Uncarboxylated osteocalcin decreases insulin-stimulated glucose uptake without affecting insulin signaling and regulators of mitochondrial biogenesis in myotubes

Uncarboxylated osteocalcin (uOC) is a circulating bone matrix protein, which has previously been shown to regulate glucose uptake and systemic metabolism. However, the cellular mechanism by which uOC acts has yet to be elucidated. C2C12 mouse myotubes were treated for 72 h with uOC (1–100 ng/mL). Cellular metabolism was analyzed using oxygen consumption and extracellular acidification rate. Metabolic gene and protein expression were measured via quantitative real-time polymerase chain reaction and Western blot, respectively. Additionally, C2C12 myotubes were treated with 10 ng/mL uOC to examine glucose uptake and activation of insulin signaling with or without insulin resistance. Finally, cellular lipid content was measured via Oil Red O and Nile Red staining. uOC treatment resulted in dose-dependent alterations of oxygen consumption with little effect on regulators of mitochondrial metabolism. Basal expression of regulators of glucose uptake were unaffected by uOC treatment. However, insulin-stimulated glucose uptake was blunted by uOC treatment with no concurrent alterations in insulin signaling. While chronic insulin treatment resulted in suppressed activation of Akt, concurrent uOC treatment was unable to prevent these detrimental effects on insulin signaling. uOC treatment had no effect on markers of lipogenesis and cellular lipid content. These findings suggest that 72-h uOC treatment may alter oxygen consumption without effect on regulators of mitochondrial biogenesis. Additionally, uOC treatment suppressed insulin-stimulated glucose uptake in cultured myotubes but had little effect on insulin signaling or regulators of cellular metabolism and was unable to mitigate insulin resistance.     

The microbiota influences the Drosophila melanogaster life history strategy

Organisms are locally adapted when members of a population have a fitness advantage in one location relative to conspecifics in other geographies. For example, across latitudinal gradients, some organisms may trade off between traits that maximize fitness components in one, but not both, of somatic maintenance or reproductive output. Latitudinal gradients in life history strategies are traditionally attributed to environmental selection on an animal's genotype, without any consideration of the possible impact of associated microorganisms (“microbiota”) on life history traits. Here, we show in Drosophila melanogaster, a key model for studying local adaptation and life history strategy, that excluding the microbiota from definitions of local adaptation is a major shortfall. First, we reveal that an isogenic fly line reared with different bacteria varies the investment in early reproduction versus somatic maintenance. Next, we show that in wild fruit flies, the abundance of these same bacteria was correlated with the latitude and life history strategy of the flies, suggesting geographic specificity of the microbiota composition. Variation in microbiota composition of locally adapted D. melanogaster could be attributed to both the wild environment and host genetic selection. Finally, by eliminating or manipulating the microbiota of fly lines collected across a latitudinal gradient, we reveal that host genotype contributes to latitude‐specific life history traits independent of the microbiota and that variation in the microbiota can suppress or reverse the differences between locally adapted fly lines. Together, these findings establish the microbiota composition of a model animal as an essential consideration in local adaptation.     

Selectivty Assessments of a Sequential Extraction Procedure for Potential Trace Metals’ Mobility and Bioavailability in Phosphate Rocks from Jordan Phosphate Mines

A modified six-step sequential extraction procedure was used to fractionate and determine the following trace metals: U⁴⁺, As⁵⁺, Cd²⁺, Cr⁺², Cu²⁺, Ni²⁺, Pb²⁺, Zn²⁺, and V⁵⁺ in three different phosphate rocks from mines in Jordan. The mean values of uranium in the samples investigated were 98 ± 6 mgkg^?1, 92 ± 3 mgkg^?1, 215 ± 6 mgkg^?1, and 159 ± 13 mgkg^?1, respectively. The sequential extraction results obtained showed that most of the U⁴⁺ in these samples was strongly bound with 87%, 93%, 97%, and 93% of the total content, respectively, remaining in the samples after the sequential extraction steps were performed. Hence, 13%–7% and 3%–7%, respectively, of the U⁴⁺ is distributed in the most labile form, indicating that the majority of the U⁴⁺ in these samples was highly incorporated within the apatite present in the samples. The aforementioned was in agreement with the XRD and SEM-EDX results obtained. The apparent mobility of U⁴⁺, As⁵⁺, Cd²⁺, Cr⁺², Cu²⁺, Ni²⁺, Pb²⁺, Zn²⁺, and V⁵⁺ (using all six extraction steps) from the Al-Abied and Al-Hasa samples was as follows: As⁵⁺ (30.17%)> Cu²⁺ (6.55%)> Zn²⁺(4.34%)> Cd²⁺ (3.84%) Cr⁺² (3.66%)> Pb²⁺ (2.57%)> V⁵⁺ (53%)> Ni²⁺ (1.71%)> U⁴⁺ (0.99%). The mobility of As⁵⁺, Cd²⁺, Cu²⁺, Cr⁺², Ni²⁺, Pb²⁺, U⁴⁺, Zn²⁺, and V⁵⁺ (using all six extraction steps) from Eshidiya samples was as follows: As⁵⁺ (17.32%)> Cr⁺² (4.84%)> Zn²⁺ (4.25%)> Pb²⁺ (4.19%)> Cu²⁺ (3.49%)> V⁵⁺ (1.42%)> Cd²⁺ (0.78) U⁴⁺ (0.09%)> Ni²⁺ (0%).     

Localization of A20 to a lysosome-associated compartment and its role in NFkappaB signaling

A20 is a tumor necrosis factor (TNF)-inducible zinc finger protein that contains both ubiquitinating and deubiquitinating activities. A20 negatively regulates NFkappaB (nuclear factor kappaB) signaling induced by TNF receptor family and Toll-like receptors, but the mechanism of A20 action is poorly defined. Here we show that a fraction of endogenous and ectopically expressed A20 is localized to an endocytic membrane compartment that is in association with the lysosome. The lysosomal association of A20 requires its carboxy terminal zinc finger domains, but is independent of its ubiquitin-modifying activities. Interestingly, A20 mutants defective in membrane association also contain reduced NFkappaB inhibitory activity. These findings suggest the involvement of a lysosome-associated mechanism in A20-dependent termination of NFkappaB signaling.     

Presence of the Chytrid Fungus Batrachochytrium dendrobatidis in Populations of the Critically Endangered Frog Mannophryne olmonae in Tobago, West Indies

The emerging infectious disease chytridiomycosis is prevalent in Central and South America, and has caused catastrophic declines of amphibian populations in the Neotropics. The responsible organism, Batrachochytrium dendrobatidis, has been recorded on three West Indian islands, but the whole of the Caribbean region is predicted to offer a suitable environment for the disease. Monitoring the spread of chytridiomycosis is thus a priority in this region, which has exceptionally high levels of amphibian endemism. PCR analysis of 124 amphibian skin swabs in Tobago (Republic of Trinidad and Tobago) demonstrated the presence of B. dendrobatidis in three widely separated populations of the frog Mannophryne olmonae, which is listed as Critically Endangered on the basis of recent population declines. Chytridiomycosis is presently endemic in this species, with a prevalence of about 20% and no associated clinical disease. Increased susceptibility to chytridiomycosis from climate change is unlikely in amphibian populations in Tobago, as this island does not have high montane environments, but remains a possibility in the sister island of Trinidad. Preventing the spread of chytridiomycosis within and between these and other Caribbean islands should be a major goal of practical conservation measures for amphibians in the region.     

Ability of the Actiwatch Accelerometer to Predict Free‐Living Energy Expenditure in Young Children

Objective: To determine whether activity counts obtained with the Actiwatch monitor are associated with total expenditure and body composition in young children. Research Methods and Procedures: Actiwatch activity monitors were tested in 29 children 4 to 6 years old under field conditions over eight days. Total energy expenditure (TEE) was assessed with the doubly labeled water (DLW) technique. Correlation analyses were used to identify variables related to energy expenditure and percentage body fat. Multiple linear regression analyses were used to examine the variance in TEE and percentage body fat explained by activity counts after adjusting for relevant covariates. Results: Both average total daily activity counts (658, 816 ± 201, 657) and the pattern of activity were highly variable among subjects. TEE was significantly related to lean body mass (r = 0.45) and age (r = 0.48; p < 0.05 for both). Activity counts alone were not associated with TEE. In multiple linear regression analyses, TEE was independently associated with only lean body mass. Percentage fat mass was independently associated with body weight, being a girl, and being white, but not with average total activity counts. Discussion: Activity counts obtained with the Actiwatch under free‐living conditions do not reflect TEE in 4‐ to 6‐year‐old children and are not correlated with percentage fat mass. Therefore, average total activity counts obtained with the Actiwatch may be of limited value in identifying children at risk for becoming obese.