Maternal diabetes induces changes in the umbilical cord gene expression.

INTRODUCTION: Since maternal diabetes may affect fetal development and the umbilical cord provides an extension of the fetal vasculature, we decided to investigate cords' biological responses to maternal diabetic milieu. METHODS: Using microarray analysis, we determined the gene expression profiles in the umbilical cords of six neonates born to type 1 diabetic mothers and in six control cords. Umbilical cord tissue was collected immediately after elective cesarean section. Expression data were confirmed by real-time polymerase chain reaction (11 genes). Additionally, the same umbilical cords were analyzed histologically. RESULTS: Two hundred eighty six genes were differentially expressed in the umbilical cords from diabetic pregnancies compared to the controls (fold change ±1.5 and P < 0.01). Maternal diabetes had a major effect on the expression of genes involved in vascular development (Bone morphogenetic protein 4, Delta-like 1, and Notch homolog 4), vessel wall integrity (Collagen type VIII alpha 1, Myocyte enhancer factor 2C, and Matrix metalloproteinase 2), and vascular function (Natriuretic peptide precursor B, Endothelin 1, Endothelin receptor B, Cyclooxygenase 1, and Phosphodiesterase 5A). Maternal diabetes was associated with thicker umbilical vein intima-media layers and larger umbilical vein and artery intima-media areas compared to the controls. DISCUSSION: Maternal diabetic environment seems to alter umbilical cord expression of genes involved in the regulation of vascular development and function with simultaneous umbilical vessel muscle layer thickening. These alterations suggest vascular phenotypic modifications, which in turn may lead to long-term vascular consequences in various tissues in infants of diabetic mothers.

Phospholipase A2 in meconium-induced lung injury.

Although the triggering mechanisms of tissue inflammation and injury in meconium-contaminated lungs are still unclear, there is increasing evidence to suggest a central role for phospholipase A(2)'s (PLA(2)). In fact, elevated PLA(2) activities together with high enzyme concentrations, especially the amount of pancreatic (group I) secretory PLA(2) (PLA(2)-I), have been detected in human meconium and in meconium-contaminated lungs. Recent data from our laboratory further indicate that human pancreatic PLA(2), introduced in high amounts within aspirated particulate meconium, is a potent inducer of lung tissue inflammatory injury. Our finding of elevated human PLA(2)-I concentrations in plasma during the first hours after intratracheal meconium administration in newborn piglets further suggests that intrapulmonary aspiration of meconium could also have systemic inflammatory and injurious effects. This, however, remains to be studied in further detail.

Study orientations of graduate entry medical students.

BACKGROUND: In order to meet the changing needs of health care, the University of Turku has introduced a graduate entry programme aimed at students with previous education and experience in health care professions. AIMS: In this study, we look at the study performance of students with different educational backgrounds with special emphasis on graduate entry students. METHODS: We surveyed the study orientations of 145 first-year medical students with different educational backgrounds in the Medical Faculty of the University of Turku, Finland. Special emphasis was placed on graduate entry students (n = 25) with previous education and work experience in health care professions. The students were characterized based on student records and the questionnaire Inventory of General Study Orientation (IGSO). RESULTS: Our results revealed that after the first year of medical studies the graduate entry students showed exceptionally strong theoretical and practical commitment to their studies with a strong work-life orientation which makes them a distinct group among medical students.

Gene expression of pulmonary cytokines after sevoflurane or thiopentone anaesthesia in pigs.

BACKGROUND: Volatile anaesthetics have diverse inflammatory effects on the lungs. They increase gene expression of some pro-inflammatory cytokines in alveolar macrophages whereas in alveolar type II cells they seem to decrease secretion and gene expression of pro-inflammatory cytokines. We have previously detected increased leukotriene C4, nitrate and nitrite concentrations in bronchoalveolar lavage fluid after sevoflurane anaesthesia. In the current study, we measured gene expression of inflammatory cytokines in the lung tissue and plasma concentrations of cytokines in pigs after thiopentone or sevoflurane anaesthesia. METHODS: Sixteen pigs were randomly selected to receive either a continuous thiopentone infusion (control group, n = 8) or sevoflurane (n = 8) at 4.0% inspiratory concentration (1.5 MAC) in air for 6 h. Tissue samples were collected at the end of the study for measurement of gene expression of inflammatory cytokines. Blood samples were collected during anaesthesia for measurement of plasma cytokine concentrations. RESULTS: Compared with thiopentone anaesthesia, lower gene expression of tumour necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) in lung tissue was observed after sevoflurane anaesthesia. Of measured cytokines IL-1beta, TNF-alpha, IL-6, IL-8 and IL-10 only plasma concentrations of IL-6 could be measured during the study without a difference between the groups. CONCLUSION: Lower gene expression of TNF-alpha and IL-1beta was found in the intact porcine lung tissue after sevoflurane anaesthesia compared with thiopentone anaesthesia. Clinical significance of this finding is unknown.

Inflammatory response and apoptosis in newborn lungs after meconium aspiration.

An important feature of meconium-instilled newborn lungs is an inflammatory response and apoptotic cell death. It was recently demonstrated by our group and supported by several other investigators in a relatively short period of time. Apoptosis exists also in healthy lungs, but in meconium-instilled lungs its level is usually dramatically higher. Apoptosis is characterized by loss of cell function, decrease in cell size, and its morphology. Apoptosis plays an important role in normal cell life, but increased levels of apoptosis induce great damage for any tissues. Apoptosis in the lungs has been greatly overlooked for the past decade, and meconium-induced apoptosis is a relatively new event and not effectively studied at the present time. This Review summarized current knowledge regarding meconium-induced inflammation and apoptosis in newborn lungs.

Intravenous immunoglobulin g attenuates pulmonary hypertension but induces local neutrophil influx in meconium aspiration in piglets.

BACKGROUND: Pulmonary hypertension and inflammation are well-identified pathogenetic features in meconium aspiration syndrome of newborns, but current approaches to their treatment or prevention are still often unsatisfactory. OBJECTIVES: To investigate the possible protective effects of human intravenous immunoglobulin G (IVIG) on the hypertensive and inflammatory lung injury in severe neonatal meconium aspiration. METHODS: Eleven newborn (10-12 days old) ventilated and catheterized piglets that received an intratracheal bolus (3 ml/kg) of a 65-mg/ml mixture of human meconium were studied for 6 h. IVIG was infused in 5 piglets 30 min before meconium administration, and 6 piglets served as controls and received the vehicle only. RESULTS: Meconium instillation induced a biphasic pulmonary hypertensive response, which was significantly diminished by IVIG pretreatment. Similarly, IVIG improved the oxygenation of the piglets, but the intrapulmonary shunt fraction or systemic hemodynamic parameters did not differ between the study groups, except of a minor decrease in the mean arterial blood pressure caused by IVIG. The blood leukocyte count was comparable in the 2 groups. The lung tissue ultrastructural and histological changes, number of apoptotic cells and phospholipase A2 activity were similar in the 2 groups. The amount of neutrophil accumulation, assessed by myeloperoxidase activity, was however significantly increased in macroscopically damaged lung tissue after IVIG administration. CONCLUSIONS: Our results thus indicate that IVIG treatment of newborns with severe meconium aspiration significantly diminishes the pulmonary hypertensive response and improves oxygenation, but the effects do not extend to protection of lung cellular injury.

Corticosteroid treatment in meconium aspiration syndrome: a solution for better outcome?

UNLABELLED: Corticosteroid treatment in severe meconium aspiration syndrome may afford, especially if started early, some improvement in oxygenation, lung function and pulmonary haemodynamics during the acute phase of the disorder. Still, the effects of corticosteroids on lung tissue perturbations and outcome of diseased infants remain unclear. CONCLUSION: Further research is needed to determine the clinical significance and optimal timing and dosing of corticosteroid treatment in severe meconium aspiration syndrome.

Pulmonary inflammatory mediators after sevoflurane and thiopentone anaesthesia in pigs.

BACKGROUND: Volatile anaesthetics have been shown to affect the release of pulmonary inflammatory mediators and exacerbate pulmonary injury after experimental aspiration. Thus, in theory, volatile anaesthetics may worsen inflammatory pulmonary injury and disease. We have previously described that no significant changes in alveolar ultrastructure are seen after sevoflurane anaesthesia. However, this does not exclude any possible physiological alterations. The aim of our study was to evaluate pulmonary inflammatory mediators in bronchoalveolar lavage (BAL) after sevoflurane and thiopentone anaesthesia in pigs with intact lungs. METHODS: Sixteen pigs were randomly selected to receive either a continuous thiopentone infusion (control group, n = 8) or sevoflurane (n = 8) at 4.0% inspiratory concentration (1.5 MAC) in air for 6 h. Bronchoalveolar lavage samples were collected at the end of the study to determine pulmonary inflammatory markers. RESULTS: Compared with thiopentone anaesthesia, significant increases in BAL leukotriene C4 (LTC4), NO3-, and NO2- levels were observed after sevoflurane anaesthesia. In addition, there was a significant decrease in total blood leukocyte count in sevoflurane-treated animals. CONCLUSION: We conclude that sevoflurane increases pulmonary LTC4, NO3-, and NO2- production in pigs, indicating an inflammatory response.

Alveolar integrity and ultrastructure in pigs remain undamaged after exposure to sevoflurane.

Previous studies have shown that both halothane and isoflurane have adverse but reversible effects on alveolar physiology. The present study was designed to test the hypothesis that also sevoflurane may affect alveolar integrity. Fifteen pigs were randomly selected to receive either thiopentone infusion (control group, n=8) or sevoflurane (n=7) at 4.0% inspiratory concentration (1.5 MAC) in air for 6 h. Tissue samples from the lungs were obtained at the end of the experiment. Both histopathological light microscopy and electron microscopy were used to assess the structural integrity of the alveoli. Pulmonary hemodynamics were comparable in both groups. Light microscopy showed no difference between the groups in the amount of alveolar macrophages, red blood cells or edema. Electron microscopy showed minor changes such as moderate local swelling of alveolar epithelium in both study groups. Alveolar type II cells were ultrastructurally unaltered in both study groups. We conclude that long-term, high concentration exposure to sevoflurane has no detrimental effect on the alveolar integrity in pigs.

Comparison of the effects of antenatal magnesium sulphate and ritodrine exposure on circulatory adaptation in preterm infants.

We examined the effects of maternal magnesium sulphate (MgSO4) and ritodrine treatments on the autonomic cardiovascular control in preterm neonates with respiratory distress syndrome during the first 2 days of life. Serial measurements of heart rate (HR), blood pressure (BP) and respirogram were performed during the first 2 days of life in 28 preterm infants below 33 weeks of gestation with antenatal exposure to MgSO4 (n = 13) or ritodrine (n = 15), and in 12 nonexposed preterm controls. Spectral analysis was used for the quantification of HR and BP variability. Although antenatal MgSO4 exposure had no effect on HR or the systolic, diastolic or mean BP, it was associated with significant decreased beat-to-beat changes in BP. In contrast, ritodrine exposure had no consistent effects on the autonomic cardiovascular control during the first 2 days of life. Our data suggest that maternal MgSO4 treatment decreases the neonatal high frequency changes in BP. This early vascular stabilizing effect of antenatal MgSO4 exposure may contribute to a lowered risk of cerebral vascular catastrophes, in the vulnerable areas of the brain, among the preterm infants with respiratory distress syndrome.

Maturational changes in ovine pulmonary metabolism of platelet-activating factor: implications for postnatal adaptation.

We recently reported that PAF acetylhydrolase (PAF-Ah) mRNA level and PAF-Ah activity in lamb lungs are up-regulated in the immediate newborn period, thereby facilitating the fall in postnatal PAF levels as well as a fall in pulmonary vascular resistance (B. O. Ibe, F. C. Sardar, and J. U. Raj, Mol Genet Metab 69:46-55, 2000). We have studied hypoxia effects on PAF synthesis and PAF-Ah activity in fetal lamb pulmonary arterial smooth muscle cells (FPASMC) and endothelial cells (FPAEC). We also studied PAF synthesis by platelets, and PAF-Ah activity in plasma of perinatal lambs at different ages. PAF synthesis (means +/- SEM, pmol/10(6) cells) by SMC in baseline was 168 +/- 27 and increased 3-fold on stimulation with A23187. Hypoxia augmented A23187-stimulated PAF synthesis by 30%. In FPAEC, baseline synthesis was 0.54 +/- 0.062 and increased 3-fold to 1.72 +/-.34. Hypoxia had no effect on PAF synthesis by EC. FPASMC produced over 300-fold more PAF than FPAEC. PAF synthesis by platelets was 47.02 +/- 7.1, 63.4 +/- 6.6, 71.5 +/- 9.9, and 62.2 +/- 5.2 for fetal, and newborn lambs <2 h, <1 day, and 6-12 days, old, respectively. PAF synthesis by platelets of <1 day-old lambs was different from that of fetal lambs. PAF-Ah activity (nmol lyso-PAF/min/mg protein) by FPASMC in normoxia was 3.41 +/- 0.38 which was 50% higher than the rate in hypoxia. Activity in FPAEC was 1.75 +/- 0.37 which was not different from hypoxia. PAF-Ah activity in fetal lamb plasma was 47.83 +/- 6.87 which was different from 155.32 +/- 12.10, the activity in plasma of newborn <1 day old. Activity in the other perinatal lambs did not differ from fetal or newborn <1 d. Our data suggest that lower pulmonary vascular PAF synthesis in normoxia together with higher PAF-Ah activity during immediate postnatal period is necessary to ensure rapid catabolism of PAF in vivo so as to facilitate postnatal adaptation of the pulmonary and systemic circulations.

Maternal magnesium sulfate treatment is associated with reduced brain-blood flow perfusion in preterm infants.

OBJECTIVE: To examine the influence of antenatally administered magnesium sulfate (MgSO4) and ritodrine on cerebral blood flow and systemic hemodynamics in preterm infants. DESIGN: Prospective, observational study. SETTING: Neonatal intensive care unit of a university central hospital. PATIENTS: Fifty-five preterm infants age <33 wks of gestation. INTERVENTIONS: Serial Doppler examinations of the brain circulation, heart rate, systemic blood pressure, and echocardiographic assessment of ductus arteriosus shunting were performed during the first week of life in infants exposed antenatally to maternal MgSO4 (n = 19) or ritodrine treatment (n = 17), and in 19 nonexposed preterm controls. MEASUREMENTS AND MAIN RESULTS: Cerebral blood flow velocity measurements were obtained from the anterior cerebral artery and internal carotid artery. Perfusion pressure and indices of resistance and blood flow in both vessels were subsequently derived. Maternal MgSO4 had no effect on neonatal cerebral blood flow velocity or resistance, but was associated with decreased (p <.05) perfusion pressure and blood flow in the anterior cerebral artery and internal carotid artery during the first day of life. Systolic blood pressure and pulse pressure were also lower (p <.05) during the whole study period in the MgSO4-exposed infants when compared with the controls. Maternal ritodrine treatment, on the other hand, had no consistent effects on either neonatal cerebral or systemic hemodynamics. CONCLUSIONS: Our data indicate that maternal MgSO4 treatment, in contrast to antenatal ritodrine, is associated with lowered cerebral perfusion in preterm infants on the first day of life.

Antenatal magnesium sulphate exposure is associated with prolonged parathyroid hormone suppression in preterm neonates.

UNLABELLED: The effects of maternal magnesium sulphate treatment on neonatal mineral status and parathyroid hormone secretory response were studied in 8 exposed and 27 control preterm infants during the first 2 wk of life. Antenatal magnesium sulphate resulted in hypermagnesaemia during the first 3-7 d of life without affecting other serum mineral concentrations. CONCLUSION: Early hypermagnesaemia was associated with hypercalciuria during the first 3 d and parathyroid hormone suppression up to the age of 2 wk in the exposed infants.

Meconium aspiration syndrome: a role for phospholipase A2 in the pathogenesis?

UNLABELLED: The pathophysiology of neonatal meconium aspiration syndrome (MAS), often resulting in severe respiratory failure, is complex and still largely unclear. Factors involved in the propagation of acute lung injury after perinatal aspiration of meconium include obstruction of the airways, ventilation/perfusion mismatch, increase of the pulmonary vascular resistance and a rapidly developing parenchymal and alveolar inflammatory reaction with associated surfactant dysfunction. CONCLUSION: Although the early pulmonary inflammatory response is believed to play a central pathogenetic role in the meconium-induced acute lung damage, its initiating mechanisms are still poorly defined. However, increasing evidence indicates a direct toxic effect of meconium.

Dexamethasone treatment attenuates pulmonary injury in piglet meconium aspiration.

To investigate the pulmonary effects of steroid treatment in neonates with meconium aspiration, 25 10- to 12-d-old piglets were studied for 6 h after an intratracheal bolus of human meconium. Dexamethasone (0.5 mg/kg) was given in two treatment schedules, either 1 h before (n = 6) or 1 h after meconium instillation (n = 8). Eight piglets served as controls. Three additional piglets were given dexamethasone without meconium instillation. Pulmonary hemodynamics and oxygenation were followed, and lung tissue samples investigated for signs of inflammation and ultrastructural injury, including apoptosis. Pulmonary artery pressure and vascular resistance increased after meconium instillation, but this rise was significantly prevented after prophylactic dexamethasone. This treatment also improved the acutely deteriorated oxygenation of the piglets after meconium insufflation. Prophylactic, but not early, dexamethasone treatment further protected the lungs from the ultrastructural changes caused by meconium instillation. Additionally, the increase of apoptotic epithelial cell deaths was significantly prevented by both dexamethasone treatments. These results show that prophylactic dexamethasone treatment significantly attenuates the early pulmonary hemodynamic deterioration and structural lung damage caused by meconium aspiration. Further studies on the apoptosis-inhibiting effect of dexamethasone administration in neonatal lungs exposed to heavy meconium are warranted.

In vivo detection of vascular adhesion protein-1 in experimental inflammation.

Vascular adhesion protein-1 (VAP-1) is an inflammation-inducible endothelial glycoprotein which mediates leukocyte-endothelial cell interactions. To study the pathogenetic significance of VAP-1 in inflammatory disorders, an in vivo immunodetection method was used to detect the regulation of luminally expressed VAP-1 in experimental skin and joint inflammation in the pig and dog. Moreover, VAP-1 was studied as a potential target to localize inflammation by radioimmunoscintigraphy. Up-regulation of VAP-1 in experimental dermatitis and arthritis could be visualized by specifically targeted immunoscintigraphy. Moreover, the translocation of VAP-1 to the functional position on the endothelial surface was only seen in inflamed tissues. These results suggest that VAP-1 is both an optimal candidate for anti-adhesive therapy and a potential target molecule for imaging inflammation.

Human meconium has high phospholipase A2 activity and induces cellular injury and apoptosis in piglet lungs.

Aspiration of meconium produces an inflammatory reaction resulting in necrotic changes in lung tissue. To further investigate the mechanisms of the meconium-induced early pulmonary injury, twenty 10-12-d-old piglets were studied for lung tissue ultrastructural and apoptotic changes and phospholipase A2 activity. Twelve piglets received an intratracheal bolus (3 mL/kg) of a 20-mg/mL (thin, n = 6) or 65-mg/mL (thick, n = 6) mixture of human meconium, and control piglets (n = 5) received the same amount of intratracheal saline. Three ventilated piglets with no aspiration were also studied. Pulmonary hemodynamics and systemic oxygenation were followed for 6 h after meconium or saline insufflation. In the control groups, the pulmonary tissue showed open alveolar spaces and intact vascular walls, whereas meconium administration resulted in severe pneumonitis, with alveolar spaces filled with inflammatory exudate. Meconium instillation additionally resulted in edematous changes in the vascular walls and alveolar epithelium, whereas type II pneumocytes were intact. The amount of apoptotic cells was increased, especially in the respiratory epithelium, and the catalytic activity of phospholipase A2 in lung tissue samples was significantly elevated after thick meconium instillation. This activity rise proved to be mainly because of human group I phospholipase A2, introduced by meconium. Our data thus show that aspiration of meconium leads to severe lung tissue inflammation with early ultrastructural changes in the pulmonary alveolar walls and is associated with apoptotic cell death in the epithelium, already during the first hours after the insult. These results further suggest that high phospholipase A2 activity, mainly introduced into the lungs within the meconium, may have an important role in the initiation of these alterations in neonatal lungs.

Nitric oxide inhalation inhibits pulmonary apoptosis but not inflammatory injury in porcine meconium aspiration.

To investigate the possible protective effects of nitric oxide (NO) inhalation in newborns with meconium aspiration, 18 10-12-d-old piglets were studied for 6h after an intratracheal bolus (3 ml/kg) of a 65-mg/ml mixture of human meconium. Twelve of the piglets were treated with continuous NO inhalation at a dose of 1 ppm (n = 6) or 10 ppm (n = 6), started 30 min before the insult. Pulmonary haemodynamics and systemic oxygenation were followed, and lung tissue samples were studied for signs of inflammation, evidence of ultrastructural injury and apoptotic cell changes. Inhalation of 10 ppm NO, in contrast to 1 ppm NO, significantly delayed the meconium-induced pulmonary pressure rise and the increase in intrapulmonary shunt fraction, and maintained better oxygenation in the piglets. Histologically and biochemically, treatment with 1 or 10 ppm NO inhalation did not protect the lungs against meconium-induced inflammatory injury. Further, ultrastructural lung tissue analysis revealed a significant amount of alveolar exudate and oedematous alveolar epithelium and endothelium after meconium instillation, also in the lungs treated with NO inhalation. However, the increase in apoptotic epithelial cell deaths, previously shown to be stimulated by intratracheal meconium, was significantly impeded after inhalation of 10 ppm. These results thus show that early continuous NO inhalation controls the rise in pulmonary artery pressure and improves the efficiency of arterial oxygenation, and further prevents the increase in epithelial apoptosis, but does not protect against early inflammatory damage caused by meconium aspiration.

Association of low-density lipoprotein oxidation to abnormal electrocardiographic late potentials.

In vivo oxidation of low-density lipoprotein is shown to be significantly related to another risk factor for coronary atherosclerosis, abnormal electrocardiographic late potentials, in clinically healthy pilots. Because both of these variables have been also associated with cardiac arrhythmogenic action, together they may improve the identification of patients at risk of ventricular tachyarrhythmias.

Meconium stimulates cyclooxygenase-2 expression in rat lungs.

Since meconium aspiration often induces an inflammatory respiratory disorder, we investigated the effects of intrapulmonary meconium on the expression of cyclooxygenase-1 and 2 in rat lungs. Suspension of human meconium was instilled intratracheally into ventilated lungs of anesthetized rats, while control rats received an equal volume of saline. The meconium lungs were ventilated either with air or 100% oxygen, and control lungs were ventilated with air. After 3 h, the lungs were removed and the amount of cyclooxygenase-1 and 2 mRNA was measured by Northern blot analysis. Cyclooxygenase-1 mRNA was clearly expressed in control rat lungs, while cyclooxygenase-2 expression was minimal. Meconium administration markedly upregulated the expression of cyclooxygenase-2 mRNA, while cyclooxygenase-1 expression remained unchanged. Increased expression of cyclooxygenase-2 was detected in rat lungs ventilated with either air or oxygen. Our data thus indicate that meconium aspiration induces pulmonary expression of cyclooxygenase-2, suggesting an important role for prostaglandins in the meconium aspiration-induced inflammation in neonatal lungs.