Emerging evidence indicates that polychlorinated biphenyls (PCBs) are involved in the development of diabetes mellitus in the obese. The purpose of this study was to determine mechanisms by which PCB 153 (2,2′,4,4′,5,5′-hexachlorobiphenyl) could influence diet-induced obesity and insulin resistance during adipogenesis. Lineage of h-ADMSCs was differentiated either as control (differentiation medium only), or with lipid vehicle modeling high fat nutrition (NuTRIflex) or lipid free vehicle (dimethylsulfoxide) for 28 days with or without PCB 153 daily co-exposure (in three concentrations 0.1, 1, and 10 μM). Gene expression analyses were performed using RT-qPCR at days 4, 10, 21, 24, 28; p rotein l evels A kt a nd phosphorylated Akt (Phospho-Akt) by Western blot at days 4, and 21. PCB 153 treatment of h-ADMSCs only in lipid vehicle was associated with down regulation of key master genes of adipogenesis: PPARγ, SREBP-1, PPARGC1B, and PLIN2 during the whole process of differentiation; and with increased Akt and decreased Phospho-Akt protein level at day 21. We have shown that PCB 153, in concentration 0.1 μM, has a potential in lipid rich environment to modulate differentiation of adipocytes. Because European and U.S. adults have been exposed to PCB 153, this particular nutrient-toxicant interaction potentially impacts human obesity and insulin sensitivity., D. Mullerova, M. Pesta, J. Dvorakova, M. Cedikova, V. Kulda, P. Dvorak, V. Bouchalová, M. Kralickova, V. Babuska, J. Kuncova, J. Langmajerova, L. Muller., and Obsahuje bibliografii
Lygus lineolaris (Palisot de Beauvois, 1818) (tarnished plant bug) is a serious pest of cotton (Gossypium hirsutum L.) in the Delta region as compared to cotton in the Hills region of the state of Mississippi in USA. The reason for this is unclear but it was hypothesized that the plant cell wall degrading polygalacturonase enzyme system in the salivary glands of L. lineolaris from the Delta could be better adapted for cotton, which is grown more predominantly in the Delta region than in the Hills region. Expression analysis of three primary polygalacturonase genes (LlPG1, LlPG2 and LlPG3) was conducted in laboratory reared and field collected populations of L. lineolaris. Assay of polygalacturonase enzyme activity was also conducted to compare wild collected populations. Initial laboratory and field data revealed gene expression differences in sex, age, region, and host plant which guided the direction of our subsequent study during 2013 and 2014. Based on the results of this study, we propose that the three genes studied may not be reflective of the entire polygalacturonase enzyme system and may not be solely responsible for the observed adaptation of L. lineolaris to cotton in the Delta region than in the Hills region. Analyses also revealed that the expression of the three targeted polygalacturonase genes was affected by the host plant from which the insects were collected and that adults had higher polygalacturonase expression than nymphs. Taken together, our results provide strong evidence for developmental stage specific and host plant based change in expression of PG genes in the salivary glands of L. lineolaris. This, however, was not reflected in total polygalacturonase enzyme activity which was not significantly different between regions, hosts, sex, or developmental stage., Daniel Fleming, Natraj Krishnan, Fred Musser., and Obsahuje bibliografii
Excessive production of oxygen free radicals has been regarded as a causative common denominator of many pathological processes in the animal kingdom. Hydroxyl and nitrosyl radicals represent the major cause of the destruction of biomolecules either by a direct reaction or by triggering a chain reaction of free radicals. Scavenging of free radicals may act preventively or therapeutically. A number of substances that preferentially react with free radicals can serve as scavengers, thus increasing the internal capacity/activity of endogenous antioxidants and protecting cells and tissues against oxidative damage. Molecular hydrogen (H2) reacts with strong oxidants, such as hydroxyl and nitrosyl radicals, in the cells, that enables utilization of its potential for preventive and therapeutic applications. H2 rapidly diffuses into tissues and cells without affecting metabolic redox reactions and signaling reactive species. H2 reduces oxidative stress also by regulating gene expression, and functions as an anti-inflammatory and anti-apoptotic agent. There is a growing body of evidence based on the results of animal experiments and clinical observations that H2 may represent an effective antioxidant for the prevention of oxidative stress-related diseases. Application of molecular hydrogen in situations with excessive production of free radicals, in particular, hydroxyl and nitrosyl radicals is relatively simple and effective, therefore, it deserves special attention., J. Slezák, B. Kura, K. Frimmel, M. Zálešák, T. Ravingerová, C. Viczenczová, Ľ. Okruhlicová, N. Tribulová., and Obsahuje bibliografii
Inflammatory changes, both in the arterial wall and adipose tissue, play a crucial role in the development of atherosclerosis. We measured the gene expression of tumor necrosis factor-alpha (TNFα), monocyte chemoattractant protein-1 (MCP-1), and interleukin 6 (IL-6) in adipose tissue (AT) of living kidney donors (LKD) and patients with peripheral arterial disease (PAD). Quantitative polymerase chain reaction (qPCR) and flow cytometry analyses were performed in subcutaneous (SAT), visceral (VAT), and perivascular adipose tissue (PVAT). Data of PAD patients showed significantly higher expression in VAT in all three genes (TNFα 5-fold, p<0.05; MCP-1 3.6-fold, p<0.05; IL-6 18.8-fold, p<0.001). The differences in PVAT and SAT were less significant. Total body pro-inflammatory status was documented by higher TNFα concentration in patients (4.86± 1.4 pg/ml) compared to LKDs (2.14±0.9 pg/ml; p<0.001), as was hsCRP (11.8±7.0 in PAD; 1.5±0.48 in LKDs; p=0.017). We found no age-dependent relationship between gene expression vs. TNFα and hsCRP concentrations in both compared groups. No effect of the atherosclerosis score on gene expression and circulating inflammatory markers within the PAD group was observed. Our results suggest that the AT of PAD patients infiltrated with macrophages produces more cytokines involved in the development of inflammation and atherosclerosis., S. Čejková, I. Králová Lesná, J. Froněk, L. Janoušek, A. Králová, J. Ždychová, R. Poledne., and Obsahuje bibliografii
Stress exposure activates the sympathoneural system, resulting in catecholamine release. Chronic stress is associated with development of numerous disorders, including cardiovascular diseases. Here we investigated the expression of mRNAs for catecholamine biosynthetic enzymes tyrosine-hydroxylase, dopamine-ß-hydroxylase and phenylethanolamine N-methyl- transferase, and for ß1- and ß2-adrenoceptors in the right and left ventricles of rats exposed to chronic unpredictable mild stress. The tyrosine-hydroxylase and dopamine-ß-hydroxylase mRNA levels were not affected by stress, whereas the phenylethanolamine N-methyltransferase mRNA levels significantly increased in both right and left ventricles. No changes in ß1-adrenoceptor mRNA levels in either right or left ventricles were observed. At the same time, stress produced a significant increase of β2-adrenoceptor mRNA levels in left ventricles. These results suggest that elevated expression of phenylethanolamine N-methyltransferase in both ventricules and ß2-adrenoceptor genes in left ventricles could provide a molecular mechanism that leads to altered physiological response, which is important for the organism coping with stress., N. Spasojevic, L. Gavrilovic, S. Dronjak., and Obsahuje bibliografii a bibliografické odkazy
Salicylic acid (SA) and nitric oxide (NO) form a new group of plant growth substances that cooperatively interact to promote plant growth and productivity. Water deficit (WD) stress is a major limiting factor for photosynthesis, which in turn limits crop yield. However, the mechanism of SA and NO in stimulating photosynthesis has not yet been elucidated. Therefore, in this study, we investigated the SA- and NO-mediated photosynthetic adaptability of maize seedlings to WD in terms of photosynthetic parameters, activities and mRNA levels of CO2 assimilation enzymes. Our results showed that SA alleviated the WD-induced reduction of photosynthetic performance. The activities of Rubisco and Rubisco activase enzymes increased significantly due to SA pretreatment. Moreover, higher transcription rates of Rbc L, ZmRCAα and ZmRCAβ mRNA further confirmed the effects of SA on CO2 assimilation. WD or SA-induced decreases or increases of CO2 assimilation ability were further decreased after c-PTIO addition., R. X. Shao, L. F. Xin, J. M. Guo, H. F. Zheng, J. Mao, X.P. Han, L. Jia, S. J. Jia, C. G. Du, R. Song, Q. H. Yang, R. W. Elmore., and Obsahuje bibliografii
In our previous work we established a T7 polymerase-driven Tetracycline-inducible protein expression system in Leishmania mexicana (Biagi, 1953). We used this system to analyse gene expression profiles during development of L. mexicana in procyclic and metacyclic promastigotes and amastigotes. The transcription of the gene of interest and the T7 polymerase genes was significantly reduced upon cell differentiation. This regulation is not locus-specific. It depends on untranslated regions flanking open reading frames of the genes analysed. In this paper, we report that the previously established conventional inducible protein expression system may not be suitable for studies on differentiation of species of Leishmania Ross, 1903 and protein expression systems might have certain limitations., Aygul Ishemgulova, Natalya Kraeva, Drahomíra Faktorová, Lucie Podešvová, Julius Lukeš, Vyacheslav Yurchenko., and Obsahuje bibliografii
Increased circulating adhesion molecules in patients with obesity play an important role in the development of endothelial dysfunction/atherosclerosis. The aim of this study was to assess the contribution of various fat depots to the production of adhesion molecules in obesity. 12 women with 1st and 2nd degree of obesity, 13 women with 3rd degree of obesity and 14 lean age-matched women were included into study. Circulating levels of vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), and E-selectin were measured by Luminex kits. mRNA expression of ICAM-1, VCAM-1, E-selectin, monocyte chemoattractant protein-1 (MCP-1), and CD68 in subcutaneous (SAT) and visceral adipose tissue (VAT) was measured by RT-PCR; ICAM-1 and VCAM-1 protein levels by Luminex kits, normalized to protein content. Obesity increased ICAM-1 and VCAM-1 mRNA expression and protein levels and CD68 mRNA expression in VAT. Expression of E-selectin and MCP-1 did not significantly differ between groups. Expression of ICAM-1 and VCAM-1 positively correlated with expression of CD68 in both adipose depots. In VAT, ICAM-1 and VCAM-1 expression and protein levels positively correlated with BMI. Obesity was associated with increased adhesion molecules mRNA expression and protein levels in VAT, but not in SAT. Increased adhesion molecules production in visceral fat may provide a novel direct link between visceral adiposity and increased risk of cardiovascular complications., L. Bošanská ... [et al.]., and Obsahuje bibliografii a bibliografické odkazy
Mangiferin is a kind of polyphenol chemical compound separated from these herbal medicines of Mangifera indica L., Anemarrhena asphodeloides Bge. and Belamcanda chinensis L., which has anti-inflammatory, anti-virus, and other physiological activities without toxic effects. Osteoarthritis (OA) is a chronic disease that is also a kind of arthritis disease in which articular cartilage or bones under the joint is damaged. In addition, artificial replacements are required in severe cases. At present, there are not too much researches on the potential biological activities of mangiferin that plays a protective role in the treatment of OA. In this study, we evaluated the protective effect of mangiferin on osteoarthritis (OA) in vitro and in vivo. First, the effect of different concentrations of mangiferin on rat chondrocytes was determined by MTT assay. Second, the effects of mangiferin on the expression levels of matrix metalloproteinase (MMP)-13, TNF-α, Collagen II, Caspase-3, and cystatin-C in interleukin-1β (IL-1β)-induced rat chondrocytes were examined by the real-time polymerase chain reaction in vitro, meanwhile the effects of mangiferin on the nuclear factor kappa-B (NF-κB) signaling pathway were also investigated by Western Blot. Finally, the antiosteoarthritic protective effect of mangiferin was evaluated in the rat model by anterior cruciate ligament transection (ACLT) combined with bilateral ovariectomy-induced OA in vivo. The results showed that the mangiferin was found to inhibit the expression of MMP-13, TNF-α, and Caspase-3 which also increased the expression of Collagen II and cystatin-C in IL-1β-induced rat chondrocytes. In addition, IL-1β-induced activation of nuclear factor kappa-B (NF-κB) and the degradation of inhibitor of κB (IκB)-α were suppressed by mangiferin. For the in vivo study in a rat model of OA, 100 μl of mangiferin was administered by intra-articular injections for rats, the results showed that the cartilage degradation was suppressed by mangiferin through Micro CT and Histological Examination. According to both in vitro and in vivo results, mangiferin has a protective effect in the treatment of OA which may be a promising therapeutic agent for OA.
To understand the pathogenesis of hypercholesterolemia in Prague hereditary hypercholesterolemic (PHHC) rat, we analyzed the response of hepatic transcriptome to dietary cholesterol in PHHC and control Wistar rats. Male PHHC and Wistar rats were fed chow (C), 5 % fat (palm kernel oil) (CF) or 1 % cholesterol + 5 % fat (CHOL) diet for three weeks. Hepatic transcriptome was analyzed using Affymetrix GeneChip arrays. No differences were found in the effect of both control diets (C and CF) on lipid metabolism and gene expression of 6500 genes. Therefore, these data were pooled for further analysis. Dietary cholesterol induced accumulation of cholesterol and triacylglycerols in the liver in both strains and hypercholesterolemia in PHHC rats. However, there were no differences in response of hepatic transcriptome to CHOL diet. On the other hand, several genes were found to be differently expressed between both strains independently of the diet. Two of those genes, Apof and Aldh1a7, were studied in more detail, and their role in pathogenesis of hypercholesterolemia in PHHC rats could not been corroborated. In conclusion, the hypercholesterolemia in PHHC rats is due to physiological response of hepatic transcriptome to dietary cholesterol in different genetic background., M. Vlachová, M. Heczková, M. Jirsa, R. Poledne, J. Kovář., and Obsahuje bibliografii