Acute inhibition of central c-Jun N-terminal kinase restores hypothalamic insulin signalling and alleviates glucose intolerance in diabetic mice.

The hypothalamus has been identified as a main insulin target tissue for regulating normal body weight and glucose metabolism. Recent observations suggest that c-Jun-N-terminal kinase (JNK)-signalling plays a crucial role in the development of obesity and insulin resistance because neuronal JNK-1 ablation in the mouse prevented high-fat diet-induced obesity (DIO) and increased energy expenditure, as well as insulin sensitivity. In the present study, we investigated whether central JNK inhibition is associated with sensitisation of hypothalamic insulin signalling in mice fed a high-fat diet for 3 weeks and in leptin-deficient mice. We determined whether i.c.v. injection of a pharmacological JNK-inhibitor (SP600125) improved impaired glucose homeostasis. By immunohistochemistry, we first observed that JNK activity was increased in the arcuate nucleus (ARC) and the ventromedial hypothalamus (VMH) in both mouse models, relative to normoglycaemic controls. This suggests that up-regulation of JNK in these regions is associated with glucose intolerance and obesity, independent of leptin levels. Acute i.c.v. injection of SP600125 ameliorated glucose tolerance within 30 min in both leptin-deficient and DIO mice. Given the acute nature of i.c.v. injections, these effects cannot be attributed to changes in food intake or energy balance. In a hypothalamic cell line, and in the ARC and VMH of leptin-deficient mice, JNK inhibition by SP600125 consistently improved impaired insulin signalling. This was determined by a reduction of phospho-insulin receptor substrate-1 [IRS-1(Ser612)] protein in a hypothalamic cell line and a decline in the number of pIRS-1(Ser612) immunoreactive cells in the ARC and VMH. Serine 612 phosphorylation of IRS-1 is assumed to negatively regulate insulin signalling. In leptin-deficient mice, in both nuclei, central inhibition of JNK increased the number of cells immunoreactive for phospho-Akt (Ser473) and phospho-GSK-3ß (Ser9), which are important markers of insulin signalling. Collectively, our data suggest that the acute inhibition of central JNK improves impaired glucose homeostasis and is associated with sensitisation of hypothalamic insulin signalling.

Did vicariance and adaptation drive cryptic speciation and evolution of brooding in Ophioderma longicauda (Echinodermata: Ophiuroidea), a common Atlanto-Mediterranean ophiuroid?

Over the last decade, cryptic speciation has been discovered in an increasing number of taxa. Species complexes are useful models for the understanding of speciation processes. Motivated by the discovery of brooding specimens in the common Atlanto-Mediterranean broadcast spawning brittle star, Ophioderma longicauda, a recent study revealed the occurrence of divergent mitochondrial lineages. We analysed 218 specimens from 23 locations spread over the geographic range of the species with partial Cytochrome c Oxidase subunit I (COI) sequences. A subset of this sample was also surveyed with the internal transcribed spacer of the ribosomal DNA cluster (nuclear ITS-1). Our study revealed six highly divergent mitochondrial lineages, and the ITS-1 data confirmed that they most likely represent a species complex. Geographic ranges, abundances and genetic structures are contrasted among the putative cryptic species. Lineages in which brooding specimens have been found form a monophyletic group and are restricted to the Eastern Mediterranean basin, an oligotrophic zone. A phylogeny-trait association analysis revealed a phylogenetic signal for low 'chlorophyll a' values (our proxy for oligotrophy). An ecological shift related to the hyper oligotrophy of the Eastern Mediterranean region is therefore likely to have played a role in the evolution of brooding. This study revealed that a complex mixture of vicariance, population expansion, adaptive divergence and possibly high local diversification rates resulting from brooding has shaped the evolution of this species complex. The dating analysis showed that these events probably occurred in the Pleistocene epoch.

Stem cell transplantation in children with infantile osteopetrosis is associated with a high incidence of VOD, which could be prevented with defibrotide.

Malignant infantile osteopetrosis (MIOP) is a rare hereditary disorder of osteoclast function, which can be reversed by hematopoietic stem cell transplantation (SCT). We observed a high incidence of hepatic veno-occlusive disease (VOD) in transplanted patients and explored the prevention of this complication by using defibrotide (DF) as a prophylaxis. Twenty children with MIOP were consecutively transplanted in our center between 1996 and 2005. Eleven of these patients were transplanted between 1996 and 2001 and experienced an overall incidence of VOD of 63.6% (7/11). VOD was severe in three patients and one patient succumbed to VOD-related multi-organ failure. Owing to this very high incidence of VOD, DF prophylaxis was initiated in nine patients consecutively transplanted between 2001 and 2005. In this group, only one patient (11.1%) was diagnosed with moderate VOD. We report here a very high risk in patients with MIOP to develop VOD after transplantation. Prophylactic DF was implemented in our current transplant protocol and reduced the VOD rate significantly in this high-risk population.

Effects of species diversity on establishment and coexistence: a phylloplane fungal community model system.

A model system was devised, evaluating the influence that species diversity (species richness) has on fungal establishment and coexistence. Seven members of the fungal phylloplane community of Vaccinium macrocarpon (American cranberry) were selected to assess how species diversity affected development and coexistence of another community member, Pestalotia vaccinii. Pestalotia was engaged in competitive interactions on 1% Malt Extract Agar (MEA) petri dishes with each of the seven individual saprotrophs (two-way interaction), in random combinations with three of the seven saprotrophs (four-way interaction), and in random combinations with five of the seven saprotrophs (six-way interaction). The saprotrophic fungi used in this study were Aspergillus sp., Alternaria alternata, Cladosporium cladosporoides, Curvularia lunata, Epicoccum purpuracens, Penicillium sp., and Pithomyces chartarum. We hypothesized that species diversity would have a significant impact on the establishment and coexistence of Pestalotia vaccinii in culture. In an effort to minimize density-dependent effects, the number of viable spores employed in the three types of interactions was kept constant. Target spore concentrations of 50 viable spores of P. vaccinii and 50 saprotroph spores were used, regardless of the number of species involved in the interaction. This proved to be a very important factor in the experiment. As our results show, species diversity had little or no effect on the establishment and coexistence of Pestalotia vaccinii; however, spore density played an extremely important role in the establishment and development of fungal propagules in our model.

Stents in the management of malignant airway obstruction.

Surgical resection is feasible in only 20% of patients with lung cancer: less than 30% of these patients survive > 5 yrs and almost 95% of them require palliative treatment. During the course of disease, 30% of lung cancers cause obstruction of the trachea and main bronchi with subsequent respiratory distress, bleeding and infection. Similar problems arise through secondary pulmonary malignancies. There are several types of central airway obstruction; this influences the modality used for their treatment. The three basic types of stenosis are endoluminal, extraluminal and a combination of both. A mainly endoluminal stenosis can be treated with various resection techniques, such as laser, electrocautery or cryotherapy; for an extraluminal compression the only option is placement of stents, which results in efficient palliation and may prolong survival. Various stent models have been developed for the treatment of inoperable airway stenoses. They consist mainly of two types: metal and silicone devices, or combinations of both (hybrid models). The choice of a specific stent depends on the nature of the airway obstruction, the endoscopist's preference and the overall costs of the procedure. The best treatment results are usually obtained using a combination of stent placement followed by tumour-specific treatment such as irradiation or chemotherapy.

Evolution of mate-choice copying: a dynamic model.

Mate-choice copying has recently been demonstrated in several species. Two, not mutually exclusive, explanations for copying have been proposed: it reduces sampling costs and/or error of mate choice. In guppies, Poecilia reticulata, and black grouse, Tetrao tetrix, young females seem most likely to copy. Therefore, copying may teach inexperienced females what attractive males look like. I developed a 2-year dynamic model, to investigate under which conditions a mate-copying strategy might first evolve. An original population of pure choosers was assumed, which was invaded by a mutant female, able to copy during her first mating season, thereby instantly improving her ability to assess male quality. Alternatively, she could either wait and learn by observing males, just as non-copiers may do, but incurring some time costs, or choose, relying on her own abilities. The degree to which copying occurred among these mutant, young, inexperienced females increased with an increasing proportion of old, experienced females in the population, and with decreasing time left until the end of the season. The model demonstrates that mate-choice copying may evolve, when young females are poor at discrimination and need to learn what high-quality males look like. Male quality proved to be unimportant for copying to evolve, as long as there are sufficient differences in quality for mate choice to be meaningful. As with previous models, time constraints are an important assumption for copying to be advantageous over non-copying. Copyright 1998 The Association for the Study of Animal Behaviour. Copyright 1998 The Association for the Study of Animal Behaviour.

Multiple regulatory steps are involved in the control of lipoprotein lipase activity in brown adipose tissue.

Lipoprotein lipase (LPL) supplies brown adipose tissue with fatty acids for nonshivering thermogenesis. In brown adipose tissue of the Djungarian hamster we studied i) the molecular mechanisms involved in cold-induced stimulation of LPL activity, ii) the adrenergic control of LPL expression, and iii) compared LPL expression in brown and white adipose tissue. i) After 8 h cold exposure we detected a 2-fold increase in LPL activity and protein level in brown adipose tissue, whereas LPL mRNA level remained unchanged. A cold-induced increase (1.5-fold) in LPL activity was observed in brown adipose tissue of hamsters treated with actinomycin D prior to 4 h cold exposure, whereas cycloheximide treatment completely abolished LPL stimulation. Thus, these data suggest that during the initial phase (< 24 h) of cold exposure the stimulation of LPL activity in brown adipose tissue is most likely due to increased translation. In contrast, during prolonged cold exposure, we detected a maximal 7-fold increase in LPL activity and a 2- to 3-fold increase in LPL mRNA level in brown adipose tissue indicating LPL regulation at the pretranslational level. Furthermore, comparison of LPL protein and activity in brown adipose tissue during prolonged (> 24 h) cold exposure provides some evidence that the active fraction of the enzyme pool in brown adipose tissue is increased in response to cold. ii) Surgical denervation and noradrenaline treatment revealed a complex role of the sympathetic innervation in the control of LPL expression in brown adipose tissue. Denervation decreased LPL mRNA level, but increased LPL activity. Noradrenaline treatment stimulated LPL activity to a similar extent as cold exposure. However, cold-induced stimulation of LPL activity was not impaired by denervation. iii) Cold exposure significantly elevated LPL mRNA content of inguinal white adipose tissue, although LPL activity was not affected. Posttranscriptional mechanisms appear to be involved in the tissue specific control of LPL expression.

Hemodilution tolerance in elderly patients without known cardiac disease.

Hemodilution tolerance is not well defined in elderly patients. In 20 patients older than 65 yr and free from known cardiovascular disease, hemodynamic variables, ST segment deviation, and O2 consumption were determined prior to and after 6 and after 12 mL/kg isovolemic exchange of blood for 6% hydroxyethyl starch. The mean age of the patients was 76 +/- 2 yr (mean +/- SEM, range 66-88 yr). During hemodilution, hemoglobin decreased from 11.6 +/- 0.4 to 8.8 +/- 0.3 g/dL (P < 0.05). With stable filling pressures, cardiac index increased from 2.02 +/- 0.11 to 2.19 +/- 0.10 L.min-1.m-2 (P < 0.05) while systemic vascular resistance decreased from 1796 +/- 136 to 1568 +/- 126 dynes.s.cm-5 (P < 0.05) and O2 extraction increased from 28.0% +/- 0.9% to 33.0% +/- 0.8% (P < 0.05) resulting in a stable O2 consumption during hemodilution. No alterations in ST segments were observed in lead II during hemodilution. In lead V5, ST segment deviation became slightly less negative during hemodilution from -0.03 +/- 0.01 to -0.02 +/- 0.01 mV (P < 0.05). The moderate decrease in hemoglobin was fully compensated by both an increase in cardiac index and in O2 extraction. Electrocardiographic signs of myocardial ischemia were not observed in this population. In conclusion, isovolemic hemodilution to a hemoglobin value of 8.8 +/- 0.3 g/dL is well tolerated in elderly patients free from known cardiac disease at the ages of 65-88 yr.

Effects of heparin on aortic versus venous smooth muscle cells: similar binding with different rates of [3H]thymidine incorporation.

The mechanisms resulting in the high incidence of stenoses of coronary venous bypass grafts are still unclear. Heparin, a potential inhibitor of cellular proliferation, neither inhibits intimal hyperplasia in animal models of vein-to-artery grafting nor prevents graft stenosis when administered to patients. We studied the effects of heparin on cultured pairs of human aortic and venous smooth muscle cells (SMC) obtained during coronary bypass surgery from patients with no history of previous restenosis or graft failure. DNA synthesis was measured as [3H]thymidine incorporation after stimulation with 10% fetal calf serum (FCS). Heparin (100 micrograms/ml) inhibited DNA synthesis of aortic SMC to 64 +/- 14% (mean +/- SEM), whereas it stimulated DNA synthesis of venous SMC to 136 +/- 23% (10% FCS alone = 100%; p = 0.01, Wilcoxon signed-rank test, n = 7). Binding studies with [3H]heparin showed no significant differences of Kd values and number of binding sites per cell between SMC derived from aorta or vein that could account for the lack of heparin inhibition of venous SMC DNA synthesis. These data suggest an inherent difference in the heparin susceptibility that may explain the failure of heparin to inhibit intimal proliferation in vein grafts.

[Isolated lung transplantation--evaluation of patients and initial results]. / Die isolierte Lungentransplantation--Evaluation von Patienten und erste Resultate.

Between November 1992 and May 1994 we performed 10 single and 5 double lung transplants in patients with end-stage lung diseases due to lymphangioleiomyomatosis (4), cystic fibrosis (3), pulmonary hypertension (3), pulmonary fibrosis (3) and chronic obstructive lung disease (2). In the 13 patients (87%) surviving for median 245 (19-567) days, FEV1 improved from median 640 ml to 1410 ml and the 12-minute walk distance from median 315 to 1100 meters. 10 patients (77%) enjoy a good or even excellent quality of life. 2 patients died 11 and 62 days postoperatively, due to multi-organ failure and invasive pulmonary aspergillosis respectively. The main postoperative problems are fungal and cytomegalovirus infections and chronic rejection in the form of bronchiolitis obliterans. In Switzerland as elsewhere, lung transplantation has become an established modality for the management of end-stage diseases of the lung and pulmonary circulation.

Expression of meta-vinculin in human coronary arteriosclerosis is related to the histological grade of plaque formation.

In the present study we demonstrate that the quantitative reduction of meta-vinculin expression parallels histological changes during the course of coronary arteriosclerosis. Immunofluorescence stainings of coronary arteries revealed that vinculin distribution resembled that of other smooth muscle-specific cytoskeletal proteins like alpha-actin, caldesmon or myosin light chain kinase in labeling smooth muscle cells brightly. Although close to arteriosclerotic plaques, the cellularity as measured by the density of nuclei was often not significantly altered. Cells of this location expressed markedly reduced amounts of vinculin, suggesting that they are smooth muscle cells of a synthetic phenotype. To determine the fractional meta-vinculin content in arteriosclerotic lesions, we performed densitometric scanning of immunoblots incubated with anti-vinculin monoclonal antibodies reacting with both meta-vinculin (150 kDa) and vinculin (130 kDa). In parallel, each tissue sample was evaluated histologically for the degree of arteriosclerotic alterations according to the morphometric atheroma score of Stratford et al. (n = 13). In type 1 lesions covering slight intimal thickening, meta-vinculin represented 36% (mean, range 35%-39%) of the total vinculin immunoreactivity. In type 2 lesions consisting of fibrous plaques of up to twice the original artery wall thickness, meta-vinculin accounted for 28% (mean, range 22%-35%) of the total vinculin content. Meta-vinculin was substantially reduced in type 3 lesions (mean 13%, range 8%-18%) which are characterized by extensive atheromatous plaques. Thus, the meta-vinculin/vinculin ratio differed significantly between early, intermediate and advanced phases of coronary arteriosclerotic plaque formation.

Effect of unilateral surgical denervation of brown adipose tissue on uncoupling protein mRNA level and cytochrom-c-oxidase activity in the Djungarian hamster.

The bilateral lobe of interscapular brown adipose tissue of the Djungarian hamster was unilaterally denervated in order to study the role of the sympathetic innervation for maintenance and cold-induced increase of non-shivering thermogenesis. Denervation decreased the noradrenaline content of brown adipose tissue to less than 9% of the intact contralateral pad. This low noradrenaline level was maintained for 1-14 days after denervation. First, to study the role of the sympathetic innervation of brown adipose tissue in the maintenance of the high thermogenic capacity characteristic of the cold acclimated state, brown adipose tissue was denervated in hamsters either kept at thermoneutrality or acclimated to 5 degrees C ambient temperature for 4 weeks. Cold-acclimated hamsters had elevated levels of uncoupling protein messenger ribonucleic acid (8.1-fold) and cytochrom-c oxidase-activity (3-fold). Denervation of brown adipose tissue decreased uncoupling protein-messenger ribonucleic acid level and cytochrom-c-oxidase-activity as compared to the intact pad in thermoneutral and in cold-acclimated hamsters. However, in cold-acclimated hamsters uncoupling protein-messenger ribonucleic acid level and cytochrom-c-oxidase-activity in denervated brown adipose tissue both were maintained on an elevated 6-fold higher level as compared to thermoneutral controls. Second, to study the role of the sympathetic innervation of brown adipose tissue in the cold-induced increase in thermogenic capacity, hamsters were denervated prior to cold acclimation and responses were measured after 3 and 14 days of cold exposure. Uncoupling protein-messenger ribonucleic acid level and cytochrom-c-oxidase-activity of intact brown adipose tissue increased after 14 days cold acclimation.(ABSTRACT TRUNCATED AT 250 WORDS)

Antiproliferative effects of a c-myc antisense oligonucleotide on human arterial smooth muscle cells.

The effects of a c-myc antisense phosphorothioate DNA oligonucleotide were assessed on the proliferation rate of human arterial smooth muscle cells (HSMCs). Compared to a control oligonucleotide the antisense oligonucleotide suppressed the proliferation of HSMCs in a concentration-dependent manner without a major cytotoxic effect. Outgrowth of HSMCs from media explants was significantly inhibited as well. Induction of c-myc expression by serum stimulation of cells was blunted by the antisense oligonucleotide, as shown by immunoblotting. These results demonstrate that c-myc expression is an essential factor for proliferation of HSMCs after growth stimulation, and they show the potential of antisense technology for modulating gene expression of HSMCs in vitro.

[Accidents with bicycles and motorcycles. The injury pattern, costs, and possibilities for prevention]. / Unfälle mit Zweiradfahrzeugen. Verletzungsmuster, Kosten, Möglichkeiten der Prävention.

This review comprises all 641 patients subjected to inpatient treatment in 1976, 1980 and 1984 at the Basel district hospital after accidents involving bicycles or motorcycles. A study of the case histories--supplemented by phone conversations--yielded the following results: Accidents involving bicycles or motor-driven bicycles were seen in all age groups, but motorcycle accidents occurred exclusively among the younger generation. Whereas motorcycle accidents mostly happened during joyrides, accidents with bicycles or mobikes mainly occurred on the way to work. The incidence rate was highest during summertime and in the rush hours at noon or in the evening. Motorcycle accidents resulted in more severe injuries, longer hospitalisation, longer periods of disability and higher costs than bicycle or mobike accidents the latter being mainly characterised by mostly slight head injuries and the former by injuries of the legs and arms.

A highly conserved cysteine in the v-Myb DNA-binding domain is essential for transformation and transcriptional trans-activation.

The v-Myb protein is nuclear, binds to DNA in a sequence-specific fashion, regulates the transcription of various reporter gene and transforms myelomonocytic cells. Cysteine is one of the most conserved residues during protein evolution and has been implicated in DNA binding, protein-protein interaction and redox regulation of various proteins. Therefore, we have now individually substituted each of the seven cysteines of v-Myb with a serine. All seven mutant proteins bound to DNA when they were expressed in E. coli. However, mutant C65S neither trans-activated transcription in vivo nor transformed myeloid cells, although it was transported into the nucleus. This cysteine is conserved in the Myb-related proteins of animals, plants, yeast and the cellular slime mold Dictyostelium discoideum. The C65S mutation and a nearby codon insertion mutation also abolished trans-activation by fusion proteins containing the v-Myb DNA-binding domain and the strong constitutive activation domain of herpes simplex virus (HSV) VP16. Because this domain of VP16 appears to activate transcription whenever it is bound upstream of an appropriate promoter, these results imply that C65 may be required for high-affinity DNA binding in vivo. In support of this hypothesis, we have also shown that, in contrast to wild-type v-Myb, mutant C65S is unable to block transcription from a reporter gene in which Myb binding sites overlap the initiation site.

Quantitative MR signal behavior in cardiac cine loops as a function of heart rate.

To evaluate the heart cycle-dependent signal intensity changes in the cardiac chambers, the aorta, and the pulmonary artery, five healthy volunteers were studied with gradient-echo magnetic resonance cine loops at different heart rates. Quantitative evaluation of signal intensity on each side of the cardiac valves showed that there were changes in signal intensity due to section-entry and spin-phase phenomena but none due to the increase in heart rate. The authors conclude that there is no heart rate-dependent signal loss in healthy persons that simulates valvular dysfunction, thus suggesting that signal intensity change can be used as an indicator for this disease, independent of heart rate.