GIS-based probability assessment of natural hazards in forested landscapes of Central and South-Eastern Europe.

We assessed the probability of three major natural hazards--windthrow, drought, and forest fire--for Central and South-Eastern European forests which are major threats for the provision of forest goods and ecosystem services. In addition, we analyzed spatial distribution and implications for a future oriented management of forested landscapes. For estimating the probability of windthrow, we used rooting depth and average wind speed. Probabilities of drought and fire were calculated from climatic and total water balance during growing season. As an approximation to climate change scenarios, we used a simplified approach with a general increase of pET by 20%. Monitoring data from the pan-European forests crown condition program and observed burnt areas and hot spots from the European Forest Fire Information System were used to test the plausibility of probability maps. Regions with high probabilities of natural hazard are identified and management strategies to minimize probability of natural hazards are discussed. We suggest future research should focus on (i) estimating probabilities using process based models (including sensitivity analysis), (ii) defining probability in terms of economic loss, (iii) including biotic hazards, (iv) using more detailed data sets on natural hazards, forest inventories and climate change scenarios, and (v) developing a framework of adaptive risk management.

Evidence for the degradation of type XI collagen by bovine intervertebral disc- and articular cartilage extracts.

Bovine intervertebral disc- and articular cartilage extracts contain a metalloproteinase system capable of degrading type XI collagen. The collagen-degrading activity is rather low in unmodified extracts but increases considerably on metalloproteinase activation. The similarity between intervertebral disc and articular cartilage in their patterns of (casein-degrading) metalloproteinases and type XI and type II collagen degradation is believed to suggest a similarity in the events underlying the degradative disorders of articular cartilage and intervertebral disc.

Effect of Rumalon (glycosaminoglycan-peptide) on the articular tissue.

The effect of glycosaminoglycan-peptide complex (GPC) (Rumalon, made by Robapharm, Switzerland) on cells of the inflammatory periarticular infiltrate and on the articular chondrocytes was studied in experimentally induced papain arthropathy by means of image cytometry and biochemistry. The GPC therapy exhibited some antiinflammatory effect as documented by the reduction of DNA proliferative activity in the inflammatory infiltrate and lowered the activity of hydrolytic enzymes and enzyme inhibitors detected in chondrocytes.

Affinity chromatography of serine proteinases from the bovine intervertebral disc on BPTI-Separon HEMA 1000.

A method for the purification of serine proteinases from the bovine intervertebral disc using affinity chromatography on basic pancreatic trypsin inhibitor (BPTI) immobilized to the hydroxyalkyl methacrylate copolymer Separon HEMA 1000 E is reported. Its advantage is the possibility of obtaining serine proteinases without an artificial alteration in relative molecular mass.

Effect of selected antirheumatic drugs on the metabolism of cartilage and synovial tissue in experimental arthropathy.

The effects of several antirheumatic drugs on the activity of degradative enzymes in normal and pathologic knee joint cartilage and on the proliferative activity of synovial tissue cells were studied. Inflammatory arthropathy was induced in rabbits by intraarticular papain administration. Elevated contents of proteoglycanase and collagenase, together with an increase in serine and cysteine proteinase inhibitors, were found in animals with papain-induced arthropathy. Inflammation also accelerated the rate of proliferation of cells present in the synovial tissue. In the treated animals, the reduction in enzyme activity, decrease in inhibitor content and decreased DNA proliferation rate were registered to a different degree. The suppression of protein synthesis by nonsteroidal antiinflammatory drugs may explain our findings. The best therapeutic results were achieved with glycosaminoglycan polysulphate (Arteparon).

[Proteinases and their inhibitors in intervertebral discs]. / K problematice proteináz a jejich inhibitoru v meziobratlové ploténce.

The authors investigated the incidence of proteinases and their inhibitors in bovine intervertebral discs. They proved the presence of proteinases of all basic enzyme classes. In some instances (in the class of cysteine proteinases) a more detailed specification of the present enzymes could be made. The authors proved also the presence of inhibitors of serine and cysteine proteinases which were characterized by relative molecular weights. The relative molecular weights were within the range of cca 7000 to cca 40,000.

The effect of the oxygen transfer rate (OSR) on the formation of cellulases by Trichoderma viride in submersion culture.

The formation of cellulases by Trichoderma viride in a medium containing cellulose as a sole source of carbon depends on the oxygen transfer rate (OSR); the OSR, on the other hand, depends on the concentration of cellulose in the medium because the concentration of cellulose strongly affects the viscosity of the medium. In the work presented here, the dependence has been determined for the oxygen transfer rate on geometric relations and viscosity in cellulose-containing media during cultivation in shaken flasks, and the oxygen transfer rate on N(Re') N(G') and N(a) during cultivation in a laboratory fermentor of 3000-mL volume. Two cellulosic materials have been compared with a different effect on viscosity: Microcrystalline beach cellulose and fibrous cellulose. It has been found that, in an applicable range of concentration, microcrystalline cellulose does not affect the oxygen transfer rate (at concentrations up to 3%). Fibrous cellulose increases the OSR during cultivation in shake flasks but decreases it during civilization in fermentors. On the basis of these results, the optimization has been carried out on the cultivation conditions in fermentors.

Biphasic kinetics of metal ion reactivation of trypsin-thiol complexes.

This report describes biphasic kinetic data obtained when trypsin was inhibited by a thiol-containing inhibitor present in Ehrlich ascites tumour cells and then subjected to addition of Hg2+, Cu2+ or Ag+. This resulted in an initial re-activation of the trypsin, followed by inhibition of the enzyme with the addition of higher concentrations of these ions. The significance of these observations is 2-fold: (i) help to elucidate the mechanism of metal ion activation of latent enzymes, and (ii) also indicate that, in certain circumstances, the concentration of added metal ion determines whether the metal acts as an activator or an inhibitor of enzyme activity.

The reversible thiol-disulphide exchange of trypsin and chymotrypsinogen with a tumour-derived inhibitor. Kinetic data obtained with fluorescein-labelled polymeric collagen fibrils and casein as substrates.

Ehrlich ascites cells contain a cytoplasmic inhibitor of both trypsin and the granule neutral protease and possess a reactive thiol which interacts with an important disulphide bond in trypsin, resulting in the formation of the trypsin-inhibitor complex. When a fixed quantity of trypsin was completely inhibited by addition of the cytoplasmic inhibitor, the trypsin could be re-activated by the addition of either trasylol-trypsin or chymotrypsinogen. Since trasyloltrypsin, chymotrypsinogen (and any derived chymotrypsin) has no ability to solubilise fluorescein-labelled peptides from the substrate, the appearance of trypsin activity was probably due to a non-enzymic exchange reaction, in which these inactive forms displaced trypsin from the trypsin-inhibitor complex. Kinetic data suggest that this displacement was a time-dependent equilibrium reaction controlled by the relative concentration of the reacting species.

The interaction of a trypsin-dependent neutral protease and its inhibitor found in tumour cells. Analysis of complex kinetic data involved in a thiol-disulphide exchange mechanism.

Ehrlich ascites tumour cells contain a granule-derived zymogen which on trypsin activation yields a collegenolytic neutral protease. The preparation of the granule fraction by subcellular fractionation procedure results in the preparation of a second fraction referred to as the post-granule supernatant fraction. The post-granule supernatant fraction contains a latent form of the granule-derived neutral protease and an excess of cytoplasmic inhibitor for this enzyme. The inhibitor of neutral protease is also capable of inhibiting trypsin and in each case the chemical mechanism of enzyme.inhibitor complex formation has been shown to be a reversible thiol-disulphide exchange. The post-granule supernatant fraction exhibited complex kinetic data when the interactions between the inhibitor, the latent enzymes and trypsin were examined simultaneously by incremental analysis. The data were interpreted and quantitatively analysed by computer analysis. It was demonstrated that the conventional types of analysis could not have provided meaningful interpretations of the experimental data provided by these complex-interacting systems.

Evidence for the inhibition of trypsin by thiols. The mechanism of enzyme-inhibitor complex formation.

Ehrlich ascites tumour cells contain a neutral protease, capable of solubilising fluorescein-labelled telopeptides from fluorescein-labelled polymeric collagen fibrils. The cells also contain an inhibitor for this enzyme and for trypsin. The enzymically inactive enzyme-inhibitor complex can be dissociated with the mercurial thiol agent, mersalyl, with the consequent regain of enzymic activity. The reactivated neutral protease and also trypsin can be inhibited by addition of thiols such as cysteine, mercaptoethanol and dithiothreitol. Trypsin can be protected from inactivation by the tumor inhibitor by addition of cystine or L-1-tosylamido-2-phenylethyl chloromethyl ketone(TosPheCH2Cl)-inactivated chymotrypsin. The evidence suggests that the inhibitor contains a reactive thiol group which exchanges with one or more significant disulphide bridges in trypsin and the neutral protease, resulting in enzyme-inhibitor complex formation and loss of activity. Similarly, thiols interact with these enzymes resulting in a corresponding loss of enzymic activity. The evidence obtained with Tos-PheCH2Cl-inactivated chymotrypsin, which reactivated previously inhibited trypsin and neutral protease, demonstrates that the active site of the enzyme is not involved in the interaction with the thiol of the inhibitor but that the significant disulphide bond in the enzyme is required for the maintenance of the active site conformation. This disulphide exchange mechanism is therefore a form of reversible allosteric control of proteolytic activity and has been shown to be distinct from the mechanism by which soya bean trypsin inhibitor interacts with trypsin.

Evidence for the presence of a trypsin inhibitor within rabbit and mouse tumour cells.

Post-granule supernatant fractions obtained from induced tumour cells in rabbits and from Ehrlich ascites tumour cells in mice have been shown to contain a protease inhibitor, inhibiting trypsin (EC 3.4.21.4) and neutral proteases located in the cytoplasm of the cells. This inhibition was found to be irreversible over the time period studied, independent of the time of enzyme incubation and independent of the extent of trypsin digestion within an insoluble substrate (within the limits of linear enzyme kinetics).