Ellen Schoof

High frequency oscillatory ventilation suppresses inflammatory response in lung tissue and microdissected alveolar macrophages in surfactant depleted piglets.

The impact of high frequency oscillatory ventilation (HFOV) compared with intermittent mandatory ventilation (IMV) on oxygenation and pulmonary inflammatory response was studied in a surfactant depleted piglet model. After establishment of lung injury by bronchoalveolar lavage, piglets either received HFOV (n =5) or IMV (control; n = 5) for eight hours. Pao2 was higher and mean pulmonary arterial pressure (MPAP) was lower with HFOV (HFOV versus control, mean ± SEM; endpoint paO2: 252 ± 73 versus 68 ± 8.4 mm Hg; p < 0.001; MPAP: 22 ± 2.3 versus 34 ± 2.5 mm Hg; p < 0.01). mRNA expression of interleukin (IL)-1β, IL-6, IL-8, IL-10, TGF-β1, Endothelin-1, and adhesion molecules (E-selectin, P-selectin, ICAM-1) in lung tissue was quantified by real time PCR normalized to β-actin and hypoxanthine-guanine-phosphoribosyl-transferase (HPRT). mRNA expression of all cytokines and adhesion molecules/HPRT was higher in controls (e.g.: HFOV versus control, mean ± SEM; IL-1β/HPRT: 1.6 ± 0.3 versus 23.1 ± 8.6 relative units (RU), p < 0.001; IL-8/HPRT: 8.5 ± 2.0 versus 63.5 ± 15.2 RU, p < 0.001). IL-8/HPRT gene expression was quantified in microdissected single cells. With HFOV, IL-8 gene expression was highly reduced in alveolar macrophages: 10 ± 3.4 copies IL-8 mRNA/copy HPRT mRNA versus 356 ± 142; p < 0.05 (bronchiolar epithelial cells: 33 ± 16 versus 208 ± 108; alveolar septum: 2.1 ± 1.3 versus 26 ± 11; bronchiolar smooth muscle cells: 1.3 ± 0.3 versus 2.8 ± 1.0; vascular smooth muscle cells: 0.7 ± 0.3 versus 1.1 ± 0.4). In conclusion, HFOV improved oxygenation, reduced pulmonary arterial pressure and attenuated pulmonary inflammatory response.

No Influence of Surgical Stress on Postoperative Leptin Gene Expression in Different Adipose Tissues and Soluble Leptin Receptor Plasma Levels

As part of the postsurgical stress response, plasma leptin levels increase in-between 12 h postoperatively. Objective: To study the kinetics of leptin gene expression in different adipose tissues before and after severe surgical trauma in children and adults. Methods: In 22 adults and 23 children with cardiac and 19 adult patients with abdominal surgery, we measured plasma leptin concentrations preoperatively, 4 and 10–17 h postoperatively and quantified leptin mRNA expression by TaqMan real-time PCR in adipose tissue taken at the beginning and the end of surgery from subcutaneous, intrathoracic, omental and mesenteric fat. Plasma-soluble leptin receptor levels were measured in 23 children after cardiosurgery. Results: Plasma leptin levels doubled between 4 and 10–17 h postoperatively in adults (p < 0.001) as well as in children (p = 0.0002) with cardiac surgery. After abdominal surgery, 10–17 h postoperatively, plasma leptin concentrations increased significantly (p < 0.05). During the operation, leptin gene expression did not change in neither of the patient groups. Plasma-soluble leptin receptor levels decreased immediately after the onset of surgery and remained unchanged thereafter. Conclusions: Leptin gene expression is not up-regulated during surgery. The measured increase in plasma leptin after surgery does not result from elevated levels of soluble leptin receptor but may follow an up-regulation of leptin gene expression later after the operation due to postsurgical metabolic changes.

Aerosolized perfluorocarbon suppresses early pulmonary inflammatory response in a surfactant-depleted piglet model

The effect of new ventilation strategies on initial pulmonary inflammatory reaction was studied in a surfactant-depleted piglet model. Sixty minutes after induction of lung injury by bronchoalveolar lavage, piglets received either aerosolized FC77 (aerosol-PFC, 10 mL/kg/h, n = 5) or partial liquid ventilation (PLV) with FC77 at functional residual capacity volume (FRC-PLV, 30 mL/kg, n = 5), or at low volume (LV-PLV, 10 mL/kg per hour, n = 5), or intermittent mandatory ventilation (control, n = 5). After 2 h, perfluorocarbon application was stopped and intermittent mandatory ventilation continued for 6 h. After a total experimental period of 8 h, animals were killed and lung tissue obtained. mRNA expression of IL-1β, IL-6, IL-8, and TGF-β in porcine lung tissue was quantified using TaqMan real-time PCR and normalized to β-actin (A) and hypoxanthine-guanine-phosphoribosyl-transferase (H). In the aerosol-PFC group, IL-1β, IL-6, IL-8, and transforming growth factor (TGF)-β mRNA expression in lung tissue was significantly lower than in the control group. Reduction was 95% for IL-1β/H (p < 0.001), 73% for IL-6/H (p < 0.05), 87% for IL-8/H (p < 0.001), and 38% for TGF-β/H (p < 0.01). A lower mRNA gene expression was also determined for IL-1β and IL-8 when the aerosol-PFC group was compared with the LV-PLV group [91% for IL-1β/H (p < 0.001), 75% for IL-8/H (p < 0.001)]. In the FRC-PLV group, mRNA expression of IL-1β was significantly lower than in the control (p < 0.05) and LV-PLV (p < 0.01) group. In a surfactant-depleted piglet model, aerosol therapy with perfluorocarbon but not LV-PLV reduces the initial pulmonary inflammatory reaction at least as potently as PLV at FRC volume.

Comparison of aerosol therapy with different perfluorocarbons in surfactant-depleted animals

Objective:The study investigates the effectiveness of aerosol treatment on gas exchange and pulmonary inflammatory reaction using perfluorocarbons with different molecular structure and vapor pressure. Design:Experimental, prospective, randomized, controlled study. Setting:Experimental laboratory at a university hospital. Subjects:Twenty anesthetized neonatal piglets assigned to four groups. Interventions:After establishment of lung injury by bronchoalveolar lavage, piglets either received aerosolized FC77 (n = 5), perfluorooctylbromide (n = 5), or FC43 (n = 5, 10 mL·kg−1·hr−1 for 2 hrs) or intermittent mandatory ventilation (control, n = 5). Thereafter, animals were supported for another 6 hrs. Measurements and Main Results:PaO2 significantly improved in the perfluorocarbon groups compared with control (p < .01). Final PaO2 (mean ± sem) was FC77, 406 ± 27 mm Hg; perfluorooctylbromide, 332 ± 32 mm Hg; FC43, 406 ± 19 mm Hg; control, 68 ± 8 mm Hg. PaCO2 and mean pulmonary arterial pressure were lower in all perfluorocarbon groups compared with control. The ratio of terminal dynamic compliance to total compliance was significantly higher in the FC77 than in the FC43, perfluorooctylbromide, and control groups. Relative gene expression of interleukin-1β, interleukin-8, P-selectin, E-selectin, and intercellular adhesion molecule-1 in lung tissue was determined by TaqMan real time polymerase chain reaction normalized to hypoxanthine-guanine-phosphoribosyl-transferase and was shown to be reduced by all perfluorocarbons. Conclusions:Aerosol treatment with all the perfluorocarbons investigated improved gas exchange and reduced pulmonary inflammatory reaction independently from molecular structure and vapor pressure of the perfluorocarbons. Although differences in vapor pressure and molecular structure may account for varying optimal dosing strategies, several different perfluorocarbons were shown to be principally suitable for aerosol treatment.

Aerosolized adrenomedullin suppresses pulmonary transforming growth factor-β1 and interleukin-1β gene expression in vivo

The effect of aerosolized adrenomedullin on interleukin-1β and transforming growth factor (TGF)-β1 mRNA and protein expression was studied in surfactant depleted piglets, receiving aerosolized adrenomedullin (adrenomedullin, n=6), aerosolized adrenomedullin plus i.v. NG-nitro-l-arginine-methylester (adrenomedullin+l-NAME, n=5), or aerosolized saline solution (control, n=6). After 8 h of aerosol interval therapy, mRNA expression of interleukin-1β and TGF-β1 in lung tissue was quantified normalized to β-actin and hypoxanthine-guanine-phosphoribosyl-transferase by real-time polymerase chain reaction (PCR). Interleukin-1β and TGF-β1 protein concentration in lung tissue was quantified by enzyme-linked immunosorbent assay (ELISA). In the adrenomedullin group, interleukin-1β and TGF-β1 mRNA expression was lower than in controls. Reduction for interleukin-1β/β-actin was 56% (p<0.001), for interleukin-1β/hypoxanthine-guanine-phosphoribosyl-transferase 60% (p<0.001), for TGF-β1/β-actin 65.5% (p<0.001), and for TGF-β1/hypoxanthine-guanine-phosphoribosyl-transferase 56.2% (p<0.001). Mean interleukin-1β protein expression was different between the groups, p<0.05 (adrenomedullin 601±61, Control 836±88 pg/mg protein). l-NAME did not antagonize adrenomedullin effect on TGF-β1 mRNA. In conclusion, aerosolized adrenomedullin reduced pulmonary inflammatory and pro-fibrotic response.

Aerosolized perfluorocarbon reduces adhesion molecule gene expression and neutrophil sequestration in acute respiratory distress

In acute respiratory distress syndrome, neutrophil migration into the lung plays a key role in the development of lung injury. To study the effect of different modes of ventilation with perfluorocarbon (FC77), intrapulmonary neutrophil accumulation and mRNA expression of E-selectin, P-selectin and intercellular adhesion molecule-1 (ICAM-1), mediating leukocyte sequestration, were measured in surfactant depleted piglets. After bronchoalveolar lavage, 20 animals either received aerosolized perfluorocarbon (Aerosol-PFC), partial liquid ventilation (PLV) with perfluorocarbon at functional residual capacity filling volume (FRC-PLV) or at low volume (LV-PLV) or intermittent mandatory ventilation (control). After 2 h of perfluorocarbon application, intermittent mandatory ventilation was continued for 6 h. In the Aerosol-PFC group, all measured adhesion molecules showed a significantly reduced gene expression compared to controls. FRC-PFC treatment was effective in significantly diminishing P-selectin and ICAM-1 mRNA expression. Relative lung tissue neutrophil counts were significantly reduced in the Aerosol-PFC and the FRC-PLV group. Treatment with aerosolized perfluorocarbon is at least as effective as partial liquid ventilation at FRC volume in reducing pulmonary adhesion molecule expression and neutrophil accumulation in acute respiratory distress syndrome.

Five-Year Follow-Up of a 13-Year-Old Boy with a Pituitary Adenoma Causing Gigantism – Effect of Octreotide Therapy

Background/Aim: In children, there is little experience with octreotide therapy for pituitary tumors, especially growth hormone (GH) producing adenomas. We report on a 13-year-old boy with gigantism due to a GH-producing pituitary adenoma caused by a Gsα mutation on the basis of McCune-Albright syndrome. Methods: At the age of 6.5 years a GH- and prolactin-producing pituitary adenoma was diagnosed. The adenoma was surgically removed. Immediately thereafter, the small adenoma residuum was treated with octreotide (2 × 100 µg/day s.c.). Results: During therapy with octreotide, the growth rate dropped to normal values; however, rose again after 2 years of treatment. The insulin-like growth factor I (IGF-I) levels remained above the 95th percentile, the GH level mostly >2 µg/l. After 5 years of octreotide therapy, GH (6.9 µg/l), IGF-I (620 µg/l), IGF-binding protein 3 (5.4 mg/l), and prolactin (17.0 ng/ml) levels were still elevated. The growth velocity was +2.4 SDS (standard deviation score), the pubertal status was mature, and the bone age was 14.3 years (prospective final height 208 cm). A magnetic resonance imaging scan showed an unchanged residual 4-mm rim of adenoma at the pituitary site. Side effects from octreotide therapy were not reported by the patient or his family. The therapy was changed to the long-acting release octreotide analog octreotide-LAR. After 1 year of treatment with octreotide-LAR, the GH level was 1.0 µg/l, and the prospective final height dropped by 10 cm. Conclusions: This case demonstrates that combined surgical and medical treatment can influence the prognosis of childhood gigantism; however, the prognosis of this rare condition remains uncertain.

Brief adrenomedullin inhalation leads to sustained reduction of pulmonary artery pressure

The effect of aerosolised adrenomedullin (ADM), a potent vasodilator peptide, on pulmonary artery pressure was studied for 24 h in a surfactant-depleted piglet model. Animals received either aerosolised ADM (50 ng·kg−1·min−1, ADM, n=6), or aerosolised normal saline solution (control, n=6). Aerosol therapy was performed for a 2 h treatment period followed by a 22 h observation period. Ventilator settings were adapted to keep arterial oxygen tension and carbon dioxide arterial tension between 13.3–14.6 kPa and 4.9–5.7 kPa, respectively. Aerosolised ADM reduced mean pulmonary artery pressure (MPAP) compared with the control group (end-point median 24 h after therapy start: ΔMPAP −14.0 versus −8.0 mmHg; 23.5 h after therapy start). After therapy start, mean systemic arterial pressure (MAP) was not significantly different between the groups (end-point median: MAP ADM 70 (61/74) versus control 72 (54/81) mmHg). Endothelin-1, a potent pulmonary vasoconstrictor, is regulated by ADM via cAMP. Twenty two hours after inhalation of aerosolised ADM, endothelin-1 mRNA in lung tissue and endothelin-1 protein expression in pulmonary arteries was reduced compared with controls (median semi-quantitative immunhistochemical score: ADM 0.21, control 0.76). Aerosolised adrenomedullin significantly reduced mean pulmonary artery pressure independently of arterial oxygen tension.

Cardiopulmonary bypass surgery does not further increase elevated serum leptin concentrations after major surgery.

Objective The objective of the present study was to examine the impact of major heart surgery with cardiopulmonary bypass (CPB) in childhood on serum leptin concentrations in relation to plasma cortisol, epinephrine, norepinephrine, and insulin. Design Controlled, prospective study. Setting Intensive care unit of a university hospital. Patients and Interventions We enrolled 20 pediatric patients undergoing open heart surgery and 20 children with major surgery not necessitating CPB (surgical control group). Leptin was measured by radioimmunoassay, cortisol and insulin were measured by chemiluminescence, and epinephrine and norepinephrine were measured by high-pressure liquid chromatography. Measurements and Main Results In the CPB group, leptin dropped from 0.4 ± 0.1 preoperatively (mean ± sem) to 0.2 ± 0.1 ng/mL intraoperatively (p < .05). It increased to 1.6 ± 0.7 ng/mL 12 hrs after surgery (p < .01) and declined thereafter. In the surgical controls, leptin rose from 0.5 ± 0.2 ng/mL before surgery to 1.8 ± 0.8 ng/mL 12 hrs after surgery (p = .001). In both groups, plasma cortisol, insulin, and epinephrine significantly increased after surgery. There was no relationship between the maximum increase of serum leptin and the other hormones. Conclusions Patients with CPB surgery and non-CPB surgery show a similar increase in serum leptin, indicating that sepsislike inflammatory syndrome does not further increase elevated leptin concentrations following major surgery. In this complex situation, serum leptin does not appear to be merely regulated by its known stimuli and suppressors.

Prolactinoma Causing Secondary Amenorrhea in a Woman with Ullrich-Turner Syndrome

This is the case report of a girl who was diagnosed as having Ullrich-Turner mosaic at the age of 12 years. She had normal pubertal development and menarche at the age of 15 years. The patient had regular menstrual cycles for 12 months before developing secondary amenorrhea. She was started on estrogen/gestagen replacement therapy by her gynecologist. Several months later a prolactinoma was diagnosed by laboratory and imaging techiques. A second-generation dopamine agonist led to almost regular cycles. Therefore, even in patients with susceptibility to ovarian failure secondary amenorrhea necessitates thorough diagnostic investigation.