Loads on an internal spinal fixation device during sitting.

Sitting is often assumed to involve high loads on the spine as well as on implants for stabilising the spine. Loads on internal spinal fixation devices were therefore measured in ten patients sitting on several types of seats, including a stool, a stool with a padded wedge, a chair, a physiotherapy ball, a knee-stool, and a bench. The patients also successively sat relaxed and erect on a stool. In addition, six of them sat on a special chair allowing different inclinations of the backrest. Implant loads were also measured for standing up and sitting down. There were only minor differences in fixator loads for sitting on the different types of seats. Sitting erect caused an average of 11% higher implant loads than sitting relaxed. Implant loads decreased with increasing inclination of the upper body while sitting on a chair with an adjustable backrest. Implant loads were about 27% higher for standing up and sitting down than for sitting.     

Evidence for an internal mechanism of copper toxicity in aquatic animals

We exposed frog (Rana pipiens) rectus abdominus muscle to 10(-6) M copper and 10(-5) M acetylcholine separately and in combination to test the hypothesis that copper is directly involved in the muscle spasms of fish dying from exposure to incipient lethal concentrations of copper. Copper alone had little effect but mixtures of copper and acetylcholine caused larger contractions than acetylcholine alone. Exposure of muscle to copper plus acetylcholine for 10-15 min resulted in spontaneous, spasmodic contractions.     

The mechanism of group I self-splicing: an internal guide sequence can be provided in trans.

We have reconstituted a group I self-splicing reaction between two RNA molecules with different functional RNA parts: a substrate molecule containing the 5' splice site and a functional internal guide sequence (IGS), and a ribozyme molecule with core structure elements and splice sites but a mutated IGS. The 5' exon of the substrate molecule is ligated in trans to the 3' exon of the ribozyme molecule, suggesting that the deficient IGS in the ribozyme can be replaced by an externally added IGS present on the substrate molecule. This result is different from catalysis mediated by proteins where it is not possible to dissect the specificity of an enzyme from its catalytic activity.     

The development of a new technology for controlled cell fixation. A methodological pilot study.

Fixation in a traditional sense means the immersion of biological material into a chemical fluid. For permanent preservation (1) the fixative is always supplied in excess of the cell sample, and (2) the process of fixation is influenced by chemical impurities of the fixative fluid. Both factors influence the subsequent staining of cells. In order to avoid these uncontrolled influences, a new technology for controlled cell fixation has to be developed, whereby freshly prepared formaldehyde gas in an "inert" gas-flow of helium was applied to thin membranes by use of a capillary flow-in technique. The amount of fixative gas supplied, adsorbed, absorbed, diffused, and desorbed after saturation of the membranes could be reliably measured with an on-line operating "inert" mass spectrometer of the Omegatron type.     

Evaluating the stability of fracture fixation systems: mechanical device for evaluation of 3-D stiffness in vitro]

Different fixation systems are used for fracture and defect treatment. A prerequisite for complication free healing is sufficient mechanical stability of the osteosynthesis. In vitro investigations offer the possibility of both analysing and assessing the pre-clinical fixation stability. Due to the complex loading environment in vivo, stiffness analysis should include a complete determination of the stiffness under standardised conditions. Based on a mathematical procedure to calculate the 3-D stiffness, a mechanical testing device for the 3-D loading of fixation systems was designed and integrated in the existing test set-up. The set-up consisted of a material testing machine to produce the necessary loads and an optical measurement device to detect the resulting inter-fragmentary movements. To validate the testing device, the 3-D stiffness matrices of different Ilizarov fixator configurations were determined and compared. The good reproducibility of the test was reflected in the small intra-individual variability of the stiffness components. A distinct direction dependence of the fixator stiffness was observed. Increasing the number of rings led to a stiffness increase of up to 50%, especially in bending. The presented testing device allows a complete standardised determination of the stiffness of different fixation systems. It considers the direction dependence of the stiffness and creates a prerequisite for a more direct implant comparison.     

A new mechanism for the aerobic catabolism of dimethyl sulfide.

Aerobic degradation of dimethyl sulfide (DMS), previously described for thiobacilli and hyphomicrobia, involves catabolism to sulfide via methanethiol (CH3SH). Methyl groups are sequentially eliminated as HCHO by incorporation of O2 catalyzed by DMS monooxygenase and methanethiol oxidase. H2O2 formed during CH3SH oxidation is destroyed by catalase. We recently isolated Thiobacillus strain ASN-1, which grows either aerobically or anaerobically with denitrification on DMS. Comparative experiments with Thiobacillus thioparus T5, which grows only aerobically on DMS, indicate a novel mechanism for aerobic DMS catabolism by Thiobacillus strain ASN-1. Evidence that both organisms initially attacked the methyl group, rather than the sulfur atom, in DMS was their conversion of ethyl methyl sulfide to ethanethiol. HCHO transiently accumulated during the aerobic use of DMS by T. thioparus but not with Thiobacillus strain ASN-1. Catalase levels in cells grown aerobically on DMS were about 100-fold lower in Thiobacillus strain ASN-1 than in T. thioparus T5, suggesting the absence of H2O2 formation during DMS catabolism. Also, aerobic growth of T. thioparus T5 on DMS was blocked by the catalase inhibitor 3-amino-1,2,4-triazole whereas that of Thiobacillus strain ASN-1 was not. Methyl butyl ether, but not CHCl3, blocked DMS catabolism by T. thioparus T5, presumably by inhibiting DMS monooxygenase and perhaps methanethiol oxidase. In contrast, DMS metabolism by Thiobacillus strain ASN-1 was unaffected by methyl butyl ether but inhibited by CHCl3. DMS catabolism by Thiobacillus strain ASN-1 probably involves methyl transfer to a cobalamin carrier and subsequent oxidation as folate-bound intermediates.     

A new device for the freeze-clamping of tissue samples

Measurement of metabolite concentrations in tissue samples involves the following procedures: Removal of the sample from the animal, temporary arrest of metabolism, extraction (including weighing, homogenization, final fixation, and neutralization) and assay. Rapid temporary fixation following the sampling of tissue is essential to prevent autolytic changes in metabolite concentrations (1,2). The freeze-clamping technique described by Wollenberger et al. (3) meets this requirement as long as the final thickness of the freeze-clamped sample is sufficiently small. For brain tissue the limit seems to be about 2 mm (4).In our laboratory we have made extensive use of the freeze-clamping tongs of Wollenberger et al., especially for small tissue samples freeze-clamped in situ. However, when in situ clamping can not be used when more than 2–3 g of tissue must be sampled, the freeze-clamping press described below has proven very useful.     

Fabrication of a new substrate for atomic force microscopic observation of DNA molecules from an ultrasmooth sapphire plate.

A new stable substrate applicable to the observation of DNA molecules by atomic force microscopy (AFM) was fabricated from a ultrasmooth sapphire (alpha-Al2O3 single crystal) plate. The atomically ultrasmooth sapphire as obtained by high-temperature annealing has hydrophobic surfaces and could not be used for the AFM observation of DNA. However, sapphire treated with Na3PO4 aqueous solution exhibited a hydrophilic character while maintaining a smooth surface structure. The surface of the wet-treated sapphire was found by x-ray photoelectron spectroscopy and AFM to be approximately 0.3 nm. The hydrophilic surface character of the ultrasmooth sapphire plate made it easy for DNA molecules to adhere to the plate. Circular molecules of the plasmid DNA could be imaged by AFM on the hydrophilic ultrasmooth sapphire plate.     

Evidence for an internal promoter in the Escherichia coli threonine operon.

Auxotrophic mutants of Halobacterium volcanii generated by chemical mutagenesis were used to demonstrate a native genetic transfer system in this extremely halophilic member of the class Archaeobacteria.