Identification of a Distal GLUT4 Trafficking Event Controlled by Actin Polymerization

The insulin-stimulated trafficking of GLUT4 to the plasma membrane in muscle and fat tissue constitutes a central process in blood glucose homeostasis. The tethering, docking, and fusion of GLUT4 vesicles with the plasma membrane (PM) represent the most distal steps in this pathway and have been recently shown to be key targets of insulin action. However, it remains unclear how insulin influences these processes to promote the insertion of the glucose transporter into the PM. In this study we have identified a previously uncharacterized role for cortical actin in the distal trafficking of GLUT4. Using high-frequency total internal reflection fluorescence microscopy (TIRFM) imaging, we show that insulin increases actin polymerization near the PM and that disruption of this process inhibited GLUT4 exocytosis. Using TIRFM in combination with probes that could distinguish between vesicle transport and fusion, we found that defective actin remodeling was accompanied by normal insulin-regulated accumulation of GLUT4 vesicles close to the PM, but the final exocytotic fusion step was impaired. These data clearly resolve multiple steps of the final stages of GLUT4 trafficking, demonstrating a crucial role for actin in the final stage of this process.     

The Association of Maternal Psychopathology and Family Socioeconomic Status with Psychological Problems in Obese Children

In an earlier study we found parental psychiatric symptoms to be a better correlate of behavioral problems in obese children than either child or parental obesity. This study sought to extend this research by assessing the association of general maternal distress, specific maternal psychopathology, family socioeconomic status and child obesity with the psychological problems of 152 children seeking treatment for obesity in two cohorts. Child psychological problems were measured using the Child Behavior Checklist/4–18 (CBCL) in each cohort. In sample 1 maternal general psychiatric symptoms were measured using the Cornell Medical Index (psychiatric subscales) and the Inventory of Interpersonal Problems whereas specific psychopathology was assessed with the Beck Depression Inventory and the Bulimia Test. In sample 2 the Symptom Checklist-90 and Binge Eating Scale provided general and specific measures of psychopathology. In the combined sample, 58% percent of the boys and 44% of the girls met criteria on at least one CBCL behavioral problem scale, with Social Problems the most prevalent problem in both boys and girls. For both samples, child obesity did not account for any variance in child psychosocial problems beyond that accounted for by maternal psychopathology and family SES. Family SES accounted for a significant increment in variance in child behavioral problems in only two scales. This study systematically replicates previous research, adding support for a broader conceptualization of factors that influence psychological problems in obese treatment-seeking children than one which focuses on child obesity.     

Short-term impacts of the 2019–20 fire season on biodiversity in eastern Australia

To address uncertainties surrounding the impacts of unprecedented 2019–20 bushfires in south-eastern Australia, we convened a symposium on field-measured impacts on biodiversity for the 2020 Ecological Society of Australia conference. Nine presentations covered a range of studies on plant species and communities, and reptiles and mammals. Here, we summarize the presentations and review other field studies from NSW, some of which are currently unpublished. The impacts were extensive, but results also estimated survival rate perhaps higher than first reported in the media. More than half of individual mammal, reptile and threatened rainforest trees may have survived the fire, though small populations remain very vulnerable to second fire in the near future. Comprehensive understanding of the impacts of these fires requires much more field study, and for the results to be interpreted in the context of the broader fire regime. The symposium was an important early step in that understanding.     

Interday variation and effect of transportation on indirect blood pressure measurements, plasma endothelin-1 and serum cortisol in Standardbred and Icelandic horses

ABSTRACT: BACKGROUND: Systemic hypertension is a prominent feature in humans with metabolic syndrome (MS) and this is partly caused by an enhanced endothelin-1 (ET-1) mediated vasoconstriction. There are indications that systemic hypertension might be a feature in equine metabolic syndrome (EMS) but if ET-1 is involved in the development of hypertension in horses is not known. Increased levels of cortisol have also been found in humans with MS but there are no reports of this in horses. Before blood pressure, plasma ET-1 and serum cortisol can be evaluated in horses with EMS, it is necessary to investigate the interday variation of these parameters on clinically healthy horses. The aims of the present study were therefore to evaluate the interday variation and influence of transportation on systemic blood pressure, plasma ET-1 and serum cortisol in healthy Standardbred and Icelandic horses, and to detect potential breed differences. METHODS: Nine horses of each breed were included in the study. Blood pressure was measured and blood samples were collected between 6 and 9 am on two separate days. Eight of the horses (four of each breed) were transported to a new stable were they stayed overnight. The next morning, the sampling procedure was repeated. RESULTS: The interday variation was higher for plasma ET-1 (37%) than for indirect pressure measurements (8-21%) and serum cortisol (18%). There were no differences in systemic blood pressure between the two breeds. The Icelandic horses had significantly lower serum cortisol and significantly higher plasma ET-1 concentrations compared to the Standardbred horses. Plasma ET-1 was significantly elevated after transportation, but systemic blood pressure and serum cortisol did not differ from the values obtained in the home environment. CONCLUSIONS: Indirect blood pressure, plasma ET-1 and serum cortisol are of interest as markers for cardiovascular dysfunction in horses with EMS. The elevated plasma ET-1 concentrations recorded after transportation was likely caused by a stress response. This needs to be considered when evaluating plasma ET-1 in horses after transportation. The differences detected in plasma ET-1 and serum cortisol between the two breeds might be related to differences in genetic setup, training status as well as management conditions.     

Protective role of fructose in the metabolism of astroglial C6 cells exposed to hydrogen peroxide

Astroglial cells represent the main line of defence against oxidative damage related to neurodegeneration. Therefore, protection of astroglia from an excess of reactive oxygen species could represent an important target of the treatment of such conditions. The aim of our study was to compare the abilities of glucose and fructose, the two monosaccharides used in diet and infusion, to protect C6 cells from hydrogen peroxide (H₂O₂)-mediated oxidative stress. It was observed using confocal microscopy with fluorescent labels and the MTT test that fructose prevents changes of oxidative status of the cells exposed to H₂O₂ and preserves their viability. Even more pronounced protective effects were observed for fructose 1,6-bis(phosphate). We propose that fructose and its intracellular forms prevent H₂O₂ from participating in the Fenton reaction via iron sequestration. As fructose and fructose 1,6-bis(phosphate) are able to pass the blood–brain barrier, they could provide antioxidative protection of nervous tissue in vivo. So, in contrast to the well-known negative effects of frequent consumption of fructose under physiological conditions, acute infusion or ingestion of fructose or fructose 1,6-bis(phosphate) could be of benefit in the cytoprotective therapy of neurodegenerative disorders related to oxidative stress.     

Enhanced detection of cytomegalovirus in shell vial cell culture monolayers by preinoculation treatment of urine with low-speed centrifugation

In an effort to reduce the number of unreadable MRC-5 cell culture monolayers (i.e., monolayers that are obscured by an opaque film or are detached from their coverslips), urine specimens were subjected to low-speed (500 g for 10 min at 4°C) centrifugation prior to their inoculation into shell vials for the detection of cytomegalovirus (CMV). A total of 182 urine specimens obtained from patients suffering from ARC or AIDS were subjected to routine centrifugation culture and to preinoculation centrifugation. Cytomegalovirus recovery among the two specimen-treatment procedures was determined by counting the number of immunofluorescent MRC 5-positive cells. Eleven of 19 positive specimens were in accord in both systems. However, routinely treated (i.e., without preinoculation centrifugation) specimens were difficult to read specifically among those specimens that were cloudy or opaque. Cytome-galovirus was recovered in 18 of the 19 positive specimens that were subjected to the preinoculation centrifugation procedure. Only 12 of 19 positive specimens were identified following direct inoculation of untreated urine into shell vials. The additional preinoculation centrifugation step is appropriate for the removal of obfuscating amorphous substances from the assay system without compromising CMV detection rates.     

Oral–aboral axis specification in the sea urchin embryo,IV: Hypoxia radializes embryos by preventing the initial spatialization of nodal activity

The oral–aboral axis of the sea urchin embryo is specified conditionally via a regulated feedback circuit involving the signaling gene nodal and its antagonist lefty. In normal development nodal activity becomes localized to the prospective oral side of the blastula stage embryo, a process that requires lefty. In embryos of Strongylocentrotus purpuratus, a redox gradient established by asymmetrically distributed mitochondria provides an initial spatial input that positions the localized domain of nodal expression. This expression is perturbed by hypoxia, leading to development of radialized embryos lacking an oral–aboral axis. Here we show that this radialization is not caused by a failure to express nodal, but rather by a failure to localize nodal activity to one side of the embryo. This occurs even when embryos are removed from hypoxia at late cleavage stage when nodal is first expressed, indicating that the effect involves the initiation phase of nodal activity, rather than its positive feedback-driven amplification and maintenance. Quantitative fluorescence microscopy of MitoTracker Orange-labeled embryos expressing nodal-GFP reporter gene revealed that hypoxia abolishes the spatial correlation between mitochondrial distribution and nodal expression, suggesting that hypoxia eliminates the initial spatial bias in nodal activity normally established by the redox gradient. We propose that absent this bias, the initiation phase of nodal expression is spatially uniform, such that the ensuing Nodal-mediated community effect is not localized, and hence refractory to Lefty-mediated enforcement of localization.     

Hormone-induced luteolysis on physiologically persisting corpora lutea in Eurasian and Iberian lynx (Lynx lynx and Lynx pardinus)

The Iberian lynx (Lynx pardinus) is the most critically endangered felid. A high reproductive success within the Iberian Lynx Conservation Breeding Program is crucial to maintaining the goal of reintroducing captive born offspring to the wild and thus increasing the population. Lynx follow a unique reproductive strategy with a monoestrous cycle and persisting CLs over many years. These persistent CLs constantly produce progesterone (on average 5 ng/mL) and are hypothesized to hinder a polyestrous cyclicity in lynx. The aim of this study was to evaluate whether artificial luteolysis can be achieved with common luteolytic drugs and if luteolysis would induce a second estrus naturally. We observed a functional regression of lynx CLs after artificial luteolysis with 2.5 μg/kg body weight PGF2α analogue (cloprostenol) administered three times every 16 hours. We could see a similar effect when combining cloprostenol with other drugs like an anti-gestagen (aglepristone) or a dopamin-agonist (prolactin-inhibitor, cabergolin) or by prolonging the cloprostenol administration to a total of 5 days. However, the sample size was too small to draw conclusions about which protocol is superior or if combining different drugs would result in a positive synergism. Neither structural regression of CLs nor subsequent spontaneous estrus induction was induced with any of these treatments. We suggest that a dose of 2.5 μg/kg body weight cloprostenol administered once daily over 3 to 5 days is sufficient for functional luteolysis in lynx. The next step would be to compare the success of estrus induction with or without the preceding artificial luteolysis.