Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is an efficient tool for large molecule analysis including proteins, carbohydrates, polymers etc., with the advantages of high sensitivity, high throughput and high speed. However, the traditional matrices, such as α-cyano-4-hydroxycinnamic acid (CHCA) and 2,5-dihydroxybenzoic acid (DHB), fail to detect the compounds with molecular weight below 500 Da due to the interference from matrix background. In this paper, we used C60 fluorine derivative as matrix for small molecule analysis by MALDI-TOF MS. With the successful detection of various small molecules from saccharides, plant extracts, fatty acids, amino acids to pesticides, C60 derivative is reported here for the first time as a novel and effective MALDI matrix for small molecule analysis in metabolism and compound characterization areas.

CM2 is the second membrane protein of influenza C virus. In the present study, to investigate the role(s) of CM2 in the virus replication, we generated recombinant influenza C viruses, rC65A and rN11A, which lack the palmitoylatioin and glycosylation sites of CM2, respectively. The rC65A virus grew as efficiently as the recombinant wild-type (rWT) virus did, whereas the rN11A grew less efficiently than the rWT virus. To study the difference more precisely, we generated influenza C virus-like particles (VLPs) lacking the CM2 glycosylation site (N11A-VLPs) and examined the VLPs and VLP-infected cells. As a result, the N11A-VLPs contain approximately 13% of the virus RNA found in wildtype VLPs (WT-VLPs), and N11A-VLP-infected HMV-II cells exhibited reduced reporter gene expression compared with that of WT-VLP-infected cells (WT : N11A = 1:0.1). Thus, supportive evidence was obtained that CM2 is involved in packaging and uncoating processes and that observed growth difference between rWT and rN11A can be attributed to the difference in the CM2 function.

Oxidative stress represents a mechanism which could lead to diabetic cataract. We exposed bovine lenses in culture conditions for two weeks to high glucose concentration (450 mg%) and investigated the damage to the lens and possible protection by special antioxidants - N-acetyl-L-cysteine (NAC) and the zinc complex of desferrioxamine (DFO), a selective chelator for iron. We monitored the optical quality of the lenses and the oxidation of the epithelium with dichlorofluorescein (DCF) assay, as well as the changes in lens proteins profile by 2D gel electrophoresis. Under high glucose changes in lens focal length, increased oxidation, and changes in lens crystalline were observed. NAC and Zn- DFO nearly completely protected the lenses; DFO showed only partial protection. The results demonstrated that antioxidants should be considered as treatment modality protecting the lens from high glucose damage. It is proposed that a combination of NAC and Zn/DFO could prove highly efficient.

Modeling and analyzing the architecture of metabolic networks using various computational strategies can be successfully used for studying their internal metabolic dynamics as well as predicting missing links in diseased networks. In the present work, we have implemented our algorithm based on structural grammars, for automated metabolic pathway reconstruction and modeling in metabolic pathways responsible for coding genes responsible for the cause of Type 1 Diabetes mellitus (T1D) in Homo sapiens. We have especially implemented our algorithm for studying the metabolic pairs responsible for the functioning of GAD1 and GAD2 genes. We have also used the algorithm for automated reconstruction of glutamate metabolism, β-alanine metabolism, taurine & hypotaurine metabolism and butanoate metabolism pathway datasets. We have also used the algorithm for missing and multiple link prediction as well as nodal point analysis for all the four metabolic pathways with 90.4-100% accuracy.

Two genetically close inbred tomato lines, one resistant to Tomato yellow leaf curl virus (TYLCV) infection (R), the other susceptible (S), showed completely different stress response upon TYLCV infection. S plants were stunted and do not yield, while R plants remained symptomless and yielded. Comparison of protein profiles and metabolites patterns in TYLCV infected R and S tomatoes revealed a completely different host stress response. S plants were characterized by higher levels of reactive oxygen species (ROS) and ROS compounds, the anti-oxidative, pathogenesis-related (PR) and wound-induced proteins were predominant. In contrast, infection of R tomatoes did not drastically activate the same host defense mechanisms as in S plants, while R homeostasis was much more effectively maintained by protein and chemical chaperones. Sources of carbon and nitrogen were less affected by TYLCV in R than in S plants, which could make R plants more balanced and more fit to sustain infection. Even though both tomato types contained comparable amounts of TYLCV at the specified stage of infection, the cellular immune responses were different. Presented results are preliminary and indicate not so much concrete data but the global tender in understanding of the cellular response to virus stress at the background of resistance and susceptibility to TYLCV.

Identification of membrane proteins that are expressed on the arteriovenous malformation (AVM) endothelium post radiosurgery is of fundamental importance in developing a new treatment of brain AVMs. We successfully optimized and then employed in vitro and in vivo biotinylation methodology of murine cerebral endothelial cell cultures (bEnd.3) and the rat model of AVM to label membrane proteins. Those membrane proteins were then captured on streptavidin resin and identified using proteomics analysis. This is the first time that proteomics has been employed in the study of AVMs.