Design and Characterization of a New pVII Combinatorial Phage Display Peptide Library for Protease Substrate Mining Using Factor VII Activating Protease (FSAP) as Model

We describe a novel, easy and efficient combinatorial phage display peptide substrate-mining method to map the substrate specificity of proteases. The peptide library is displayed on the pVII capsid of the M13 bacteriophage, which renders pIII necessary for infectivity and efficient retrieval, in an unmodified state. As capture module, the 3XFLAG was chosen due to its very high binding efficiency to anti-FLAG mAbs and its independency of any post-translational modification. This library was tested with Factor-VII activating protease (WT-FSAP) and its single-nucleotide polymorphism variant Marburg-I (MI)-FSAP. The WT-FSAP results confirmed the previously reported Arg/Lys centered FSAP cleavage site consensus as dominant, as well as reinforcing MI-FSAP as a loss-of-function mutant. Surprisingly, rare substrate clones devoid of basic amino acids were also identified. Indeed one of these peptides was cleaved as free peptide, thus suggesting a broader range of WT-FSAP substrates than previously anticipated.     

Experimental methods in hepatology. Guidelines of the German Association for the Study of the Liver (GASL). Liver perfusion--technique and applications

Purified cell-envelope polyphosphatase as well as polyphoshatase activities of cytosol and isolated vacuoles, of nuclei and mitochondria of the yeast Saccharomyces cerevisiae were compared. The polyphosphatases of cell envelope and cytosol are similar, the polyphosphatases of nuclei, vacuoles and mitochondria differ in their kinetic properties, substrate specificity, requirements in divalent cations and in some effector actions both from these and from each other.     

Increased concentrations of octachlorodibenzo-p-dioxin in cases with breast cancer--results from a case-control study

Organochlorines are persistent and highly lipophilic environmental contaminants which bioaccumulate in the food chain. Some of these chemicals, 2,2-bis(p-chlorophenyl)-1,1,1-trichloroethane (DDT) and polychlorinated biphenyls (PCBs), have been suggested to be of significance in the aetiology of breast cancer. 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is an anti-oestrogen in animal studies and should be thus lower the risk of breast cancer. The other isomers of polychlorinated dibenzo-p-dioxins (PCDDs) or the chemically related polychlorinated dibenzofurans (PCDFs) have not been tested regarding carcinogenesis of the breast. The purpose of this study was to investigate whether PCDDs or PCDFs influence the risk for breast cancer. Consecutive patients who underwent surgery for a breast disease between 1993 and 1995 were recruited for the study. Cases were 22 patients with infiltrative breast cancer and controls were 19 patients operated for a benign breast disease during the same time period. Approximately 10 g of breast tissue free from tumour was taken from the specimen and frozen until analysis. Fat was extracted, cleaned and analysed with a high-resolution gas chromatograph coupled to a high-resolution mass spectrometer. Median concentrations of octachlorinated dibenzo-p-dioxin (OCDD) were 598 (170-14,880) and 396 (103-1,847) pg/g lipid in the cases and in the controls, respectively. In a multivariate logistic regression analysis controlling for other risk factors for breast cancer increased odds ratio (OR) was obtained for OCDD: 401-1000 pg/g lipid yielded OR 3.8, 95% confidence interval (CI) 0.4-39, > 1000 pg/g lipid gave OR 5.2, CI 0.4-72. When the lipid OCDD variable was examined as a continuous risk factor there was a 1.09 (9%), CI 0.95-1.25, increase in the adjusted OR for breast cancer per 100 unit (pg/g lipid) increase in OCDD. No differences were found between cases and controls for the other six tested PCDDs. Mean concentration of TCDD was in the cases 3.6 (1.0-7.9) and in the controls 3.3 (1.1-6.3) pg/g lipid. For PCDFs no significant differences were found between cases and controls. The results were not changed if oestrogen or progesterone receptor status, S-phase fraction and DNA ploidy were considered. Breast tissue concentration of OCDD was increased in cancer patients, whereas the concentrations of other PCDDs and PCDFs were equal in cases and controls.     

An 8-bit, 200 MSPS Folding and Interpolating ADC

An 8-bit, 200 MSPS folding and interpolating analog-to-digitalconverter, ADC, has been implemented in a 1.2 µmBiCMOS-process. It achieves 7.5 effective bits with a power dissipationof 575mW. The active area is 4mm². The implementationand measured results are presented. A simple analytical modelfor the interpolation-induced nonlinearity in a folding and interpolatingADC using sinusoidal folding is presented. The bowing of thereference ladder due to interaction with the input stages isanalyzed, and analytical models are derived.     

ALUM SHALE BITUMEN MATURATION AND MIGRATION: IMPLICATIONS FOR GOTLAND'S OIL

Thermal maturation of Swedish Alum Shale kerogen and bitumen has been determined from core samples from Eastern and Central Sweden. In samples from Eastern Sweden (Öland and Gotland), the kerogen and bitumen are thermally immature with respect to petroleum generation. In some areas of Central Sweden (Närke, ÖstergÖtland, and Kinnekulle in VästergÖtland), the kerogen is immature, whereas the bitumen is marginally mature to mature. In other areas of Central Sweden (Halleberg-Hunneberg in VästergÖtland), the kerogen is supermature and the bitumen mature. This suggests that bitumen from a mature source-rock has migrated into the Alum Shales of Central Sweden. Migration in Central Sweden is further evidenced by the occurrence of obviously-migrated bituments in vugs and voids in the organicpoor Ordovician limestone overlying the Alum Shale in Central Sweden, and in concretions within the Alum Shale itself. Based on biomarker distributions of extracted bitumen, Alum Shale kerogen pyrolysate and obviously-migrated oils, and the fact that the Alum Shale in most of the areas studied is the only petroleum source-rock extant, it is suggested that the migrated bitumen in Central Sweden is from the Alum Shale itself. Bitumen has migrated from areas where the Alum Shale is in close proxmity to Permo-Carboniferous intrusions, such as Halleberg-Hunneberg, into nearby areas such as Närke and ÖstergÖtland, where there is no evidence of intrusion and the indigenous organic matter is thermally immature. Other areas, where Alum Shales were associated with intrusions and consequently sourced oil, may have been eroded away. There are producing wells on the island of Gotland, where the Alum Shale is also thermally immature. It is therefore assumed that heating which was responsible for generating Gotland's oil was very localized (such as by an intrusion) or that the oil has migrated from a thermally moremature, distant area. On the basis of reservoir rock porosity, and the fact that the Alum Shale of Gotland contains no migrated component, localized heating is favored.     

Solubility of some transition metal oxides in cryolite-alumina melts: Part II. Solubility of TiO2

The solubility of TiO₂ in cryolite-alumina melts at 1020 °C was measured; it decreased with increasing alumina concentration up to ∼3.5 wt pct total oxide and then increased at higher alumina concentrations. The solubility was found to be 3.1 wt pct Ti in cryolite, and 2.7 wt pct Ti in an alumina-saturated melt. Modeling indicated that the most probable titanium species are TiOF₂ and Na₂TiO₃, which coexist in the solution; the former dominates at low alumina concentrations and the latter at high alumina concentrations. Additional unknown amounts of fluoride may also be associated with these species. Determination of the solubility of TiO₂ in alumina-saturated melts as a function of temperature showed that the solubility increased from 1.9 wt pct Ti at 975 °C to 2.8 wt pct Ti at 1035 °C, the apparent partial molar enthalpy of dissolution of TiO₂ being 88±4 kJ mol⁻¹.     

Long-term low-dose calcitriol treatment in predialysis chronic renal failure: can it prevent hyperparathyroid bone disease?

In an uncontrolled open study 13 patients with moderate to preterminal renal failure were treated with low doses (average 0.36 micrograms/day) of calcitriol up to the time of renal transplantation, which was performed before dialysis had been initiated. A transiliac bone biopsy was obtained both at the start and at the end of the treatment period, the latter coinciding with renal transplantation. All patients who started calcitriol treatment at a creatinine clearance (Ccr) above 30 ml/min had normal bone histology at the time of transplantation, but this was not observed when calcitriol treatment was started at Ccr below 30 ml/min. The study suggests that full benefit of calcitriol at the bone level is obtained only if prophylactic administration is started early in the course of renal failure.