‐convulsive treatment after Tx. Sensorineural hearing loss was documented in six patients. The mean intelligence quotient (IQ) was 87, and 6–24% showed impairment in neuropsychological tests. The children attending a special school had been more premature, but had not had a greater number of pre‐ or neonatal complications. They had experienced a greater number of hypertensive crises (p = 0.002) and seizures (p = 0.03), mainly during dialysis, but the number of septic infections and the mean serum aluminum levels were not significantly greater than in the children with normal school performance. In these previously lethal diseases, the overall neurodevelopmental outcome is reassuring. However, it is of crucial importance to further minimize the risk factors prior to Tx.
Diagnosis and treatment of Wilson's disease
Authors: Schilsky Michael L.
Published: March 2003

Abstract: Wilson's disease (WD) has moved on from being a recognized syndrome that was uniformly fatal to a curative disease for which the genetic basis has been discovered. Most pediatric patients present with hepatic manifestations, but some may have neurologic or psychiatric features. Clinical and biochemical screening, including liver biopsy for hepatic copper analysis, remain the standard for diagnosis, but haplotype analysis for siblings is now available and should be considered for family screening when possible. Lifelong medical therapy remains the mainstay of treatment, but treatment preferences are changing from penicillamine to alternative agents such as trientine and zinc. OLT remains lifesaving for those with fulminant WD and those in whom initial medical therapy fails. The future will probably see the application of rapid and accurate molecular diagnostic testing for this disorder and new therapeutic modalities such as hepatocyte transplantation, gene replacement therapy, and gene modification.
Peripheral blood as a stem cell source for hematopoietic cell transplantation in children: is the effort in vein?
Authors: Lipton Jeffrey M.
Published: March 2003

Abstract: Since 1968 HSCT utilizing bone marrow as a stem cell source has become an accepted treatment modality for a variety of immunologic, hematologic and malignant disorders. However, with widespread use it became apparent that BM donation is inconvenient, uncomfortable and not without risk. These observations led to a search for a more easily and safely acquired hematopoietic stem cell source. Recent experience suggests that peripheral blood may serve as an alternative to marrow. Indeed if the current trend continues PBSC will soon replace BM in adults as the preferred stem cell source for both HLA‐matched and unrelated HSCT. Furthermore adults who have experienced both seem to prefer PBSC to BM donation. Recently a number of small trials support the feasibility of PBSC harvest from HLA‐matched minor sibling donors. With regard to the recipient, data indicate more rapid engraftment, an acceptable incidence of acute and chronic GVHD, decreased infection and an increased survival for patients with malignancies (suggesting an increased GVT/L effect) when PBSC are utilized. These observations based almost entirely on the adult experience are far from definitive when children are considered. The relatively lower risk and severity of acute and chronic GVHD, the more frequent use of HSCT for non‐malignant disorders and the diminished role of GVT/L in the treatment of typical malignancies are factors particular to pediatric HSCT. In addition the sense that mobilization and harvest of PBSC may pose unique and significant risk for the young donor suggest to some that PBSC may not posses sufficient advantage to warrant their use in the pediatric transplant setting. Thus both the available adult‐derived data and experience suggest clinical equipoise with regard to the choice of stem cell product, under certain circumstances, when children are considered. This circumstance strongly supports the need for a comprehensive study evaluating the safety and efficacy of PBSC vs. BM HSCT in children.
Plasma pharmacokinetics of high‐dose oral busulfan in children and adults undergoing bone marrow transplantation
Authors: Bostrom Bruce , Enockson Karen , Johnson Amy , Bruns Alyssa , Blazar Bruce
Published: March 2003

Abstract: We have analyzed the plasma pharmacokinetics of busulfan in 272 patients receiving high‐dose oral busulfan and intravenous cyclophosphamide in conjunction with allogeneic or autologous bone marrow transplantation. The patients ranged in age from 2 months to 59 yr (mean 10, median 12 yr) and had the following diagnoses: thalassemia or sickle cell anemia (n = 74); leukemia or myelodysplasia (n = 112); inborn errors of metabolism (n = 41) or immunodeficiency (n = 45). Plasma specimens were collected following the first dose for each patient which ranged from 1 to 4 mg/kg (mean ± SD, 1.21 ± 0.41, median 1.15). Busulfan was quantitated using ultraviolet absorbance detection after derivatization and HPLC separation. Pharmacokinetic parameters were derived by modeling the raw data to fit first‐order single compartment kinetics. The kinetic parameters showed wide interpatient variability independent of age and diagnosis. There was a statistically significant correlation of age with the following parameters: area under the curve (AUC); maximal concentration; minimum concentration; clearance; volume of distribution and absorption half‐time. The coefficients of determination (i.e. correlation coefficient squared) were low ranging from 0.04 to 0.12 implying only a small part (i.e. 4–12%) of the variance was explained by age. Although busulfan pharmacokinetics are age‐related most of the variability is not explained by age or diagnosis.
Genetic variation and hematopoietic stem cell transplantation: expansion of the paradigm
Authors: Savage Sharon A. , Chanock Stephen
Published: March 2003

Abstract:  Genetic variation has been the mainstay of hematopoietic stem cell transplantation since the first transplants were attempted. A significant expansion of genetic knowledge is under way, as the draft sequence of the human genome is annotated. Hematopoietic stem cell transplantation is a field that will greatly benefit from this new knowledge, but the manner in which it is applied is daunting. Variation within key molecules related to hematopoietic stem cell transplant in combination with the current knowledge of human leukocyte antigen variation will serve to improve donor‐recipient matches and clinical outcome.
Sucrose accumulation in sweet sorghum stem internodes in relation to growth
Authors: Hoffmann‐Thoma Gudrun , Hinkel Karin , Nicolay Peter , Willenbrink Johannes
Published: March 2003

Sweet sorghum (Sorghum bicolor L. Moench) stems of different cultivars (NK 405. Keller and Tracy) reveal a different pattern of sucrose accumulation with respect to in‐ternodal sugar content and distribution. The onset of sucrose storage is not necessarily associated with the reproductive stage of the plant, as was hitherto assumed, but obviously occurs after cessation of internodai elongation as was postulated for the sugarcane stem. For at least two of the three cultivars, ripening is an internode to internode process beginning at the lowermost culm parts. Intensive growth of the internodes, combined with a high hexose content in stern parenchyma, shows a strong positive correlation (r |Mg 0.94) to the activity of sucrose synthase (SuSy; EC 2.4.13), but not to invertase (EC 3.2.1.26) which is not present as soluble (neutral and acid) or cell wall‐bound, salt‐extractable enzyme in the three culsivars investigated. Sucrose synthase measured in sucrose cleavage and synthesis direction reveals divergent activity rates and sensitivity towards exogenously applied Mg2+ ions and pH. SuSy activity is connected to the increase of internodai sucrose content in so far as (1) its decline is a prerequisite for the onset of sucrose accumulation and (2) it remains at a constant low level during sucrose storage. Sucrose phosphate synthase (SPS; EC 2.4.1.14) activity in the sorghum stem is low compared to SuSy and uniformly distributed over all inter‐nodes. Only source leaves of sorghum show a considerable SPS activity, but neither stem nor leaf SPS reveal a positive correlation to the increase of internodai sucrose content. Sucrose phosphate phosphatase (SPP; EC 3.1.3.24) amounts lo only 24–30% of the respective SPS activity but follows the same distribution pattern. None of the enzymes under study proves to be responsible for the extent of sucrose storage in the stem, so other phenomena such as transport processes within the stern tissue require further investigation.
Genetically stable cell lines of cucumber for the large‐scale production of diploid somatic embryos
Authors: Kreuger Marc , van der Meer Wiert , Postma Erik , Abbestee Rob , Raaijmakers Natasja , Holst Gerrit‐Jan van
Published: March 2003

For the initiation of embryogenic cucumber (Cucumis sativus L.) cell lines, from excised radicles, directly in liquid medium, the culture regime, explant density and type and concentration of hormones were adjusted so that pro‐embryogenic masses (PEMs) were formed within about 8 weeks. The established cucumber cell lines were maintained tor several years without loss of embryogenic and genetic stability. The ploidy level of somatic embryos from different cucumber eell lines was either diploid or tetraploid and depended on the ploidy level of Ihe cell line. Cucumber cell lines that produced only diploid embryos were obtained by selecting completely diploid explant material and growing it in the dark during the initiation phase. Mixoploid explains could lead to tetraploid or mixoploid ceil lines. Isolation and additional selection and subculturing of single PEMs resulted in either completely diploid or tetraploid cell lines, indicating that all cells of individual PEMs are either diploid or tetraploid. The ernbryogenic cucumber cell Imes, differing only in ploidy level, were indistinguishable in growth rate and embryogetiic potential and were genetically stable over several years.
Ultraweak light emission, free radicals, chilling and light sensitivity
Authors: Hideg Éva , Olof Björn Lars
Published: March 2003

Ultraweak light emission (UWLE) was measured from leaves of various chilling‐sensitive (Lycopersicon esculentum, Cucumis sativus and Phaseolus vulgaris) and ‐tolerant (Pisum sativum and Vicia faba) plants after exposure to low (4‐7°C) temperature in the light. UWLE increased upon chilling treatment combined with illumination with 200 µmol m−2 s−1 PAR in all plants, by about 30% in tolerant and by more than 100% in sensitive plants. It increased more when applied together with 400 µmol m−2 s−1 PAR: by 90‐100% and by 250‐280% in chilling‐tolerant and ‐sensitive plants, respectively. Free radical production was detected by spin‐trapping EPR spectroscopy in thylakoid membranes isolated from the chilling‐treated Lycopersicon esculentum and Vicia faba leaves. After 12 h chilling at 200 µmol m−2 s−1 PAR, free radical production was approximately 3 times greater in the former than in the latter species. The same ratio was approximately 6 if chilling was carried out at 400 µmol m−2 s−1 PAR, indicating the role of photooxidative stress in chilling injury.

Our results also confirm that the stress‐induced increase in UWLE is an indicator of free radical production and offers the possibility of using UWLE for monitoring the effect of chilling on temperature‐sensitive plants in an early stage.
Growth habit of Lotus tenuis shoots and the influence of photosynthetic photon flux density, sucrose and endogenous levels of gibberellins A1 and A3
Authors: Clúa Ariel , Bottini Rubén , Brocchi Gustavo N. , Bogino Judith , Luna Virginia , Montaldi Edgardo R.
Published: March 2003

The tropic response of Lotus tenuis shoots under conditions of full sunlight (FS) or artificial shade (AS) has been studied in relation to their endogenous levels of gibberellins (GAs) and sucrose. Single‐node explants with one axillary branch each were incubated in darkness with different concentrations of sucrose, GA3, B9 or their combinations, or whole plants were treated with FS, AS, AS + uniconazole, or AS + cimectacarb. High sucrose concentration supplied via xylem turned the axillary branch growth diagravitropic. GA3 reversed the effect of sucrose, both in shoot sections and whole plants, turning the axillary branch growth ortotropic and diminishing the sucrose levels at the shoot bases. High irradiance induced a prostrate growth of the shoots. Artificial shade increased both internode and branch length and induced an upward curvature of the shoots. This was correlated with a lower sucrose concentration and increased content of GA1 and GA3. Cimectacarb and uniconazole reversed the AS effects, although the sucrose content in the shoots remained low. These results are direct evidence that gravitropism of L. tenuis shoots under different light intensities, but of similar light quality, is controlled by the levels of GA1 and GA3 and sucrose.
Characterisation of glutathione transferases and glutathione peroxidases in pea (Pisum sativum)
Authors: Edwards Robert
Published: March 2003

Glutathione peroxidases (GPOXs) and glutathione transferases, also termed glutathione S‐transferases (GST, EC 2.5.1.18), with activities toward a range of xenobiotic substrates including herbicides, have been characterized in etiolated pea (Pisum sativum L. cv. Feltham's First) seedlings. Crude extracts showed high activity toward a range of GST substrates including 1‐chloro‐2,4‐dinitrobenzene (GSTC activity) and the herbicide fluorodifen (GSTF) but low activities toward chloroacetanilides and atrazine. Treatment of the pea seedlings with the herbicide safener dichlormid selectively increased the activity of GSTC and the GST which detoxified atrazine. This induction was restricted to the roots and was not observed with any of the other GST or GPOX activities. In contrast, treatment with CuCl2 increased GPOX activity in the root but had no effect on any GST activity, while treatment of epicotyls with elicitors of the phytoalexin response increased GST activity toward ethacrynic acid, but had no effect on other GST or GPOX activities.

The major enzymes with GSTC, GSTF and GPOX activities were purified from pea epicotyls 3609‐fold, 1431‐fold and 1554‐fold, respectively. During purification by hydrophobic interaction chromatography and affinity chromatography using S‐hexyl‐glutathione as ligand all three activities co‐eluted but could be partially resolved by anion exchange chromatography and gel filtration chromatography. Both GSTC and GPOX had a molecular mass of 48 kDa and their activities were associated with a similar 27.5‐kDa subunit but distinct 29‐kDa subunits. GSTF could be resolved into two isoenzymes with molecular masses of 49.5 and 54 kDa. GSTF activity was associated with a unique 30‐kDa subunit in addition to 27.5‐ and 29‐kDa peptides, suggesting that the two isoenzymes were composed of differing subunits. These results demonstrate that peas contain multiple GST isoenzymes some of which have GPOX activity and that the various activities are differentially responsive to biotic and abiotic stress.