Soil eutrophication in seabird colonies affects cell wall composition: Implications for the conservation of rare plant species.

Seabird colonies exert a strong influence on coastal ecosystems, increasing soil nitrogen bioavailability and modifying plant communities. Previous studies have evidenced that increased N in soils leads to changes in plant cell wall composition; however, this effect has not been assessed in seabird colonies. The main objective of this study was to determine the influence of seabird colonies on nitrogen, cellulose and lignin content in cell walls. For this purpose, analyses were performed on droppings, soils and three native plant species (Armeria pubigera, Armeria pungens and Corema album) growing in yellow-legged gull colonies. The results showed that N excreted by yellow-legged gull is assimilated by plants, increases N content in plant tissues and reduces cellulose and lignin synthesis, therefore potentially altering plant resistance against phytoparasites.

Energetic cost of walking in fossil hominins.

OBJECTIVE: Many biomechanical studies consistently show that a broader pelvis increases the reaction forces and bending moments across the femoral shaft, increasing the energetic costs of unloaded locomotion. However, a biomechanical model does not provide the real amount of metabolic energy expended in walking. The aim of this study is to test the influence of pelvis breadth on locomotion cost and to evaluate the locomotion efficiency of extinct Pleistocene hominins. MATERIAL AND METHODS: The current study measures in vivo the influence of pelvis width on the caloric cost of locomotion, integrating anthropometry, body composition and indirect calorimetry protocols in a sample of 46 subjects of both sexes. RESULTS: We show that a broader false pelvis is substantially more efficient for locomotion than a narrower one and that the influence of false pelvis width on the energetic cost is similar to the influence of leg length. Two models integrating body mass, femur length and bi-iliac breadth are used to estimate the net and gross energetic costs of locomotion in a number of extinct hominins. The results presented here show that the locomotion of Homo was not energetically more efficient than that of Australopithecus and that the locomotion of extinct Homo species was not less efficient than that of modern Homo sapiens. DISCUSSION: The changes in the anatomy of the pelvis and lower limb observed with the appearance of Homo ergaster probably did not fully offset the increased expenditure resulting from a larger body mass. Moreover, the narrow pelvis in modern humans does not contribute to greater efficiency of locomotion.

Carrying loads: Validating a portable tri-axial accelerometer during frequent and brief physical activity.

OBJECTIVES: The aim of this study is to evaluate the validity of the SenseWear Armband™ (SWA, Model MF-SW, BodyMedia Inc.) as a tool to assess daily brief, light- to moderate-intensity activities. DESIGN: A total of 41 volunteers were recruited (27 males, 14 females) to perform several trials, including a resting metabolic rate test and a number of walking trials while carrying different loads in a backpack. METHODS: Energy expenditure during trials was measured using both the SWA™ mini device, with the indirect calorimetry method (MasterScreen CPX and Oxycon Mobile JAEGER™ devices) used for validated comparative measurement. RESULTS: The SWA™ mini shows agreement with indirect calorimetry in all trials. However, the SWA™ mini over-estimated expenditure in all participants. Individual assessment estimates with the SWA™ mini also exhibited random errors. The variations in energy expenditure (EE) resulting from increased carried loads during the trials were not statistically significant when EE was measured with the SWA™ mini. Furthermore, Metabolic Equivalent of Task (MET) calculation, highly dependent on estimated energy expenditure per unit time, also was likely overestimated. In contrast, the SWA™ mini provided estimates of the resting metabolic rate with a small error. CONCLUSIONS: The SWA™ mini is not a valid device for estimating energy expenditure in brief light- or moderate activities.

Microencapsulation of insulin using a W/O/W double emulsion followed by complex coacervation to provide protection in the gastrointestinal tract.

Microcapsules containing insulin were prepared using a combination of a W/O/W double emulsion and complex coacervation between WPI (used as a hydrophilic emulsifier) and CMC or SA with further spray drying of the microcapsules in order to provide protection in the gastrointestinal tract. The microcapsules prepared exhibited high encapsulation efficiency and showed the typical structure of a double emulsion. After spray drying of these microcapsules, the integrity of the W/O/W double emulsion was maintained and the biological residual activity remained high when using the combination of 180 °C inlet air temperature and 70 °C outlet air temperature. The microcapsules exhibited low solubility at pH 2 and high solubility at pH 7 so they might protect insulin at acid pH values in the stomach and release it at intestinal pH values. The microcapsules developed in this study seem to be a promising oral delivery vehicle for insulin or other therapeutic proteins.

Cloning and expression pattern of a gene encoding an alpha-xylosidase active against xyloglucan oligosaccharides from Arabidopsis.

An alpha-xylosidase active against xyloglucan oligosaccharides was purified from cabbage (Brassica oleracea var. capitata) leaves. Two peptide sequences were obtained from this protein, the N-terminal and an internal one, and these were used to identify an Arabidopsis gene coding for an alpha-xylosidase that we propose to call AtXYL1. It has been mapped to a region of chromosome I between markers at 100.44 and 107.48 cM. AtXYL1 comprised three exons and encoded a peptide that was 915 amino acids long, with a potential signal peptide of 22 amino acids and eight possible N-glycosylation sites. The protein encoded by AtXYL1 showed the signature regions of family 31 glycosyl hydrolases, which comprises not only alpha-xylosidases, but also alpha-glucosidases. The alpha-xylosidase activity is present in apoplastic extractions from Arabidopsis seedlings, as suggested by the deduced signal peptide. The first eight leaves from Arabidopsis plants were harvested to analyze alpha-xylosidase activity and AtXYL1 expression levels. Both increased from older to younger leaves, where xyloglucan turnover is expected to be higher. When this gene was introduced in a suitable expression vector and used to transform Saccharomyces cerevisiae, significantly higher alpha-xylosidase activity was detected in the yeast cells. alpha-Glucosidase activity was also increased in the transformed cells, although to a lesser extent. These results show that AtXYL1 encodes for an apoplastic alpha-xylosidase active against xyloglucan oligosaccharides that probably also has activity against p-nitrophenyl-alpha-D-glucoside.

Changes in Dehydrodiferulic Acids and Peroxidase Activity against Ferulic Acid Associated with Cell Walls during Growth of Pinus pinaster Hypocotyl.

Hydroxycinnamic acids associated with hypocotyl cell walls of dark-grown seedlings of Pinus pinaster Aiton were extracted with 1 N NaOH and identified by gas chromatography-mass spectrometry. The main hydroxycinnamic acid found was ferulic acid. Diferulic acid dehydrodimers were also found, with the 8,8-coupled isomer (compound 11) being the dehydrodiferulate present in the highest amount. However, the 5,5-coupled isomer, commonly referred to referred to as diferulic acid, was not detected. Two truxillic acids, 4-4[prime]-dihydroxy-3-3[prime]-dimethoxy-[alpha]-truxillic acids I and II, were tentatively identified. The 8,8-coupled dehydrodiferulic acid (compound 11) was the phenolic acid that showed the most conspicuous changes with hypocotyl age as well as along the hypocotyl axis. Peroxidase activity against ferulic acid was found in the apoplastic fluid as well as being ionically and covalently bound to the cell walls. The peroxidase activity increased with hypocotyl age as well as from the subapical toward the basal region of the hypocotyls. A key role in the cell-wall stiffening of 8,8 but not 5,5 dimerization of ferulic acid catalyzed by cell-wall peroxidases is proposed.

Glucose represses formation of delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-valine and isopenicillin N synthase but not penicillin acyltransferase in Penicillium chrysogenum.

The content of alpha-aminoadipyl-cysteinyl-valine, the first intermediate of the penicillin biosynthetic pathway, decreased when Penicillium chrysogenum was grown in a high concentration of glucose. Glucose repressed the incorporation of [14C]valine into alpha-aminoadipyl-cysteinyl-[14C]valine in vivo. The pool of alpha-aminoadipic acid increased sevenfold in control (lactose-grown) penicillin-producing cultures, coinciding with the phase of rapid penicillin biosynthesis, but this increase was very small in glucose-grown cultures. Glucose stimulated homocitrate synthase and saccharopine dehydrogenase activities in vivo and increased the incorporation of lysine into proteins. These results suggest that glucose stimulates the flux through the lysine biosynthetic pathway, thus preventing alpha-aminoadipic acid accumulation. The repression of alpha-aminoadipyl-cysteinyl-valine synthesis by glucose was not reversed by the addition of alpha-aminoadipic acid, cysteine, or valine. Glucose also repressed isopenicillin N synthase, which converts alpha-aminoadipyl-cysteinyl-valine into isopenicillin N, but did not affect penicillin acyltransferase, the last enzyme of the penicillin biosynthetic pathway.

Carbon catabolite repression of penicillin biosynthesis by Penicillium chrysogenum.

The addition of glucose to batch cultures of Penicillium chrysogenum AS-P-78 reduced the biosynthesis of penicillin. This regulatory effect was also observed in penicillin biosynthesis by nitrogen-limited resting cells when cultures were previously grown in high concentrations of glucose. The effect of glucose was concentration-dependent in the range of 28-140 mM. Incorporation of L-[U-14C]valine into penicillin in nitrogen-limited resting cultures was reduced by 70% when cells were grown on 140 mM glucose, as compared with that grown on lactose. It was not affected when the sugar was added to the resting cell system, in which penicillin biosynthesis took place without growth. Fructose, galactose and sucrose exerted the regulatory effect to the same extent as glucose (64 to 70%). Lactose did not exert suppression of penicillin biosynthesis. Penicillin-synthesizing activity in control cultures with lactose reached a peak at 24 hours of incubation and decreased slowly thereafter, as studied with resting cell cultures in which further protein synthesis was blocked with cycloheximide. Glucose repressed the formation of penicillin-synthesizing enzymes, but had no effect on the activity of these enzymes. These results suggest that glucose represses but does not inhibit penicillin biosynthesis.

Inhibition and repression of homocitrate synthase by lysine in Penicillium chrysogenum.

Homocitrate synthase in the first enzyme of the lysine biosynthetic pathway. It is feedback regulated by L-lysine. Lysine decreases the biosynthesis of penicillin (determined by the incorporation of [14C]valine into penicillin) by inhibiting and repressing homocitrate synthase, thereby depriving the cell of alpha-aminoadipic acid, a precursor of penicillin. Lysine feedback inhibited in vivo the biosynthesis and excretion of homocitrate by a lysine auxotroph, L2, blocked in the lysine pathway after homocitrate. Neither penicillin nor 6-aminopenicillanic acid exerted any effect at the homocitrate synthase level. The molecular mechanism of lysine feedback regulation in Penicillium chrysogenum involved both inhibition of homocitrate synthase activity and repression of its synthesis. In vitro studies indicated that L-lysine feedback inhibits and represses homocitrate synthase both in low- and high-penicillin-producing strains. Inhibition of homocitrate synthase activity by lysine was observed in cells in which protein synthesis was arrested with cycloheximide. Maximum homocitrate synthase activity in cultures of P. chrysogenum AS-P-78 was found at 48 h, coinciding with the phase of high rate of penicillin biosynthesis.

Lysine regulation of penicillin biosynthesis in low-producing and industrial strains of Penicillium chrysogenum.

The inhibitory effect of L-lysine on penicillin biosynthesis by Penicillium chrysogenum has been compared in a low-producing strain (Wis. 54-1255) and a high-producing strain (ASP-78). Lysine inhibited total penicillin synthesis to a similar extent in both strains. However, in the high-producing strain the onset of penicillin synthesis occurred even at a high lysine concentration, whereas in the low-producing strain lysine had to be depleted before penicillin production commenced.