Elevated hypothalamic beacon gene expression in Psammomys obesus prone to develop obesity and type 2 diabetes

Objective: To investigate hypothalamic beacon gene expression at various developmental stages in genetically selected diabetes-resistant and diabetes-prone Psammomys obesus. In addition, effects of dietary energy composition on beacon gene expression were investigated in diabetes-prone P. obesus. Methods: Hypothalamic beacon gene expression was measured using TaqmanÔ fluorogenic PCR in 4-, 8- and 16-week-old animals from each genetically selected line. Results: Expression of beacon was elevated in the diabetes-prone compared with diabetes-resistant P. obesus at 4 weeks of age despite no difference in body weight between the groups. At 8 weeks of age, hypothalamic beacon gene expression was elevated in diabetes-prone animals fed a high-energy diet, and was correlated with serum insulin concentration. Conclusion: P. obesus with a genetic predisposition for the development of obesity and type 2 diabetes have elevated hypothalamic beacon gene expression at an early age. Overexpression of beacon may contribute to the development of obesity and insulin resistance in these animals.

Effect of elevated lipid concentrations on human skeletal muscle gene expression

Dietary fatty acids regulate the abundance and activity of various proteins involved in the regulation of fat oxidation by functioning as regulators of gene transcription. To determine whether the transcription of key lipid metabolic proteins necessary for fat metabolism within human skeletal muscle are regulated by acute elevations in circulating free fatty acid (FFA) concentrations, 7 healthy men underwent 3 randomized resting infusions of Intralipid (20%) with heparin sodium, saline and heparin sodium, or saline only for 5 hours. These infusions significantly elevated plasma FFA concentrations by 15-fold (to 1.67 ± 0.13 mmol/L) in the Intralipid infusion trial, with modest elevations observed in the saline and heparin sodium and saline alone infusion groups (0.67 ± 0.09 and 0.49 ± 0.087 mmol/L, P < .01 both vs Intralipid infusion). Analysis of messenger RNA (mRNA) concentration demonstrated that pyruvate dehydrogenase kinase isoform 4 (PDK4) mRNA, a key negative regulator of glucose oxidation, was increased in all trials with a 24-fold response after Intralipid infusion, 15-fold after saline and heparin infusion, and 9-fold after saline alone. The PDK4 increases were not significantly different between the 3 trials. The mRNA concentration of the major uncoupling protein within skeletal muscle, uncoupling protein 3, was not elevated in parallel to the increased plasma FFA as similar (not, vert, similar2-fold) increases were evident in all trials. Additional genes involved in lipid transport (fatty acid translocase/CD36), oxidation (carnitine palmitoyltransferase I), and metabolism (1-acylglycerol-3-phosphate O-acyltransferase 1, hormone-sensitive lipase, and peroxisomal proliferator-activated receptor-γ coactivator-1α) were not altered by increased circulating FFA concentrations. The present data demonstrate that of the genes analyzed that encode proteins that are key regulators of lipid homeostasis within skeletal muscle, only the PDK4 gene is uniquely sensitive to increasing FFA concentrations after increased plasma FFA achieved by intravenous lipid infusion.

Comparison of the color stability and lipid oxidative stability of fresh vacuum packaged lamb muscle containing elevated omega-3 and omega-6 fatty acid levels from dietary manipulation

A series of three experiments were conducted with second cross ([Merino×Border Leicester]×Poll Dorset) wether lambs to evaluate the effects of dietary treatments on manipulation of muscle long-chain (LC) omega-3 fatty acids (FA) on the color stability and oxidative stability of fresh and vacuum packaged lamb. At the end of 7-, 6- and 6-week experimental periods for experiments (Exp.) 1–3 respectively, lambs were slaughtered at a commercial abattoir. At 24 h post-mortem, muscle longissimus lumborum (LL) and longissimus thoracis (LT) were removed and evaluated for color and lipid oxidative stability under specified commercial storage and display condition. Of the dietary supplements used, fish meal and fish oil moderately (P<0.01) and markedly (P<0.001) increased muscle omega-3 FA content, while both protected canola seed (P<0.001) and protected sunflower meal protein significantly (P<0.02) increased muscle omega-6 FA content or ratio of omega-6/omega-3 of the longissimus muscle. In all experiments, the substantial increase (P<0.001) in muscle LC omega-3 and omega-6 FA had no consistent significant effect on color values (redness (a*), yellowness (b*) and lightness (L*)) for fresh and vacuum packaged lamb over a 6-day display period. Lipid oxidation, determined by the levels of thiobarbituric acid reactive substances (TBARS) indicated the enrichment of muscle polyunsaturated fatty acid (PUFA) levels in lambs did not produce significant differences resulting either from main treatment effects or for treatment×day×type interactions (where type was fresh and vacuum packaged). Present results demonstrated the color and lipid oxidative stability of lamb longissimus muscle during refrigerated display was not affected by enhanced levels of omega-3 and omega-6 FA due to dietary treatments.

Comparison of the color stability and lipid oxidative stability of fresh and vacuum packaged lamb muscle containing elevated omega-3 and omega-6 fatty acid levels from dietary manipulation

A series of three experiments were conducted with second cross ([Merino×Border Leicester]×Poll Dorset) wether lambs to evaluate the effects of dietary treatments on manipulation of muscle long-chain (LC) omega-3 fatty acids (FA) on the color stability and oxidative stability of fresh and vacuum packaged lamb. At the end of 7-, 6- and 6-week experimental periods for experiments (Exp.) 1–3 respectively, lambs were slaughtered at a commercial abattoir. At 24 h post-mortem, muscle longissimus lumborum (LL) and longissimus thoracis (LT) were removed and evaluated for color and lipid oxidative stability under specified commercial storage and display condition. Of the dietary supplements used, fish meal and fish oil moderately (P<0.01) and markedly (P<0.001) increased muscle omega-3 FA content, while both protected canola seed (P<0.001) and protected sunflower meal protein significantly (P<0.02) increased muscle omega-6 FA content or ratio of omega-6/omega-3 of the longissimus muscle. In all experiments, the substantial increase (P<0.001) in muscle LC omega-3 and omega-6 FA had no consistent significant effect on color values (redness (a*), yellowness (b*) and lightness (L*)) for fresh and vacuum packaged lamb over a 6-day display period. Lipid oxidation, determined by the levels of thiobarbituric acid reactive substances (TBARS) indicated the enrichment of muscle polyunsaturated fatty acid (PUFA) levels in lambs did not produce significant differences resulting either from main treatment effects or for treatment×day×type interactions (where type was fresh and vacuum packaged). Present results demonstrated the color and lipid oxidative stability of lamb longissimus muscle during refrigerated display was not affected by enhanced levels of omega-3 and omega-6 FA due to dietary treatments.

Comparison of the color stability and lipid oxidative stability fo fresh and vacuum packaged lamb muscle containing elevated omega-3 and omega-6 fatty acid levels from dietary manipulation

A series of three experiments were conducted with second cross ([Merino×Border Leicester]×Poll Dorset) wether lambs to evaluate the effects of dietary treatments on manipulation of muscle long-chain (LC) omega-3 fatty acids (FA) on the color stability and oxidative stability of fresh and vacuum packaged lamb. At the end of 7-, 6- and 6-week experimental periods for experiments (Exp.) 1–3 respectively, lambs were slaughtered at a commercial abattoir. At 24 h post-mortem, muscle longissimus lumborum (LL) and longissimus thoracis (LT) were removed and evaluated for color and lipid oxidative stability under specified commercial storage and display condition. Of the dietary supplements used, fish meal and fish oil moderately (P<0.01) and markedly (P<0.001) increased muscle omega-3 FA content, while both protected canola seed (P<0.001) and protected sunflower meal protein significantly (P<0.02) increased muscle omega-6 FA content or ratio of omega-6/omega-3 of the longissimus muscle. In all experiments, the substantial increase (P<0.001) in muscle LC omega-3 and omega-6 FA had no consistent significant effect on color values (redness (a*), yellowness (b*) and lightness (L*)) for fresh and vacuum packaged lamb over a 6-day display period. Lipid oxidation, determined by the levels of thiobarbituric acid reactive substances (TBARS) indicated the enrichment of muscle polyunsaturated fatty acid (PUFA) levels in lambs did not produce significant differences resulting either from main treatment effects or for treatment×day×type interactions (where type was fresh and vacuum packaged). Present results demonstrated the color and lipid oxidative stability of lamb longissimus muscle during refrigerated display was not affected by enhanced levels of omega-3 and omega-6 FA due to dietary treatments.

Total arsenic accumulation in yabbies (Cherax Destructor Clark) exposed to elevated arsenic levels in Victorian gold mining areas, Australia

Arsenic is a proven carcinogen often found at high concentrations in association with gold and other heavy metals. The freshwater yabby, Cherax destructor Clark (Decapoda, Parastacidae), is a ubiquitous species native to Australia's central and eastern regions, with a growing international commercial market. However, in this region of Australia, yabby farmers often harvest organisms from old mine tailings dams with elevated environmental arsenic levels. Yabbies exposed to elevated environmental arsenic were found to accumulate and store as much as 100 μg/g arsenic in their tissues. The accumulation is proportional to the concentration of arsenic in the sediment and is high enough to be of concern for people who eat the yabbies. A comparison of arsenic levels in wild and lab-fed animals also was performed. Although there was no significant difference in the level of arsenic in the various organs of the wild animals, the animals purchased from a yabby farm showed a significantly higher arsenic concentration in their hepatopancreas (3.7 ± 0.9 μg/g) compared to other organs (0.6–1.8 μg/g). Furthermore, after a 40-d exposure to food containing 200 to 300 μg/g inorganic arsenic, arsenate (As[V])-exposed animals showed a significant increase in tissue-specific arsenic accumulation, whereas arsenite (As[III])-exposed animals showed a lower, nonsignificant increase in As uptake, primarily in the hepatopancreas. These results have important implications for yabby growers and consumers alike.

Elevated intracardiac angiotensin II leads to cardiac hypertrophy and mechanical dysfunction in normotensive mice

Introduction
Angiotensin II (Ang II) is known to induce cardiac growth and modulate myocardial contractility. It has been reported that elevated levels of endogenous Ang II contribute to the development of cardiac hypertrophy in hypertensives. However, the long-term functional effects of cardiac exposure to Ang II in normotensives is unclear.

A recently developed transgenic mouse (TG1306/1R), in which cardiac-specific overproduction of Ang II produces primary hypertrophy, provides a new experimental model for investigation of this phenotype. The aim of the present study was to use this model to investigate whether there is a functional deficit in primary hypertrophy that may predispose to cardiac failure and sudden death. We hypothesised that primary cardiac hypertrophy is associated with mechanical dysfunction in the basal state.

Methods
Normotensive heterozygous TG1306/1R mice harbouring multiple copies of a cardiac-specific rat angiotensinogen gene were studied at age 30—40 weeks and compared with age-matched wild-type littermates. Left ventricular function was measured ex vivo in bicarbonate buffer-perfused, Langendorffmounted hearts ( at a perfusion pressure of 80 mmHg, 37°C) using a fluid-filled PVC balloon interfaced to a pressure transducer and digital data acquisition system.

Results
There was no difference in the mean (±SEM) intrinsic heart rate of TG1306/1R and wild-type control mice (357.4±11.8 vs. 367.5±20.9 bpm, n=9 & 7). Under standardised end-diastolic pressure conditions, TG1306/1R hearts exhibited a significant reduction in peak developed pressure (132.2±9.4 vs. 161.5±3.1 mmHg, n=9 & 7, p<0.05) and maximum rate of pressure development (3566.7±323.7 vs. 4486.3±109.4 mmHg, n=9 & 7, p<0.05). TG1306/1R mice show a significant correlation between incidence of arrhythmia and increasing heart size (Spearman's correlation coefficient 0.61).

Conclusion
These data demonstrate that chronic in vivo exposure to elevated levels of intra-cardiac Ang II is associated with significant contractile abnormalities evident in the ex vivo intact heart. Our findings suggest that endogenous overproduction of cardiac Ang II, independent of changes in blood pressure, is sufficient to induce ventricular remodelling that culminates in impaired cardiac function which may precede failure.

Elevated dietary sodium intake exacerbates myocardial hypertrophy associated with cardiac-specific overproduction of angiotensin II

Introduction/hypothesis
Cardiac hypertrophy is an independent risk factor predictive of cardiovascular disease and is significantly associated with morbidity and mortality. The mechanism by which angiotensin II (Ang II) and dietary sodium exert additive effects on the development of cardiac hypertrophy is unclear. The goal of this study was to evaluate the hypothesis that, where there is a genetic predisposition to Ang II-dependent hypertrophy, there is also an increased susceptibility to sodium-induced hypertrophy mediated by AT₁-receptor expression.

Methods
Diets of low sodium (LS, 0.3% w:w) and high sodium (HS, 4.0% w:w) content were fed to adult (age 25 weeks) control wild-type mice (WT) and to weeks) control wild-type mice (WT) and to transgenic mice exhibiting cardiac specific overexpression of angiotensinogen (TG). At the conclusion of a 40-day dietary treatment period, cardiac tissue weights were compared and the relative expression levels of Ang II receptor subtypes (AT_1A and AT₂) were evaluated using RT-PCR.

Results
WT and TG mice fed HS and LS diets maintained comparable weight gains during the treatment period. The normalised heart weights of TG mice were elevated compared to WT, and the extent of the increase was greater for mice maintained on the HS diet treatments (WT 12% vs. TG 41% increase in cardiac weight index). While a similar pattern of growth was observed for ventricular tissues, the atrial weight parameters demonstrated an additional significant effect of dietary sodium intake on tissue weight, independent of animal genetic type. No differences in the relative (GAPDH normalised) expression levels of AT_1A- and AT₂-receptor mRNA were observed between diet or animal genetic groups.

Conclusion
This study demonstrates that, where there is a pre-existing genetic condition of Ang II-dependent cardiac hypertrophy, the pro-growth effect of elevated dietary sodium intake is selectively augmented. In TG and WT mice, this effect was evident with a relatively short dietary treatment intervention (40 days). Evaluation of the levels of Ang II receptor mRNA further demonstrated that this differential growth response was not associated with an altered relative expression of either AT_1A- or AT₂-receptor subtypes. The cellular mechanistic bases for this specific Ang II-dietary sodium interaction remain to be elucidated.

Plastic flow properties and microstructural evolution in an ultrafine-grained Al-Mg-Si alloy at elevated temperatures

An AA6082 alloy was subjected to eight passes of equal channel angular pressing at 100 °C, resulting in an ultrafine grain size of 0.2 to 0.4 µm. The tensile deformation behavior of the material was studied over the temperature range of 100 °C to 350 °C and strain rate range of 10¯⁴ to 10¯¹ s¯¹. The evolution of microstructure under tensile deformation was investigated by analyzing both the deformation relief on the specimen surface and the dislocation structure. While extensive microshear banding was found at the lower temperatures of 100 °C to 150 °C, deformation at higher temperatures was characterized by cooperative grain boundary sliding and the development of a bimodal microstructure. Dislocation glide was identified as the main deformation mechanism within coarse grains, whereas no dislocation activity was apparent in the ultrafine grains.

Stability of oxide films formed on mild steel in turbulent flow conditions of alkaline solutions at elevated temperatures

In the industries involving alkaline solutions in different process streams, the nature and stability of oxide films formed on the metallic surfaces determine the rates of erosion–corrosion of the equipment. In the present study the characteristics of the oxide films formed on AISI 1020 steel in a 2.75 M sodium hydroxide solution at temperatures up to 175°C, have been investigated by employing electrochemical techniques of cyclic voltammetry and chronoamperometry. The experiments were carried out in an autoclave system based upon a ‘rotating cylinder electrode’ geometry to determine the effects of turbulence on the stability of the films. The results suggest that little protection is afforded in the active region (at about −0.8 V_SHE). In the passive region at low potentials (−0.6 V to −0.4 V_SHE), it appears the films are compact and more stable, and therefore provide good protection. At higher potentials (>−0.4 V_SHE) in the passive region, the results suggest that film formation and dissolution occur simultaneously and the increase in temperature and turbulence causes a breakdown of the passive film resulting in a situation similar to nonprotective magnetite growth.

Composite cell components for elevated temperature all-solid-state Li-ion batteries

Application of Li-ion batteries with liquid electrolytes at elevated temperatures (above 60°C) is limited due to the decomposition of the electrolyte. Stable solid state electrolytes can solve this problem, but the conductivity of these electrolytes are relatively low, the interfacial contacts with the electrodes are poor, and the charge transfer kinetics in the electrodes are limited. Solutions for these problems by using composite electrodes and electrolytes have been investigated and the results are described. A new concept for making all-solid-state Li-ion batteries that can be applied in the temperature range between room temperature and about 150°C will be presented.

Cordycepin-hypersensitive growth links elevated polyphosphate levels to inhibition of poly(A) polymerase in Saccharomyces cerevisiae

To identify genes involved in poly(A) metabolism, we screened the yeast gene deletion collection for growth defects in the presence of cordycepin (3′-deoxyadenosine), a precursor to the RNA chain terminating ATP analog cordycepin triphosphate. Δpho80 and Δpho85 strains, which have a constitutively active phosphate-response pathway, were identified as cordycepin hypersensitive. We show that inorganic polyphosphate (poly P) accumulated in these strains and that poly P is a potent inhibitor of poly(A) polymerase activity in vitro. Binding analyses of poly P and yeast Pap1p revealed an interaction with a k_D in the low nanomolar range. Poly P also bound mammalian poly(A) polymerase, however, with a 10-fold higher k_D compared to yeast Pap1p. Genetic tests with double mutants of Δpho80 and other genes involved in phosphate homeostasis and poly P accumulation suggest that poly P contributed to cordycepin hypersensitivity. Synergistic inhibition of mRNA synthesis through poly P-mediated inhibition of Pap1p and through cordycepin-mediated RNA chain termination may thus account for hypersensitive growth of Δpho80 and Δpho85 strains in the presence of the chain terminator. Consistent with this, a mutation in the 3′-end formation component rna14 was synthetic lethal in combination with Δpho80. Based on these observations, we suggest that binding of poly P to poly(A) polymerase negatively regulates its activity.