Relationship between circulating VCAM-1, ICAM-1, E-selectin and MMP9 and the extent of coronary lesions

OBJECTIVES: Inflammatory molecules play a role in the development of atherosclerosis, which is the primary origin of cardiovascular disorders. However, to the best of our knowledge, no study has attempted to investigate the relationship between these circulating molecules and the prediction of cardiovascular risk. The present study aimed to investigate the relationships of vascular cell adhesion molecule-1, intercellular adhesion molecule-1, E-selectin and matrix metalloproteinase 9 serum concentrations with the extent of coronary lesions. METHODS: Seventy-four individuals who were undergoing coronary angiography for the first time for diagnostic purposes were enrolled in this study. The extent of the coronary lesion was assessed using the Friesinger Index, and subjects were classified into four groups: no lesions, minor lesions, intermediate lesions and major lesions. Serum biochemical parameters and serum concentrations of vascular cell adhesion molecule-1, intercellular adhesion molecule-1, E-selectin and matrix metalloproteinase 9 were analyzed. RESULTS: The vascular cell adhesion molecule-1 concentration was higher than 876 ng/mL in individuals with intermediate and major lesions (

Involvement of eukaryotic translation initiation factor 5a (eif5a) in skeletal muscle stem cell differentiation.

A proteína eIF5A apresenta um resíduo exclusivo de aminoácido chamado hipusina formado por modificação pós-traducional envolvendo espermidina como substrato. Neste estudo, observamos que a expressão de eIF5A é intensificada ao longo da diferenciação de células-tronco progenitoras de fibras musculares (células satélites) e que a inibição da hipusinação com GC7 bloqueia a diferenciação. Associado a esse bloqueio encontramos aumento do consumo de glicose e produção de lactato, diminuição da descarboxilação de glicose e palmitato, redução da proliferação celular e alteração do perfil traducional, efeitos que podem estar envolvidos na inibição do programa de diferenciação. Em seguida, o músculo tibial anterior de ratos foram criolesados e após severa supressão da expressão de eIF5A (período agudo de lesão) a mesma foi retomada ao longo da regeneração, chegando a quantidades superiores ao encontrado em músculos não lesados. Verificamos que a L-arginina, um supressor parcial do fenótipo distrófico e precursor de espermidina, reverte parcialmente o efeito de GC7.eIF5A protein contains an exclusive amino acid residue named hypusine produced by a post-translational modification involving spermidine as substrate. In this study, we observed that eIF5A expression is raised during muscle fiber stem cells (satellite cells) differentiation and the hypusination inhibition by GC7 abolished the differentiation process. In association with this blockage, an increase in glucose consumption and lactate production, a decrease in glucose and palmitate decarboxylation, a reduction in cell proliferation and an alteration in translational profile were observed. These changes may be involved in the inhibition of the differentiation induced by GC7. The rat tibialis anterior muscle was injured and a marked reduction of eIF5A expression (acute injury period) was found. The expression of eIF5A was reestablished during regeneration, reaching higher levels than that observed in non injured muscle. We also verified that L-arginine, a partial suppressor of muscle dystrophic phenotype condition and precursor of spermidine, partially abolished the GC7 effects

Revealing the translation control by transcriptome analysis

Taking the eukaryotic translation initiation factor 5A (eIF5A) asmodel, some aspects of different points of gene expression controland transcriptome studies are discussed. A parallel with proteomicstudies is presented, as well as the optimization of thetranscriptome analysis using polysome profile assay. The polysomeprofile assay application reveals the translational control of mRNAsnot identified by traditional differential analysis of transcriptomes,which is widely employed to study diseases, such as tumors

Titanium-protein nanocomposites as new biomaterials produced by high-pressure torsion

Abstract The development of new biomaterials with outstanding mechanical properties and high biocompatibility has been a significant challenge in the last decades. Nanocrystalline metals have provided new opportunities in producing high-strength biomaterials, but the biocompatibility of these nanometals needs to be improved. In this study, we introduce metal-protein nanocomposites as high-strength biomaterials with superior biocompatibility. Small proportions of bovine serum albumin (2 and 5 vol%), an abundant protein in the mammalian body, are added to titanium, and two nanocomposites are synthesized using a severe plastic deformation process of high-pressure torsion. These new biomaterials show not only a high hardness similar to nanocrystalline pure titanium but also exhibit better biocompatibility (including cellular metabolic activity, cell cycle parameters and DNA fragmentation profile) compared to nano-titanium. These results introduce a pathway to design new biocompatible composites by employing compounds from the human body

A Genome-wide Screen for Neurospora crassa Transcription Factors Regulating Glycogen Metabolism

Transcription factors play a key role in transcription regulation as they recognize and directly bind to defined sites in promoter regions of target genes, and thus modulate differential expression. The overall process is extremely dynamic, as they have to move through the nucleus and transiently bind to chromatin in order to regulate gene transcription. To identify transcription factors that affect glycogen accumulation in Neurospora crassa, we performed a systematic screen of a deletion strains set generated by the Neurospora Knockout Project and available at the Fungal Genetics Stock Center. In a wild-type strain of N. crassa, glycogen content reaches a maximal level at the end of the exponential growth phase, but upon heat stress the glycogen content rapidly drops. The gene encoding glycogen synthase (gsn) is transcriptionally down-regulated when the mycelium is exposed to the same stress condition. We identified 17 deleted strains having glycogen accumulation profiles different from that of the wild-type strain under both normal growth and heat stress conditions. Most of the transcription factors identified were annotated as hypothetical protein, however some of them, such as the PacC, XlnR, and NIT2 proteins, were biochemically well-characterized either in N. crassa or in other fungi. The identification of some of the transcription factors was coincident with the presence of DNA-binding motifs specific for the transcription factors in the gsn 5'-flanking region, and some of these DNA-binding motifs were demonstrated to be functional by Electrophoretic Mobility Shift Assay (EMSA) experiments. Strains knocked-out in these transcription factors presented impairment in the regulation of gsn expression, suggesting that the transcription factors regulate glycogen accumulation by directly regulating gsn gene expression. Five selected mutant strains showed defects in cell cycle progression, and two transcription factors were light-regulated. The results indicate that there are connections linking different cellular processes, such as metabolism control, biological clock, and cell cycle progression. Molecular & Cellular Proteomics 10: 10.1074/mcp.M111.007963, 1-13, 2011.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

Insulin action on protein synthesis and its association with eIF5A expression and hypusination

The hormone insulin plays a central role in the metabolism of carbohydrates, lipids, and proteins. In relation to protein metabolism, insulin stimulates amino acid uptake and activates protein synthesis in responsive cells by modulation of signal transduction pathways, such as associated to Akt/PkB, mTOR, S6Ks, 4E-BP1, and several translation initiation/elongation factors. In this context, there is no information on direct cellular treatment with insulin and effects on eukaryotic translation initiation factor 5A (eIF5A) regulation. The eIF5A protein contains an exclusive amino acid residue denominated hypusine, which is essential for its activity and synthesized by posttranslational modification of a specific lysine residue using spermidine as substrate. The eIF5A protein is involved in cellular proliferation and differentiation processes, as observed for satellite cells derived from rat muscles, revealing that eIF5A has an important role in muscle regeneration. The aim of this study was to determine whether eIF5A expression and hypusination are influenced by direct treatment of insulin on L6 myoblast cells. We observed that insulin increased the content of eIF5A transcripts. This effect occurred in cells treated or depleted of fetal bovine serum, revealing a positive insulin effect independent of other serum components. In addition, it was observed that hypusination follows the maintenance of eIF5A protein content in the serum depleted cells and treated with insulin. These results demonstrate that eIF5A is modulated by insulin, contributing the protein synthesis machinery control, as observed by puromycin incorporation in nascent proteins461587596FAPESP – Fundação de Amparo à Pesquisa Do Estado De São Paulo2010/18095-0; 2013/20504-3; 2013/23620-4; 2014/27154-0; 2017/21914-

Potential contribution of translational factors to triiodo-l-hyronine-induced Insulin synthesis by pancreatic beta cells

Thyroid hormones (THs) are known to regulate protein synthesis by acting at the transcriptional level and inducing the expression of many genes. However, little is known about their role in protein expression at the post-transcriptional level, even though studies have shown enhancement of protein synthesis associated with mTOR/p70S6K activation after triiodo-l-thyronine (T3) administration. On the other hand, the effects of TH on translation initiation and polypeptidic chain elongation factors, being essential for activating protein synthesis, have been poorly explored. Therefore, considering that preliminary studies from our laboratory have demonstrated an increase in insulin content in INS-1E cells in response to T3 treatment, the aim of the present study was to investigate if proteins of translational nature might be involved in this effect. INS-1E cells were maintained in the presence or absence of T3 (10(-6) or 10(-8) M) for 12 hours. Thereafter, insulin concentration in the culture medium was determined by radioimmunoassay, and the cells were processed for Western blot detection of insulin, eukaryotic initiation factor 2 (eIF2), p-eIF2, eIF5A, EF1A, eIF4E binding protein (4E-BP), p-4E-BP, p70S6K, and p-p70S6K. It was found that, in parallel with increased insulin generation, T3 induced p70S6K phosphorylation and the expression of the translational factors eIF2, eIF5A, and eukaryotic elongation factor 1 alpha (eEF1A). In contrast, total and phosphorylated 4E-BP, as well as total p70S6K and p-eIF2 content, remained unchanged after T3 treatment. Considering that (i) p70S6K induces S6 phosphorylation of the 40S ribosomal subunit, an essential condition for protein synthesis; (ii) eIF2 is essential for the initiation of messenger RNA translation process; and (iii) eIF5A and eEF1A play a central role in the elongation of the polypeptidic chain during the transcripts decoding, the data presented here lead us to suppose that a part of T3-induced insulin expression in INS-1E cells depends on the protein synthesis activation at the post-transcriptional level, as these proteins of the translational machinery were shown to be regulated by T3226637642FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP08/56446-

Influence of Cardiorespiratory Fitness on PPARG mRNA Expression Using Monozygotic Twin Case Control

The influence of cardiorespiratory fitness (VO2max) on anthropometric variables and PPARG mRNA expression was investigated. Monozygotic twin pairs aged 11-18 years were grouped into discordant (D) and concordant (C) high and low VO2max groups. VO2max was determined by progressive maximal exercise test on treadmill with gas exchange analysis. Body mass (BM), BMI, waist circumference (WC), triceps (TR), and subscapular (SB) skinfold thicknesses were measured. Twins from the discordant group had differences in VO2max values (D-high = 45.9 +/- 10.0 versus D-low = 32.4 +/- 10.6mL.kg(-1).min(-1), P = 0.025), while no differences were found in the concordant group (C-high = 42.4 +/- 9.2 versus C-low = 38.8 +/- 9.8mL.kg(-1).min(-1), P = 0.952). In discordant group, VO2max was negatively correlated with TR + SB (r = -0.540, P = 0.021) and positively correlated with PPARG expression in leukocytes (r = 0.952, P = 0.001). Moreover, PPARG expression was directly correlated with BM (e = -0.714, P = 0.047) and height (r = -0.762, P = 0.028). In concordant twins, VO2max was inversely correlated with BM (r = -0.290, P = 0.027), BMI (r = -0.472,P = 0.001), WC (r = -0.426, P = 0.001), and TR + SB (r = -0.739, P = 0.001). Twins D-high had 1.78-fold greater PPARG expression when compared with twins D-low (P = 0.048). In conclusion, the cardiorespiratory fitness may modulate PPARG expression in childhood and adolescence, independently of the genetic background.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Low-cost hybrid scaffolds based on polyurethane and gelatin

The production of scaffolds using synthetic and natural polymer combination has been widely studied for numerous tissue engineering applications, as it results in a material with superior properties, combining availability, processability, and the strength and resilience of synthetic polymers with the high biocompatibility of natural polymers. In the present study, fibrous membranes composed of polyurethane (PU) and gelatin (Gel) were fabricated by rotary jet spinning and were posteriorly characterized for their morphological, chemical composition, thermal stability, hydrophilic properties as well as cell viability. Viscosity measurements were taken to achieve the critical concentration of the polymeric solution (9% wt/v), and the production of fibers at different rotational speeds (3,000, 6,000, 9,000 and 12,000 rpm) was performed to evaluate the effect of rotational speed on fiber diameter and morphology, as observed in scanning electron microscopy analyses. Continuous and bead-free fibers were achieved at 6,000 rpm with an average diameter of 12.5 μm. Chemical composition characterization showed the characteristic functional groups of both polymers and the absence of the organic solvent, and thermogravimetric analysis showed thermal stability of the membrane up to 314 °C. Additionally, the water contact angle proved the membrane hydrophilic nature (81.3°). Cell viability assays exhibited cytocompatibility with endothelial cells for 24, 48 and 72 h. The results demonstrate that the PU–Gel combination with the rotary jet spinning process is promising to obtain low-cost scaffolds with interesting properties for numerous tissue engineering applications, and, thus, should be further studied9477777785FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP2017/13273-6We would like to express our thanks to the Information Technology Center (CTI Renato Archer -Electronic Packaging staff) and assistance with the use of open installations of bioprocesses at LNBR – Brazilian Biorenewables National Laboratory (CNPEM/MCTIC) and Lubrizol. This work was supported by grant #2017/13273-6, São Paulo Research Foundation (FAPESP