Relationships between conformation of β-lactoglobulin in solution and gel states as revealed by attenuated total reflection Fourier transform infrared spectroscopy

Attenuated total reflection Fourier transform infrared spectroscopy (ATR FT-IR) has been used to compare the structure of β-lactoglobulin, the major component of whey proteins, in solution and in its functional gel state. To induce variation in the conformation of β-lactoglobulin under a set of gelling conditions, the effect of heating temperature, pH, and high pressure homogenization on the conformation sensitive amide I band in the infrared spectra of both solutions and gels has been investigated. The results showed that gelification process has a pronounced effect upon β-lactoglobulin secondary structure, leading to the formation of intermolecular hydrogen-bonding β-sheet structure as evidenced by the appearance of a strong band at 1614 cm⁻¹ at the expense of other regular structures. These results confirm that this structure may be essential for the formation of a gel network as it was previously shown for other globular proteins. However, this study reveals, for the first time, that there is a close relationship between conformation of β-lactoglobulin in solution and its capacity to form a gel. Indeed, it is shown that conditions which promote predominance of intermolecular β-sheet in solution such as pH 4, prevent the formation of gel in conditions used by increasing thermal stability of β-lactoglobulin. On the basis of these findings, it is suggested that by controlling the extent of intermolecular β-structure of the protein in solution, it is possible to modify the ability of protein to form a gel and as a consequence to control the properties of gels.     

A zymogel enhances the self-cleaning abilities of the skin of the pilot whale (Globicephala melas)

Enzyme activity in the stratum corneum of the pilot whale Globicephala melas was investigated employing colorimetric enzyme screening assays combined with NATIVE PAGE, size exclusion chromatography (SEC) and histochemical staining procedures. Applying different substrates, several enzymes were detected. The histochemical demonstration of some selected hydrolytic enzymes enriched in the stratum corneum showed high extracellular accumulation. As demonstrated by size exclusion chromatography, high molar mass aggregates were built up from a glycoprotein-rich 20-30-kD fraction. Using NATIVE PAGE experiments under non-reducing conditions, a selection of five degrading enzymes was recovered within the above-reported aggregates. Activity of extracellular aggregate-attached enzymes in the superficial layer of the stratum corneum exhibited no remarkable decrease potentially resulting from self-degradation. We thus conclude that due to their enclosure within the microenvironment of aggregates, a zymogel is formed and autolysis of the stratum corneum is reduced. With respect to the skin surface, the zymogel with hydrolytic activities covering major parts of it enhances the self-cleaning abilities of the skin of the pilot whale based on physical pre-requisites by hydrolyzing adhesive glycoconjugates of settling biofouling organisms considered as primary steps in fouling.     

Effects of liming on soil chemical characteristics and grass growth in laboratory and long-term field-amended soils

Summary The influence of liming on soil solution composition was compared in two laboratory amended soils and one field amended soil. In the laboratory study, soil solutions were sampled by miscible displacement at intervals of 1 and 10 weeks after liming. In addition to increases in pH and Ca, there were large reductions in the concentrations of Mg, K, Na, Si and Mn. Solution concentration of free Al decreased with liming; however, organically complexed Al increased, as did soluble organic matter. Liming also stimulated mineralization of N as indicated by increased solution NO₃ levels. The field amended soils were obtained from a long-term cutting trial investigating the effects of lime on pasture. Despite the passage of a 16-year interval since application, the effects of lime on soil solution characteristics were still clearly evident and generally consistent with those observed in the laboratory study. Estimated leaching losses of Ca from limed soil were relatively low, amounting to 12%, 27% and 44% of the 4.2, 8.4 and 12.5 t lime ha⁻¹ applied, respectively. The results suggest that, in Eastern Ireland, a lime treatment would maintain and elevated pH and would influence the avialability and mobility of plant nutrients for some decades following application.     

Gel-forming structures and stages of red algal galactans of different sulfation levels

Sol-gel transition processes of algal galactans were studied using cryofixation method in combination with freeze-drying and scanning electron microscopy (SEM) techniques. The structures formed in successive stages of gelling process upon cooling were rapidly frozen at defined temperature points and viewed by SEM. It was established that in the case of both types of gelling galactans investigated, a fine honeycomb-like network exists for a wide range of solution temperatures. The formation and structure of this network depends on the structural type, gelling stage, and concentration of the galactan in solution. The honeycomb suprastructures exist also in carrageenan and agarose sols (at temperatures considerably exceeding the gelling temperatures). An additional helical network formed showed different behaviour in the case of carrageenan and agar-type polysaccharides. In the gel-formation process, tightening of the network takes place in both types of galactan gels; the honeycomb structures persist in carrageenan (furcellaran) but not in agarose gels.     

Long photophase is not a sufficient stimulus to reduce thermogenic capacity in winter-acclimatized short-tailed field voles (Microtus agrestis) during long-term cold acclimation

The thermogenic capacity of brown adipose tissue in winter- and summer-acclimatized short-tailed field voles (Microtus agrestis) was investigated by examining changes in mass of brown adipose tissue, the ratio of white adipose tissue to brown adipose tissue, the concentration of the uncoupling protein (thermogenin) in whole depots (μg) and in mitochondrial mass (μg·mg^-1) and the activity of cytochrome c oxidase in the depots (mmol·min^-1). The concentration of thermogenin in winter-acclimatized voles (n=8), per brown adipose tissue depot and per mitochondrial mass, was significantly higher than in summer-acclimatized voles (n=6). There was no significant difference in the level of cytochrome c oxidase activity between these two groups. Four groups of winter-acclimatized voles (n=6 in each group) were exposed to 5°C for 10, 20, 50 and 100 days in a 14L:10D photoperiod. Body mass, brown adipose tissue mass, white adipose tissue mass and basal metabolic rate were significantly positively related to the length of time cold exposed up to 100 days. There was a significant inverse relationship between the ratio of white to brown adipose tissue mass and the duration of cold exposure. There was no significant relationship between thermogenin concentration, either per depot or in mitochondrial mass of brown adipose tissue, with the length of time cold exposed. The level of cytochrome c oxidase activity increased significantly from control levels to a maximum after 10 days in the cold but decreased from 10 days onwards. In winter-acclimatized M. agrestis, a 14L:10D photoperiod is not a sufficient stimulus to reduce thermogenic capacity during cold acclimation. Indeed, some changes in the indirect parameters reflecting thermogenesis, notably the increase in basal metabolic rate and the decrease in the ratio of white to brown adipose tissue mass, indicated that despite the long photophase the thermogenic capacity was slightly further enhanced during the cold acclimation.     

Release of membrane proteins in relation to heat injury of spinach chloroplasts

Thylakoids isolated from spinach leaves ( Spinacia oleracea L. cvs. Monatol and Montako) were exposed to supraoptimal temperatures that inactivated their photochemical reactions. Membrane injury was accompanied by release of a small amount of membrane proteins. When sucrose was present during high-temperature treatment, thylakoids were partially protected and release of membrane proteins was less pronounced than in the absence of sugar. From thylakoids, which were isolated from heat-damaged spinach leaves, less protein was released when heated again after the isolation procedure, indicating that protein release also takes place during heat inactivation in vivo . Sodium dodecyl sulfate gel electropherograms of thylakoids demonstrated that heat inactivation of the lamellae was not accompanied by significant changes in the pattern of the proteins, which remained in the membranes. The same was found when thylakoids were solubilized with Triton X-100 before and after heat damage. It is suggested that the protein release that occurs during heat treatment is a consequence of irreversible alterations in the membrane structure; these changes may be responsible for thermal damage of chloroplast membranes.