Coenzyme Q Metabolism Is Disturbed in High Fat Diet-Induced Non Alcoholic Fatty Liver Disease in Rats

Oxidative stress is believed to be a major contributory factor in the development of non alcoholic fatty liver disease (NAFLD), the most common liver disorder worldwide. In this study, the effects of high fat diet-induced NAFLD on Coenzyme Q (CoQ) metabolism and plasma oxidative stress markers in rats were investigated. Rats were fed a standard low fat diet (control) or a high fat diet (57% metabolizable energy as fat) for 18 weeks. The concentrations of total (reduced + oxidized) CoQ9 were increased by >2 fold in the plasma of animals fed the high fat diet, while those of total CoQ10 were unchanged. Reduced CoQ levels were raised, but oxidized CoQ levels were not, thus the proportion in the reduced form was increased by about 75%. A higher percentage of plasma CoQ9 as compared to CoQ10 was in the reduced form in both control and high fat fed rats. Plasma protein thiol (SH) levels were decreased in the high fat-fed rats as compared to the control group, but concentrations of lipid hydroperoxides and low density lipoprotein (LDL) conjugated dienes were unchanged. These results indicate that high fat diet-induced NAFLD in rats is associated with altered CoQ metabolism and increased protein, but not lipid, oxidative stress

Individually ventilated cages – Microbiological containment testing

Modern biomedical research projects need “high quality” laboratory animals in order to obtain repeatability and homogeneity of the experimental data and to reduce the number of animals. It is, thus necessary to use animals standardised with respect to genetic, microbiological and pathological conditions. Environmental control, by eliminating the chemical, physical and microbial contaminants, can guarantee more standardised housing conditions and, at the same time, highly reduce the biological risk for the personnel working with the animals in accordance with the European and Italian legislation. Among the physical parameters temperature and humidity have to be considered the most important for their influence on animal behavior and metabolism. The optimal values for temperature range between 20 and 23 °C and for humidity between 40 and 60 %; in fact it has been widely demonstrated that when these two parameters are too high, they enhance the growth of moulds and the life span of microorganisms and that most bacteria and fungi, that colonize man and animals, grow at temperatures between 25 and 40 °C.Microbiological quality assurance of laboratory animals aims to produce animals that meet requirements for microbiological quality, and to maintain the same quality throughout the experiment. Outbreaks of infectious disease in laboratory animals have to a great extent adversely affected their use in biomedical research. Some microorganisms occurring in laboratory animals can also affect man (zoonosis) and this risk is also present in animals which are used as a source of sera and vaccines for use in man. Due to the introduction of specified pathogen free (SPF) animals outbreaks of infectious diseases have been in part replaced by more subtle microbial interference in the outcome of animal experiments. Plenty of viruses, mycoplasma, bacteria and parasites that may affect biomedical research exist for all laboratory animal species (Boot, 1996).The Service for Quality and Assurance of Animal Experimentation routinely controls the animal housing environment. For several years it has been focusing its activity on monitoring microbiological conditions and even in the animal housing it has considered the indication of Whittard (Table 1), as acceptable levels of contamination. With regard to this problem we have analyzed the count of environmental microorganisms in the animal facility rooms and the isolating capacity of a ventilated cage system (IVC, Techniplast Gazzada, Varese, Italy) which has been conceived to protect, by filtering the supply and exhaust circulating air, both the housed animals and the working personnel. The experiment has been performed in a conventional animal facility to verify the possibility to use animals in mixed conditions i.e. animals housed in normal conditions and animals housed in IVC

Applicazioni della PCR e PCR in situ nella diagnosi di infezioni batteriche e virali da biopsie fissate in formalina e incluse in paraffina

In situ PCR, amplification of target DNA sequences in fixed cells, is a very useful molecular biology tecnique with potential to combine the high sensitivity of tube PCR with the precise anatomical localization of the targeted bsequences. It allows the study of low copy viral or bacterial DNA. In this study we document the utility of directin situ PCR with single primer pair by applying it to 3 infectious agents in different model systems: Borrelia burgdorferi in 5 Eritema migrans lesions and 55 primitive cutaneous B cell lymphomas, Chlamydia pneumoniae in 200 autoptic atheromasic lesions, and Papilloma virus in 20 CIN 1 (mild cervical dysplasia). In situ PCR seems to be a very promising tecnique; however, the prerequisite for the success of in situ PCR is conditioned by optimal standardization of the key variables which, on the other hand, are influenced by tissue composition

Localizzazione e valutazione dell’espressione di Chlamydophila pneumoniae mediante RT-PCR in situ

Chlamydophila pneumoniae, an obligate intracellular gram negative bacterium, is involved in a wide spectrum of symptomatic respiratory tract diseases. However more recently it has been reported to be a pathogenic agent in the mechanism leading to atherosclerosis. In the present study the presence of Chlamydophila pneumoniae was assessed, using nested PCR and in situ PCR, while the viability of the microorganism was investigated using RT in situ PCR.The results obtained demonstrated that Chlamydophila pneumoniae was present and alive in the tissues examined.The global concordance of results in the three techniques used was 100%. RT in situ PCR can be considered a precious tool to detect bacterial mRNA in formalin fixed paraffin embedded samples provided an optimal standardization of the key variables is achieved

Metabolic Heterogeneity Evidenced by MRS among Patient-Derived Glioblastoma Multiforme Stem-Like Cells Accounts for Cell Clustering and Different Responses to Drugs

Clustering of patient-derived glioma stem-like cells (GSCs) through unsupervised analysis of metabolites detected by magnetic resonance spectroscopy (MRS) evidenced three subgroups, namely clusters 1a and 1b, with high intergroup similarity and neural fingerprints, and cluster 2, with a metabolism typical of commercial tumor lines. In addition, subclones generated by the same GSC line showed different metabolic phenotypes. Aerobic glycolysis prevailed in cluster 2 cells as demonstrated by higher lactate production compared to cluster 1 cells. Oligomycin, a mitochondrial ATPase inhibitor, induced high lactate extrusion only in cluster 1 cells, where it produced neutral lipid accumulation detected as mobile lipid signals by MRS and lipid droplets by confocal microscopy. These results indicate a relevant role of mitochondrial fatty acid oxidation for energy production in GSCs. On the other hand, further metabolic differences, likely accounting for different therapy responsiveness observed after etomoxir treatment, suggest that caution must be used in considering patient treatment with mitochondria FAO blockers. Metabolomics and metabolic profiling may contribute to discover new diagnostic or prognostic biomarkers to be used for personalized therapies