Multi-scale permeability of murine bone measured by nanoindentation

The determination of lacunar-canalicular permeability is essential to understand the mechano-transduction mechanism of bone. Murine models are widely used to investigate skeletal growth and regulation, but the value of lacunar-canalicular permeability is still unclear. To address this question, a poroelastic analysis based on nanoindentation data was used to calculate the lacunar-canalicular permeability of wild type C57BL/6 mice of 12 months. Cross-sections of three tibiae were indented using spherical fluid cell indenter tips of two sizes. Results suggest that the value of lacunar-canalicular intrinsic permeability of B6 female murine tibia is in the order of 10 -24 m2. The distribution of the values of intrinsic permeability suggests that with larger contact sizes, nanoindentation alone is capable of capturing the multi-scale permeability of bone. Multi-scale permeability of bone measured by nanoindentation will lead to a better understanding of the role of fluid flow in mechano-transduction. © 2013 American Society of Civil Engineers.

Effect of dietary linolenic acid/linoleic acid ratio on growth performance, hepatic fatty acid profiles and intermediary metabolism of juvenile yellow catfish Pelteobagrus fulvidraco

The present study aimed to evaluate the effect of dietary linolenic acid (LNA)linoleic acid (LA) ratio on growth performance, hepatic fatty acid profile and intermediary metabolism of juvenile yellow catfish Pelteobagrus fulvidraco. Six isonitrogenous and isolipidic diets were formulated to contain incremental levels of LNA from 0 to 5% at the expense of corn oil (rich in LA), resulting in six dietary treatments with LNA to LA ratios ranging from 0.35 to 14.64. The experiment continued for 7 weeks. Best growth and feed intake were obtained in the fish fed the diets containing the LNA/LA ratios of 1.17 and 2.12 (P<0.05). In contrast, feed conversion ratio was the lowest for fish fed the diets containing the LNA/LA ratios of 1.17 and 2.12 (P<0.05). Dietary LNA to LA ratios significantly influenced viscerosomatic index and hepatosomatic index (P<0.05), but not condition factor (P>0.05). Body composition was also significantly influenced by dietary LNA to LA ratios (P<0.05). Generally, liver FA compositions reflected dietary FA profiles. Declining LA and increasing LNA contents in liver were observed with the increasing dietary LNA/LA ratios (P<0.05). Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) increased with the increasing LNA to LA ratios, suggesting that yellow catfish could elongate and desaturate C18 polyunsaturated fatty acids into highly unsaturated fatty acids. As a consequence, the n-6 fatty acids (FA) declined, and total n-3 FA and n-3/n-6 ratios increased with the dietary ratios of LNA/LA (P<0.05). Dietary LNA to LA ratios significantly influenced several enzymatic activities involved in liver intermediary metabolism (P<0.05), such as lipoprotein lipase, hepatic lipase, pyruvate kinase, succinate dehydrogenase, malic dehydrogenase and lactate dehydrogenase, suggesting that dietary LNA/LA ratios had significant effects on nutrient metabolism in the liver. To our knowledge this is the first demonstration of the effects of dietary LNA to LA ratios on the enzymatic activities of liver in fish, which provides information on diet quality and utilization, and can also be used as an indicator of the nutritional status of this fish. (C) 2009 Elsevier B.V. All rights reserved.

Comparative studies on photosynthesis and phosphate metabolism of Cylindrospermopsis raciborskii with Microcystis aeruginosa and Aphanizomenon flos-aquae

The physiological differences for three bloom-forming cyanobacteria (Cylindrospermopsis raciborskii, Microcystis aeruginosa, and Aphanizomenon flos-aquae) were investigated. In comparison with M. aeruginosa and A. flos-aquae, C. raciborskii exhibited a significantly higher concentration of carotenoids, higher values in maximum photosynthesis rate (P-m), apparent photosynthetic efficieny (a), and maximum electron transport rate (ETRmax) during the growth period. In addition, higher extracellular alkaline phosphatase activities and lower light compensation point (I-c) were also detected in C raciborskii (p < 0.05, ANOVA). Therefore, it is suggested that the higher photosynthetic activities, more effective uptake and utilization to phosphate, and low light requirements might play important roles in the occurrence and invasive behavior of C. raciborskii. Crown Copyright (C) 2009 Published by Elsevier B.V. All rights reserved.

Age-related changes in mouse bone permeability.

The determination of lacunar-canalicular permeability is essential for understanding local fluid flow in bone, which may indicate how bone senses changes in the mechanical environment to regulate mechano-adaptation. The estimates of lacunar-canalicular permeability found in the literature vary by up to eight orders of magnitude, and age-related permeability changes have not been measured in non-osteonal mouse bone. The objective of this study is to use a poroelastic approach based on nanoindentation data to characterize lacunar-canalicular permeability in murine bone as a function of age. Nine wild type C57BL/6 mice of different ages (2, 7 and 12 months) were used. Three tibiae from each age group were embedded in epoxy resin, cut in half and indented in the longitudinal direction in the mid-cortex using two spherical fluid indenter tips (R=238 μm and 500 μm). Results suggest that the lacunar-canalicular intrinsic permeability of mouse bone decreases from 2 to 7 months, with no significant changes from 7 to 12 months. The large indenter tip imposed larger contact sizes and sampled larger ranges of permeabilities, particularly for the old bone. This age-related difference in the distribution was not seen for indents with the smaller radius tip. We conclude that the small tip effectively measured lacunar-canalicular permeability, while larger tip indents were influenced by vascular permeability. Exploring the age-related changes in permeability of bone measured by nanoindentation will lead to a better understanding of the role of fluid flow in mechano-transduction. This understanding may help indicate alterations in bone adaptation and remodeling.

Carbon and nitrogen metabolism of an eutrophication tolerative macrophyte, Potamogeton crispus, under NH4+ stress and low light availability

Submersed macrophytes in eutrophic lakes often experience high NH4+ concentration and low light availability in the water column. This study found that an NH4+-N concentration of 1 mgL(-1) in the water column apparently caused physiological stress on the macrophyte Potamogeton crispus; L The plants accumulated free amino acids (FAA) and lost soluble carbohydrates (SC) under NH4+ stress. These stressful effects of NH4+ were exacerbated under low light availability. Shading significantly increased NH4+ and FAA contents and dramatically decreased SC and starch contents in the plant shoots. At an NH4+-N concentration of 1 mg L-1 in the water column, neither growth inhibition nor NH4+ accumulation was observed in the plant tissues of P. crispus under normal light availability. The results showed that 1 mg L-1 NH4+-N in the water column was not toxic to P. crispus in a short term. To avoid NH4+ toxicity. active NH4+ transportation out of the cell may cost energy and thus result in a decline of carbohydrate. When NH4+ inescapably accumulates in the plant cell, i.e. under NH4+ Stress and shading, NH4+ is scavenged by FAA synthesis. (c) 2009 Published by Elsevier B.V.

Metabolism traits of 'all-fish' growth hormone transgenic common carp (Cyprinus carpio L.)

Transgenic animals with improved qualities have the potential to upset the ecological balance of a natural environment. We investigated metabolic rates of 'all-fish' growth hormone (GH) transgenic common carp under routine conditions and during starvation periods to determine whether energy stores in transgenic fish would deplete faster than controls during natural periods of starvation. Before the oxygen uptake was measured, the mean daily feed intake of transgenic carp was 2.12 times greater than control fish during 4 days of feeding. The average oxygen uptake of GH transgenic fish was 1.32 times greater than control fish within 96 h of starvation, but was not significantly different from controls between 96 and 144 h of starvation. At the same time, GH transgenic fish did not deplete energy reserves at a faster rate than did the controls, as the carcass energy contents of the two groups following a 60-d starvation period were not significantly different. Consequently, we suggest that increased routine oxygen uptake in GH transgenic common carp over that of control fish may be mainly due to the effects of feeding, and not to an increase in basal metabolism. GH transgenic fish are similar to controls in the regulation of metabolism to normally distribute energy reserves during starvation. (c) 2008 Published by Elsevier B.V.

The accumulation and metabolism of astaxanthin in Scenedesmus obliquus (chlorophyceae)

The biosynthesis and metabolism of astaxanthin in coenobium alga Scenedesmus obliquus were investigated using a two-stage culture. The first stage was for the analysis of biosynthesis and accumulation of astaxanthin in alga cells which were cultured under induction conditions (incubation at 30 degrees C and illumination of 180 mu mol m(-2) s(-1)) for 48 h. The composition of the secondary carotenoids in algal cells was analyzed and seven ketocarotenoids were identified. The results implied that S. obliquus synthesized astaxanthin from beta-carotene through three possible pathways. In the second stage, the cultures were transferred to normal conditions (incubation at 25 C and illumination of 80 mu mol m(-2) s(-1)) for 72 h. Algal cells accumulated more chlorophyll and biosynthesis of secondary carotenoids terminated, the content of secondary carotenoids decreased from 59.48 to 6.57%. The results inferred that accumulation and metabolism of astaxanthin could be controlled by cultivated conditions which also could lead the mobilization of secondary carotenoids to support the algal cell growth. The results also implied that presumed conversions from astaxanthin to lutein or antheraxanthin could be modulated by culturing conditions. (C) 2008 Published by Elsevier Ltd.

The role of glutathione metabolism in tolerance of tobacco BY-2 suspension cells to microcystin-RR

This study was undertaken to investigate the role of the glutathione-involved detoxifying mechanism in defending the tobacco BY-2 suspension cells against microcystin-RR (MC-RR). Analysis showed that exposure of the cells to different concentrations of MC-RR (0.1, 1 and 10 mu g/mL) for 0-6 days resulted in a time and concentration-dependent decrease in cell viability and increase in reactive oxygen species (ROS) content. Reduced glutathione (GSH) and total glutathione (tGSH) content as well as glutathione reductase (GR), glutathione peroxidase (GPX) and glutathione-S-transferase (GST) activities significantly increased after 3-4 days exposure in the highest two concentration treated groups, while decreased until reaching the control values except for GPX at day 6. Oxidized glutathione (GSSG) content markedly increased compared with control in high concentration MC-RR treated group after 6 days exposure. The GSH/GSSG ratio was much higher than control in 10 mu g/mL MC-RR treated group at day 4, but after 6 days exposure, the ratios in all treated groups were lower than that of the control group.

Effects of ammonium on growth, nitrogen and carbohydrate metabolism of Potamogeton maackianus A. Benn

Growth, nitrogen and carbohydrate metabolism in relation to eutrophication were studied for a submerged plant Potamogeton maackianus, a species common in East Asian shallow lakes. The plants were grown in six NH4+-N concentrations (0.05, 0.50, 1.00, 3.50, 5.00 and 10.00 mg/L) for six days. NH4+-N levels in excess of 0.50 mg/L inhibited the plant growth. The relationships between external NH4+-N availability and total nitrogen (TN), protein-N, free amino acid-N (FAA-N) and NH4+-N in plant tissues, respectively, conformed to a logarithmic model suggesting that a feedback inhibition mechanism may exist for ammonium uptake. The response of starch to NH4+-N was fitted with a negative, logarithmic curve. Detailed analysis revealed that the influx NH4+-N had been efficiently incorporated into organic-N and eventually stored as protein at the expense of starch accumulation. These data suggest that this species may be able to tolerate high levels of ammonium when dissolved oxygen is sufficient.

Age-related changes in mouse bone permeability

The determination of lacunar-canalicular permeability is essential for understanding local fluid flow in bone, which may indicate how bone senses changes in the mechanical environment to regulate mechano-adaptation. The estimates of lacunar-canalicular permeability found in the literature vary by up to eight orders of magnitude, and age-related permeability changes have not been measured in non-osteonal mouse bone. The objective of this study is to use a poroelastic approach based on nanoindentation data to characterize lacunar-canalicular permeability in murine bone as a function of age. Nine wild type C57BL/6 mice of different ages (2, 7 and 12 months) were used. Three tibiae from each age group were embedded in epoxy resin, cut in half and indented in the longitudinal direction in the mid-cortex using two spherical fluid indenter tips (R=238 μm and 500 μm). Results suggest that the lacunar-canalicular intrinsic permeability of mouse bone decreases from 2 to 7 months, with no significant changes from 7 to 12 months. The large indenter tip imposed larger contact sizes and sampled larger ranges of permeabilities, particularly for the old bone. This age-related difference in the distribution was not seen for indents with the smaller radius tip. We conclude that the small tip effectively measured lacunar-canalicular permeability, while larger tip indents were influenced by vascular permeability. Exploring the age-related changes in permeability of bone measured by nanoindentation will lead to a better understanding of the role of fluid flow in mechano-transduction. This understanding may help indicate alterations in bone adaptation and remodeling. © 2013 Elsevier Ltd.

Effects of ytterbium ion on the growth, metabolism and membrane fluidity of Tetrahymena thermophila

Power-time curves and metabolic properties of Tetrahymena thermophila BF5 exposed to different Yb3+ stop levels were studied by ampoule method of isothermal calorimetry at 28 degrees C. Metabolic rate (r) decreased significantly while peak time (PT) increased with the increase of Yb3+ stop. These results were mainly due to the inhibition of cell growth, which corresponded to the decrease of cell number obtained by cell counting. Compared with cell counting, calorimetry was sensible, easy to use and convenient for monitoring the toxic effects of Yb3+ stop on cells and freshwater ecosystem. It was also found that cell membrane fluidity decreased significantly under the effects of Yb3+ stop, which indicated that Yb3+ could be membrane active molecules with its effect on cell membranes as fundamental aspect of its toxicity.

Meat and bone meal replacement in diets for juvenile gibel carp (Carassius auratus gibelio): effects on growth performance, phosphorus and nitrogen loading

A 11-week growth trial was conducted in a flow-through system with juvenile gibel carp Carassius auratus gibelio to evaluate the effects of gradual replacement of fish meal (FM) by meat and bone meal (MBM) on growth performance, phosphorus (P) and nitrogen (N) loading. Six isonitrogenous (crude protein: 410 g kg(-1)) and isoenergetic (gross energy: 18 kJ g(-1)) diets were formulated. FM was used as the control protein. In the other five diets, 20, 40, 60, 80 and 100% FM protein was substituted with MBM20, MBM40, MBM60, MBM80, MBM100, respectively. Total P content in the diets ranged from 16.0 to 28.3 g kg(-1) and the available P was 5.0-6.6 g kg(-1). The results showed that the best growth was achieved with fish fed on the control diet and MBM20. Final body weight, weight gain, feed efficiency, protein retention efficiency and energy retention efficiency decreased with increased dietary MBM. No significant differences were found in the feeding rate and hepatosomatic index between the groups. Apparent digestibility coefficient (ADC) of dry matter, protein and P decreased with increase in dietary MBM, while there were no significant differences in the ADC of energy. P and N retention decreased linearly while P and N loading increased linearly with the increased dietary MBM levels. No significant differences were observed in the activity of alkaline phosphatase, aspartate aminotransferase and alanine aminotransferase, as well as pyruvate kinase in liver or in serum. Total superoxide dismutase activity in MBM20 was significantly higher than that of MBM100.

Microcalorimetric investigation of the toxic action of Cr(VI) on the metabolism of Tetrahymena thermophila BF5 during growth

Tetrahymena thermophila BF5 produce heat by metabolism and movement. Using a TAM air isothermal microcalorimeter, the power-time curves of the metabolism of T thermophila BF5 during growth were obtained and the action on them by the addition of Cr(VI) were studied. The morphological change with Cr(VI) coexisted and biomass change during the process of T thermophila BF5 growth were studied by light microscope. Chromium has been regarded as an essential trace element for life. However, hexavalent chromium is a known carcinogen, mutagen, cytotoxicant and strong oxidizing agent. Cr(VI) of different concentration have different effects on T thermophila BF5 growth with the phenomenon of low dose stimulation (0-3 x 10(-5) mol L-1) and high dose inhibition (3 x 10(-5) to 2.4 x 10(-4) mol L-1). The relationship between the growth rate constant (k) and c is a typical U-shaped curve, which is a characteristic of hormesis. T thermophila BF5 cannot grow at all when the concentration of Cr(VI) is up to 2.4 x 10(-4) mol L-1. The microscopic observations agree well with the results obtained by means of microcalorimetry. And T thermophila BF5 had obviously morphological changes by the addition of Cr(VI). (c) 2006 Elsevier B.V. All rights reserved.

Effects of salt stress on carbohydrate metabolism in desert soil alga Microcoleus vaginatus Gom.

The effects of salt stress on carbohydrate metabolism in Microcoleus vaginatus Gom., a cyanobacterium isolated from desert algal crusts, were investigated in the present study. Extracellular total carbohydrates and exopolysaccharides (EPS) in the culture medium produced by M. vaginatus increased significantly during the growth phase and reached a maximum during the stationary phase. The production of extracellular carbohydrates also significantly increased under higher salt concentrations, which was attributed to an increase in low molecular weight carbohydrates. In the presence of NaCl, the production of cellular total carbohydrates decreased and photosynthetic activity was impaired, whereas cellular reducing sugars, water-soluble sugars and sucrose content and sucrose phosphate synthase activity increased, reaching a maximum in the presence of 200 mmol/L NaCl. These parameters were restored to original levels when the algae were transferred to a non-saline medium. Sodium and K+ concentrations of stressed cells decreased significantly and H+-ATPase activity increased after the addition of exogenous sucrose or EPS. The results suggest that EPS and sucrose are synthesized to maintain the cellular osmotic equilibrium between the intra- and extracellular environment, thus protecting algal cells from osmotic damage, which was attributed to the selective exclusion of cellular Na+ and K+ by H+-ATPase.