bluuebunny/crossref_metadata_2025 huggingface dataset:
Intragraft cytokine mRNA expression in rejecting and non‐rejecting vascularized xenografts
Authors: Lorant Tomas , Krook Henrik , Wilton Jacob , Olausson Michael , Tufveson Gunnar , Korsgren Olle , Johnsson Cecilia
Published: June 2003

Abstract: Background: The aim of the present study was to further investigate the characteristics of both graft‐infiltrating cells and splenocytes during acute vascular rejection (AVR), cell‐mediated rejection and non‐rejection of vascularized concordant xenografts, by analysing both proinflammatory [interleukin‐1β (IL‐1β) and tumour necrosis factor (TNF‐α)] and more specific [(IL‐2, IL‐4, IL‐10, IL‐12p40 and interferon‐γ (IFN‐γ)] cytokines. A parallel investigation was made of the antibody response of IgM and IgG to the xenografts.

Methods: Mouse hearts were heterotopically transplanted to the neck vessels of recipient rats. Grafts, spleens and sera were collected from untreated (AVR) and cyclosporin A (CyA) treated animals on day 2 after transplantation. Organs from rats treated with 15‐deoxyspergualin (DSG) or CyA and DSG in combination were harvested on both day 2 and day 8. Grafts from DSG‐treated rats undergo cell‐mediated rejection and stop beating on day 9 and forth, while CyA + DSG treatment results in long‐term graft survival. Real‐time quantitative reverse transcriptase polymerase chain reaction (RT‐PCR) was applied for analysis of intragraft and splenic cytokine messenger RNA (mRNA) expression. The phenotypes of the graft infiltrating cells were characterized by immunohistochemistry. The antibody response was investigated by means of immunofluorescence, haemagglutination and flow cytometry.

Results: All the studied cytokines (IL‐1β, IL‐2, IL‐4, IL‐10, IL‐12p40, IFN‐γ and TNF‐α) were up‐regulated in the grafts from rejecting untreated (day 2) and DSG‐treated animals (day 8) in comparison with grafts from CyA + DSG treated animals (day 8). On day 2 under immunosuppression with CyA, DSG or CyA + DSG no or low cytokine mRNA levels were found. The mRNA levels of IL‐2, IL‐4 and IFN‐γ in the spleens were suppressed under both DSG‐ and CyA + DSG treatment on day 8. Immunofluorescence showed deposits of both IgM and IgG in grafts from untreated, CyA‐treated (day 2) and DSG‐treated (day 8) animals, while CyA + DSG treatment diminished these deposits on both day 2 and day 8. No circulating antibodies were identified in either group.

Conclusion: We hereby conclude that both AVR on day 2 and cell‐mediated rejection on day 8 (under DSG treatment) in a concordant cardiac mouse‐to‐rat xenotransplantation model are associated with an increase of proinflammatory cytokines, T helper 1 (Th1)‐associated cytokines as well as IL‐10, while immunosuppression with CyA + DSG diminishes the levels of all examined cytokines. Grafts undergoing AVR or cellular rejection are subjected to deposits of both IgM and IgG, although circulating donor specific antibodies are undetectable in serum.
Methods for the exclusion of circoviruses and gammaherpesviruses from pigs
Authors: Tucker A. W. , McNeilly F. , Meehan B. , Galbraith D. , McArdle P. D. , Allan G. , Patience C.
Published: June 2003

Abstract: The use of porcine tissues is being developed as a means to alleviate the shortage of allogeneic tissues and organs available for transplantation. To reduce the possibility of a microorganism of pigs being inadvertently transferred to the recipient of the xenograft, recommendations have been published on the microbiological specifications for organ source pigs. The porcine circoviruses (PCV1 and PCV2) and porcine lymphotropic herpesviruses (PLHV1 and PLHV2) are two infectious agents of pigs which are considered to be of significance for the microbiological safety of xenotransplantation. To ensure the exclusion of these microorganisms from animals destined for use under clinical conditions, reliable breeding methodologies are required. We investigated the efficiency of established derivation procedures for the removal of PCV and PLHV. In comparison with conventionally reared pigs, caesarian and barrier derived animals showed a markedly reduced prevalence of PCVs and PLHVs. Our results indicate that the derivation of animals free of both of these microorganisms is achievable and will enhance the microbiological safety of xenotransplantation.
Inhibition of baboon platelet aggregation in vitro and in vivo by the garlic derivative, ajoene
Authors: Teranishi Katsuhito , Apitz‐Castro Rafael , Robson Simon C. , Romano Egidio , Cooper David K. C.
Published: June 2003

Abstract: Background: The infusion of pig growth factor‐mobilized peripheral blood leukocytes (containing 1 to 2% progenitor cells) (pPBPC) into baboons is associated with a thrombotic microangiopathy, which results from a direct effect of these pig cells on platelet aggregation. Ajoene is a synthetic derivative of garlic that inhibits aggregation of human platelets induced by all known agents. To assess its potential use in models of xenotransplantation, this agent was tested for its effect on baboon platelet aggregation in vitro and in vivo. Methods: In vitro studies: Baboon platelet aggregation assays, using adenosine diphosphate (ADP) (20 or 40 μm) or collagen (12.5 μg/ml), were performed after incubation with ajoene (0 to 150 μg/ml) or dipyridamole (0 to 200 μg/ml). Platelets were also incubated with pPBPC (5 × 106 cells) without or with ajoene in the absence of a known agonist. In vivo studies: Baboons received either a single intravenous dose of ajoene (10 to 25 mg/kg) or dipyridamole (0.8 mg/kg), or repeated doses of both agents at 2 to 3 h intervals. Platelet‐rich plasma was obtained for platelet aggregation assays at time points up to 4 h post‐drug administration. Results: In vitro, platelet aggregation was inhibited by 95% (ADP assay) and 89% (collagen assay) by ajoene at concentrations of ≥75 μg/ml. Dipyridamole had no effect at concentrations of <100 μg/ml, but inhibited aggregation almost completely at higher concentrations. Ajoene inhibited the aggregation caused by pPBPC by 33 to 50%. In vivo, platelet aggregation was completely inhibited for 2 h by ajoene at 25 mg/kg. Dipyridamole at 0.8 mg/kg reduced aggregation by 20% for 15 min, but the effect was lost by 60 min. In combination, the two agents prolonged inhibition marginally. Repeated doses of both agents at 2 h intervals maintained complete inhibition of aggregation, but did not do so when the interval between doses was extended to 2.5 or 3 h. Combined therapy was not associated with any bleeding complications. Conclusions: Although ajoene is a powerful inhibitor of platelet aggregation, the need for repeated administration and its partial effect on pPBPC‐induced platelet aggregation would suggest that it may be of only limited value in preventing the thrombotic microangiopathy that develops when pPBPC are infused into baboons. However, it would seem worthy of further investigation when used in combination with other agents.
Approaching virus safety in xenotransplantation: a search for unrecognized herpesviruses in pigs
Authors: Chmielewicz Barbara , Goltz Michael , Lahrmann Karl‐Heinz , Ehlers Bernhard
Published: June 2003

Abstract: The identification of porcine viruses so far unrecognized is required to minimize virus‐related risks associated with xenotransplantation. We used a pan‐herpes consensus polymerase chain reaction assay to search for unrecognized porcine species of the Herpesviridae. The assay targets conserved regions of the herpesvirus DNA polymerase (DPOL) gene, using primers that were modified to diminish the assay's recognition capacity for the highly prevalent porcine lymphotropic herpesviruses 1, 2 and 3 (PLHV‐1, ‐2, ‐3), without substantially lowering the universal detection capacity of the assay. Analysis of 495 porcine blood and tissue samples from 294 animals, including 35 samples from 20 immunosuppressed pigs, resulted in the amplification of 128 herpesviral DPOL sequences. Sequence analysis attributed 127 of the amplimers to the known porcine herpesviruses (PLHV‐1, ‐2, ‐3; porcine cytomegalovirus; pseudorabiesvirus). In none of the pig samples analyzed here, evidence was obtained for the presence of additional novel porcine herpesvirus species. Therefore we conclude that pigs bred for the purpose of xenotransplantation pose a negligible risk of transmitting presently unrecognized herpesviruses to organ recipients.
Pathology of xenograft rejection: a commentary
Authors: Schuurman Henk‐Jan , Cheng Jane , Lam Tuan
Published: June 2003

Abstract: Trends in solid organ xenograft pathology are presented, with the focus on pig‐to‐nonhuman primate models. A simplified classification of rejection is followed, including hyperacute rejection (HAR), acute humoral xenograft rejection (AHXR), and acute cellular xenograft rejection (ACXR). The main components in HAR are natural xenoreactive antibodies in combination with complement activation. This is evident from the prevention of HAR in recipients in whom either antibodies or complement activation is depleted or inhibited. However, these strategies generally fail to prevent AHXR, which occurs later. AHXR is a multifactorial process in which natural and elicited antibodies may play roles, possibly in conjunction with complement, coagulation factors, and white blood cells. A main target appears to be the microvasculature which, in kidney grafts, is associated with a glomerular thrombotic microangiopathy. It is not clear to what extent species‐specific physiologic disparities in complement and coagulation processes may play a role, separate from antibody‐initiated processes. As rejection of solid organ xenografts is currently from AHXR, ACXR has not yet received close attention. In addition to intragraft rejection events, systemic complications following host–graft interactions have emerged, including (often fatal) consumptive coagulopathy and immune complex disease. It is anticipated that rejection processes will change when pigs with new genetic modifications become available. For instance, the precise role of natural antibodies to Galα1,3Gal will be able to be distinguished from other factors when pigs that lack the target antigen are available, and their organs can be evaluated in large animal xenotransplantation models.
Microsurgical access flap and enamel matrix derivative for the treatment of periodontal intrabony defects: a controlled clinical study
Authors: Wachtel Hannes , Schenk Günther , Böhm Sonja , Weng Dietmar , Zuhr Otto , Hürzeler Markus B.
Published: June 2003

Objectives: Application of the guided tissue regeneration (GTR) principle and utilization of enamel matrix derivative (EMD) have both been shown to result in periodontal regeneration. While clinical investigations have demonstrated that the use of a microsurgical concept in combination with the GTR technique positively affects the percentage of primary closure and the amount of tissue preservation, no such information is available for EMD‐treated periodontal defects. It was the aim of the present investigation to assess the clinical effect of the microsurgical access flap and EMD treatment with an emphasis on the evaluation of early wound healing.

Material and Methods: Eleven patients displaying at least one pair of intrabony periodontal defects with an intrabony component of ≥3 mm participated in the study. At baseline and at 6 and 12 months after surgery, the following clinical parameters were assessed by a blinded examiner: oral hygiene status (API), gingival inflammation (BOP), probing pocket depth (PPD), clinical attachment level (CAL) and gingival recession (GR). Defects were randomly assigned to test or control treatment, which both consisted of a microsurgical access flap procedure designed for maximum tissue preservation. The exposed root surfaces of the test sites were conditioned with a 24% EDTA gel followed by EMD (Emdogain®) application. Primary flap closure was achieved by a 2‐layered suturing technique. Postoperative healing was evaluated by a newly introduced early wound‐healing index (EHI) at 1 and 2 weeks after surgery.

Results: Both test and control treatment resulted in a statistically significant mean CAL gain of 2.8 and 2.0 mm at 6 months, and 3.6 and 1.7 mm at 12 months, respectively (p<0.05). Differences in CAL gain between the two treatment modalities were statistically significant at both time points (p<0.05). Additional GR values after 12 months averaged 0.3 and 0.4 mm for test and control sites, respectively, and did not reach statistical significance (p≥0.05). Two weeks after surgery, primary closure was maintained in 89% of the test sites and in 96% of the control sites.

Conclusion: Both treatment modalities using the microsurgical flap procedure resulted in a high percentage of primary flap closure and maximum tissue preservation. In terms of PPD reduction and CAL gain, the combination with EMD application appeared to be superior to the microsurgical access flap alone.
Deproteinized bovine bone and gentamicin as an adjunct to GTR in the treatment of intrabony defects: a randomized controlled clinical study
Authors: Stavropoulos A. , Karring E.S. , Kostopoulos L. , Karring T.
Published: June 2003

Objectives: To evaluate whether Bio‐Oss® used as an adjunct to guided tissue regeneration (GTR) improves the healing of 1‐ or 2‐wall intrabony defects as compared with GTR alone, and to examine whether impregnation of Bio‐Oss® with gentamicin may have an added effect.

Material and methods: Sixty patients, with at least one interproximal intrabony defect with probing pocket depth (PPD) 7 mm and radiographic evidence of an intrabony component (IC) 4 mm, were treated at random with either a resorbable membrane (GTR), a resorbable membrane in combination with Bio‐Oss® impregnated with saline (DBB−), a resorbable membrane in combination with Bio‐Oss® impregnated with gentamicin (DBB+), or with flap surgery (RBF).

Results: All treatment modalities resulted in statistically significant clinical improvements after 1 year. Defects treated with GTR alone presented a probing attachment level (PAL) gain of 2.9 mm, a residual PPD (PPD12) of 4.9 mm, a radiographic bone level (RBL) gain of 3.1 mm, and a residual IC (IC12) of 2.7 mm. GTR combined with Bio‐Oss® did not improve the healing outcome (PAL gain: 2.5 mm; PPD12: 4.9 mm; RBL gain: 2.8 mm; IC12: 3.3 mm). Impregnation of the Bio‐Oss® with gentamicin 2% mg/ml resulted in clinical improvements (PAL gain: 3.8 mm; PPD12: 4.2 mm; RBL gain: 4.7 mm; IC12: 2.1 mm), superior to those of the other treatment modalities, but the difference was not statistically significant. Defects treated with only flap surgery showed the most inferior clinical response (PAL gain: 1.5 mm; PPD12: 5.1 mm; RBL gain: 1.2 mm; IC12: 4.2 mm) of all groups.

Conclusion: The results failed to demonstrate an added effect of Bio‐Oss® implantation in combination with GTR on the healing of deep interproximal 1‐ or 2‐wall, or combined 1‐ and 2‐wall intrabony defects compared with GTR alone. Local application of gentamicin, on the other hand, improved the treatment outcome but not to an extent that it was statistically significant.
Treatment of peri‐implant infections: a literature review
Authors: Roos‐Jansåker Ann‐Marie , Renvert Stefan , Egelberg Jan
Published: June 2003

Objectives: The purpose of the present paper is to review available information on the treatment of peri‐implant mucositis and peri‐implantitis.

Materials and Methods: The results of animal research and human studies are presented. Proposed strategies for the treatment of peri‐implantitis presented in the literature are also included.

Results: Most of the information accessible at this time derives from case reports. The reports provide evidence that efforts to reduce the submucosal infection may result in short‐term improvements of the peri‐implant lesion. They also indicate that regenerative procedures in intrabony peri‐implant defects can result in the formation of new bone.

Conclusions: Several uncertainties remain regarding the treatment of peri‐implantitis. Properly conducted long‐term follow‐ups of consecutively treated cases would seem to be a realistic avenue for accumulation of more information. This may assist in establishing the predictability, magnitude and stability of improvements that can be achieved.
Efficacy of a 0.5% chlorhexidine gel on the control of gingivitis in Brazilian mentally handicapped patients
Authors: Pannuti C. M. , Saraiva M. C. , Ferraro A. , Falsi D. , Cai S. , Lotufo R. F. M.
Published: June 2003

Objective: To evaluate the effectiveness of a 0.5% chlorhexidine gel on the reduction of interdental gingival bleeding.

Material and Methods: Forty‐three mentally handicapped subjects, living in an institution (Casas André Luiz, Brazil), were enrolled in a double‐blind randomised clinical trial. The subjects were randomly divided into two groups: test (chlorhexidine gel) and control (placebo gel). The gel was applied with trays, twice a day, over a period of 8 weeks. The interdental bleeding index (IBI) was recorded before and after the period in which the gel was used.

Results: Statistical analysis showed that baseline IBI scores were similar (p=0.82) for test and control groups (55.6% and 54.5%, respectively). After 8 weeks, a statistically significant (p<0.001) decrease in IBI means took place in the test group (33.2%). The control group presented a small increase (60.6%) in IBI scores. The difference between groups was significant at the end of the study (p<0.001).

Conclusion: The results indicated that the use of a 0.5% chlorhexidine gel was effective in reducing interdental gingival bleeding in special patients.
Modulation of androgen metabolism by phenytoin, oestradiol and tamoxifen in human gingival fibroblasts
Authors: Soory M. , Tilakaratne A.
Published: June 2003

Objectives: The aim of this investigation is to study androgen metabolism in gingival fibroblasts in response to phenytoin, oestradiol and the antioestrogen tamoxifen, in order to establish the possible role of hormones in the aetiopathogenesis of phenytoin‐induced gingival overgrowth.

Materials and Methods: Six cell lines of human gingival fibroblasts were established in monolayer culture in Eagle's minimum essential medium. Duplicate incubations were performed independently with radiolabelled testosterone and 4‐androstenedione, respectively (14C‐T/14C‐4‐A), with optimal concentrations of phenytoin, oestradiol and tamoxifen alone and in combination. At the end of a 24‐h incubation period, the medium was solvent extracted for steroid metabolites, which were separated by thin layer chromatography and quantified using a radioisotope scanner.

Results: The substrates were metabolised mainly to the diols, 5α‐dihydrotestosterone (DHT) and 4‐androstenedione or testosterone, with the two substrates used. The trends were that phenytoin and oestradiol significantly elevated the yields of the androgens DHT, diols and 4‐A/testosterone from both substrates while tamoxifen inhibited the stimulatory effects of oestradiol and phenytoin alone and in combination (n=6; p<0.01, one‐way anova).

Conclusion: Specific hormone‐mediated activity in response to phenytoin could contribute to the pathogenesis of gingival overgrowth, which can be decreased by the anti oestrogen tamoxifen.
Short‐term epidemic dynamics in the Cakile maritima–Alternaria brassicicola host–pathogen association
Authors: Thrall Peter H. , Burdon J. J. , Bock Clive H.
Published: June 2003

Studies combining within‐ and among‐population processes are crucial for understanding ecological and co‐evolutionary dynamics in host–pathogen interactions. We report on work over an 18‐month period involving multiple beach populations of the plant Cakile maritima and its fungal pathogen Alternaria brassicicola along the south‐east coast of Australia.

Results from permanent transects replicated on several beaches show that disease incidence and prevalence vary significantly with plant age, class and distance from the sea, as well as time during the season. Plant density is also positively related to disease levels.

Results from three subregions indicate that disease persistence depends on survival of infected plants behind the foredunes of protected beaches. Population extinction was more likely on beaches with greater wind exposure and sea access, with the latter also related to colonization consistent with the dispersal of Cakile seeds via ocean currents.

Although disease dynamics during the epidemic were similar across subregions, the severity of the epidemic varied significantly among these areas, suggesting that large‐scale environmental factors may influence the timing and development of the epidemic.

Estimates of synchrony in disease dynamics indicated that populations within a subregion were significantly correlated with respect to epidemic development. There was, however, no evidence for spatial synchrony in disease dynamics based on among‐population covariances in disease prevalence and interbeach distances. Populations within a subregion were thus often at quite different phases of the epidemic at any given time.
Absence of association between delayed type hypersensitivity to tuberculin and atopy in children in The Gambia
Authors: Ota M. O. C. , Van Der Sande M. A. B. , Walraven G. E. L. , Jeffries D. , Nyan O. A. , Marchant A. , McAdam K. P. W. J.
Published: June 2003

Background An inverse association between delayed type hypersensitivity to tuberculin and atopy has been observed in children, suggesting that exposure to mycobacteria may influence the immune response to allergens.

Objective To investigate the relationship between tuberculin responses and atopy in children living in three different environments in The Gambia.

Methods In this cross‐sectional study a total of 507 school‐aged children were recruited from rural, urban poor or urban affluent communities. They were assessed for skin responses to five common allergens and tuberculin, presence of bacille Calmette–Guérin (BCG) scar, presence of intestinal parasites, and total serum IgE. Atopy was defined as the presence of a skin prick test response ≥ 3 × 3 mm to at least one allergen.

Results The overall prevalence of atopy was 33% but there was a significant variation among the three study groups. The prevalence of atopy was 22% in urban poor, 36% in urban affluent, and 43% in rural children. Controlling for potential confounding factors, children in the rural community had a significantly higher odds ratio, 3.3 (95% onfidence interval 1.8–6.0) of being atopic than children from the urban poor community. No association between atopy and tuberculin response or BCG scar was observed in any of the three groups. Serum IgE levels were higher among children of the urban poor group but were not associated with tuberculin response or BCG scar in any of the groups.

Conclusion Environmental factors have an important influence on the development of atopy in children in The Gambia but delayed type hypersensitivity to tuberculin is not a protective factor.
Pranlukast, a cysteinyl leukotriene receptor 1 antagonist, attenuates allergen‐specific tumour necrosis factor alpha production and nuclear factor kappa B nuclear translocation in peripheral blood monocytes from atopic asthmatics
Authors: Tomari S. , Matsuse H. , Machida I. , Kondo Y. , Kawano T. , Obase Y. , Fukushima C. , Shimoda T. , Kohno S.
Published: June 2003

Background The cysteinyl leukotriene receptor 1 (cysLTR1) antagonists are useful for oral treatment of bronchial asthma. The underlying mechanism of cysLTR1 antagonists on inhibition of inflammatory cytokine production is yet to be determined.

Objective The present study was designed to determine the effect of pranlukast, a cysLTR1 antagonist, on production of inflammatory cytokines by allergen‐stimulated peripheral blood monocytes (PBM) from atopic asthmatics.

Methods PBM were obtained from normal control (n = 10) and Dermatophagoides farinae (Der f) allergen‐sensitized atopic asthmatics (n = 12), and were cultured in the presence of Der f allergen. The production of TNF‐α and nuclear‐translocation of nuclear factor kappa B (NF‐κB) was determined. In atopic asthmatics, pranlukast, tacrolimus or dexamethasone was added before stimulation by Der f. The additive effect of pranlukast and dexamethasone was also determined.

Results PBM from atopic asthmatics cultured with Der f exhibited a significant increase in TNF‐α production and nuclear translocation of NF‐κB compared with normal control (P < 0.01). Pranlukast, tacrolimus and dexamethasone significantly inhibited production of TNF‐α and nuclear‐translocation of NF‐κB in PBM of atopic asthmatics (P < 0.01). An additive effect of pranlukast on low‐dose dexamethasone was also demonstrated. However, LTD4 did not induce TNF‐α production or NF‐κB nuclear translocation.

Conclusion Our results suggest that pranlukast may inhibit TNF‐α production via suppression of NF‐κB activation through pathways distinct from cysLTR1 antagonism.
Lipoteichoic acid from Staphylococcus aureus enhances allergen‐specific immunoglobulin E production in mice
Authors: Matsui K. , Nishikawa A.
Published: June 2003

Background Our previous study demonstrated that lipoteichoic acid (LTA) from Staphylococcus aureus induced T helper type 2 (Th2)‐prone dermatitis resembling that seen in atopic dermatitis (AD) patients in mice sensitized percutaneously with an allergen. However, the effects of LTA on allergen‐specific IgE production in such sensitized mice have not been elucidated.

Objective The purpose of this study was to determine the effects of LTA from S. aureus on allergen‐specific IgE production in mice sensitized percutaneously with a house dust mite antigen (MA).

Methods Mice were sensitized with a single topical application of MA and/or LTA to barrier‐disrupted abdominal skin. One to 5 weeks later, MA‐specific IgE antibodies in sera from sensitized mice were detected by an enzyme‐linked immunosorbent assay (ELISA). Expression of B7.1 (CD80), B7.2 (CD86) and CD40L molecules by CD40‐positive (CD40+) and CD4‐positive (CD4+) cells in the lymph nodes of sensitized mice were analysed by flow‐cytometry (FACS).

Results Simultaneous sensitization with MA and LTA increased IgE production 3 weeks later, significantly more than sensitization with MA alone. FACS analysis of CD40+ cells in the lymph nodes from sensitized mice showed that simultaneous sensitization with MA and LTA did not enhance CD80‐ or CD86‐expression by antigen‐presenting cells such as B lymphocytes and dendritic cells more than sensitization with MA alone. However, analysis of CD4+ cells in the lymph nodes showed that simultaneous sensitization with MA and LTA increased the number of CD40L‐expressing Th cells more than sensitization with MA alone.

Conclusion These results suggest that LTA enhances allergen‐specific IgE production by a mechanism associated with up‐regulation of CD40L‐expressing Th cells and this might explain the role of skin colonization with S. aureus in AD patients.
Caesarean section increases the risk of hospital care in childhood for asthma and gastroenteritis
Authors: Håkansson S. , Källén K.
Published: June 2003

Objective To investigate if caesarean section (CS) increases the risk for childhood asthma and gastroenteritis with reference made to children born with vaginal delivery (VD).

Methods Retrospective study of data from linked Swedish medical service registers – Medical Birth Registry (MBR) and Hospital Discharge Registry (HDR). Data were obtained from women without any background/perinatal morbidity noted, and from children without any neonatal complications. Children that had reached at least 1 year of age and were found in the HDR were considered as cases, whereas children not found in the HDR or hospitalized for other causes than asthma or gastroenteritis were defined as controls. Odds ratios (OR) stratified for year of birth, maternal age, parity and smoking in early pregnancy were calculated. Investigations were made comparing the risk for in hospital treatment for asthma or gastroenteritis in CS children and in VD siblings of CS children. The overall inpatient morbidity in CS and VD children were also investigated.

Results The OR for asthma in CS children was 1.31 [95% confidence interval (CI) 1.23–1.40]. The same OR, 1.31, was found for gastroenteritis (95% CI 1.24–1.38). The OR for CS children having experienced both asthma and gastroenteritis was further increased (1.74, 95% CI 1.36–2.23). The risk for asthma in VD siblings of CS children was not significantly increased, whereas VD siblings experienced a slightly increased risk for gastroenteritis. CS children had an increased overall in hospital morbidity when compared to VD children.

Conclusion There is a significant increase of the risk for developing symptoms of asthma and/or gastroenteritis that motivates admission for hospital care in CS children older than 1 year. It is speculated that a disturbed intestinal colonization pattern in CS children may be a common pathogenic factor.
Neonatal head circumference, neonatal weight, and risk of hayfever, asthma and eczema in a large cohort of adolescents from Sheffield, England
Authors: Katz K. A. , Pocock S. J. , Strachan D. P.
Published: June 2003

Background Neonatal physical characteristics, including head circumference and birth weight, have been hypothesized to be markers of in utero thymic development. Greater head circumference and lower birth weight have been linked in previous studies to subsequent development of asthma, and greater birth weight has been associated with subsequent development of eczema.

Objective To investigate potential associations between neonatal head circumference and weight and hayfever, asthma and eczema in a cohort of adolescents from Sheffield, England.

Methods Responses to a questionnaire inquiring about physician‐diagnosed hayfever, asthma and eczema among adolescents in Sheffield, England, were linked to previously recorded measurements of weight at birth and at 1 month and head circumference at 1 month. Logistic regression methods were used to relate diagnoses to neonatal measurements and potential confounders.

Results The cohort consisted of 10 809 adolescents, of whom 16.5% reported hayfever, 18.0% asthma, and 16.2% eczema. After adjusting for sex, age at the time of the questionnaire, maternal age and gestational age at birth, number of older and younger siblings, time since birth of next older sibling, neonatal sickness, type of neonatal feeding, and maternal and paternal educational backgrounds, hayfever was the only disease associated with neonatal measurements. Comparing the highest with the lowest fifths of distributions, lifetime prevalence of hayfever was positively associated with neonatal head circumference (adjusted odds ratio 1.23, 95% CI 1.03 to 1.47) and with birth weight (1.17, 0.99 to 1.39). Hayfever was inversely related to the ratio of head circumference to birth weight (0.89, 0.75 to 1.05) and to gestational age. The associations with head circumference and birth weight were not substantially altered by further adjustment for gestational age.

Conclusion Greater neonatal head circumference may be associated with an increased risk of hayfever, but the inverse relationship between hayfever prevalence and the ratio of head circumference to birth weight challenges the prior hypothesis that greater head circumference relative to body mass reflects abnormal thymic development in utero, increasing the likelihood of allergic sensitization.
Pranlukast inhibits NF‐κB activation in human monocytes/macrophages and T cells
Authors: Ichiyama T. , Hasegawa S. , Umeda M. , Terai K. , Matsubara T. , Furukawa S.
Published: June 2003

Background Pranlukast is a leukotriene 1 (LT1) receptor antagonist and is effective against bronchial asthma. Pranlukast inhibits contraction of the tracheal muscle, and thereby antagonizes the binding of LTC4, LTD4 and LTE4. However, the action of pranlukast on monocytes/macrophages and T cells is unknown.

Objective We examined whether or not pranlukast inhibits TNF‐α‐induced activation of nuclear transcription factor NF‐κB, a factor that is essential for the expression of proinflammatory cytokines, on human monocytic 1.3% dimethylsulphoxide (DMSO)‐differentiated U‐937 cells, which have cysteinyl LT1 (CysLT1) receptors on their membranes, and T cells (Jurkat), which do not.

Methods We examined whether or not LTC4, LTD4 or LTE4 induced NF‐κB activation in 1.3% DMSO‐differentiated U‐937 cells by Western blotting. The inhibitory effects of pranlukast and MK‐571, which is an LTD4 receptor‐selective antagonist, on TNF‐α‐induced NF‐κB activation was evaluated by Western blotting and flow cytometry, and those on lipopolysaccharide (LPS)‐induced interleukin‐6 (IL‐6) production in peripheral blood mononuclear cells (PBMC) were evaluated by enzyme‐linked immunosorbent assaying.

Results LTC4, LTD4 or LTE4 did not induce NF‐κB activation in 1.3% DMSO‐differentiated U‐937 cells. Western blotting demonstrated that 10−5 M pranlukast inhibits NF‐κB activation in 1.3% DMSO‐differentiated U‐937 and Jurkat cells by about 40% & 30%, respectively. Flow cytometry demonstrated that pranlukast and MK‐571 inhibit NF‐κB activation in 1.3% DMSO‐ differentiated U‐937 and Jurkat cells in a dose‐related manner. Moreover, 10−5 M pranlukast and MK‐571 inhibited LPS‐induced IL‐6 production in PBMC by about 65% and 15%, respectively.

Conclusion Pranlukast and MK‐571 partially inhibited NF‐κB activation in 1.3% DMSO‐ differentiated U‐937 and Jurkat cells, and IL‐6 release in PBMC. These findings are consistent with the idea that, independently of CysLT1 receptor antagonism, micromolar concentrations of pranlukast suppress the production of proinflammatory cytokines via inhibition of NF‐κB activation in monocytes/macrophages and T cells, but the contribution of this effect to the anti‐inflammatory activity of pranlukast at oral therapeutic doses in asthmatic patients is unclear.
Safety of nasal budesonide in the long‐term treatment of children with perennial rhinitis
Authors: Möller C. , Ahlström H. , Henricson K.‐Å. , Malmqvist L.‐Å. , Åkerlund A. , Hildebrand H.
Published: June 2003

Background Intranasal budesonide is an efficacious treatment for perennial allergic rhinitis. Long‐term effects on safety, particularly in children, need further investigation.

Objective To investigate the long‐term safety of intranasal budesonide in children.

Methods In an open trial, 78 children (5–15 years) with perennial rhinitis were treated with intranasal budesonide pressurized metered dose inhaler 200 µg twice daily (delivered daily dose 256 µg) for 12 months; 43 children stayed in the study for 12 additional months and were switched to aqueous suspension (400 µg delivered daily dose) for 6 months. Statural growth, bone age, ophthalmologic and rhinoscopic status, cortisol and biochemical analyses in blood and urine were monitored during the first and second years, and adverse events (AEs) were continuously recorded.

Results No significant effects on statural growth and bone age, compared with reference values, were observed. Morning plasma cortisol and 24‐h urinary cortisol were not changed during treatment. Patients reported 195 AEs, most commonly nasal dryness (30%), blood‐tinged secretions (21%) and, among non‐nasal AEs, headache (13%). Rhinoscopy revealed no signs of mucosal atrophy, ulceration, or candidiasis but some nasal dryness. No treatment‐related ophthalmological or biochemical aberrations were found. Reduction of blood eosinophils and nasal symptom scores, compared with pre‐treatment values, indicated the efficacy of budesonide treatment.

Conclusion Long‐term treatment for 1–2 years with intranasal budesonide 256–400 µg daily in children with perennial rhinitis revealed no negative effects on growth or endogenous cortisol production. Local side‐effects were mild and patient symptoms decreased.
Age‐related changes in blood capillary endothelium of human dental pulp: an ultrastructural study
Authors: Espina A. I. , Castellanos A. V. , Fereira J. L.
Published: June 2003

Aim  To describe the ultrastructural changes that occur in pulpal blood capillaries as a result of ageing.

Methodology  Thirty samples of healthy dental pulps were obtained from functional human permanent teeth. Two age groups were examined: young (10–17 years) and old (>60 years). The teeth were extracted under local anaesthesia using mepivacaine without adrenaline (Scandonest 3%, Septodont, Saint‐Maur des Fossés, France) and split longitudinally in a bench press. The pulps were gently removed, immersed in fixative solution, sectioned and processed by conventional transmission electron microscopic techniques. Micrographs were taken from the endothelium, and the whole capillary area of each vessel was examined.

Results  In young pulps, the endothelial cell layer was characterized by the presence of numerous pinocytotic vesicles and microvesicles, RER cisterns, free ribosomes, a small Golgi complex, centrioles, microtubules, microfilaments and mitochondria. In the endothelial cell cytoplasm of older pulpal vessels, pinocytotic vesicles and microvesicles, as well as microfilaments, were more numerous. In addition, lipid‐like vacuoles, monogranular glycogen granules and extensive Golgi complexes with dilated cisterns were also present. Weibel‐Palade bodies were observed in both age groups without showing variations related with age.

Conclusions  The results obtained in capillaries of aged pulpal tissue suggest that the endothelium experiences morphological changes that could be associated with advancing age.
Immunolocalization of bone extracellular matrix proteins (type I collagen, osteonectin and bone sialoprotein) in human dental pulp and cultured pulp cells
Authors: Garcia J. M. Q. , Martins M. D. , Jaeger R. G. , Marques M. M.
Published: June 2003

Aim  To simultaneously analyse the expression of type I collagen, osteonectin and bone sialoprotein (BSP) in human dental pulp of different ages.

Methodology  Cultured dental pulp fibroblasts (FP1 cell line), pulps from dental germs with incomplete root formation (n = 4) and pulps of erupted teeth with total root formation (n = 4) were used. Bone proteins were searched by immunohistochemistry and immunofluorescence using polyclonal antibodies and compared among the three groups assessed.

Results  Immunohistochemistry detected the three proteins in dental pulp tissue, as it labelled extracellular matrix, predentine and odontoblasts. The BSP label was weaker, when compared to both type I collagen and osteonectin. The presence of type I collagen was more evident in pulps from erupted teeth, when compared to germ dental pulps. On the other hand, a strong expression of osteonectin in germ dental pulps was observed.

Conclusions  Regardless of the degree of maturation, dental pulps present type I collagen, osteonectin and BSP in the extracellular matrix (ECM) and in the odontoblastic layer. Thus, the results suggest that these proteins are related to the production and mineralization of dentine.
Interactions of ethylenediamine tetraacetic acid with sodium hypochlorite in aqueous solutions
Authors: Grawehr M. , Sener B. , Waltimo T. , Zehnder M.
Published: June 2003

Aim  To evaluate interactions of ethylenediamine tetraacetic acid (EDTA) with sodium hypochlorite (NaOCl).

Methodology  Solutions consisting of 8.5% EDTA and 0.5% NaOCl were compared to a 1 : 1 (w/w) mixture of 17% EDTA and 1% NaOCl for their calcium‐chelating, tissue‐dissolving, and antimicrobial properties. Amounts of available chlorine were determined in the EDTA/NaOCl solutions with an iodine/thiosulphate titration method. Calcium chelation capacity was titrated with a pure calcium solution using a murexide indicator. Weight loss of porcine palatal mucosal specimens incubated in the test solutions was measured over time. Antimicrobial potential of pure solutions and the combination was recorded using an agar diffusion test in plates incubated with Enterococcus faecalis or Candida albicans.

Results  The presence of hypochlorite had little effect on the calcium‐chelating ability or on the antimicrobial potential of EDTA. Available chlorine content decreased to 0.06% in the combined EDTA–NaOCl solution compared to 0.50% in an equivalent NaOCl mixture with deionized water. The EDTA–NaOCl solution did not dissolve more tissue than an equivalent pure EDTA solution at any time (anova, P > 0.05).

Conclusions  Ethylenediamine tetraacetic acid retained its calcium‐complexing ability when mixed with NaOCl, but EDTA caused NaOCl to lose its tissue‐dissolving capacity and virtually no free chlorine was detected in the combinations. Clinically, this suggests that EDTA and NaOCl should be used separately. In an alternating irrigating regimen, copious amounts of NaOCl should be administered to wash out remnants of the EDTA.
Comparison of antibacterial and toxic effects of various root canal irrigants
Authors: Önçağ Ö. , Hoşgör M. , Hilmioğlu S. , Zekioğlu O. , Eronat C. , Burhanoğlu D.
Published: June 2003

Aim  To compare the antibacterial properties and toxicity of 5.25% sodium hypochlorite (NaOCl), 2% chlorhexidine gluconate and 0.2% chlorhexidine gluconate plus 0.2% cetrimide (Cetrexidin®; Vebas, San Giuliano, Milan, Italy).

Methodology  The antibacterial effects of the irrigants in vitro were examined after 5 min and 48 h in freshly extracted human teeth with single roots, whose canals were infected by Enterococcus faecalis ATCC 29212. In a separate in vivo study, bacterial culture samples were collected before treatment from the infected root canals of deciduous teeth containing necrotic pulp tissue. Irrigants were used to clean the canals which were then left empty for 48 h. Aerobic/facultative anaerobic and anaerobic bacterial growth were compared before and 48 h after irrigation. Finally, the toxic effects of the irrigants were assessed by injecting them into the subcutaneous tissues of rats. The inflammatory reactions that occurred 2 h, 48 h and 2 weeks after the injections were evaluated.

Results  In the laboratory study, the 2% chlorhexidine gluconate and Cetrexidin® were significantly more effective on E. faecalis than the 5.25% NaOCl at 5 min (P < 0.05). Similarly, in the in vivo study, 2% chlorhexidine gluconate and Cetrexidin® were significantly more effective on anaerobic bacteria than the 5.25% NaOCl at 48 h (P < 0.05). At the end of 2 weeks, the toxicity of the NaOCl solution was greater than that of the other irrigants (P < 0.05).

Conclusions  Cetrexidin® and 2% chlorhexidine gluconate were more effective, and had more residual antibacterial effects and lower toxicity than 5.25% NaOCl solution.
Constitutive nuclear localization and initial cytoplasmic apoptotic activation of endogenous caspase‐3 evidenced by confocal microscopy
Authors: Ramuz Olivier , Isnardon Daniel , Devilard Elisabeth , Charafe‐Jauffret Emmanuelle , Hassoun Jacques , Birg FranÇoise , Xerri Luc
Published: June 2003

Summary.  The localization of caspases and their substrates in different cellular compartments may be one way to regulate apoptosis. Caspase‐3‐dependent proteolysis of inhibitor caspase‐activated deoxyribonuclease (ICAD) activates caspase‐activated deoxyribonuclease (CAD), which induces apoptotic internucleosomal DNA degradation. The nuclear localization of ICAD, pro‐ and active‐caspase‐3 molecules remains a controversial issue. Using a combination of immunodetection of endogenous molecules and confocal microscopy, we analysed the kinetics of the procaspase‐3 and CAD activation induced by FAS triggering in Jurkat cells. Through a semi‐quantitative image analysis, we showed a constitutive nuclear localization of pro‐caspase 3 and ICAD in non‐apoptotic cells. FAS stimulation induced 7A6 apoptotic antigen expression, which could be related to three different sequential patterns of nuclear chromatin organization. Active‐caspase‐3 first appeared in the cytoplasm and was next observed in the nucleus. Simultaneously, the amount of ICAD located in the nucleus decreased, whereas the amount of ICAD located in the cytoplasm remained unchanged. Thus, our experiments using in situ immunodetection of endogenous molecules show that the ICAD cleavage by the active‐caspase‐3 probably takes place in the nucleus. These results provide new perspectives about the subcellular compartmentation and traffic of caspases during the apoptotic process.
Pathogenesis of axonal dystrophy and demyelination in αA‐crystallin‐expressing transgenic mice
Authors: Van Rijk A.F. , Sweers M.A.M. , Merkx G.F.M. , Lammens M. , Bloemendal H.
Published: June 2003

Summary.  We recently described a transgenic mouse strain overexpressing hamster αA‐crystallin, a small heat shock protein, under direction of the hamster vimentin promoter. As a result myelin was degraded and axonal dystrophy in both central nervous system (especially spinal cord) and peripheral nervous system occurred. Homozygous transgenic mice developed hind limb paralysis after 8 weeks of age and displayed progressive loss of myelin and axonal dystrophy in both the central and peripheral nervous system with ongoing age. Pathologically the phenotype resembled, to a certain extent, neuroaxonal dystrophy. The biochemical findings presented in this paper (activity of the enzymes superoxide dismutase, catalase and transglutamase, myelin protein zero expression levels and blood sugar levels) confirm this pathology and exclude other putative pathologies like Amyothrophic Lateral Sclerosis and Hereditary Motor and Sensory Neuropathy. Consequently, an excessive cytoplasmic accumulation of the transgenic protein or a disturbance of the normal metabolism are considered to cause the observed neuropathology. Therefore, extra‐ocular αA‐crystallin‐expressing transgenic mice may serve as a useful animal model to study neuroaxonal dystrophy.
Interleukin‐4‐expressing plasmid DNA inhibits reovirus type‐2‐triggered autoimmune insulitis in DBA/1 J suckling mice
Authors: Hayashi T. , Yasutomi Y. , Hasegawa K. , Sasaki Y. , Onodera T.
Published: June 2003

Summary.  In this study we have examined the effect of systemic administration of T helper (Th) 2 cytokines on reovirus type‐2 (Reo‐2)‐triggered Th1‐mediated autoimmune insulitis with impaired glucose tolerance (IGT) in DBA/1J suckling mice. We have demonstrated clearly that the systemic administration of both interleukin (IL)‐4‐expressing plasmid DNA (pIL‐4) and recombinant IL‐4 (rIL‐4) inhibited the development of insulitis with IGT in a dose dependent manner as compared to untreated groups in Reo‐2‐infected DBA/1J suckling mice. The inhibitory effects of IL‐4 on the development of insulitis with IGT and the advantages of pIL‐4 as compared to rIL‐4 in this model are discussed.