Early postnatal illness severity scores predict neurodevelopmental impairments at 10 years of age in children born extremely preterm.

OBJECTIVE: A neonatal illness severity score, The Score for Neonatal Acute Physiology-II (SNAP-II), predicts neurodevelopmental impairments at two years of age among children born extremely preterm. We sought to evaluate to what extent SNAP-II is predictive of cognitive and other neurodevelopmental impairments at 10 years of age. STUDY DESIGN: In a cohort of 874 children born before 28 weeks of gestation, we prospectively collected clinical, physiologic and laboratory data to calculate SNAP-II for each infant. When the children were 10 years old, examiners who were unaware of the child's medical history assessed neurodevelopmental outcomes, including neurocognitive, gross motor, social and communication functions, diagnosis and treatment of seizures or attention deficit hyperactivity disorder (ADHD), academic achievement, and quality of life. We used logistic regression to adjust for potential confounders. RESULTS: An undesirably high SNAP-II (⩾30), present in 23% of participants, was associated with an increased risk of cognitive impairment (IQ, executive function, language ability), adverse neurological outcomes (epilepsy, impaired gross motor function), behavioral abnormalities (attention deficit disorder and hyperactivity), social dysfunction (autistic spectrum disorder) and education-related adversities (school achievement and need for educational supports. In analyses that adjusted for potential confounders, Z-scores ⩽-1 on 11 of 18 cognitive outcomes were associated with SNAP-II in the highest category, and 6 of 18 were associated with SNAP-II in the intermediate category. Odds ratios and 95% confidence intervals ranged from 1.4 (1.01, 2.1) to 2.1 (1.4, 3.1). Similarly, 2 of the 8 social dysfunctions were associated with SNAP-II in the highest category, and 3 of 8 were associated with SNAP-II in the intermediate category. Odds ratios and 95% confidence intervals were slightly higher for these assessments, ranging from 1.6 (1.1, 2.4) to 2.3 (1.2, 4.6). CONCLUSION: Among very preterm newborns, physiologic derangements present in the first 12 postnatal hours are associated with dysfunctions in several neurodevelopmental domains at 10 years of age. We are unable to make inferences about causality.

The new 6-unit individualized curriculum for pediatric residents: the perspective of neonatology fellowship program directors.

OBJECTIVE: Starting in 2013, all pediatric residents entering fellowship must be provided six educational units whose structure is to be determined by their individual career plans. We sought to determine whether (1) neonatology fellowship program directors (PDs) consistently identify certain weaknesses among incoming fellows and (2) neonatology fellowship PDs agree on the most beneficial activities in which pediatric residents should participate to improve preparation for entry into neonatology fellowships. STUDY DESIGN: We sent a 21-question survey focused on the structure and implementation of the 6-unit curriculum to all members of the Organization of Neonatology Training Program Directors. RESULTS: Sixty-seven percent of PDs responded. Seventy-five percent cited insufficient procedural skills as the primary weakness of incoming fellows. More than 80% rated additional training in clinical neonatology, including procedural and resuscitation training, as 'beneficial' or 'highly beneficial'. In contrast, fewer than 40% of PDs gave the same positive ratings to activities broadly focused on scholarship. CONCLUSIONS: The results of the survey may help guide pediatric residency programs as they undertake development of these new curricular initiatives for individual residents entering neonatology.

Bronchopulmonary dysplasia and inflammatory biomarkers in the premature neonate.

Bronchopulmonary dysplasia (BPD) is the most common, serious sequela of premature birth. Inflammation is a major contributor to the pathogenesis of BPD. Often initiated by a pulmonary fetal inflammatory response, lung inflammation is exacerbated by mechanical ventilation and exposure to supplemental oxygen. In response to these initiators of injury, a complex interaction occurs between proteins that attract inflammatory cells (ie, chemokines), proteins that facilitate the transendothelial migration of inflammatory cells from blood vessels (ie, adhesion molecules), proteins that promote tissue damage (ie, pro-inflammatory cytokines and proteases), and proteins that modulate the process (eg, anti-inflammatory cytokines, binding proteins and receptor antagonists). In addition, during recovery from inflammatory injury, growth factors and other substances that control normal lung growth and mediate repair influence subsequent lung structure. In this review, we discuss the role of each aspect of the inflammatory process in the development of BPD. This discussion will include data from measurements of biomarkers in samples of fluid aspirated from the airways of human infants relevant to each phase of inflammation. Despite their limitations, these measurements provide some insight into the role of inflammation in the development of BPD and may be useful in identifying infants at risk for the disease.

Interleukin-10 high producer allele and ultrasound-defined periventricular white matter abnormalities in preterm infants: a preliminary study.

OBJECTIVES: Inflammation plays a role in prematurity, in neonatal disorders of the brain, lung, eye, bowel, and in developmental disability among preterm infants. We initiated a pilot study in preterm children to determine the prevalence of single nucleotide polymorphisms (SNPs) in the infection/inflammation-associated genes for interleukin (IL)-10 (- 1082 G/A), IL-1beta (+ 3953 C/T), tumor necrosis factor (TNF)-alpha (- 308 G/A) and toll-like receptor 4 (TLR-4) (Asp299Gly) and whether these SNPs affect the risk for neonatal disorders. STUDY DESIGN: We genotyped 73 children >/= 2 years of age whose gestational age at birth was < 32 weeks, and explored the associations between genotypes and neonatal disorders and developmental status at age 2 + years. RESULTS: Infants homozygous for the high IL-10 producer - 1082 G-allele (n = 15) were significantly less likely to develop ultrasound-defined periventricular echodensities. A non-significant, but prominent, risk reduction for bronchopulmonary dysplasia, high-grade retinopathy, cerebral palsy, and developmental delay at age 2 + years was present. Polymorphisms in the IL-1beta, TNF-alpha, and TLR-4 genes were too infrequent in our pilot sample to allow for reasonable analysis. CONCLUSION: Infants homozygous for the IL-10 high producer - 1082 G allele might be at reduced risk for prematurity-associated disorders.

ErbB receptor regulation by dexamethasone in mouse type II epithelial cells.

Glucocorticoids stimulate foetal surfactant synthesis. Therefore, they are used in impending pre-term birth. One mechanism of action on surfactant synthesis is through the induction of neuregulin (NRG) secretion by foetal lung fibroblasts. The direct effects on signalling pathways, and specifically on erbB receptors in foetal type II cell surfactant synthesis, are less well understood. The present authors studied the effect of known promoters of foetal surfactant synthesis (namely dexamethasone and mature (i.e. NRG-containing) fibroblast-conditioned medium (FCM)) on erbB receptor activation, protein content and dimerisation patterns in foetal mouse lung type II cells. Dexamethasone inhibited surfactant synthesis in immature type II cells at day (d)16 of gestation, while the mature FCM had stimulatory effects. Both treatments directly stimulated surfactant synthesis in more mature (d17) cells. At this gestational day, dexamethasone had only a small effect on phosphorylation, but it stimulated the protein levels of all four erbB receptors. Dexamethasone effects were distinct from those of mature FCM, which stimulated both protein content and phosphorylation of all erbB receptors and of the signalling intermediate phospholipase Cgamma. Dexamethasone modulated erbB receptor dimerisation patterns, such that erbB2 became the main dimerisation partner for erbB4. In conclusion, dexamethasone signalling involves erbB receptors in foetal type II cells, in a manner similar to, but distinct from, neuregulin-containing fibroblast-conditioned medium signalling.

[Impact of thoracic epidural analgesia on revenue for G-DRG M01B, OPS-301 5-604.0 (radical retropubic prostatectomy)]. / Betriebswirtschaftliche Auswirkungen der thorakalen Epiduralanästhesie bei urologischen Operationen. Dargestellt am Beispiel der G-DRG M01B, OPS-301 5-604.0 (radikale retropubische Prostatektomie).

INTRODUCTION: Using the surgical procedure OPS 5-604.0 (radical retropubic prostatectomy) as an example, our study identifies revenue-relevant patient characteristics and describes the impact of the perioperative application of thoracic epidural analgesia (TEA). METHODS: Factors affecting duration of stay were determined in 460 patients undergoing OPS 5-604.0 in the year 2001 and 2002 using multifactorial regression analysis. Preoperative parameters served as factors for matched-pair analysis of the effects of TEA. RESULTS: Characteristics significantly affecting length of postoperative hospital stay were ASA status, age, preoperative haemoglobin concentration, postoperative tachycardia, number of transfused packed red cells, wound infection and surgical revision. Based on identical matching criteria 27 pairs (with/without TEA) could be formed. While the induction time in the TEA group was 8+/-18 min longer (p=0.04), emergence was briefer by 3+/-9 min (p=0.045). Neither anaesthesia presence time nor anaesthesia costs or total costs of surgery differed significantly between the pairs. However, duration of epidural postoperative pain therapy was longer with TEA but in contrast, the postoperative length of hospital stay after TEA was reduced. Assuming a continuous demand for OPS 5-604.0 procedures, TEA enables 32 more procedures to be carried out per year with an increased yield on turnover of 2.7%. CONCLUSION: At first sight combined anaesthesia procedures require more human resources and material, however, as a result of shortened hospital stay and optimized pain therapy patient satisfaction increases and a substantial potential for increased revenue is gained.

Inhibition of phosphoinositide-specific phospholipase C results in the induction of pathogenesis-related genes in soybean.

The inositol 1,4,5-trisphosphate (IP3) content is decreased in soybean cells following infection with Pseudomonas syringae pv. glycinea (Psg). In this investigation, a differential display approach was applied to isolate soybean genes that are transcriptionally up-regulated by the inhibition of phosphoinositide-specific phospholipase C (PI-PLC) activity and to study if the transcription of those genes is altered following Psg infection. Four genes, transcriptionally activated following treatment with the PI-PLC-specific inhibitor U-73122, were cloned. Three of the four genes were induced following infection with Psg. The transcripts of a hydrolase homologue (GmHy) were induced in the incompatible but not compatible soybean-Psg interaction. The transcripts of a putative ascorbate oxidase gene (GmAO) were induced in both compatible and incompatible interactions. GmHy and GmAO may represent new classes of pathogenesis-related genes. In addition to these two novel genes, homologues of PR-10 and polygalacturonase inhibitor protein (GmPR10 and GmPGIP, respectively) were identified. These two genes have previously been reported as pathogenesis-related. Transcripts of GmPR-10, but not GmPGIP, were induced in both compatible and incompatible soybean-Psg interactions. Induction of these genes, except for GmPGIP, following inhibition of PI-PLC by either the U-73122 treatment or bacterial infection suggests that PI-PLC may negatively regulate the expression of defence genes.

The matrix metalloproteinase gene GmMMP2 is activated in response to pathogenic infections in soybean.

Matrix metalloproteinases (MMPs) play an important role in host defense responses against pathogens in mammals where their activities lead to the production of antimicrobial peptides. We have identified a novel soybean (Glycine max) metalloproteinase gene, GmMMP2, that is transcriptionally up-regulated in infected tissues. The deduced amino acid sequence indicates that this gene belongs to the MMP family. It is a preproprotein containing an N-terminal signal peptide, a cysteine switch, a zinc-binding catalytic motif, and a C-terminal transmembrane domain. The GmMMP2 expressed in and purified from Escherichia coli exhibited an in vitro enzymatic activity in digesting myelin basic protein. All plant metalloproteinases reported so far have no known functions. However, they have been suggested to be involved in extracellular cell matrix degradation during development or senescence. Our investigations demonstrate that the GmMMP2 transcript levels were rapidly increased in compatible and incompatible interactions of soybean tissues with the oomycete pathogen Phytophthora sojae or the bacterial pathogen Pseudomonas syringae pv. glycinea. In agreement with the GmMMP2 activation, a metalloproteinase activity was gradually increased in suspension-cultured cells following the bacterial infection. GmMMP2 was also activated in response to wounding and dehydration. However, GmMMP2 activation did not correlate with the oxidative burst leading to the hypersensitive response cell death or the tissue senescence progress that involves programmed cell death. Our investigations suggest that GmMMP2 may be involved in a novel defense response of soybean against pathogenic infections.

Fetal growth restriction is not associated with a reduced risk for bilateral spastic cerebral palsy in very-low-birthweight infants.

OBJECTIVE: To evaluate the influence of confounding and sampling bias on the relationship between fetal growth restriction in a very-low-birthweight-defined cohort (VLBW, < or =1500 g) and bilateral spastic cerebral palsy (BSCP) at early school-age. METHODS: Three hundred twenty-four of 407 long-term survivors of a regional cohort of VLBW newborns were followed until age 6 years. We categorized as small for gestational age (SGA) all infants whose birthweight Z-score was below -2 relative to published reference values. Uni- and multivariable logistic regression models were fit to estimate the risk of BSCP associated with SGA in the total sample, in subsamples defined by gestational age, and in a gestational age-matched case-control sample. RESULTS: In the total sample, no child below 28 weeks was SGA, and no child above 32 weeks had an appropriate birthweight for gestational age (AGA). The prevalence of BSCP was 14% in AGA and 2% in SGA infants. In both uni- and multivariable logistic regression analyses of the total sample, SGA was associated with a prominently reduced risk of BSCP (odds ratios range from 0.1 to 0.2, all 95% confidence limits exclude 1.0). However, analyses performed in samples defined by different gestational age cutoffs (24--31 weeks, 28--31 weeks) and in a sample using three gestational age-matched controls per BSCP-case did not show a protection by growth restriction (odds ratios range from 0.8 to 2.2, all 95% confidence limits include 1.0). CONCLUSIONS: In VLBW-defined samples, the apparent protective effect of SGA for BSCP can be explained, at least in part, by the highly skewed distribution of SGA over the available gestational age range. From this follows that study cohorts should be defined by gestational age and not by birthweight. In distorted samples like this one, even controlling for gestational age does not reduce the illusion of a reduced cerebral palsy risk for growth restricted infants. Only restriction of the sample by gestational age and/or matching for gestational age reveals the absence of this apparent protective effect.

Androgen regulation of signaling pathways in late fetal mouse lung development.

During lung development there is tension between positive and negative regulators of fibroblast-epithelial communication controlling type II cell differentiation. A clinical consequence of imbalance of this tension is the increased risk for respiratory distress syndrome in male infants. We hypothesized that chronic intrauterine androgen exposure alters fetal lung fibroblast maturation by down-regulating epidermal growth factor receptor (EGF-R) activity and by up-regulating transforming growth factor-beta receptor (TGFbeta-R) activity, leading to an inhibition of surfactant protein B (SP-B) and -C (SP-C) gene expression in type II cells. We treated pregnant mice with dihydrotestosterone (DHT; 2 mg/day) or vehicle for 7 days, starting on gestational day 11. On day 18, EGF binding, EGF-R phosphorylation, TGFbeta-R binding, and TGFbeta1-induced cell proliferation were studied in sex-specific fibroblast cultures. SP-B and -C messenger RNA levels were measured in whole lungs. Chronic DHT treatment reduced both EGF binding (females to 78+/-8% and males to 65+/-9% of controls) and EGF-induced EGF-R phosphorylation. TGFbeta-R binding was increased (females to 173+/-39% and males to 280+/-64% of controls), and TGFbeta-induced cell proliferation was increased in female cells (231+/-57% of controls). SP-B and -C messenger RNA expression was reduced to 55+/-10% and 75+/-4%, respectively. We conclude that chronic DHT exposure beginning early in lung development alters the balance of growth factor signaling that regulates lung maturation.

Antisense-mediated depletion of potato leaf omega3 fatty acid desaturase lowers linolenic acid content and reduces gene activation in response to wounding.

Fatty acid omega3 desaturases act on membrane lipids to catalyse the formation of trienoic fatty acids, the most abundant in plant tissues being alpha-linolenic acid. This fatty acid is a precursor of jasmonic acid, a plant growth regulator involved in the control of wound-induced gene activation in plants and in the induction of tuberization in potato. We isolated a potato omega3 desaturase cDNA, possibly encoding a plastidial isoform, and used it to investigate its expression pattern throughout plant development and in response to wounding. Plastidial omega3 desaturase gene transcripts accumulate rapidly upon wounding, preceding the jasmonate-dependent induction of the wound-responsive proteinase inhibitor II gene. We generated transgenic potato plants constitutively expressing an antisense RNA to this plastidial omega3 desaturase. Selected transgenic lines in which the cognate omega3 desaturase mRNA is largely depleted show a marked reduction, of up to 60%, in trienoic acids in leaves and tubers. In these lines, a corresponding reduction in jasmonate content and proteinase inhibitor II expression is observed upon wounding. Our results indicate that a reduction in omega3 desaturase mRNA levels compromises the wound-induced activation of proteinase inhibitor II, suggesting that wound-induced synthesis of linolenic acid is required for jasmonic acid production. The antisense-mediated depletion of fatty acid omega3 desaturases is a viable alternative for reducing trienoic fatty acid content in plant species in which a mutant screening approach is not applicable.

Pulmonary function at school-age in surfactant-treated preterm infants.

A follow-up study was conducted in 40 children who had been enrolled in a prospective randomized study of exogenous surfactant therapy for respiratory distress syndrome (RDS) (n = 22; S) or placebo (n = 18; P) to determine long-term pulmonary sequelae of surfactant treatment in premature infants with RDS. At follow-up, mean (SD) age was 6.63 (0.18) and 6.55 (0.23) years for S and P, respectively. Complete lung function tests (LFT) were attempted in all patients. Satisfactory data were obtained in 17/22 surfactant-treated and in 12/18 control children. There was no significant difference between groups for any of the parameters measured. Mean (SD) functional residual capacity (FRC) was 92% (16%) and 90% (21%) predicted, mean (SD) airway resistance (R(aw,exp)) was 122% (25%) and 127% (61%), and mean (SD) forced expiratory volume in 1 s (FEV1) was 104% (12%) and 99% (17%) predicted for S and P. Only maximal expiratory flow at 25% vital capacity (L/s) was significantly below the predicted range in S and P groups, with 74% (23%) and 77% (28%), respectively. To test bronchial hyperreactivity, a simple standardized running test was performed: 4 children in S and 5 in P showed a significant response as defined by clinical airway obstruction or changes in FEV1 and/or R(aw), with no significant difference between groups. Although we found no major abnormalities in lung function and no difference between S and P at early school-age, lack of cooperation during lung function tests makes further follow-up necessary.

Transforming growth factor beta 1 binding and receptor kinetics in fetal mouse lung fibroblasts.

TGFbeta1 inhibits fetal lung maturation in vitro. As TGFbeta1 is present in fetal lung, mechanisms must exist to overcome this inhibition and allow late gestation maturation to progress. We studied the ontogeny of TGFbeta1 binding, and TGFbeta receptor kinetics and subtypes in primary cultures of fetal mouse lung fibroblasts from Day 16 to Day 18 of gestation. TGFbeta1 specific binding in fetal lung fibroblasts declined with advancing gestation. The decrease occurred earlier, and was more pronounced in female fibroblasts (50% decrease) than in the male fibroblasts (29% decrease). Dihydrotestosterone treatment of Day 18 female fibroblasts resulted in a dose-dependent increase in TGFbeta1 binding. Scatchard analysis revealed a decline in receptor number with advancing gestation (Day 16 female Bmax: 7.3 x 10(-16); Day 18 female Bmax: 5.5 x 10[-16]) whereas binding affinities remained constant. Affinity labeling followed by chemical cross-linking and autoradiography showed the three known TGFbeta receptor subtypes at both Days 16 and 18 of gestation. The relative abundance of nonsignaling Type III receptors in comparison to signaling Type II and Type I receptors was increased at Day 18 versus Day 16. Incorporation of [3H]thymidine into DNA after treatment with TGFbeta1 changed from Day 16 to Day 18, consistent with changes previously reported between fetal and adult lung fibroblasts. We conclude that as fetal mouse lung maturation progresses, TGFbeta receptor number decreases in fibroblasts, the relative proportion of nonsignaling versus signaling receptor types increases, and the response to TGFbeta1 stimulation changes. These changes may contribute to overcoming TGFbeta1 inhibition of lung maturation.

Regulation of the epidermal growth factor receptor in fetal rat lung fibroblasts during late gestation.

Lung epithelial cell differentiation is predominantly regulated by mesenchymal-epithelial cell communication. We have previously shown that epidermal growth factor (EGF) positively influences this process, and that EGF receptor (EGF-R) binding in fetal rat lung fibroblasts peaks on d18-19 of gestation, just before the onset of augmented surfactant synthesis. This regulation of EGF-R in late gestation fetal lung fibroblasts may control the timing of mesenchymal-epithelial cell communication leading to surfactant synthesis. Hormones and growth factors exert positive and negative influences on lung development, but whether they regulate the EGF-R is unknown. We hypothesized that positive [EGF, cortisol, retinoic acid (RA)] and negative [transforming growth-factor-beta1 (TGF-beta1), dihydrotestosterone (DHT)] regulators of lung cell development regulate the EGF-R in the fetal lung. We studied EGF-R binding and protein abundance in sex-specific fetal rat lung fibroblasts cultured at d17, d19, and d21. EGF-R binding was significantly elevated after RA (both sexes d17 and d19, females d21) and after DHT (females d19) treatment. EGF and cortisol had minimal or inhibitory effects on EGF-R binding. Western blot analysis showed that the observed changes in EGF-R binding were associated with similar changes in EGF-R protein. We conclude that factors that affect lung maturation continue to regulate EGF-R in a developmental, sex-specific manner during late gestation.

Expression and activity of epidermal growth factor receptor in late fetal rat lung is cell- and sex-specific.

Epidermal growth factor (EGF) augments late fetal lung maturation by advancing the ontogeny of fetal lung development and by stimulating surfactant synthesis. Previous studies have indicated that fibroblastalveolar epithelial cell communications mediate surfactant synthesis in the fetal lung and EGF acts through such a mechanism. We investigated the hypothesis that is differential activity and expression of the epidermal growth factor receptor (EGF-R) in fetal lung fibroblasts during the canalicular stage of lung development mediates EGF effects. To test this hypothesis, we examined fetal rat lung fibroblasts (FLFs) and type II cells of late gestation (canalicular and saccular stages; 17-22 days) by EGF-R binding techniques, SDS-PAGE, and Western blot analysis. Specific EGF binding increased 181% in day 18 female FLFs, with male FLFs exhibiting a similar increase on day 19. In contrast, specific EGF binding was low in type II cells, did not increase during late gestation, and there were no sex-specific differences. SDS-PAGE and Western blot analysis revealed a predominant 170-kDa EGF-R band in fibroblasts that increased with gestation (peak = 19 days), and was stronger in females. Immunoprecipitation of EGF-treated cells demonstrated the tyrosine kinase activity of the identified receptor. In contrast, type II cells showed minimal signal that did not increase until day 21 of gestation. We also examined whole fetal lung sections by immunohistochemistry to determine cell-specific expression of the EGF-R in vivo. Immunohistochemistry revealed specific EGF-R staining in columnar and cuboidal epithelia of small conducting airways and in mesenchyme of epithelial-mesenchymal borders (including subepithelial mesenchyme). In contrast, alveolar epithelia showed minimal staining, while subalveolar mesenchyme EGF-R staining peaked at day 19 of gestation. We conclude that cell-specific and sex-specific differences in EGF-R binding and EGF-R immunolocalization appears in the fetal lung at a developmental stage that is critical for alveolar epithelial cell differentiation. The results suggest a role for EGF-R activation in late fetal alveolar epithelial cell maturation, which is mediated through mesenchymal-epithelial cell communication.

Abscisic acid and jasmonic acid activate wound-inducible genes in potato through separate, organ-specific signal transduction pathways.

Mechanical damage to leaf tissue causes an increase in abscisic acid (ABA) which in turn activates the biosynthesis of jasmonic acid (JA). The resulting higher endogenous JA levels subsequently activate the expression of wound-inducible genes. This study shows that JA induces the expression of different sets of genes in roots and leaves of potato plants. When roots of intact plants were treated with JA, high levels of proteinase inhibitor II (pin2), cathepsin D inhibitor, leucine aminopeptidase and threonine deaminase mRNAs accumulated in the systemic leaves. However, in the treated roots, very low, if any, expression of these genes could be detected. In contrast, a novel, root-specific pin2 homologue accumulated in the JA-treated root tissue which could not be detected in leaves, either systemic or those directly treated with JA. Application of okadaic acid and staurosporine revealed that a protein phosphorylation step is involved in the regulation of this differential response. In leaves, a protein phosphatase is required for the JA-induced expression of pin2 and the other genes analysed. This phosphatase activity is not necessary for the JA-induced expression of a pin2 homologue in roots, suggesting the existence of different transduction pathways for the JA signal in these organs. The requirement of a protein phosphatase activity for JA-mediated gene induction has enabled identification of a JA-independent pathway for ABA induction of pin2 and the other wound-inducible genes. This alternative pathway involves a protein kinase, and appears to be selective for wound-inducible genes. Our data suggest the presence of a complex, organ-specific transduction network for regulating the effects of the plant hormones ABA and JA on gene expression upon wounding.

Serum levels of sCD23 and sCD25 in children with asthma and in healthy controls.

To investigate whether markers of lymphocyte activation are useful markers of disease activity in childhood asthma, we studied serum levels of soluble CD25 (receptor for IL-2) and soluble CD23 (low-affinity receptor for IgE) in 178 children (aged 2-18 years) suffering from mild to moderate asthma (mean asthma severity score: 2, range: 1-4), and in 175 healthy age-matched controls. Levels of sCD23 and sCD25 were inversely related to age. sCD23 was lower in patients with asthma (means per age group: 4.93-2.29 micrograms/l; controls: 6.92-4.11 micrograms/l, P < 0.05), while sCD25 tended to be higher (1601-597 kU/ml, controls: 1350-661 kU/ml, P = NS). sCD25 correlated significantly with asthma severity score (r = 0.41; P < 0.01) and MEF25 (maximum expiratory flow at 25% of vital capacity, r = -0.43; P < 0.05) in children < 10 years, while sCD23 correlated with asthma severity (r = 0.28; P < 0.05) in children > 10 years. On follow-up, levels of sCD25 normalized with clinical improvement. In children with nonatopic asthma, levels of sCD25 were significantly higher than in atopic patients. Our observations provide further evidence of the role of T-cell activation in asthma. Monitoring of lymphocyte activation markers, particularly levels of sCD25, may be useful in the follow-up of asthmatic children.

Promoter elements involved in environmental and developmental control of potato proteinase inhibitor II expression.

The proteinase inhibitor II (pin2) gene family exhibits two different modes of expression. It is, on the one hand, constitutively expressed in flowers of potato and tomato plants. and in potato tubers. On the other hand, its expression is induced in the plant foliage by mechanical wounding. To define cis-regulatory elements involved in pin2 promoter activity, deletion analysis of a potato pin2 promoter has been performed in stably and transiently transformed potato and tobacco plants. Two different elements, a quantitative enhancer and a regulatory element, are required for promoter activity. While functional promoter elements required for pin2 activity in tubers and wounded leaves could not be separated, its expression in flowers is mediated by different cis-acting sequences. Induction of pin2 expression in leaves by treatment with the plant growth regulators abscisic acid and jasmonic acid, and the general metabolite sucrose, depends on the presence of the regulatory element involved in expression in tubers and wounded leaves. Thus, pin2 expression in tubers and wounded leaves apparently results from the action of similar hormonal signals on closely linked promoter elements, while a different signal pathway leads to its constitutive expression in flowers.