Online ISSN: 2515-8260

Keywords : therefore

Autophagic flux without a block differentiates varicella from herpes simplex virus infection

Charles Grose

European Journal of Molecular & Clinical Medicine, 2015, Volume 2, Issue 4, Pages -

Varicella-zoster virus (VZV) is a herpesvirus that causes a characteristic vesicular exanthem in humans with primary infection (varicella) or reactivation (zoster). We have previously observed that vesicular cells are filled with autophagosomes that are easily detectable by confocal microscopy after immunolabeling for the LC3 protein. Through a 3D imaging software program called Imaris we have quantitated autophagosomes as greater than 100 per cell. Similarly, we have assessed autophagy in VZV-infected monolayers after inoculation by the traditional method with infected cells at a ratio of one infected to 8 uninfected cells. Again, autophagosomes are easily detected, but their count is lower than that observed in human skin cells. As an additional control, we enumerated the autophagosomes in the Severe Combined Immuno-Deficient (SCID) Mouse model of VZV infection. In this model, human skin is inserted under the skin of the mouse and subsequently inoculated with VZV-infected cells. Again, autophagy was abundant in the VZV-infected skin and minimal in the mock-infected skin sample. Subsequently, we investigated autophagy following infection with sonically prepared cell free virus in cultured cells. After cell free virus inoculation, autophagy was detected in a majority of infected cells at all time points, but was less than that seen after an infected-cell inoculum. Finally, we investigated VZV-induced autophagic flux by two different methods (radiolabeling proteins and a dual-colored LC3 plasmid);

Metabolic phenotyping in mouse and man: Mind the differences!

Denise Sonntag; Guido Krebiehl; Torben Friedrich

European Journal of Molecular & Clinical Medicine, 2015, Volume 2, Issue 2, Pages -

Metabolic phenotyping comprises the quantification of endogenous metabolites in biofluids, cells, and tissues. It provides insights into normal as well as aberrant metabolic pathways and biological processes, which is important for the understanding of disease phenotypes. It also allows the identification of biological markers, which can serve as early disease indicators and therapeutic markers for the evaluation of treatment effects. As metabolic markers are not species-restricted, the concept of metabolic phenotyping is highly applicable for translational research. Species independence allows the use of established animal and cell culture models for various diseases within a preclinical context. However, differences in the metabolic set-up of study organisms compared to humans needs to be taken into consideration to prevent misleading conclusions from otherwise valid experimental designs. To determine species-related metabolic differences, a targeted metabolomics approach was applied using a mass spectrometry platform for the quantification of a predefined set of endogenous metabolites, i.e. amino acids, biogenic amines, phosphatidylcholines, sphingomyelins, hexoses, steroid hormones and others. Results from this species comparison on the metabolic level will be presented.