bluuebunny/crossref_metadata_2025 huggingface dataset:
Terminal deoxynucleotidyl transferase is down‐regulated by AP‐1‐like regulatory elements in human lymphoid cells
Authors: Peralta‐Zaragoza Oscar , Recillas‐Targa Félix , Madrid‐Marina Vicente
Published: January 2004

Terminal deoxynucleotidyl transferase (TdT) is a template‐independent DNA polymerase that catalyses the incorporation of deoxyribonucleotides into the 3′‐hydroxyl end of DNA templates and is thought to increase junctional diversity of antigen receptor genes. TdT is expressed only on immature lymphocytes and acute lymphoblastic leukaemia cells and its transcriptional expression is tightly regulated. We had previously found that protein kinase C (PKC) activation down‐regulates TdT expression. PKC‐activation induces the synthesis of the Fos and Jun proteins, known as the major components of activation protein 1 (AP‐1) transcriptional factor implicated in transcriptional control. Here we report the identification of several DNA–protein interactions within the TdT promoter region in non‐TdT expressing human cells. Sequence analysis revealed the presence of a putative AP‐1‐like DNA‐binding site, suggesting that AP‐1 may play a relevant role in TdT transcriptional regulation. Using a different source of nuclear extracts and the AP‐1–TdT motif as a probe we identified several DNA‐protein retarded complexes in electrophoretic mobility shift assays. Super‐band shifting analysis using an antibody against c‐Jun protein confirmed that the main interaction is produced by a nuclear factor that belongs to the AP‐1 family transcription factors. Our findings suggest that the TdT gene expression is down‐regulated, at least in part, through AP‐1‐like transcription factors.
Regulation of tumour necrosis factor‐related apoptosis‐inducing ligand (TRAIL) and TRAIL receptor expression in human neutrophils
Authors: Kamohara Hidenobu , Matsuyama Wataru , Shimozato Osamu , Abe Koichiro , Galligan Carole , Hashimoto Shin‐Ichi , Matsushima Kouji , Yoshimura Teizo
Published: January 2004

Tumour necrosis factor (TNF)‐related apoptosis‐inducing ligand (TRAIL) is a member of the TNF superfamily, which is capable of inducing apoptosis in many cell types, including tumour and virus‐infected cells, but rarely in normal cells. Expression of TRAIL mRNA and TRAIL receptors has previously been detected in neutrophils; however, the expression of TRAIL protein and the regulation of TRAIL and TRAIL receptor expression in these cells remain unknown. Here we report, for the first time, that neutrophils constitutively express TRAIL protein on their cell surface and that the TRAIL protein is shed during culture. TNF‐α is a down‐regulator of TRAIL expression, whereas IFN‐γ up‐regulates the expression of TRAIL. Neutrophils did not express a detectable level of TRAIL‐R1 or ‐R4, but constitutively expressed a low, but substantial, level of TRAIL‐R2 and a high level of TRAIL‐R3. Although the level of TRAIL‐R2 was not significantly altered during culture under different experimental conditions, ≈ 30% of TNF‐α‐treated cells rapidly lost their high‐level TRAIL‐R3 expression, whereas the majority of IFN‐γ‐treated cells retained a high level of TRAIL‐R3 expression. Anti‐TRAIL neutralizing antibody significantly inhibited neutrophil apoptosis during cultures in medium alone, or in the presence of TNF‐α or IFN‐γ. Thus, our study identified human neutrophils as a cellular source of TRAIL and suggests that neutrophil‐derived TRAIL may play a role in immune surveillance. Our results also suggest a role for the TRAIL/TRAIL receptor system in neutrophil apoptosis.
Altered T‐dependent antigen responses and development of autoimmune symptoms in mice lacking E2A in T lymphocytes
Authors: Pan Lihua , Bradney Curtis , Zheng Biao , Zhuang Yuan
Published: January 2004

E2A has been shown to be an important transcription factor downstream of the T‐cell receptor (TCR) signal during T‐cell development. The TCR signal is known to elicit different cellular responses at different stages of T‐cell development. Whether E2A is still required for normal TCR signalling in mature T cells is unknown. Here we examined T‐cell function after disruption of the E2A gene exclusively in the T‐cell lineage. The conditional E2A‐deficient mice show enhanced humoral immunity to a T‐dependent antigen. We further show that E2A is involved in regulating TCR‐induced T‐cell proliferation events. However, E2A seems to play opposite roles in naïve and effector T cells. In the absence of E2A, TCR‐induced proliferation is increased in naïve T cells and decreased in effector T cells. At older ages, these mice frequently develop antinuclear antibodies and proteinuria. Our studies suggest that E2A regulates T‐cell function and the loss of E2A may promote age‐dependent autoimmune diseases.
Toll‐like receptor 2 (TLR2) and TLR4 are present inside human dendritic cells, associated with microtubules and the Golgi apparatus but are not detectable on the cell surface: integrity of microtubules is required for interleukin‐12 production in response to internalized bacteria
Authors: Uronen‐Hansson Heli , Allen Jennifer , Osman Mohamed , Squires Ginette , Klein Nigel , Callard Robin E.
Published: January 2004

The activation of dendritic cells (DCs) by microbes is mediated by pattern recognition receptors including the Toll‐like receptors (TLR). Bacterial lipopolysaccharide acts via TLR4 whereas peptidoglycan and lipoprotein responses are mediated by TLR2. It is generally accepted that TLR binding to microbes occurs at the cell surface but this has not been directly demonstrated for human DCs. We show here that TLR2 and TLR4 are expressed inside DCs in an abundant tubulovesicular pattern with a focus of intense staining adjacent to the nucleus. In contrast, there was no detectable expression on the cell surface. TLR2 and TLR4 were readily found both intracellularly and on the surface of monocytes. They were shown to be closely associated with the Golgi complex and colocalized with α‐tubulin, displaying a high focal concentration at the microtubule organizing centre. Alignment of TLR2 and TLR4 with microtubules was observed, suggesting that microtubules serve as transport tracks for TLR vesicles. Depolymerization of the microtubule network disrupted the intracellular expression of TLR2 and TLR4 and profoundly inhibited interleukin‐12 (IL‐12) production in response to Neisseria meningitidis but did not prevent phagocytosis. These data are consistent with the bacterial signalling through TLR2 and TLR4 required for IL‐12 production occurring inside DCs after phagocytosis.
Central and peripheral autoantigen presentation in immune tolerance
Authors: Pugliese Alberto
Published: January 2004

Recent studies in both humans and experimental rodent models provide new insight into key mechanisms regulating tolerance to self‐molecules. These recent advances are bringing about a paradigm shift in our views about tolerance to self‐molecules with tissue‐restricted expression. There is, indeed, mounting evidence that selected antigen‐presenting cells (APCs) have the ability to synthesize and express self‐molecules, and that such expression is critical for self‐tolerance. Insulin is a key hormone produced exclusively by pancreatic β‐cells and a critical autoantigen in type 1 diabetes. It provides an excellent example of a molecule with tissue‐restricted expression that is expressed ectopically by APCs. The fact that APCs expressing insulin have been demonstrated in both thymus and peripheral lymphoid tissues suggests that they may play a role in insulin presentation in both the central and peripheral immune system. Experimental mice, in which insulin expression was altered, provide functional data that help to dissect the role of insulin presentation by APCs of the immune system. This review addresses recent literature and emerging concepts about the expression of self‐molecules in the thymus and peripheral lymphoid tissues and its relation to self‐tolerance.
Limited role of the Toll‐like receptor‐2 in resistance to Mycobacterium avium
Authors: Gomes M. Salomé , Flórido Manuela , Cordeiro João V. , Teixeira C. Miguel , Takeuchi Osamu , Akira Shizuo , Appelberg Rui
Published: January 2004

The role of the Toll‐like receptor (TLR)‐2 in the generation of protective immunity to Mycobacterium avium was evaluated using gene‐disrupted mice. TLR‐2−/− mice were more susceptible than wild‐type C57Bl/6 mice to M. avium strains that were able to proliferate in vivo before the development of protective immunity and mycobacteriostasis. In contrast, the elimination of non‐virulent strains was not affected by the mutation. The generation of interferon‐γ (IFN‐γ)‐producing T cells and the expression of the interleukin‐12 p40 gene were reduced in TLR‐2‐deficient mice as compared to C57Bl/6 mice early during infection with M. avium strain 2447. The generation of protective CD4+ T cells was also compromised in the mutated mice as compared with the controls. Our data show that TLR‐2 is required for optimal immunity against certain virulent M. avium strains.
Fluvastatin treatment inhibits leucocyte adhesion and extravasation in models of complement-mediated acute inflammation
Authors: FISCHETTI F , CARRETTA R , BOROTTO G , DURIGUTTO P , BULLA R , MERONI P L , TEDESCO F
Published: January 2004

Complement activation plays a relevant role in the development of tissue damage under inflammatory conditions, and clinical and experimental observations emphasize its contribution to inflammatory vasculitides. Statins have recently been shown to reduce cardiovascular morbidity independently of plasma cholesterol lowering and in vitro studies support a direct anti-inflammatory action of these drugs. The aim of this study was to verify the in vivo effect of fluvastatin on complement-mediated acute peritoneal inflammation. The effect of oral treatment with fluvastatin was investigated in normo-cholesterolaemic rats that received intraperitoneal injection of either yeast-activated rat serum (Y-act RS) or lipopolysaccharide to induce peritoneal inflammation monitored by the number of PMN recruited in peritoneal fluid washes. In addition, vascular adherence and extravasation of leucocytes were evaluated by direct videomicroscopy examination on mesentery postcapillary venules topically exposed to Y-act RS. The number of PMN in the peritoneal washes of rats treated with fluvastatin was 38% lower than that of untreated animals (P < 0·05) 12  h after LPS injection, and was even lower (56%) in rats treated with Y-act RS already 8 h after injection (P < 0·02). Firm adhesion to endothelium and extravasation of leucocytes evaluated under direct videomicroscopy observation were significantly inhibited in fluvastatin treated rats (77% and 72%, respectively; P < 0·01), 120 min after treatment with Y-act RS. Our results demonstrate that fluvastatin inhibits in vivo complement-dependent acute peritoneal inflammation and suggest a role for statins in preventing the inflammatory flares usually associated with complement activation in chronic diseases, such as SLE or rheumatoid arthritis.
Qualitative and quantitative abnormalities in splenic dendritic cell populations in NOD mice
Authors: VASQUEZ A C , FEILI-HARIRI M , TAN R J , MOREL P A
Published: January 2004

The phenotype and function of splenic DC populations from diabetes-prone NOD mice were chara-cterized and compared to DC from diabetes-resistant strains in the presence or absence of Flt3 ligand (FL) treatment. NOD mice were found to have significantly fewer CD8α+ DC than both B10.BR and C57BL/6 mice, and this defect was reversed by FL treatment. Freshly isolated CD8α+ and CD8α– DC from all three strains were found to express similar levels of costimulatory molecules and this was similar in both FL-treated and untreated animals. IL-12 p40 production was significantly lower in purified CD11c+ DC from NOD mice compared to DC from C57BL/6 or B10.BR mice. CD8α+ DC isolated from NOD mice produced lower levels of IL-12p40 than CD8α+ DC from C57CBL/6 and this was dependent on the nature of the stimulus given. In contrast both CD8α+ and CD8α– DC from FL-treated mice produced high levels of IL-12p40 following activation, but only the CD8α– DC produced IL-12p70. Functionally, freshly isolated CD8α– DC were more stimulatory than CD8α+ DC in a primary allogeneic mixed lymphocyte reaction. However, DC maturation resulted in increased T cell stimulatory capacity for both DC subsets, and this pattern was seen in all strains. These results demonstrate significant differences in phenotype and function of splenic NOD CD8α+ DC, and further suggest that FL treatment may reverse some of these abnormalities.
Stimulation of innate immunity by susceptible and multidrug-resistant Pseudomonas aeruginosa: an in vitro and in vivo study
Authors: GIAMARELLOS-BOURBOULIS E J , PLACHOURAS D , TZIVRA A , KOUSOULAS V , BOLANOS N , RAFTOGIANNIS M , GALANI I , DONTAS I , DIONYSSIOU-ASTERIOU A , GIAMARELLOU H
Published: January 2004

In attempt to investigate the stimulatory effect of Pseudomonas aeruginosa on innate immunity and to correlate it to its level of resistance to antimicrobials, 20 isolates were applied; 8 isolates were susceptible and 12 multidrug-resistant. Genetic diversity was defined by PFGE. Human monocytes of two healthy volunteers were in vitro stimulated by the isolates for the production of pro-inflammatory (TNF-α, IL-1β, IL-6, IL-8 and IL-12) and anti-inflammatory cytokines (IL-10), of malondialdehyde and of procalcitonin. Cytokines were estimated by EIA, malondialdehyde by the thiobarbiturate assay and procalcitonin by an immunochemiluminometric assay. Survival of 48 Wistar rats was recorded after induction of sepsis by the intraperitoneal injection of three susceptible and three multidrug-resistant isolates. To test whether comparative effect of the latter isolates on survival correlates with any difference of monocyte-mediated release of pro-inflammatory mediators, monocytes of two rats were in vitro stimulated for the production of TNF-α and of malondialdehyde. In vitro stimulation of human monocytes by the susceptible isolates elicited elevated production of malondiadeheyde, of IL-1β and of IL-6 compared to stimulation by multidrug-resistant isolates. Similar differences were found for TNF-α and IL-8, but they were not statistically significant. Production of IL-10 and IL-12 was not detected after stimulation with any isolate. Levels of procalcitonin were similar after induction with either susceptible or multidrug-resistant isolates. Mean survival of animals was 7·56, 21·80 and 55·20 h, respectively, after challenge by the susceptible isolates and 28·89, 61·8 and more than 120 h, respectively, after challenge by the multidrug-resistant isolates. Differences of survival were accompanied by greater rodent monocyte-release of TNF-α and malondialdehyde after stimulation by the susceptible isolates compared to multidrug-resistant ones. It is concluded that considerable differences are encountered on the stimulation of human monocytes by susceptible and resistant isolates of Pseudomonas aeruginosa. These results correlate with in vivo evidence and might influence decision on therapeutics.
Detection of HLA class II-dependent T helper antigen using antigen phage display
Authors: SOMASUNDARAM R , SATYAMOORTHY K , CAPUTO L , YSSEL H , HERLYN D
Published: January 2004

Major histocompatibility complex (MHC) class II-dependent antigens not only activate CD4+ T helper (Th) cells, but also cytolytic T lymphocytes and effector cells of the innate immune system. These antigens therefore are candidate vaccines against cancer and infectious agents. We have developed a novel approach using a model antigen, tetanus toxoid (TT), which provides the basis for the establishment of a novel strategy of cloning Th antigens. In the TT model system, a cDNA library encoding part of the TT light chain which contained a TT-associated Th epitope recognized by TT-specific Th clones was displayed on a phage vector (TT-phage) and presented to TT-specific Th cells by autologous Epstein-Barr virus-transformed B cells (APC). These TT-phages were able to specifically activate two different TT-specific CD4+ Th cell lines as demonstrated both in [3H]thymidine incorporation and cytokine release assays. Th cell stimulation by TT-phages was significant at a ratio of one TT-phage in 50 irrelevant phages. The described approach provides the basis for the development of a novel strategy of cloning MHC class II-dependent Th antigens, using available Th cells. This strategy has several potential advantages over existing antigen cloning methods or biochemical peptide isolation.
Expression of CTLA-4 in nonhuman primate lymphocytes and its use as a potential target for specific immunotoxin-mediated apoptosis: results ofin vitrostudies
Authors: PALMISANO G L , TAZZARI P L , COZZI E , BOLOGNESI A , POLITO L , SEVESO M , ANCONA E , RICCI F , CONTE R , STIRPE F , FERRARA G B , PISTILLO M P
Published: January 2004

T-cell-mediated immunoregulation is one of the main mechanisms implicated in induction and maintenance of transplantation tolerance. In this regard, deletion or modulation of xeno/alloantigen-specific T cells, as well as blocking of their interactions with other cell populations, are currently being pursued for tolerance induction in humans as well as nonhuman primates. In order to investigate whether cytotoxic T-lymphocyte antigen-4 (CTLA-4) may represent a suitable target for a T cell depletion approach in nonhuman primate models, we analysed CTLA-4 expression in peripheral blood mononuclear cells (PBMCs) from nonhuman primates and the potential role of two anti-CTLA-4 saporin-conjugated immunotoxins. The analysis was performed in PBMCs from 8 cynomolgus monkeys from Philippines and from Mauritius both at protein level by flow cytometry and at transcriptional level by RT-PCR. In addition, the apoptotic role of the immunotoxins was investigated. The results showed that CTLA-4 was expressed at variable levels depending on the origin of the cynomolgus monkeys and the resting or activated cell condition. CTLA-4 was not expressed on resting Mauritius PBMCs and showed a lower up-regulation upon PMA/PHA activation compared to the Philippines PBMCs that expressed CTLA-4 also before activation. Two CTLA-4 RNA transcripts (672 and 550 bp) were detected with levels variations after cell stimulation. Two anti-CTLA-4 immunotoxins induced in vitro apoptosis of activated PBMCs from both sources of cynomolgus monkeys. This is the first report that documents CTLA-4 expression both at protein and transcriptional level by nonhuman primate PBMCs and provides novel perspectives of xeno/allograft rejection immunotherapy based on CTLA-4 targeting.
IL-12 enhances the generation of tumour antigen-specific Th1 CD4 T cells duringex vivoexpansion
Authors: KNUTSON K L , DISIS M L
Published: January 2004

CD4+ T cells are essential for the immune response against cancer. Vaccination against cancer will likely only be effective at preventing growth of micrometastatic disease while adoptive T cell therapy will be better suited for eradication of bulky pre-existing disease (Knutson et al. Expert Opin Biol Ther 2002; 2:55–66). Problems with the use of adoptive T cell therapy include lack of CD4+ T cell help, low frequency of antigen-specific T cells, and lack of effective ex vivo expansion techniques. In this study, we focused on improving ex vivo expansion of CD4+ T helper cells. The effects of IL-12, along with IL-2, on the ex vivo generation of HER-2/neu antigen-specific T cells were examined. Patients were immunized with a peptide-based vaccine that contained a helper epitope, p776–790, derived from the intracellular domain of HER-2/neu. While T cell immunity to p776–790, assessed by proliferation assays, could be readily measured in short-term cultures, cell line generation by multiple in vitro stimulation with peptide and IL-2 as the only added cytokine resulted in loss of antigen-specific proliferation. The inclusion of IL-12, along with IL-2, restored antigen-specific proliferation in a dose-dependent fashion. The resulting p776–790-specific T cells responded readily to antigen by proliferating and producing type I cytokines (IFN-γ and TNF-α). The increased proliferative response of the cultures was due in part to an increase in the number of HER-2/neu-specific T cells. These results suggest that IL-12 is an important cytokine for ex vivo recovery and maintenance of antigen-specific CD4+ T lymphocytes that would otherwise be lost by using IL-2 alone in combination with antigen. Furthermore, these results have important implications for ex vivo expansion of CD4+ T cell for use in anti-tumour adoptive immunotherapy.
Increased Toll-like receptor 4 expression in infants with respiratory syncytial virus bronchiolitis
Authors: GAGRO A , TOMINAC M , KRŠULOVIĆ-HREŠIĆ V , BAĆE A , MATIĆ M , DRAŽENOVIĆ V , MLINARIĆ-GALINOVIĆ G , KOSOR E , GOTOVAC K , BOLANČA I , BATINICA S , RABATIĆ S
Published: January 2004

The fusion protein of the respiratory syncytial virus (RSV) binds to the pattern recognition receptors, TLR4 and CD14, and initiates innate immunity response to the virus. The aim of the study was to investigate the expression of TLR4 on peripheral blood lymphocytes and monocytes in peripheral blood of infants in both acute and convalescent phase of RSV bronchiolitis (n = 26). In addition, TNF-α expression in lipopolysaccharide-stimulated monocytes was also assessed. The results showed TLR4 to be expressed predominantly by monocytes in both sick infants and controls. During the acute phase of infection monocytes up-regulated TLR4 in eight infants, which returned to the levels recorded in controls 4–6 weeks from infection. There was no difference in the percentage of TNF-α secreting monocytes. Of the clinical parameters tested, minimal oxygen saturation was found to correlate negatively with this expression in the group of infants with increased TLR4. Additional studies are under way to correlate this finding with the outcome of the immune response to RSV.
Low plasma level of adiponectin is associated with stavudine treatment and lipodystrophy in HIV-infected patients
Authors: LINDEGAARD B , KELLER P , BRUUNSGAARD H , GERSTOFT J , PEDERSEN B K
Published: January 2004

This study tested the hypothesis that in patients with HIV-associated lipodystrophy, adiponectin levels were related to insulin resistance, TNF-α and IL-6 and treatment with nucleoside analaogues. HIV seropositive men undergoing highly active antiretroviral treatment were enrolled into three predetermined clinical groups: lipodystrophy with central fat accumulation (n = 12); lipodystrophy without central fat accumulation (n = 15); no lipodystrophy (n = 15). HIV-negative healthy men served as controls (n = 12). Both lipodystrophic groups had a low percentage of limb fat compared to the two control groups. Patients with lipodystrophy with fat accumulation had increased truncal fat compared with controls. Levels of adiponectin did not correlate with either TNF-α or IL-6. Low levels of adiponectin were found in both lipodystrophic groups and were associated with current or previous treatment with stavudine. Furthermore, the adiponectin level correlated with the percentage of limb fat. Patients with lipodystrophy with fat accumulation were more insulin resistant, measured by HOMA-IR, compared with controls. However, HOMA-IR did no correlate to adiponectin or other cytokines. In conclusion, the finding of no difference between the two lipodystrophic groups with regard to adiponectin, indicates that low levels of adiponectin reflects fat atrophy, whereas the insulin resistance was best explained by increased truncal fat mass.
Cellular immunity induced by the recombinant Plasmodium falciparum malaria vaccine, RTS,S/AS02, in semi-immune adults in The Gambia
Authors: PINDER M , REECE W H H , PLEBANSKI M , AKINWUNMI P , FLANAGAN K L , LEE E A M , DOHERTY T , MILLIGAN P , JAYE A , TORNIEPORTH N , BALLOU R , MCADAM K P M J , COHEN J , HILL A V S
Published: January 2004

Vaccination of malaria-naive humans with recombinant RTS,S/AS02, which includes the C-terminus of the circumsporozoite protein (CS), has been shown to induce strong T cell responses to both the whole protein antigen and to peptides from CS. Here we show that strong T cell responses were also observed in a semi-immune population in The Gambia, West Africa. In a Phase I study, 20 adult male volunteers, lifelong residents in a malaria-endemic region, were given three doses of RTS,S/AS02 at 0, 1 and 6 months. Responses to RTS,S, hepatitis B surface antigen and peptides from CS were tested using lymphocyte proliferation, interferon (IFN)-γ production in microcultures, and IFN-γ ex vivo and cultured ELISPOT, before and after vaccination. Cytotoxic responses were tested only after vaccination and none were detected. Before vaccination, the majority of the volunteers (15/20) had detectable responses in at least one of the tests. After vaccination, responses increased in all assays except cytotoxicity. The increase was most marked for proliferation; all donors responded to RTS,S after the third dose and all except one donor responded to at least one peptide after the second or third dose. There was a lack of close association of peptide responses detected by the different assays, although in microcultures IFN-γ responses were found only when proliferative responses were high, and responses by cultured ELISPOT and proliferation were found together more frequently after vaccination. We have therefore identified several peptide-specific T cell responses induced by RTS,S/AS02 which provides a mechanism to investigate potentially protective immune responses in the field.
Inter‐observer reliability of radiological signs of necrotising enterocolitis in a population of high‐risk newborns
Authors: Di Napoli Anteo , Di Lallo Domenico , Perucci Carlo A. , Schifano Patrizia , Orzalesi Marcello , Franco Francesco , De Carolis Maria Pia
Published: January 2004

Radiological examination of the abdomen is critical to the diagnosis of necrotising enterocolitis (NEC). Previous studies on the reproducibility of radiological findings have been limited in size and based only on infants with the disease. We conducted a study among a sample of high‐risk infants with and without the diagnosis of NEC: (1) to measure the degree of inter‐observer agreement of NEC radiological diagnosis and signs and (2) identify a profile of radiological signs which led the observers to make a diagnosis of NEC.

We collected 297 X‐rays from a sample of 57 newborns admitted in 1999 to neonatal intensive care and neonatal surgery units in Rome, Italy. Three specialists in paediatric radiology examined the films independently and without any clinical information about patients. The analyses were conducted on a total of 891 forms filled in by the observers. Kappa values were calculated to measure the inter‐observer reliability. To identify the profiles of radiological signs, a multidimensional analysis, binary segmentation, was carried out. The reproducibility of radiographic signs was 0.55 (P < 0.01) for diffuse gaseous intestinal distention, 0.22 (P < 0.01) for bowel wall thickening, 0.10 (P < 0.01) for presence of portal venous gas and 0.29 (P < 0.01) for pneumatosis intestinalis. The agreement for radiographic diagnosis suspected/confirmed of NEC was 0.31 (P < 0.01). Among the 23 possible combinations of radiographic signs, the three radiologists indicated four profiles that produced a diagnosis of NEC containing, respectively, two, three, four and five signs.

Our study found, in a large sample of radiographs selected from a population of infants with and without NEC, a poor reliability for NEC diagnosis and individual radiological signs among three expert radiologists. Clinical information and the presence of more than one radiological sign can reduce the margin of observer's error that inevitably exists when dealing with a diagnosis as difficult as NEC.
Repeated validation of parental self‐reported smoking during pregnancy and infancy: a prospective cohort study of infants at high risk for allergy development
Authors: Christensen Anne E. , Tobiassen Mette , Jensen Tina K. , Wielandt Hanne , Bakketeig Leiv , Høst Arne
Published: January 2004

Exposure to environmental tobacco smoke (ETS) during fetal life and infancy is closely related to the smoking habits of the parents. Estimates of exposure to ETS require valid and detailed information on changes in cigarette smoking over time. The objective was to test the validity of self‐reported smoking among parents during pregnancy and early childhood in a cohort of children at high risk for allergy development by measurement of exhaled carbon monoxide (CO). The cohort comprised 117 families enrolled from the general population of pregnant women at admission to antenatal care.

Data on parental tobacco smoking were obtained by interview and exhaled CO was measured (Micro‐Smokerlyzer®) in parents twice during pregnancy and when the child was 6 and 18 months old. The median (range) exhaled CO levels were 3 (0–10) parts per million (ppm) for non‐smokers and 15 (1–39) ppm for smokers (P < 0.0005). A receiver operating characteristic (ROC) analysis was performed at each examination. The areas under the ROC curve were high for both mothers (between 0.88 and 0.99) and fathers (between 0.87 and 0.89), indicating exhaled CO as a good diagnostic tool for determining smoking status. Comparing the ROC areas obtained for mothers from late pregnancy and during infancy with the area from early pregnancy showed no statistical differences (P = 0.21, 0.43 and 0.44 respectively) and the same was true for fathers during infancy (P = 0.81). The level of 8 ppm was used as the cut‐off between smokers and non‐smokers, based on data from a pilot study. Using CO as a diagnostic tool for smoker status showed very high specificity (between 97 and 100%), indicating that very few persons claiming to be non‐smokers had CO levels higher than 8 ppm.

In conclusion, the validity of interview‐obtained self‐reported smoking among parents during pregnancy and early childhood was high. Repeated interviews and CO measurements in a prospective study design did not change the validity, indicating a low risk of information bias. A structured interview combined with measurement of exhaled CO is a valid and reliable method for estimating ETS exposure to the fetus and young infant.
Growth, development and health from early fetal life until young adulthood: the Generation R Study
Authors: Hofman Albert , Jaddoe Vincent W. V. , Mackenbach Johan P. , Moll Henriette A. , Snijders Rosalinde F. M. , Steegers Eric A. P. , Verhulst Frank C. , Witteman Jacqueline C. M. , Büller Hans A.
Published: January 2004

In this paper the Generation R Study is presented. This study examines growth, development and health in urban children from fetal life until young adulthood. With an integrated approach of epidemiological, clinical and basic research, it focuses on four primary areas of research: (1) growth and physical development; (2) behavioural and cognitive development; (3) diseases in childhood; and (4) health and health care for pregnant women and children. The general aims of the study are:

1. to describe normal and abnormal growth, development and health from fetal life until young adulthood in a multiethnic population‐based cohort;

2. to identify biological, social and environmental determinants of normal and abnormal growth, development and health from fetal life until adulthood;

3. to examine the utilisation and effectiveness of current strategies for prevention and early identification of groups at risk.

Eventually, this study will contribute to the development of strategies for optimising health and health care for pregnant women and children. The Generation R Study is a prospective population‐based cohort study in Rotterdam, the Netherlands. In this urban setting, 10 000 children will be examined from early fetal life until young adulthood. Data are collected by physical examinations, questionnaires, interviews, ultrasound and biological samples. The study entered its pilot phase to test the logistics in December 2001. Full participant recruitment and complete data collection started in 2002.
Risks of adverse pregnancy outcomes among Inuit and North American Indian women in Quebec, 1985–97
Authors: Luo Zhong‐Cheng , Wilkins Russell , Platt Robert W. , Kramer Michael S. ,   for the Fetal and Infant Health Study Group of the Canadian Perinatal Surveillance System
Published: January 2004

We used Statistics Canada's linked stillbirth, live birth and infant death files to assess the risks of adverse pregnancy outcomes among Inuit and North American Indian vs. other ethnic women in Quebec, 1985–97 (1 125 462 singleton births). Mother tongue was used to define ethnicity, with the largest French language group as the reference. Main  outcome  measures  are  adjusted  odds  ratios  (AOR)  for  preterm  birth,  small‐for‐gestational‐age (SGA), stillbirth, neonatal and postneonatal death controlled for maternal age, education, marital status, parity, infant sex, community size, and community‐level random effects using multilevel logit models.

Inuit women had higher risks of preterm birth (AOR = 1.49, 95% CI [1.25, 1.78]) and immaturity‐related  infant  mortality  (AOR = 3.03  [1.36,  6.74]),  while  Indian  women did not. Infants of Inuit (AOR = 0.39 [0.31, 0.49]) and Indian (AOR = 0.27 [0.24, 0.31]) women had substantially lower risks of SGA. Elevated risks of stillbirth were observed among Indian women [AOR = 1.53 (1.09, 2.15)], and of postneonatal death among both Inuit (AOR = 4.45 [2.74, 7.22]) and Indian (AOR = 1.86 [1.28, 2.70]) infants. Both Inuit and Indian infants had much higher risks of sudden infant death syndrome (SIDS) and infection‐related mortality. Although the absolute risks of adverse outcomes declined from 1985–87 to 1995–97, the relative disparities between aboriginal and non‐aboriginal women changed little over this period. We conclude that Inuit and Indian women have different risk profiles for adverse pregnancy outcomes, and that prevention of preterm birth among Inuit women, and of SIDS and infection‐related infant mortality in both aboriginal groups, are important targets for future research and intervention.
New evidence for the Theory of the Stork
Authors: Höfer Thomas , Przyrembel Hildegard , Verleger Silvia
Published: January 2004

Data from Berlin (Germany) show a significant correlation between the increase in the stork population around the city and the increase in deliveries outside city hospitals (out‐of‐hospital deliveries). However, there is no correlation between deliveries in hospital buildings (clinical deliveries) and the stork population. The decline in the number of pairs of storks in the German state of Lower Saxony between 1970 and 1985 correlated with the decrease of deliveries in that area. The nearly constant number of deliveries from 1985 to 1995 was associated with an unchanged stork population (no statistical significance). However, the relevance of the stork for the birth rate in that part of Germany remains unclear, because the number of out‐of‐hospital deliveries in this area is not well documented. A lack of statistical information on out‐of‐hospital deliveries in general is a severe handicap for further proof for the Theory of the Stork.
Association of head circumference at birth among sibling pairs
Authors: Samuelsen Sven O. , Stene Lars C. , Bakketeig Leiv S.
Published: January 2004

The objective of the study was to estimate and compare the correlation coefficients of head circumference and weight at birth among sibling pairs. Pairs of singleton siblings were ascertained among children born in Norway to the same mother between 1978 and 1997. Head circumference, birthweight and other perinatal factors were registered in the Medical Birth Registry of Norway. Head circumference measurements were obtained for first‐ and second‐born in 287 448 sibling pairs. The correlation coefficient of head circumference among first‐ and second‐born siblings was 0.343 (standard error 0.002) compared with 0.477 (standard error 0.002) for birthweight. These results were similar for later‐born sibling pairs and only slightly influenced by other factors such as sex, fatherhood and time between pregnancies. Sensitivity analyses showed that substantial measurement error is required to explain the lower correlation for head circumference. In conclusion, the sibling correlation of head circumference was consistently weaker than that of birthweight.
Racial differences in leading causes of infant death in the United States
Authors: Muhuri Pradip K. , MacDorman Marian F. , Ezzati‐Rice Trena M.
Published: January 2004

We used linked birth/infant death records of over 23 million singletons belonging to six birth cohorts (1989–91 and 1995–97) and examined changes in race differentials in the overall and cause‐specific infant mortality risks across time in the United States. Results show that infant mortality declined for all races during the time period, with disproportionately greater declines among non‐Hispanic American Indians (AIs). Among the leading causes of infant death, declines in mortality from sudden infant death syndrome (SIDS), respiratory distress syndrome (RDS) and congenital anomalies contributed the most to the overall decline in infant mortality in the 1995–97 cohorts, compared with the 1989–91 cohorts. Disproportionately greater reductions in mortality resulting from SIDS and congenital anomalies led to more rapid mortality declines among non‐Hispanic AIs than for other races. There are disturbing findings that infants of almost every race experienced increases in mortality from newborn affected by maternal complications of pregnancy (maternal complications) and that none of the race groups experienced a significant decline in mortality from disorders resulting from short gestation/low birthweight.
Measurement of activated clotting time in children – comparison of the Celite i‐STAT® ACT with the Medtronic ACT® II
Authors: Hug M. I. , Di Bernardo S. , Berger F. , Bauersfeld U. , Weiss M.
Published: January 2004

Background:  To evaluate a recently introduced blood‐saving method for the measurement of activated clotting time (ACT), the Celite i‐STAT® ACT, by comparing the values obtained with those from the widely used Medtronic ACT® II device.

Methods:  In a prospective clinical study, we compared ACT values from the i‐STAT® device with the Medtronic ACT® II device in 60 paediatric, interventional, cardiac catheterization procedures necessitating prophylactic heparinization. Blood samples were pair‐analyzed using two i‐STAT® analyzers and one Medtronic ACT® II device with double‐tube‐cartridges before and after heparinization. Data were compared using Bland‐Altman bias analysis, Student's t‐test and simple regression analysis.

Results:  Bias and precision between the Medtronic ACT® II and the i‐STAT® ACT values was −5.4 ± 26.3. The i‐STAT® and Medtronic ACT® II values were similar before heparinization (P = 0.22), but i‐STAT® ACT values became significantly longer than Medtronic ACT® II values after heparinization (P = 0.021). The increase of ACT after heparinization was greater (median 86.3; range 40–187 s) in the i‐STAT® analyzer than in the Medtronic ACT® II analyzer (median 73.0; range 19–235.5 s). Increase in ACT values was inversely affected by intraoperative haemoglobin concentration in the Medtronic ACT® II analyzer (P = 0.001) but not in the i‐STAT® analyzer.

Conclusion:  Activated clotting times obtained from the Celite i‐STAT® ACT and the Medtronic ACT® II demonstrated poor agreement. The technical principles are quite different and the two methods showed diverse susceptibility to intraoperative haemoglobin concentration. Users have to be aware of technique‐specific ACT target ranges and their confounders, which need to be provided by the manufacturers.
Fetal alcohol syndrome: Diagnosis, epidemiology, and developmental outcomes
Authors: O'Leary CM
Published: January 2004

Abstract:  In Australia the issue of fetal alcohol syndrome (FAS) has not been the subject of policy development or of extensive research. There is a lack of knowledge, both in the general community and by health professionals, of the nature of the risks associated with heavy alcohol consumption during pregnancy and the factors that increase this risk. This paper reviews the literature surrounding FAS with the aim of providing the reader an understanding of the diagnostic features and epidemiology of FAS and of the developmental sequelae associated with this syndrome.
Slight and mild hearing loss in primary school children
Authors: Wake M , Poulakis Z
Published: January 2004

Abstract:  Although slight/mild sensorineural hearing loss affects about 3% of the school‐aged population, with many more children having such impairments at single frequencies or in only one ear, little is known about its impacts on language, learning, and quality of life. This annotation explores what is known about prevalence and impact of this condition and argues for large‐scale research to better address these issues.
Perinatal mortality 1982−2001 at Vila Central Hospital, Vanuatu
Authors: Grace RF , Everard LB
Published: January 2004

Objective:  To determine the perinatal mortality at Vila Central Hospital (VCH) and thereby make an estimate of Vanuatu's perinatal mortality.

Methods:  Labour floor and nursery records from VCH, Vanuatu's only tertiary referral hospital, were examined and collated for the period 1982−2001. Death certificate books were also examined. The perinatal mortality rate was calculated as the sum of stillbirths (infants = 20 weeks and/or = 500 g) and neonatal mortality (deaths <28 days). Premature infants were recorded for infants less than 37 weeks gestation and small‐for‐gestational‐age for infants less than 2500 g.

Results:  The mean perinatal mortality rate for the period was 27/1000, (range 15−36/1000). The mean stillbirth rate was 15/1000, (range 8−22/1000). The neonatal mortality was 12/1000, (range 4−18/1000). Premature babies accounted for 37/1000 births, (range 11−80/1000). Small‐for‐gestational‐age infants were recorded in 45/1000 births, (range 16−83/1000). The mean twinning rate was 12/1000. Seven spontaneous triplets, two quadruplets and two pairs of siamese twins were born. Sepsis, meconium aspiration and birth asphyxia were the commonest causes of neonatal death.

Conclusion:  Accurate data collection is difficult in Vanuatu. There is the risk of under‐ascertainment however, the figure of 27/1000 represents the current best possible estimate of perinatal mortality at VCH over the last 20 years. VCH is Vanuatu's premier hospital and it is likely that the national figure for perinatal mortality is higher, probably in the range of 37−39/1000. This places Vanuatu's perinatal mortality at a level 30 years or more behind Australia. These figures represent the largest account of hospital based perinatal data collected from any Pacific island nation.
Memory function in childhood epilepsy syndromes
Authors: Nolan MA , Redoblado MA , Lah S , Sabaz M , Lawson JA , Cunningham AM , Bleasel AF , Bye AME
Published: January 2004

Objective:  Children with epilepsy are at risk of specific cognitive deficits. We aimed to compare and characterize the memory function of children with childhood absence epilepsy (CAE), frontal lobe epilepsy (FLE) and temporal lobe epilepsy (TLE).

Methods:  Epilepsy syndrome was identified by clinical data, seizure semiology, interictal and ictal electroencephalogram (EEG). Seventy children aged 6−18 years with CAE, FLE or TLE had neuropsychological assessment including memory function. After adjusting for epilepsy variables, neuropsychological results of the syndrome groups and normative data were compared.

Results:  Children from all three syndrome groups were at risk of memory difficulties. The duration of epilepsy correlated negatively with memory function. Children with TLE had the worst memory function, significantly lower in verbal memory tasks than children with CAE (P = 0.02) and children with FLE (P = 0.01). The performance of children with TLE was significantly below the normed mean across all verbal and most visual tasks. Compared to the normed means, children with FLE had results that were statistically lower in some verbal and visual tasks, and children with CAE were lower in two visual tasks only.

Conclusions:  This study demonstrates memory dysfunction in three common childhood epilepsy syndromes. Children with TLE had the greatest impairment, children with FLE had memory difficulties not previously reported, and children with CAE had subtle memory deficits. Qualitative differences were also evident. Longer duration of intractable epilepsy was associated with reduced memory ability. Memory function and its potential impact on academic achievement are vital considerations when managing children with epilepsy.
Clinical profile of children with nephrotic syndrome not on glucorticoid therapy, but presenting with infection
Authors: Alwadhi RK , Mathew JL , Rath B
Published: January 2004

Objective:  This is a prospective, hospital based study over a period of one year, to examine the clinical profile of nephrotic children not on glucocorticoid therapy, but presenting with infection.

Methods:  Sixty‐eight children with 76 episodes of nephrotic syndrome were enrolled during the study year. They were examined for evidence of infection using clinical and investigative criteria. The children were monitored daily for proteinuria and improvement of infection. After the infection was controlled, prednisolone therapy was started in those who were not already in remission.

Results:  Of the 76 episodes, eight were excluded from analysis as they had developed infection while they were on glucocorticoid therapy. Of the remaining 68 nephrotic episodes in 60 children, there was evidence of infection in 57 episodes (83.8%). Upper respiratory infection was the most common (28.0%) followed by urinary tract infection (22.8%), peritonitis (15.8%), pneumonia (14.0%), acute invasive diarrhoea (10.5%) and empyema (5.3%). Children with infection had significantly lower serum albumin and higher serum cholesterol compared to non‐infected children. Of the 57 episodes with infection, remission occurred with control of infection alone in nine episodes (15.8%) and glucocorticoid therapy was not required.

Conclusion:  Infection is widely prevalent among Indian children presenting with episodes of nephrotic syndrome, even when they are not on glucocorticoid therapy. Some children may go into remission by control of infection alone. Among the infections, upper respiratory and urinary tract infections are the most common.