Development, preclinical evaluation and validation of a novel quick vascular closure device for transluminal, cardiac and radiological arterial catheterization.

Following percutaneous coronary intervention, vascular closure devices (VCDs) are increasingly used to reduce time to ambulation, enhance patient comfort, and reduce potential complications compared with traditional manual compression. Newer techniques include complicated, more or less automated suture devices, local application of pads or the use of metal clips and staples. These techniques often have the disadvantage of being time consuming, expensive or not efficient enough. The VCD failure rate in association with vascular complications of 2.0-9.5%, depending on the type of VCD, is still not acceptable. Therefore, the aim of this study is to develop a self-expanding quick vascular closure device (QVCD) made from a bioabsorbable elastic polymer that can be easily applied through the placed introducer sheath. Bioabsorbable block-co-polymers were synthesized and the chemical and mechanical degradation were determined by in vitro tests. The best fitting polymer was selected for further investigation and for microinjection moulding. After comprehensive haemocompatibility analyses in vitro, QVCDs were implanted in arterial vessels following arteriotomy for different time points in sheep to investigate the healing process. The in vivo tests proved that the new QVCD can be safely placed in the arteriotomy hole through the existing sheath instantly sealing the vessel. The degradation time of 14 days found in vitro was sufficient for vessel healing. After 4 weeks, the remaining QVCD material was covered by neointima. Overall, our experiments showed the safety and feasibility of applying this novel QVCD through an existing arterial sheath and hence encourage future work with larger calibers.

Changes in ventral respiratory column GABAaR ε- and δ-subunits during hibernation mediate resistance to depression by EtOH and pentobarbital.

During hibernation in the 13-lined ground squirrel, Ictidomys tridecemlineatus, the cerebral cortex is electrically silent, yet the brainstem continues to regulate cardiorespiratory function. Previous work showed that neurons in slices through the medullary ventral respiratory column (VRC) but not the cortex are insensitive to high doses of pentobarbital during hibernation, leading to the hypothesis that GABA(A) receptors (GABA(A)R) in the VRC undergo a seasonal modification in subunit composition. To test whether alteration of GABA(A)R subunits are responsible for hibernation-associated pentobarbital insensitivity, we examined an array of subunits using RT-PCR and Western blots and identified changes in ε- and δ-subunits in the medulla but not the cortex. Using immunohistochemistry, we confirmed that during hibernation, the expression of ε-subunit-containing GABA(A)Rs nearly doubles in the VRC. We also identified a population of δ-subunit-containing GABA(A)Rs adjacent to the VRC that were differentially expressed during hibernation. As δ-subunit-containing GABA(A)Rs are particularly sensitive to ethanol (EtOH), multichannel electrodes were inserted in slices of medulla and cortex from hibernating squirrels and EtOH was applied. EtOH, which normally inhibits neuronal activity, excited VRC but not cortical neurons during hibernation. This excitation was prevented by bicuculline pretreatment, indicating the involvement of GABA(A)Rs. We propose that neuronal activity in the VRC during hibernation is unaffected by pentobarbital due to upregulation of ε-subunit-containing GABA(A)Rs on VRC neurons. Synaptic input from adjacent inhibitory interneurons that express δ-subunit-containing GABA(A)Rs is responsible for the excitatory effects of EtOH on VRC neurons during hibernation.

Skin melanoma in Saarland: incidence, survival and mortality 1970-1996.

Over the past few decades, the incidence of cutaneous malignant melanoma has been rising in both sexes in almost all developed countries, notably those with fair-skinned populations. Detailed population-based time trend analyses of skin melanoma incidence and survival in Germany accounting for stage have not been published until now. We analysed skin melanoma data from the population-based Saarland Cancer Registry in Germany from 1970 to 1996. Incidence rates were age-standardized. We estimated 5-year disease-specific survival rates and evaluated the effects of sex, age, calendar period and staging on the prognosis in Cox's proportional hazards models. From 1970-1972 to 1994-1996, melanoma incidence increased 170% from 2.4 to 6.5 per 100 000 person-years among men and 150% from 2.4 to 6.0 per 100 000 person-years among women. Mortality rates peaked in 1988-1990. After 1988-1990, mortality rates declined among women and remained roughly constant among men. The increase in the incidence of localized melanoma and T1-T2 melanoma respectively is driving the overall incidence trend. The improvement of survival over time is most likely due to earlier detection of skin melanoma. In the 1990s, about 30% of all newly diagnosed skin melanoma had stage T3 or T4, implying that further improvement in survival by earlier detection is feasible.

Primary structure and topological analysis of a skeletal muscle-specific junctional sarcoplasmic reticulum glycoprotein (triadin).

The primary amino acid sequence for a highly abundant junctional sarcoplasmic reticulum glycoprotein (triadin) has been deduced from the cDNA sequence. Based on both biochemical analysis and the predicted amino acid sequence we suggest that this protein is an intrinsic membrane glycoprotein containing a single transmembrane domain that separates the protein into cytoplasmic and luminal domains. The cytoplasmic domain is proposed to contain the amino-terminal 47 amino acids. The remainder of the protein including the carboxyl terminus is proposed to be found within the lumen of the sarcoplasmic reticulum and contains an extremely high concentration of basic residues. Protease analysis of intact triads was consistent with the topological predictions. Western and Northern blots suggest that the protein is specifically expressed in skeletal muscle and not cardiac muscle or brain. The abundance and localization of this protein suggest that it plays an important regulatory or structural role in excitation-contraction coupling in skeletal muscle.

Biochemical characterization of ultrastructural localization of a major junctional sarcoplasmic reticulum glycoprotein (triadin).

Monoclonal antibodies were used to identify a 94-kDa protein that was greatly enriched in traids and junctional face membranes (9.3 +/- 0.2%) but not detected in the transverse tubular and nonjunctional sarcoplasmic reticulum membranes. The 94-kDa protein is a hydrophobic glycoprotein based on endoglycosidase H sensitivity, concanavalin A binding, and labelling with a hydrophobic probe. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the absence and presence of reducing agents suggests that this protein is present as a population of multimeric structures containing a variable number of the 94-kDa subunits. Immunofluorescent staining of serial transverse sections of skeletal muscle shows staining of all fiber types with preferential staining of type II fast fibers. Specific immunofluorescence staining in longitudinal sections of skeletal muscle is confined to the interface between the A- and I-bands where the triad structures are localized. Immunocolloidal gold labeling revealed the 94-kDa glycoprotein to be localized over a region of the junctional sarcoplasmic reticulum where the ryanodine receptor/Ca2+ release channel is localized. The distribution and high abundance of the 94-kDa glycoprotein in the junctional membrane suggest that it performs a structural or functional role in the storage or release of calcium from the junctional sarcoplasmic reticulum in skeletal muscle.