Mice with Whole-Body Disruption of AMPK-Glycogen Binding Have Increased Adiposity,Reduced Fat Oxidation and Altered Tissue Glycogen Dynamics

The AMP-activated protein kinase (AMPK), a central regulator of cellular energy balance and metabolism, binds glycogen via its β subunit. However, the physiological effects of disrupting AMPK-glycogen interactions remain incompletely understood. To chronically disrupt AMPK-glycogen binding, AMPK β double knock-in (DKI) mice were generated with mutations in residues critical for glycogen binding in both the β1 (W100A) and β2 (W98A) subunit isoforms. We examined the effects of this DKI mutation on whole-body substrate utilization, glucose homeostasis, and tissue glycogen dynamics. Body composition, metabolic caging, glucose and insulin tolerance, serum hormone and lipid profiles, and tissue glycogen and protein content were analyzed in chow-fed male DKI and age-matched wild-type (WT) mice. DKI mice displayed increased whole-body fat mass and glucose intolerance associated with reduced fat oxidation relative to WT. DKI mice had reduced liver glycogen content in the fed state concomitant with increased utilization and no repletion of skeletal muscle glycogen in response to fasting and refeeding, respectively, despite similar glycogen-associated protein content relative to WT. DKI liver and skeletal muscle displayed reductions in AMPK protein content versus WT. These findings identify phenotypic effects of the AMPK DKI mutation on whole-body metabolism and tissue AMPK content and glycogen dynamics.     

Neuroprotection Mediated by Human Blood Plasma in Mouse Hippocampal Slice Cultures and in Oxidatively Stressed Human Neurons

Neuroprotection from oxidative stress is critical during neuronal development and maintenance but also plays a major role in the pathogenesis and potential treatment of various neurological disorders and neurodegenerative diseases. Emerging evidence in the murine system suggests neuroprotective effects of blood plasma on the aged or diseased brain. However, little is known about plasma-mediated effects on human neurons. In the present study, we demonstrate the neuroprotective effect mediated by human plasma and the most abundant plasma–protein human serum albumin against oxidative stress in glutamatergic neurons differentiated from human neural crest-derived inferior turbinate stem cells. We observed a strong neuroprotective effect of human plasma and human serum albumin against oxidative stress-induced neuronal death on the single cell level, similar to the one mediated by tumor necrosis factor alpha. Moreover, we detected neuroprotection of plasma and human serum albumin against kainic acid-induced excitatory stress in ex vivo cultured mouse hippocampal tissue slices. The present study provides deeper insights into plasma-mediated neuroprotection ultimately resulting in the development of novel therapies for a variety of neurological and, in particular, neurodegenerative diseases.     

Angiopoietin-like Proteins in Colorectal Cancer—A Literature Review

Colorectal cancer (CRC) is one of the most common types of malignancy, with an annual incidence of about 10% of the total number of new cases. Despite well-developed screening tests, mortality from this type of cancer remains unchanged. Therefore, it is important to search for more accurate markers that are useful in the detection of colorectal cancer (especially in its early stages), and treatment. Angiopoietin-like proteins (ANGPTLs) are a family of eight proteins with a diversity of applications, including pro- and anti-angiogenic properties. Consequently, we performed an extensive search of the literature, pertaining to our investigation, via the MEDLINE/PubMed database. Based on the available literature, we summarize that some of those proteins are characterized by increased or decreased concentrations during the course of CRC. We can also assume that some ANGPTLs can inhibit the development of CRC, while others induce its progress. Moreover, some factors are dependent on the stage or histological type of the tumor, the presence of hypoxia, or metastases. Most importantly, some ANGPTLs can be useful in anti-cancer therapy. Therefore, further studies on ANGPTLs as potential markers of CRC should be continued.     

The Obscure Effect of Tribulus terrestris Saponins Plus Inulin on Liver Morphology,Liver Fatty Acids,Plasma Glucose,and Lipid Profile in SD Rats with and without Induced Type 2 Diabetes Mellitus

Diabetes is a predictor of nonalcoholic fatty liver disease (NAFLD). There are data suggesting that Tribulus terrestris (TT) saponins act as antidiabetic agents and protect against NAFLD. The effect of saponins may be increased by fermentable fibers such as inulin. The aim of the present study was to investigate the influence of TT saponins and TT saponins plus inulin on the plasma lipid profile and liver fatty acids of rats with induced diabetes mellitus type 2 (T2DM). The study was performed on 36 male Sprague–Dawley rats divided into two main groups: control and diabetic. Animals of the diabetic (DM) group were fed a high-fat diet and injected with streptozotocin (low doses). Animals of the control group (nDM) were on a regular diet and were injected with buffer. After the injections, the animals were split into subgroups: three non-diabetic (nDM): (i) control (c-C); (ii) saponin-treated rats (C-Sap); (iii) rats treated with saponins + inulin (C-Sap + IN), and three diabetic subgroups (DM): (iv) control (c-DM); (v) saponin-treated rats (DM-Sap); (vi) rats treated with saponins + inulin (DM-Sap + IN). Liver fatty acids were extracted and analyzed by gas chromatography, and plasma glucose and lipids were measured. The study showed significant changes in liver morphology, liver fatty acids, plasma lipid profile, and plasma glucose. In summary, supplementation with TT saponins or saponins with inulin for one month decreased the level of steatosis in rats with induced type 2 diabetes. Moreover, there were favorable effects on the plasma lipid profile in the rats. However, additional supplementation with inulin had a negative effect on liver morphology (with a microvesicular type of steatosis) in the non-diabetes group. Moreover, supplementation with inulin had a negative effect on plasma glucose in both diabetic and non-diabetic rats. These data show that a diet enriched with fermentable fibers reveals different effects in different organisms, and not all sources and forms of fiber are beneficial to health.     

Protease stability in bovine milk under combined thermal-high hydrostatic pressure treatment

At atmospheric pressure, inactivation of protease from B. subtilis in raw milk and pasteurized milk (with and without homogenization) was studied in a temperature range of 50–80 °C. Thermal inactivation followed a first order kinetic model in the temperature range tested. Temperature dependence of the first order inactivation rate constants could be accurately described by the Arrhenius equation, allowing Ea values to be calculated. Different milk systems did not show differences in enzyme thermo stability.The combined thermal (40, 50 and 60 °C)-high hydrostatic pressure (300–450 and 600 MPa) effect on protease activity was studied. Protease was very resistant to high pressures. Pressure stability was higher in raw milk than in pasteurized milk; homogenization appeared to have a protective effect on the enzyme. The separate effects of pressure and temperature on enzyme inactivation were related to changes in L?-values and milk appearance.A very pronounced antagonistic effect between high temperature and pressure was observed, i.e. at temperatures where thermal inactivation at atmospheric pressure occurs rapidly, application of pressure up to 600 MPa exerted a protective effect.Industrial relevanceHigh hydrostatic pressure (HHP) is an emerging technology that has been successfully applied as a minimal process for a variety of foods. Although the potential for the use of HHP treatment as an alternative method to heat treatment of milk was proposed almost a century ago, the suitability of this innovative technology to extend the shelf-life of milk hinges not only on its ability to inactivate pathogenic vegetative microorganisms but also on its effectiveness to inactivate indigenous and endogenous enzymes. This work examines the combined effects of temperature, pressure and homogenization on the protease (exogenous enzyme from B. subtilis) activity in milk. Inactivation of protease could extend the shelf life of milk.     

Finding the daredevils: development of a Sensation Seeking Scale for children that is relevant to physical risk taking

OBJECTIVE: There were two objectives in this research. First, to develop and evaluate the criterion validity of a questionnaire measure of sensation seeking in children, by examining how scores on this instrument relate to various indices of physical risk taking in children 7-12 years of age. Second, to develop both a parent-report and child-report version of the instrument. METHODS: Drawing on the literature, items tapping five potential aspects of sensation seeking were developed, with parents and children responding to comparable items. For each of these five subscales, internal reliability estimates were computed separately for the parent-report and child-report versions. To establish criterion validity, subscale scores were related to three indices of children's risk taking commonly used in research: actual risk taking, intentions-to-risk take, self-reports about risk taking on a standardized questionnaire. RESULTS: On both the child-report and parent-report versions, the same three of the five subscales yielded acceptable internal reliability scores and significantly related to the various indices of risk taking. CONCLUSIONS: This new measure adequately assesses aspects of sensation seeking relevant to children's risk taking. Because individuals high in sensation seeking engage in greater risk taking and are at increased risk of injury, this instrument may prove particularly useful to aid in identifying those children most likely to engage in injury-risk behaviors so that targeted interventions can be applied to this group.     

Analysis of peptide MS/MS spectra from large-scale proteomics experiments using spectrum libraries

A widespread proteomics procedure for characterizing a complex mixture of proteins combines tandem mass spectrometry and database search software to yield mass spectra with identified peptide sequences. The same peptides are often detected in multiple experiments, and once they have been identified, the respective spectra can be used for future identifications. We present a method for collecting previously identified tandem mass spectra into a reference library that is used to identify new spectra. Query spectra are compared to references in the library to find the ones that are most similar. A dot product metric is used to measure the degree of similarity. With our largest library, the search of a query set finds 91% of the spectrum identifications and 93.7% of the protein identifications that could be made with a SEQUEST database search. A second experiment demonstrates that queries acquired on an LCQ ion trap mass spectrometer can be identified with a library of references acquired on an LTQ ion trap mass spectrometer. The dot product similarity score provides good separation of correct and incorrect identifications.     

Natural revegetation in the vicinity of the former lead smelter in Zerjav, Slovenia

The response of plant communities to pollution associated with the lead smelter in Zerjav, Slovenia, was investigated on spatial and temporal scales. In 2001, the total concentrations of contaminating metals in the soil measured at the most polluted plot were 59000 mg kg(-1) Pb, 180 mg kg(-1) Cd, and 3300 mg kg(-1) Zn. A negative correlation between the concentration of metals in the soil and plant biodiversity parameters along the gradient of pollution in 2001 was detected. Plant species lists were compiled in 2001 for plots located at different distances from the emission source and compared to that of 1981. In the period from 1981 to 2001, smelter emissions were reduced, and plant species richness increased at all examined plots. Among the successful survivals were some metal hyperaccumulators (Minuartia gerardii, Thlaspi praecox, and Biscutella laevigata). Of special interest were plants that survived the period of highest pollution. We believe that these species can be used in metal-degraded environments for natural revegetation to immobilize heavy metals. The ecosystem in the surroundings of the former smelter is presently recovering. Our results suggest that high metal concentrations in soil are a potential limiting factor for revegetation.