Stbd1-deficient mice display insulin resistance associated with enhanced hepatic ER-mitochondria contact

Starch binding domain-containing protein 1 (STBD1) is an endoplasmic reticulum (ER)-resident, glycogen-binding protein. In addition to glycogen, STBD1 has been shown to interact with several proteins implicated in glycogen synthesis and degradation, yet its function in glycogen metabolism remains largely unknown. In addition to the bulk of the ER, STBD1 has been reported to localize at regions of physical contact between mitochondria and the ER, known as Mitochondria-ER Contact sites (MERCs). Given the emerging correlation between distortions in the integrity of hepatic MERCs and insulin resistance, our study aimed to delineate the role of STBD1 in vivo by addressing potential abnormalities in glucose metabolism and ER-mitochondria communication associated with insulin resistance in mice with targeted inactivation of Stbd1 (Stbd1KO). We show that Stbd1KO mice at the age of 24 weeks displayed reduced hepatic glycogen content and aberrant control of glucose homeostasis, compatible with insulin resistance. In line with the above, Stbd1-deficient mice presented with increased fasting blood glucose and insulin levels, attenuated activation of insulin signaling in the liver and skeletal muscle and elevated liver sphingomyelin content, in the absence of hepatic steatosis. Furthermore, Stbd1KO mice were found to exhibit enhanced ER-mitochondria association and increased mitochondrial fragmentation in the liver. Nevertheless, the enzymatic activity of hepatic respiratory chain complexes and ER stress levels in the liver were not altered. Our findings identify a novel important role for STBD1 in the control of glucose metabolism, associated with the integrity of hepatic MERCs

A parametric model for the changes in the complex valued conductivity of a lung during tidal breathing

Classical homogenization theory based on the Hashin-Shtrikman coated ellipsoids is used to model the changes in the complex valued conductivity (or admittivity) of a lung during tidal breathing. Here, the lung is modeled as a two-phase composite material where the alveolar air-filling corresponds to the inclusion phase. The theory predicts a linear relationship between the real and the imaginary parts of the change in the complex valued conductivity of a lung during tidal breathing, and where the loss cotangent of the change is approximately the same as of the effective background conductivity and hence easy to estimate. The theory is illustrated with numerical examples, as well as by using reconstructed Electrical Impedance Tomography (EIT) images based on clinical data from an ongoing study within the EU-funded CRADL project. The theory may be potentially useful for improving the imaging algorithms and clinical evaluations in connection with lung EIT for respiratory management and monitoring in neonatal intensive care units

Chitosan and Oregano Oil Treatments, Individually or in Combination, Used To Increase the Shelf Life of Vacuum-Packaged, Refrigerated European Eel () Fillets.

We investigated the impact of chitosan and oregano essential oil (EO) individually or in combination on the quality of eel fillets in vacuum packaging (VP) and stored under refrigeration (4°C). Treatments studied were (i) control eel fillets stored in VP (E), (ii) eel fillets treated with 0.3% (v/w) oregano EO and stored in VP (E-OR), (iii) eel fillets treated with 2.0% (w/v) chitosan and stored in VP (E-CH), and (iv) eel fillets treated with 2.0% (w/v) chitosan and 0.3% (v/w) oregano EO and stored in VP (E-CH-OR). Treatments E-CH-OR and E-CH significantly reduced counts of mesophilic bacteria, and yeasts and molds during storage. Use of chitosan alone or in combination with oregano EO led to a significant reduction in concentrations of trimethylamine nitrogen and total volatile basic nitrogen in fillets, which led to lower concentrations of thiobarbituric acid reactive substances compared with the control samples. The eel samples in the E-CH and E-CH-OR groups were sensorially acceptable during the entire refrigerated storage period of 18 days. Presence of chitosan in the E-CH and E-CH-OR fillets did not negatively affect the taste of the fillets. E-CH fillets received a higher taste score than did E-CH-OR fillets probably because of the distinct and "spicy" lemon taste of chitosan, which was well received by the sensory panel. Based on overall sensory data (based on mean sensory scores of odor and taste), the shelf life was 6 days for the control fillets, 10 days for the E-OR fillets, and >18 days for the E-CH and E-CH-OR fillets stored in VP at 4°C. Overall, chitosan-treated eel fillets had lower microbial loads and a longer shelf life compared with the controls. Chitosan-treated eel fillets were preferred over oregano-treated fillets. Chitosan alone or in combination with oregano could be used as a preservative treatment and shelf-life extender for other seafoods

A Naturally Derived Watercress Flower-Based Phenethyl Isothiocyanate-Enriched Extract Induces the Activation of Intrinsic Apoptosis via Subcellular Ultrastructural and Ca²⁺ Efflux Alterations in an In Vitro Model of Human Malignant Melanoma

The aim of the current study was to (i) extract isolated fractions of watercress flowers enriched in polyphenols, phenethyl isothiocyanate and glucosinolates and (ii) characterize the anticancer mode of action of non-lethal, sub-lethal and lethal concentrations of the most potent extract fraction in primary (A375) and metastatic (COLO-679) melanoma cells as well as non-tumorigenic immortalized keratinocyte (HaCaT) cells. Cytotoxicity was assessed via the Alamar Blue assay, whereas ultrastructural alterations in mitochondria and the endoplasmic reticulum were determined via transmission electron microscopy. Mitochondrial membrane depolarization was determined using Mito-MP dye, whereas apoptosis was evaluated through the activation of caspases-3, -8 and -9. Among all extract fractions, the phenethyl isothiocyanate-enriched one (PhEF) possessed significant cytotoxicity against A375 and COLO-679 cells, while HaCaT cells remained relatively resistant at sub-lethal and lethal concentrations. Additionally, ultrastructural subcellular alterations associated with apoptosis were observed by means of increased mitochondrial area and perimeter, decreased cristae density and a shorter distance of the endoplasmic reticulum to the mitochondria, all taking place during “early” time points (2–4 h) of exposure. Moreover, PhEF induced mitochondrial membrane depolarization associated with “late” time points (24 h) of exposure, thereby leading to the activation of intrinsic apoptosis. Finally, the inhibition of cytosolic Ca2+ efflux reduced levels of caspases-9 and -3 activity, suggesting the involvement of Ca2+ efflux in modulating the activation of intrinsic apoptosis. To conclude, our data demonstrate an association of “early” ultrastructural alterations in mitochondria and the endoplasmic reticulum with the “late” induction of intrinsic apoptosis via the modulation of Ca2+ efflux

Immobilized Ag-nanoparticles (iNPs) for environmental applications: Elucidation of immobilized silver-induced inhibition mechanism of Escherichia coli

Although silver nanoparticles (AgNPs) appear to be promising for certain medical/pharmaceutical applications, they present significant disadvantages when it comes to environmental applications due to the need for recovery of the metal (Ag) which is considered hazardous for the environment. The present study examines the antimicrobial properties and mechanism of action of immobilized (on Al2O3) silver nanoparticles’ (1 wt% Ag-iNPs) regarding the treatment of E. coli microbial solutions. Antimicrobial experiments were conducted in semi-batch mode using a three-phase continuous flow stirred tank reactor. Every treated sample was taken from the outlet of the reactor for the time intervals of 0, 5 and 25 min. To ensure that the bactericidal property of Ag-iNPs is not attributed to the dissolution of surface silver (as free Ag+), a suitable Ag+ scavenger was used as already described in our earlier studies. Regarding the molecular analysis performed under this study, the regulation of key enzymes involved in bactericidal activity of E. coli, was examined, after their treatment either with immobilized silver nanoparticles (Ag-iNPs) or AgNO3. Specifically, 50 mL sample for each case was centrifuged (10 min at 10,000 rpm) and the pellet (≈109 cells) was immediately subjected to total RNA extraction. For real-time RT-qPCR analyses, 1 μg of total RNA was converted into cDNA. It was found that PldA gene, which encodes outer membrane's phospholipase A (OMPLA), was up-regulated after 5 and 25 min of treatment with Ag-iNPs. OMPLA's activation appears to be the initial step of Ag-iNPs bactericidal mechanism that ultimately leads to the creation of holes on the outer membrane (OM), irreversibly disturbing the cells’ respiration cycle. In addition, after treatment with Ag-iNPs, Blue copper oxidase CueO (encoded by cueO gene) was found to be over-produced and appears to play a key-role in the oxidative transfer of silver from the surface of Ag-iNPs to the cell. This leads to the Ag-induced displacement of copper from its native protein sites such as CusS/CusR, a fact that causes the release of labile copper and ROS inside the cell. In addition, TEM analysis of bacteria treated with Ag-iNPs revealed severe morphological changes (“holes”) on bacterial outer membrane, indicating that there is an intermediate step in the proposed antimicrobial mechanism which takes place on the surface of Ag-iNPs