Differential regulation of human 3β-hydroxysteroid dehydrogenase type 2 for steroid hormone biosynthesis by starvation and cyclic AMP stimulation: studies in the human adrenal NCI-H295R cell model

Human steroid biosynthesis depends on a specifically regulated cascade of enzymes including 3β-hydroxysteroid dehydrogenases (HSD3Bs). Type 2 HSD3B catalyzes the conversion of pregnenolone, 17α-hydroxypregnenolone and dehydroepiandrosterone to progesterone, 17α-hydroxyprogesterone and androstenedione in the human adrenal cortex and the gonads but the exact regulation of this enzyme is unknown. Therefore, specific downregulation of HSD3B2 at adrenarche around age 6-8 years and characteristic upregulation of HSD3B2 in the ovaries of women suffering from the polycystic ovary syndrome remain unexplained prompting us to study the regulation of HSD3B2 in adrenal NCI-H295R cells. Our studies confirm that the HSD3B2 promoter is regulated by transcription factors GATA, Nur77 and SF1/LRH1 in concert and that the NBRE/Nur77 site is crucial for hormonal stimulation with cAMP. In fact, these three transcription factors together were able to transactivate the HSD3B2 promoter in placental JEG3 cells which normally do not express HSD3B2. By contrast, epigenetic mechanisms such as methylation and acetylation seem not involved in controlling HSD3B2 expression. Cyclic AMP was found to exert differential effects on HSD3B2 when comparing short (acute) versus long-term (chronic) stimulation. Short cAMP stimulation inhibited HSD3B2 activity directly possibly due to regulation at co-factor or substrate level or posttranslational modification of the protein. Long cAMP stimulation attenuated HSD3B2 inhibition and increased HSD3B2 expression through transcriptional regulation. Although PKA and MAPK pathways are obvious candidates for possibly transmitting the cAMP signal to HSD3B2, our studies using PKA and MEK1/2 inhibitors revealed no such downstream signaling of cAMP. However, both signaling pathways were clearly regulating HSD3B2 expression.

Structure/affinity studies in the bicyclo-DNA series: Synthesis and properties of oligonucleotides containing bcen-T and iso-tricyclo-T nucleosides

We present the synthesis of the two novel nucleosides iso-tc-T and bcen-T, belonging to the bicyclo-/tricyclo-DNA molecular platform. In both modifications the torsion around C6’–C7’ within the carbocyclic ring is planarized by either the presence of a C6’–C7’ double bond or a cyclopropane ring. Structural analysis of these two nucleosides by X-ray analysis reveals a clear preference of torsion angle γ for the gauche orientation with the furanose ring in a near perfect 2’-endo conformation. Both modifications were incorporated into oligodeoxynucleotides and their thermal melting behavior with DNA and RNA as complements was assessed. We found that the iso-tc-T modification was significantly more destabilizing in duplex formation compared to the bcen-T modification. In addition, duplexes with complementary RNA were less stable as compared to duplexes with DNA as complement. A structure/affinity analysis, including the already known bc-T and tc-T modifications, does not lead to a clear correlation of the orientation of torsion angle γ with DNA or RNA affinity. There is, however, some correlation between furanose conformation (N- or S-type) and affinity in the sense that a preference for a 3’-endo like conformation is associated with a preference for RNA as complement. As a general rule it appears that Tm data of single modifications with nucleosides of the bicyclo-/tricyclo-DNA platform within deoxyoligonucleotides are not predictive for the stability of fully modified oligonucleotides.

Vocational identity trajectories: Differences in personality and development of well-being

This person-centred study investigated the longitudinal patterns of vocational identity development in relation to personality, the development of well-being, gender, nationality and the attended school track among two cohorts of Swiss adolescents in 8th or 9th grade (N = 269) and in 11th or 12th grade (N = 230). The results confirmed the existence of four identity statuses, namely, achievement, foreclosure, moratorium and diffusion. Forty-two per cent of students showed progressive patterns of identity development, while 37% remained in their identity status over time. Students with different statuses and status change patterns differed significantly in their personality traits. Higher neuroticism related to the emergence of identity exploration over time, while conscientiousness related to maintaining or achieving a sense of identity commitment in terms of achievement or foreclosure. Controlling for the effects of socio-demographics and personality traits, students who reached or maintained a state characterized by identity clarity and commitment showed a relative increase in life satisfaction, while those entering a state of identity crisis or exploration showed a decrease in life satisfaction. Copyright © 2011 John Wiley & Sons, Ltd.

The long-term development of avalanche risk in settlements considering the temporal variability of damage potential

Recent studies on the avalanche risk in alpine settlements suggested a strong dependency of the development of risk on variations in damage potential. Based on these findings, analyses on probable maximum losses in avalanche-prone areas of the municipality of Davos (CH) were used as an indicator for the long-term development of values at risk. Even if the results were subject to significant uncertainties, they underlined the dependency of today's risk on the historical development of land-use: Small changes in the lateral extent of endangered areas had a considerable impact on the exposure of values. In a second step, temporal variations in damage potential between 1950 and 2000 were compared in two different study areas representing typical alpine socio-economic development patterns: Davos (CH) and Galtür (A). The resulting trends were found to be similar; the damage potential increased significantly in number and value. Thus, the development of natural risk in settlements can for a major part be attributed to long-term shifts in damage potential.

Myelography in the Age of MRI: Why We Do It, and How We Do It

Myelography is a nearly ninety-year-old method that has undergone a steady development from the introduction of water-soluble contrast agents to CT myelography. Since the introduction of magnetic resonance imaging into clinical routine in the mid-1980s, the role of myelography seemed to be constantly less important in spinal diagnostics, but it remains a method that is probably even superior to MRI for special clinical issues. This paper briefly summarizes the historical development of myelography, describes the technique, and discusses current indications like the detection of CSF leaks or cervical root avulsion.

Bedouin in the Negev and Environmental Studies: Current and Future Research Prospects

Environmental aspects are increasingly being integrated in Negev Bedouin studies by both, NGO activists and scholars. We will present these recent works and discuss new concepts and methodologies of environmental studies with potential relevance in the field of Negev Bedouin studies. We will then identify research areas where environmental and development approaches converge or diverge with mainstream social sciences on this specific field of research. While most of the Bedouin population in southern Israel lives in urban centers in the Northern Negev, a large part of Bedouin people live in unrecognized clusters of houses in remote areas. Extensive livestock rearing is an important source of livelihood at least for non-urbanized Bedouin, the latter forming the lowest economic strata of the Israeli spectrum of incomes. Numerous stressors affect this Bedouin community enduring uncertain livelihood and access to land. The erratic precipitations from year to year and long-term changes in precipitation trends are a source of great uncertainty. With a significant price increase for feeding supplements to compensate for dry years, livestock rearing has become a harsher source of livelihood. Land scarcity for grazing adds to the difficulty in ensuring enough income for living. Studies in the last 15 years have described several livelihood strategies based on a livestock rearing semi-nomadic economy in the Negev. A number of other analyses have shown how Bedouin herders and governmental agencies have found agreements at the advantage of both, the agencies and the herders. New concepts such as transformability, resilience and adaptation strategies are important tools to analyze the capacity of vulnerable communities to cope with an ever increasing livelihood uncertainty. Such research concepts can assist in better understanding how Bedouin herders in the Negev may adapt to climate and political risks.

Spastin in the human and mouse central nervous system with special reference to its expression in the hippocampus of mouse pilocarpine model of status epilepticus and temporal lobe epilepsy

In the present in situ hybridization and immunocytochemical studies in the mouse central nervous system (CNS), a strong expression of spastin mRNA and protein was found in Purkinje cells and dentate nucleus in the cerebellum, in hippocampal principal cells and hilar neurons, in amygdala, substantia nigra, striatum, in the motor nuclei of the cranial nerves and in different layers of the cerebral cortex except piriform and entorhinal cortices where only neurons in layer II were strongly stained. Spastin protein and mRNA were weakly expressed in most of the thalamic nuclei. In selected human brain regions such as the cerebral cortex, cerebellum, hippocampus, amygdala, substania nigra and striatum, similar results were obtained. Electron microscopy showed spastin immunopositive staining in the cytoplasma, dendrites, axon terminals and nucleus. In the mouse pilocarpine model of status epilepticus and subsequent temporal lobe epilepsy, spastin expression disappeared in hilar neurons as early as at 2h during pilocarpine induced status epilepticus, and never recovered. At 7 days and 2 months after pilocarpine induced status epilepticus, spastin expression was down-regulated in granule cells in the dentate gyrus, but induced expression was found in reactive astrocytes. The demonstration of widespread distribution of spastin in functionally different brain regions in the present study may provide neuroanatomical basis to explain why different neurological, psychological disorders and cognitive impairment occur in patients with spastin mutation. Down-regulation or loss of spastin expression in hilar neurons may be related to their degeneration and may therefore initiate epileptogenetic events, leading to temporal lobe epilepsy.

The impact of input fluctuations on the frequency-current relationships of layer 5 pyramidal neurons in the rat medial prefrontal cortex

The role of irregular cortical firing in neuronal computation is still debated, and it is unclear how signals carried by fluctuating synaptic potentials are decoded by downstream neurons. We examined in vitro frequency versus current (f-I) relationships of layer 5 (L5) pyramidal cells of the rat medial prefrontal cortex (mPFC) using fluctuating stimuli. Studies in the somatosensory cortex show that L5 neurons become insensitive to input fluctuations as input mean increases and that their f-I response becomes linear. In contrast, our results show that mPFC L5 pyramidal neurons retain an increased sensitivity to input fluctuations, whereas their sensitivity to the input mean diminishes to near zero. This implies that the discharge properties of L5 mPFC neurons are well suited to encode input fluctuations rather than input mean in their firing rates, with important consequences for information processing and stability of persistent activity at the network level.

Percutaneous aortic valve replacement: valvuloplasty studies in vitro

Valvuloplasty of the aortic valve is currently used in selected patients for severe calcified aortic valve disease, but clinical effectiveness is low and complication rate remains high. In this study, the total particle load after valvuloplasty and the embolization risk of calcific debris into the coronary arteries was analyzed in an in vitro model.

Inbreeding and selection on sex ratio in the bark beetle Xylosandrus germanus

Background Local Mate Competition (LMC) theory predicts a female should produce a more female-biased sex ratio if her sons compete with each other for mates. Because it provides quantitative predictions that can be experimentally tested, LMC is a textbook example of the predictive power of evolutionary theory. A limitation of many earlier studies in the field is that the population structure and mating system of the studied species are often estimated only indirectly. Here we use microsatellites to characterize the levels of inbreeding of the bark beetle Xylosandrus germanus, a species where the level of LMC is expected to be high. Results For three populations studied, genetic variation for our genetic markers was very low, indicative of an extremely high level of inbreeding (FIS = 0.88). There was also strong linkage disequilibrium between microsatellite loci and a very strong genetic differentiation between populations. The data suggest that matings among non-siblings are very rare (3%), although sex ratios from X. germanus in both the field and the laboratory have suggested more matings between non-sibs, and so less intense LMC. Conclusions Our results confirm that caution is needed when inferring mating systems from sex ratio data, especially when a lack of biological detail means the use of overly simple forms of the model of interest.

Delayed development of pneumothorax after pulmonary radiofrequency ablation

Acute pneumothorax is a frequent complication after percutaneous pulmonary radiofrequency (RF) ablation. In this study we present three cases showing delayed development of pneumothorax after pulmonary RF ablation in 34 patients. Our purpose is to draw attention to this delayed complication and to propose a possible approach to avoid this major complication. These three cases occurred subsequent to 44 CT-guided pulmonary RF ablation procedures (6.8%) using either internally cooled or multitined expandable RF electrodes. In two patients, the pneumothorax, being initially absent at the end of the intervention, developed without symptoms. One of these patients required chest drain placement 32 h after RF ablation, and in the second patient therapy remained conservative. In the third patient, a slight pneumothorax at the end of the intervention gradually increased and led into tension pneumothorax 5 days after ablation procedure. Underlying bronchopleural fistula along the coagulated former electrode track was diagnosed in two patients. In conclusion, delayed development of pneumothorax after pulmonary RF ablation can occur and is probably due to underlying bronchopleural fistula, potentially leading to tension pneumothorax. Patients and interventionalists should be prepared for delayed onset of this complication, and extensive track ablation following pulmonary RF ablation should be avoided.