Westernized diet is characterized by a high content of saturated fatty acids (SFA) and a low level of omega-<em>3</em> polyunsaturated fatty acids (PUFA), often accompanied by an imbalance in the omega-6/omega-<em>3</em> PUFA ratio. Since increased intake of SFA and n-6 PUFA is considered as a cardiovascular disease risk factor, this study was conducted to determine whether a three-month dietary supplementation of high-fat diets (HFDs) with saturated fatty acids and a significant proportion of various n-6 and n-<em>3</em> PUFA ratios would affect the architecture and <em>protein</em> expression patterns of the murine heart.
Therefore, three HFD (<i>n</i> = 6) feeding groups: rich in SFA, dominated by PUFA with the n-6/n-<em>3</em>-14:1, and n-6/n-<em>3</em>-5:1, ratios were compared to animals fed standard mouse chow. For this purpose, we performed two-dimensional electrophoresis with MALDI-ToF mass spectrometry-based identification of differentially expressed cardiac <em>proteins</em>, and a histological examination of cardiac morphology. The results indicated that mice fed with all HFDs developed signs of hypertrophy and cardiac fibrosis. Animals fed SFA-rich HFD manifested the most severe cardiac hypertrophy and fibrosis lesions, whereas less pronounced changes were observed in the group of animals that ingested the highest amount of omega-<em>3</em> FA.
In general, all HFDs, regardless of FA composition, evoked a comparable pattern of cardiac <em>protein</em> changes and affected the following biological processes: lipid metabolism and FA β-oxidation, glycolysis, <em>TCA</em> cycle, respiratory chain, myocardium contractility, oxidative stress and PUFA eicosanoid metabolism. However, it should be noted that three <em>proteins</em>, namely IDH<em>3</em>A, LDHB, and AK1, were affected differently by various FA contents. High expression of these myocardial <em>proteins</em> found in the group of animals fed a HFD with the highest n-<em>3</em> PUFA content could be closely related to the observed development of hypertrophy.
LC/MS-Based Global Metabolomic Identification of Serum Biomarkers Differentiating Hepatocellular Carcinoma from Chronic Hepatitis B and Liver Cirrhosis
Chronic hepatitis B virus (CHB) infection is one of the primary risk factors associated with the development of hepatocellular carcinoma (HCC). Despite having been extensively studied, diagnosing early-stage HCC remains challenging, and diagnosed patients have a poor (<em>3</em>-5%) survival rate. Identifying new approaches to detect changes in the serum metabolic profiles of patients with CHB and liver cirrhosis (LC) may provide a valuable approach to better detect HCC at an early stage when it is still amenable to treatment, thereby improving patient prognosis and survival. In the present study, we, therefore, employed a liquid chromatography-mass spectrometry (LC-MS)-based approach to evaluate the serum metabolic profiles of <em>3</em>0 CHB patients, 29 LC patients, and <em>3</em>0 HCC patients. We then employed appropriate statistical methods to identify those metabolites that were best able to distinguish HCC cases from LC and CHB controls.
A mass-based database was then used to putatively identify these metabolites. We then confirmed the identities of a subset of these metabolites through comparisons with the MS/MS fragmentation patterns and retention times of reference standards. The serum samples were then reanalyzed to quantify the levels of these selected metabolites and of other metabolites that have previously been identified as potential HCC biomarkers. Through this approach, we observed clear differences in the metabolite profiles of the CHB, LC, and HCC patient groups in both positive- and negative-ion modes.
Reduced Fragmentation of IGFBP-2 and IGFBP-<em>3</em> as a Potential Mechanism for Decreased Ratio of IGF-II to IGFBPs in Cerebrospinal Fluid in Response to Repeated Intrathecal Administration of Triamcinolone Acetonide in Patients With Multiple Sclerosis
Multiple sclerosis (MS) is a chronic autoimmune disease of the brain and spinal cord causing a wide range of symptoms such as impaired walking capability, spasticity, fatigue, and pain. The insulin-like growth factor (IGF) system has regulatory functions for the induction of inflammatory pathways in experimental encephalomyelitis. We have therefore assessed expression and regulation of the IGF system on the level of IGFs and IGFBPs in serum and cerebrospinal fluid (CSF) in the course of four repeated triamcinolone acetonide (<em>TCA</em>) administrations in two female and four male MS patients.
Sample series of 20 treatment cycles were analyzed. IGF-I and IGF-II were quantified by ELISAs, and IGFBPs were analyzed by quantitative Western ligand (qWLB) and Western immunoblotting (WIB) in order to differentiate intact and fragmented IGFBPs. The ratios of fragmented to intact IGFBP-2 and -<em>3</em> were calculated in serum and CSF. Finally, the ratios of IGF-I and IGF-II to the total IGF-binding activity, quantified by qWLB, were determined as an indicator of IGF-related bioactivity.
After the fourth <em>TCA</em> administration, the average level of IGF-I was increased in serum (p < 0.001). The increase of IGF-I concentrations in serum resulted in an increased ratio of IGF-I to IGFBPs in the circulation. By contrast in CSF, fragmentation of IGFBP-2 and IGFBP-<em>3</em> and the ratio of IGF-II to intact IGFBPs were decreased at the fourth <em>TCA</em> administration (p < 0.01).
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Furthermore, reduced fragmentation of IGFBP-<em>3</em> in CSF was accompanied by increased concentrations of intact IGFBP-<em>3</em> (p < 0.001). We conclude that reduced fragmentation of IGFBPs and concomitant reduction of IGF-II to IGFBP ratios indicate regulation of bioactivity of IGF-II in CSF during repeated intrathecal <em>TCA</em> administration in MS patients.