Effect of Protein Intake on Bone Mineralization during Weight Loss: A 6‐Month Trial

Objective: The long‐term effect of dietary protein on bone mineralization is not well understood. Research Methods and Procedures: Sixty‐five overweight (body mass index, 25 to 29.9 kg/m²) or obese (≥30 kg/m²) subjects were enrolled in a randomized, placebo‐controlled, 6‐month dietary‐intervention study comparing two controlled ad libitum diets with matched fat contents: high protein (HP) or low protein (LP). Body composition was assessed by DXA. Results: In the HP group, dietary‐protein intake increased from 91.4 g/d to a 6‐month intervention mean of 107.8 g/d (p < 0.05) and decreased in the LP group from 91.1 g/d to 70.4 g/d (p < 0.05). Total weight loss after 6 months was 8.9 kg in the HP group, 5.1 kg in the LP group, and none in the control group. After 6 months, bone mineral content (BMC) had declined by 111 ± 13 g (4%) in the HP group and by 85 ± 13 g (3%) in the LP group (not significant). Loss of BMC was more positively correlated with loss of body fat mass (r = 0.83; p < 0.0001) than with loss of body weight. Six‐month BMC loss, adjusted for differences in fat loss, was greater in the LP group than in the HP group [difference in LP vs. HP, 44.8 g (95% confidence interval, 16 to 73.8 g); p < 0.05]. Independent of change in body weight and composition during the intervention, highprotein intake was associated with a diminished loss of BMC (p < 0.01). Discussion: Body‐fat loss was the major determinant of loss of BMC, and we found no adverse effects of 6 months of high‐protein intake on BMC.     

Effects of hyperthermia on cerebral blood flow and metabolism during prolonged exercise in humans.

The development of hyperthermia during prolonged exercise in humans is associated with various changes in the brain, but it is not known whether the cerebral metabolism or the global cerebral blood flow (gCBF) is affected. Eight endurance-trained subjects completed two exercise bouts on a cycle ergometer. The gCBF and cerebral metabolic rates of oxygen, glucose, and lactate were determined with the Kety-Schmidt technique after 15 min of exercise when core temperature was similar across trials, and at the end of exercise, either when subjects remained normothermic (core temperature = 37.9 degrees C; control) or when severe hyperthermia had developed (core temperature = 39.5 degrees C; hyperthermia). The gCBF was similar after 15 min in the two trials, and it remained stable throughout control. In contrast, during hyperthermia gCBF decreased by 18% and was therefore lower in hyperthermia compared with control at the end of exercise (43 +/- 4 vs. 51 +/- 4 ml. 100 g(-1). min(-1); P < 0.05). Concomitant with the reduction in gCBF, there was a proportionally larger increase in the arteriovenous differences for oxygen and glucose, and the cerebral metabolic rate was therefore higher at the end of the hyperthermic trial compared with control. The hyperthermia-induced lowering of gCBF did not alter cerebral lactate release. The hyperthermia-induced reduction in exercise cerebral blood flow seems to relate to a concomitant 18% lowering of arterial carbon dioxide tension, whereas the higher cerebral metabolic rate of oxygen may be ascribed to a Q(10) (temperature) effect and/or the level of cerebral neuronal activity associated with increased exertion.     

Paradoxical selective feeding on a low-nutrient diet: why do mangrove crabs eat leaves?

Sesarmid crabs dominate Indo West-Pacific mangroves, and consume large amounts of mangrove litter. This is surprising, since mangrove leaves have high tannin contents and C/N ratios that far exceed 17, normally taken as the maximum for sustainable animal nutrition. This paradox has led to the hitherto untested hypothesis that crabs let leaves age in burrows before consumption, thereby reducing tannin content and C/N ratio. We excavated burrows of Neosarmatium meinerti within high-shore Avicennia marina mangroves, and investigated whether burrow leaves had C, N or C/N values significantly different from those of senescent leaves. Leaves were found in <45% of burrows, mostly only as small fragments, and N concentrations and C/N ratios of burrow leaves never varied significantly from senescent leaves. The leaf-ageing hypothesis was therefore not supported. In the field N. meinerti and Sesarma guttatum fed on sediment in 76% and 66-69% of observations, respectively, and on leaves in <10% of observations. Sediments from two A. marina mangroves had a mean C/N ratio of 19.6. Our results, and the literature, show that mangrove leaves are unlikely to fulfil the N requirements of crabs, whether or not leaf ageing takes place. Sediment detritus could be a richer source of N, as shown by lower C/N ratios and regular ingestion by crabs. By fragmenting leaves crabs may be elevating the nutritional quality of the substrate detritus.     

Changes in the fish community and water quality during seven years of stocking piscivorous fish in a shallow lake

SUMMARY 1. Piscivores (annual stocking of 1000 individuals ha^?1 of 0+ pike and a single stocking of 30 kg ha^?1 of large 20–30 cm perch) were stocked in seven consecutive years in a shallow eutrophic lake in Denmark. The stocking programme aimed at changing food‐web structure by reducing zooplanktivorous and benthivorous fish, with resultant effects on lower trophic levels and ultimately water quality. 2. The fish community and water quality parameters (Secchi depth, concentrations of total phosphorus, chlorophyll a and suspended solids) were monitored between 1996 and 2000 and relationships were evaluated between predatory fish and potential prey and between zooplanktivorous or benthivorous fish and water quality parameters. In addition, potential consumption of piscivorous fishes was calculated. 3. The density of fish feeding on larger zooplankton or benthos (roach >15 cm, crucian carp >15 cm) declined distinctly during the study period. This effect was attributed to predation by large (>50 cm) pike. Based on scale readings, we cautiously suggest that the stocking of 0+ pike boosted the adult pike population to produce an unexpected impact in later years. Conversely, no direct impact of stocked 0+ pike was detected on 0+ roach. 4. A major decline in the recruitment strength of 0+ roach was observed in 2000. A combination of (i) the indirect effect of large pike preying on adult roach, with negative effect on roach reproduction and (ii) the direct predation effect of 0+ pike and/or 1+ and 2+ perch recruited in the lake, provides the most likely explanation of this phenomenon. 5. A marked increase in Secchi depth in 2000 and declining trends in suspended solids, chlorophyll‐a and total phosphorus concentrations were observed. These changes may also be attributable to changes in the fish community, although the relationships were not straightforward. 6. This 7‐year study indicates that piscivorous fish may be a significant structuring force in shallow eutrophic lakes, suggesting that stocking piscivores can increase predation pressure on cyprinids. However, the general lack of impact of 0+ pike points to the need of refining current stocking practices in several countries across Europe.     

Purification of full-length recombinant human and rat type 1 11β-hydroxysteroid dehydrogenases with retained oxidoreductase activities

11β-Hydroxysteroid dehydrogenase type 1 (11β-HSD1) is a membrane-bound glycoprotein localized in the endoplasmic reticulum. This enzyme has a key role in regulating local tissue glucocorticoid concentration, acting in vivo predominantly as an oxidoreductase. Previous attempts to purify the native enzyme have yielded a protein without reductase activity. To facilitate detailed studies on its structure and regulation, we have developed a method to purify the full-length human and rat 11β-HSD1 with retention of their natural oxidoreductase activities. This procedure involved recombinant expression of these histidine-tagged enzymes in the yeast Pichia pastoris; large-scale culturing in a fermentor; and single-step purification by metal affinity chromatography. Both enzymes were 90–95% pure and exhibited dehydrogenase and reductase activities with K_M values in agreement with those reported in the literature.     

Estimations of the importance of plant resources extracted by inhabitants of the Peruvian Amazon flood plains

Extractions/applications of plant-resources were investigated in Mestizo communities in the Jenaro Herrera district, Peruvian Amazon. Knowledge differences between genders on forest resource uses were also examined. Two methods were applied: (1) a forest survey based on structured interviews of ten informants on potential uses of 334 pre-selected trees and lianas in permanent sample plots, and interviews of 20 male and female informants concerning potential uses of 85 trees and lianas; and (2) a household survey recording extracted materials in or near the dwellings of 73 families. The conclusions are: (i) most forest products are extracted from a limited number of species, whereas a large number of species is mentioned as important/potentially useful for identical purposes; (ii) coherence exists between extracted species and their abundance in easily accessible forest formations; (iii) knowledge of forest resources is gender correlated; and (iv) a combination of methods for collection of information on use and extraction of plant resources provides more detailed and profound results than using each method separately.     

Improved nitrogen removal by application of new nitrogen-cycle bacteria

In order to meet increasingly stringentEuropean discharge standards, new applicationsand control strategies for the sustainableremoval of ammonia from wastewater have to beimplemented. In this paper we discuss anitrogen removal system based on the processesof partial nitrification and anoxic ammoniaoxidation (anammox). The anammox process offersgreat opportunities to remove ammonia in fullyautotrophic systems with biomass retention. Noorganic carbon is needed in such nitrogenremoval system, since ammonia is used aselectron donor for nitrite reduction. Thenitrite can be produced from ammonia inoxygen-limited biofilm systems or in continuousprocesses without biomass retention. Forsuccessful implementation of the combinedprocesses, accurate biosensors for measuringammonia and nitrite concentrations, insight inthe complex microbial communities involved, andnew control strategies have to be developed andevaluated.     

Selectivity by moose vs the spatial distribution of aspen: a natural experiment

Patch use theory predicts that herbivores perceive food as patches and spend more time in high quality patches, i.e. feeding sites providing a high net rate of intake of energy and/or limiting nutrients. The herbivores should accordingly not discriminate among food items in such high quality patches, and food items should thus be eaten in proportion to availability. In contrast, classical diet theory assumes food selection to take place at the level of individual plants, and predicts that the forager should concentrate on the most profitable item until availability drops below some critical threshold.
Here we address how the spatial distribution of European aspen Populus tremula , a highly preferred browse species, affects the selectivity by moose Alces alces at the patch and the tree level. The study was performed in a managed boreal forest landscape in coastal northern Sweden, where aspen occurs highly aggregated almost exclusively in discrete patches. We compared moose' selectivity for aspen and browsing intensity on aspen ramets and other browse species in aspen patches versus at randomly located sites.
Random sites and aspen stands were utilised equally by moose in terms of overall use of forage. There was no difference in total coverage of forage species and relative moose density. Selectivity for aspen was stronger at random sites than at aspen sites, and the browsing intensity on aspen was similar. We conclude that moose did not perceive aspen stands as discrete patches, and used aspen ramets more in accordance with diet theory. These findings agree with the idea that large generalist herbivores strive to maintain a mixed and balanced diet, causing rare species to be over-utilised (negative frequency-dependent food selection). By such selective feeding, moose may reinforce the aggregated distribution of aspen in the managed boreal forest landscape.     

Geographic trends in mangrove crab abundance in East Africa

The aim of this work was to determine the abundance of crabs inmangrove communities along a latitudinal gradient along the eastern coastof Africa from 4°S to 32°S. Surveys were made atMombasa (Kenya), Zanzibar (Tanzania), Maputo (Mozambique) and in theTranskei (South Africa). Crabs were estimated at three designated levelsin the mangroves by visual census using a common protocol, and numberswere converted to biomass.Even after standardising the selection of sites and methods of censusthere was still extensive variability in the data, emphasising the complexheterogeneity of mangrove ecosystems. Lunar phase (full versus new moonsprings) did not have a consistent effect on results, but shore height hadseveral effects. Total crab biomass was similar in the two lower shore strataexamined, but about twice as high at the top-Avicennia level. Theratio of grapsid biomass:ocypodid biomass also changed with height: fromnear unity in the lower mangrove, to 0.14 in the middle strata, but to 15at the top.There was no consistent latitudinal trend in total crab numbers, but totalcrab biomass increased from north to south. In addition there was aconsistent and marked change in the grapsid biomass:ocypodid biomassratio: this swung from 0.65 at Mombasa to 6.8 in the Transkei. This hasimplications for the transfer of primary production through the food chain. Grapsids are important macrophagous feeders on the leaves and other partsof mangroves, whereas ocypodids are microphagous deposit feeders.     

Ser-474 is the major target of insulin-mediated phosphorylation of protein kinase B beta in primary rat adipocytes.

The mechanism of activation for protein kinase B (PKB), an important target for insulin signaling, has been scarcely investigated in primary cells. In this study, we have characterized the insulin-induced phosphorylation and activation of PKB beta in primary rat adipocytes. Insulin stimulation resulted in a translocation of PKB beta from cytosol to membranes, and phosphorylation and activation of PKB beta. Phosphoamino acid analysis and phosphopeptide mapping demonstrated that the phosphorylation occurred mainly on serines, also when using calyculin A, and that these were localized within one major phosphopeptide. Radiosequencing showed that the radioactivity was released in Cycle No. 7. In addition, the peptide was specifically immunoprecipitated from a tryptic digest of PKB beta using the anti-phospho-PKB (Ser-473) antibody. Taken together, these results show that rat adipocyte PKB beta mainly is phosphorylated on Ser-474 in response to insulin stimulation, in contrast to previous studies in human embryonic kidney (HEK) 293 cells demonstrating, in addition, phosphorylation of Thr-309.