Activation of the aryl hydrocarbon receptor by clozapine induces preadipocyte differentiation and contributes to endothelial dysfunction.

BACKGROUND: The superior therapeutic benefit of clozapine is often associated with metabolic disruptions as obesity, insulin resistance, tachycardia, higher blood pressure, and even hypertension. AIMS: These adverse vascular/ metabolic events under clozapine are similar to those caused by polycyclic aromatic hydrocarbons (PAHs), and clozapine shows structural similarity to well-known ligands of the aryl hydrocarbon receptor (AhR). Therefore, we speculated that the side effects caused by clozapine might rely on AhR signaling. METHODS: We examined clozapine-induced AhR activation by luciferase reporter assays in hepatoma HepG2 cells and we proved upregulation of the prototypical AhR target gene Cyp1A1 by realtime-PCR (RT-PCR) analysis and enzyme activity. Next we studied the physiological role of AhR in clozapine's effects on human preadipocyte differentiation and on vasodilatation by myography in wild-type and AhR-/- mice. RESULTS: In contrast to other antipsychotic drugs (APDs), clozapine triggered AhR activation and Cyp1A1 expression in HepG2 cells and adipocytes. Clozapine induced adipogenesis via AhR signaling. After PGF2α-induced constriction of mouse aortic rings, clozapine strongly reduced the maximal vasorelaxation under acetylcholine in rings from wild-type mice, but only slightly in rings from AhR-/- mice. The reduction was also prevented by pretreatment with the AhR antagonist CH-223191. CONCLUSION: Identification of clozapine as a ligand for the AhR opens new perspectives to explain common clozapine therapy-associated adverse effects at the molecular level.

Very high expander processing of maize on animal performance, digestibility and product quality of finishing pigs and broilers.

The present study investigated the effect of hydrothermic maize processing and supplementation of amino acids (AA) in two experiments. In total, 60 barrows and 384 broilers were fed four diets including either unprocessed (T1), or hydrothermically processed maize, that is short- (T2), or long-term conditioned (LC) (T3), and subsequently expanded maize of the same batch. Assuming a higher metabolizable energy (ME) content after processing, the fourth diet (T4) contains maize processed as treatment T3, but AA were supplemented to maintain the ideal protein value. Performance, digestibility and product quality in both species were assessed. Results show that in pigs receiving T4 the average daily feed intake was lower compared with the other treatments, whereas no difference was observed in broilers. The T3 improved the feed conversion rate compared with T1 (P<0.10) for both species. In contrast, average daily gain (ADG) (1277 g/day for T2 and 1267 g/day for T3 v. 971 g/day for T1) was only altered in pigs. The hydrothermic maize processing increased the apparent total tract digestibility (ATTD) of dry matter, starch and ether extract after acid hydrolysis. This may be a consequence of higher ATTD of gross energy in the finishing phase for both animal species, suggesting a higher ME content in diets with processed maize. The higher ME content of diets with processed maize is supported also by measurements of product quality. Supplementation of AA in T4 enhanced the loin depth in pigs as well as the amount of breast meat in broilers. Further effects of processing maize on meat quality were the reduced yellowness and antioxidative capacity (P<0.10) for broilers, likely due to the heat damage of xanthophylls and tocopherols. Processing also increased springiness and chewiness (P<0.10) of the broilers breast meat, whereas the loin meat of pigs showed a decreased lightness and yellowness (P<0.10) in meat when hydrothermic processed maize was used (for T2, T3 and T4). LC processed maize (T3) showed the lowest springiness in pork, however the supplementation of AA in T4 did not show differences between the treatments. Shown results demonstrated positive effects of hydrothermic processing of maize on animal performance and digestibility in both species. However, effects on carcass characteristics and product quality differed. The negative effects on product quality could be partly compensated with the AA supplementation, whereas a change in meat colour and reduced antioxidative capacity was observed in all groups fed hydrothermic maize processing.

Prospective study of bacteraemia in acute haemorrhagic diarrhoea syndrome in dogs.

In dogs with idiopathic acute haemorrhagic diarrhoea syndrome (AHDS), a serious loss of intestinal mucosal barrier integrity occurs. However, the incidence of bacterial translocation in dogs with idiopathic AHDS is not known. Thus, the objectives of this prospective study were to identify the incidence of bacteraemia, to evaluate the frequency of septic events and the influence of bacteraemia on various clinical and laboratory parameters, duration of hospitalisation and survival of dogs with idiopathic AHDS. The study included 87 dogs with idiopathic AHDS. Twenty-one healthy dogs served as control group. To evaluate clinical significance of bacterial translocation, blood culture results were compared between patients and controls. Clinical and laboratory parameters were compared between patients with positive and negative blood cultures. There was no significant difference in either incidence of bacteraemia between patients with idiopathic AHDS (11 per cent) and controls (14 per cent) or in severity of clinical signs, laboratory parameters, duration of hospitalisation or mortality between blood culture-positive and culture-negative dogs with idiopathic AHDS. The results of this study suggest that the incidence of bacteraemia in dogs with idiopathic AHDS is low and not different from that of healthy control dogs. Bacteraemia does not influence the clinical course or survival and thus antibiotic treatment is not indicated to prevent sepsis.

Short-term morbidities in moderate and late preterm infants.

OBJECTIVE: To determine (1) the association between neonatal morbidity and gestational age and (2) the impact of pre-existing maternal medical conditions, pregnancy and birth complications on neonatal outcome in moderate and late preterm infants (32-36 completed weeks). METHODS: Retrospective single-centre cohort study including all moderate and late preterm infants without congenital anomalies born at the Children's and Maternity Hospital Linz, Austria, between January 2007 and June 2010. Stepwise regression analysis was used to determine significant associations between morbidities, maternal and perinatal complications and the gestational age. RESULTS: Of 870 infants included the incidence of neonatal morbidities increased from 24% at 36 weeks to 43% at 35 weeks', 55% at 34 weeks', 75% at 33 weeks' and 93% at 32 weeks' gestation. Infants at 32 weeks had a 4-fold (RR: 3.88; 95% CI: 1.87-8.06) increased risk compared with those at 36 weeks, and infants of 32 weeks were 16 times (RR: 16.01; 95% CI: 9.82-26.09) more likely to be admitted to the NICU than infants of 36 weeks'. Hyperbilirubinemia (29%) and respiratory morbidity (14.3%) were the most common neonatal diagnoses. Intrauterine growth restriction, preeclampsia, preterm premature rupture of the membranes, lack of antenatal steroid administration, antepartum hemorrhage, multiple pregnancy and male gender were all associated with any kind of neonatal morbidity, admission rate to the NICU and length of hospital stay (p<0.05). CONCLUSION: Nearly half of all infants suffered from any morbidity, and several risk factors were identified being significantly associated with NICU admission rate and length of hospitalization.

Thrombospondin-1 triggers macrophage IL-10 production and promotes resolution of experimental lung injury.

Mononuclear phagocyte recognition of apoptotic cells triggering suppressive cytokine signaling is a key event in inflammation resolution from injury. Mice deficient in thrombospondin (TSP)-1 (thbs1⁻/⁻), an extracellular matrix glycoprotein that bridges cell-cell interactions, are prone to lipopolysaccharide-induced lung injury and show defective macrophage interleukin (IL)-10 production during the resolution phase of inflammation. Reconstitution of IL-10 rescues thbs1⁻/⁻ mice from persistent neutrophilic lung inflammation and injury and thbs1⁻/⁻ alveolar macrophages show defective IL-10 production following intratracheal instillation of apoptotic neutrophils despite intact efferocytosis. Following co-culture with apoptotic neutrophils, thbs1⁻/⁻ macrophages show a selective defect in IL-10 production, whereas prostaglandin E2 and transforming growth factor beta 1 responses remain intact. Full macrophage IL-10 responses require the engagement of TSP-1 structural repeat 2 domain and the macrophage scavenger receptor CD36 LIMP-II Emp sequence homology (CLESH) domain in vitro. Although TSP-1 is not essential for macrophage engulfment of apoptotic neutrophils in vivo, TSP-1 aids in the curtailment of inflammatory responses during the resolution phase of injury in the lungs by providing a means by which apoptotic cells are recognized and trigger optimal IL-10 production by macrophages.

MATH/BTB CRL3 receptors target the homeodomain-leucine zipper ATHB6 to modulate abscisic acid signaling.

Being sessile organisms, plants need rapid and finely tuned signaling pathways to adapt their growth and survival over their immediate and often adverse environment. Abscisic acid (ABA) is a plant hormone crucial for both biotic and abiotic stress responses. In this study, we highlight a function of six Arabidopsis MATH-BTB proteins in ABA signaling. MATH-BTB proteins act as substrate-binding adaptors for the Cullin3-based ubiquitin E3 ligase. Our genetic and biochemical experiments demonstrate that the MATH-BTB proteins directly interact with and target for proteasomal degradation the class I homeobox-leucine zipper (HD-ZIP) transcription factor ATHB6, which was previously identified as a negative regulator of ABA responses. Reducing CUL3(BPM) function leads to higher ATHB6 protein accumulation, reducing plant growth and fertility, and affects stomatal behavior and responses to ABA. We further demonstrate that ABA negatively regulates ATHB6 protein turnover, a situation reminiscent to ABI5, another transcription factor involved in ABA signaling.

Complex regulation of the TIR1/AFB family of auxin receptors.

Auxin regulates most aspects of plant growth and development. The hormone is perceived by the TIR1/AFB family of F-box proteins acting in concert with the Aux/IAA transcriptional repressors. Arabidopsis plants that lack members of the TIR1/AFB family are auxin resistant and display a variety of growth defects. However, little is known about the functional differences between individual members of the family. Phylogenetic studies reveal that the TIR1/AFB proteins are conserved across land plant lineages and fall into four clades. Three of these subgroups emerged before separation of angiosperms and gymnosperms whereas the last emerged before the monocot-eudicot split. This evolutionary history suggests that the members of each clade have distinct functions. To explore this possibility in Arabidopsis, we have analyzed a range of mutant genotypes, generated promoter swap transgenic lines, and performed in vitro binding assays between individual TIR1/AFB and Aux/IAA proteins. Our results indicate that the TIR1/AFB proteins have distinct biochemical activities and that TIR1 and AFB2 are the dominant auxin receptors in the seedling root. Further, we demonstrate that TIR1, AFB2, and AFB3, but not AFB1 exhibit significant posttranscriptional regulation. The microRNA miR393 is expressed in a pattern complementary to that of the auxin receptors and appears to regulate TIR1/AFB expression. However our data suggest that this regulation is complex. Our results suggest that differences between members of the auxin receptor family may contribute to the complexity of auxin response.

[Medical prescriptions to premature and newborn infants in an Austrian neonatal intensive care unit]. / Verschreibepraxis von Arzneimitteln bei Früh- und Neugeborenen auf einer österreichischen neonatologischen Intensivstation.

BACKGROUND: In December 2006, the European Commission issued the Regulation 1901/2006 with the aim to improve the development of medicines for children, because currently many of the pediatric drugs are used outside their authorization. The aim of the present study was to establish a status quo of the medical prescriptions to patients admitted to a neonatal intensive care unit. METHOD: The records of all patients admitted during a three-month period were analyzed retrospectively. Study endpoints were all prescribed medications, including dosage, route and form of administration. Medications were categorized in three groups: licenced, unlicenced and off-label medications. RESULTS: A total of 81 patients with a total of 84 admissions (3 re-admissions) had a median length of stay of 6 days (range: 1-101 days) in the intensive care unit. Of the 84 admissions, 20 were attributable to heart surgery conditions and 11 to extremely low birth weight (less than 1 000 g). A total of 748 prescriptions of 82 different medications were registered with an average of 6 (range: 1-38) different drugs per stay. 364 prescriptions (49%) were licenced, 250 (34%) were off-label, and 134 (18%) were unlicenced. During 77% of all 84 admissions, we used at least one off-label or unlicenced drug. CONCLUSION: The data of our study demonstrate that the availability of licenced drugs for neonates is greatly limited. The results are consistent with studies in other European countries. This study underlines once more the need of adequate testing of medicines in children to assess their safety and efficacy. Whether the new European regulation will achieve this goal remains to be seen.

Disease progression in HIV-1 infected children and adolescents--results of a German-Austrian cohort study.

OBJECTIVE: The Paediatric Working Group AIDS (PAAD) initiated a prospective cohort study in order to investigate disease progression in HIV- infected children and adolescents and the effect of antiretroviral treatment regimes. PATIENTS AND METHODS: Between 1998 and 2003, paediatric centres documented HIV-infected patients under clinical care using a questionnaire for basic data and annual follow up. Main outcome measures were: use of antiretroviral therapy, adverse events, disease progression and change of therapeutic regimes. RESULTS: 174 HIV- infected paediatric patients were followed up in 12 centres in Germany and Austria between 1998 and 2003. Initially 54 (31%) patients had no antiretroviral therapy, 35 (20%) received a two-drug regimen (ART) and 85 patients (49%) a highly active antiretroviral therapy (HAART>or=3 drugs). After an observation period of 5 years, 8 patients (4%) had no therapy, 17 (10%) were on ART and 134 patients on HAART (77%). The number of patients with salvage therapy (>or=4 drugs) increased from 5 (3%) to 15 patients (9%). 72 of 166 treated patients (43%) had no change of their drug regimes, 68 patients (41%) had one change and 26 patients (16%)>or=2 changes. Main reasons for changes were increased viral load (49%), immunologic deterioration (21%) and adverse events (14%). During the follow up period no patient died. According to the CDC classification, disease progression was seen in 48 of 174 patients (28%), of whom 20 had deteriorations of clinical categories (A, B, C) and 28 of immunologic categories. Using Kaplan-Meier curves, the mean time from study onset until change of clinical categories was 61 months for patients on HAART, 26 months for patients on ART and 14 months for patients without ART. CONCLUSION: In paediatric patients with HIV infection, disease progression has declined substantially by introduction of HAART. Superiority of HAART compared with ART was demonstrated. Non-adherence as well as other reasons for treatment failure have to be studied more carefully.

Early and mid-term outcome of the arterial switch operation in 114 consecutive patients : A single centre experience.

BACKGROUND: The arterial switch operation (ASO) has become the treatment of choice in patients with simple or complex transposition of the great arteries (TGA). The purpose of this study was to assess early and mid-term outcome after ASO in a single centre. PATIENTS AND METHODS: Between 1995 and December 2005, 114 consecutive patients underwent an ASO at our institution, performed by one single surgeon. Patients charts, surgical reports and echocardiograms were retrospectively reviewed. Patients were analyzed in 3 different groups: Group I consisted of 77 neonates with TGA and intact ventricular septum, group II of 13 patients with TGA and ventricular septal defect which had to be closed surgically and group III of 24 patients with various forms of TGA in a complex setting. The patient's median weight was 3.23 kg (1.65-8.30). Twenty-five neonates were born preterm, 18 were diagnosed prenatally. Median follow-up time was 20.7 months (0.3-128.6). RESULTS: The thirty day mortality was 1.75% (2/114), late mortality 0.88% (1/112) accounting for an overall mortality of 2.63%. There was only one early coronary event and so far no late events. Ten of 111 survivors required reoperation, all but 1 from group III. Prevalence of supravalvular pulmonary stenosis was 4.7%. Freedom from reoperation at 5 years of follow-up time was 87.5%. One patient required permanent pacing, no other late arrhythmias occurred. In our series the only risk factor for increased mortality and morbidity was a body weight of less than 2500 g at the time of operation. No better outcome could be demonstrated in the prenatally diagnosed patients. CONCLUSION: The ASO can be performed safely and with low mortality and morbidity even in patients with complex TGA. Follow-up of these patients is required to detect residual problems like supravalvular pulmonary stenosis, coronary problems, arrhythmias and aortic valve dysfunction.

Basal level of anti-Müllerian hormone is associated with oocyte quality in stimulated cycles.

BACKGROUND: Serum anti-Müllerian hormone (AMH) levels provide a powerful means for predicting ovarian response, which is reflected not only by the size of the primordial follicle pool but also by the quality of the oocytes. Considering a mutual interdependence between AMH-expressing somatic cells and gametes, this prospective morphological study was set up to evaluate whether extreme AMH levels represent diminished oocyte quality and developmental incompetence. METHODS: A total of 141 consecutive ICSI patients were subdivided into three groups using the 25th and 75th percentiles of the serum AMH levels (cycle day 3). In these three groups, morphology of all oocytes and fertilization rate, embryo quality and blastocyst formation were evaluated, and FSH, LH and estradiol (E(2)) levels were also measured. RESULTS: Cycle cancellation rate was correlated with AMH levels (P < 0.05). AMH groups 1 (<1.66 ng/ml) and 3 (>4.52 ng/ml) showed oocytes of lower quality [dark central granulation, aggregation of smooth endoplasmic reticulum (sER)] compared with the median group 2 (1.66-4.52 ng/ml). Basal serum FSH did not allow for adequate prognosis in terms of gamete appearance. Fertilization and further cleavage up to blastocyst stage was not affected by AMH levels. CONCLUSIONS: AMH seems to be superior to FSH in predicting both oocyte number and quality.

Vasodilatory shock after surgery for aortic valve endocarditis: use of low-dose vasopressin.

This is the case report of a 13-year-old male who developed vasopressor-resistant hypotension after cardiac surgery for endocarditis. As norepinephrine resulted in aggravation of the preexisting ventricular arrhythmia, vasopressin was used to maintain blood pressure. The vasopressin continuous infusion was started at 0.00002 units/kg/min and titrated up to 0.0003 U/kg/min. This low dose led to resolution of hypotension without causing side effects. As the appropriate indication and dose of vasopressin is not established, the cautious use of vasopressin in children is recommended.

Life threatening cardiopulmonary failure in an infant following protamine reversal of heparin after cardiopulmonary bypass.

Life threatening cardiopulmonary failure following protamine reversal of heparin after cardiopulmonary bypass (CPB) was reported to occur in adults but rarely in children. Atrial septal defect closure was performed in a 6-week-old infant erroneously suspected to suffer from right atrial thrombosis in addition. Protamine administration after CPB led to critical pulmonary hypertension and severe haemorrhagic pulmonary oedema resulting in severe hypoxia. Inhaled nitric oxide, together with high frequency oscillation ventilation supplemented by intravenous prostacycline, enabled complete recovery of cardiopulmonary and neurological function. Life threatening cardiovascular compromise after intravenous protamine can occur even in young infants which then require challenging paediatric critical care.

[Emergencies in pediatric cardiology]. / Kinderkardiologische Notfälle.

Congenital heart defects, not recognized antenatally, may lead to emergency situation immediately after birth. Fast recognition of clinical symptoms and the use of correct diagnostic tools are life-saving under these circumstances. Principles of cardio-pulmonary resuscitation will be presented in this article as well as diagnosis and treatment of hypoxemic spells, pulmonary hypertensive crises and (usually postoperative) pericardial effusion.

[A case of neonatal 49,XXXXY syndrome]. / 49,XXXXY syndroma újszülöttkori esete.

The authors present the case of 49,XXXXY syndrome diagnosed in neonatal age. During the investigation of ambiguous genitalia was identified sex chromosome anomaly. The authors deal with possible pathogenesis of the syndrome and its clinical symptoms in details. They emphasize the importance of both testosterone substitution with pubertal timing, and early intellectual and social development.

D- and L-lactate determination in dairy products: presentation of a fast automated analysis system.

A flow-injection system is presented, which is versatile, automated and has an integrated enzyme system. Its efficiency at determining D- and L-lactate separately in dairy products is compared with the officially used reference method (DIN 10335). By means of a mathematical elimination of matrix effects, the pretreatment steps can be clearly reduced. As a result of the immobilization of the enzymes, an appropriate automation software and an adaptation of the system developed to the given medium, the analysis could be performed within 5 min and was distinctly cheaper than alternative methods. The values determined in 15 different dairy products deviated from those of the reference method by 3% at a maximum.

The Sequence of Change within the Photosynthetic Apparatus of Wheat following Short-Term Exposure to Ozone.

The basis of inhibition of photosynthesis by single acute O(3) exposures was investigated in vivo using analyses based on leaf gas exchange measurements. The fully expanded second leaves of wheat plants (Triticum aestivum L. cv Avalon) were fumigated with either 200 or 400 nanomoles per mole O(3) for between 4 and 16 hours. This reduced significantly the light-saturated rate of CO(2) uptake and was accompanied by a parallel decrease in stomatal conductance. However, the stomatal limitation, estimated from the relationship between CO(2) uptake and the internal CO(2) concentration, only increased significantly during the first 8 hours of exposure to 400 nanomoles per mole O(3); no significant increase occurred for any of the other treatments. Analysis of the response of CO(2) uptake to the internal CO(2) concentration implied that the predominant factor responsible for the reduction in light-saturated CO(2) uptake was a decrease in the efficiency of carboxylation. This was 58 and 21% of the control value after 16 hours at 200 and 400 nanomoles per mole O(3), respectively. At saturating concentrations of CO(2), photosynthesis was inhibited by no more than 22% after 16 hours, indicating that the capacity for regeneration of ribulose bisphosphate was less susceptible to O(3). Ozone fumigations also had a less pronounced effect on light-limited photosynthesis. The maximum quantum yield of CO(2) uptake and the quantum yield of oxygen evolution showed no significant decline after 16 hours with 200 nanomoles per mole O(3), requiring 8 hours at 400 nanomoles per mole O(3) before a significant reduction occurred. The photochemical efficiency of photosystem II estimated from the ratio of variable to maximum chlorophyll fluorescence and the atrazine-binding capacity of isolated thylakoids demonstrated that photochemical reactions were not responsible for the initial inhibition of CO(2) uptake. The results suggest that the apparent carboxylation efficiency appears to be the initial cause of decline in photosynthesis in vivo following acute O(3) fumigation.

Analysis of light-induced reduction of the photochemical capacity in field-grown plants. Evidence for photoinhibition?

It was tested whether field-grown plants (Phaseolus vulgaris, Zea mays and Helianthus annuus) reflect photoinhibitory effects under natural conditions. Attached leaves were used for determination of the photochemical capacity of Photosystem II (Fv/Fm) by means of a portable fluorimeter (PSM, BioMonitor, S.). For a more qualitative description of Fv/Fm, the modifications of the absolute values F0, Fm as well as of the half-rise time of Fm (T/2) were also considered. By comparing artificially shaded and 'sun exposed' plants, the direct influence of light on the photochemical capacity was investigated. Under low natural light conditions the differences of the photochemical capacity between shaded and 'sun exposed' leaves were negligible in all three species. On a day with full sunlight a decline of Fv/Fm was observable at noon-time in the 'sun exposed' leaves of all three species, although the absolute values differed between the species compared. Additionally, the extent of the recovery of Fv/Fm was varying. Both phenomena could be due to differences in the photosynthetic apparatus (e.g., C3-C4, ontogenetic stage, sun-shade type), to self-shading phenomena (comparing leaf layers of Zea and Helianthus) or to differences in the activity of repair mechanisms possibly caused by other environmental factors (vapour pressure deficit = VPD, drought and temperature phenomena).Nevertheless, the results of the shading experiments and the comparison of species lead to the conclusion that primarily light-induced reduction of the photochemical capacity appears at noon in leaves exposed to full sunlight, a partial restoration of Fv/Fm takes place till the evening. Artifically shaded plants show only a slight alteration of the photochemical capacity.