28 research outputs found
Looking at the role of disordered E-tails of IGF-1
No full textIntrinsically disordered proteins (IDPs) encompass signalling and regulatory functions and altered expression of IDPs is associated with many diseases and imbalance in signalling pathways, transcriptional regulation, and splicing. Interest in insulin-like growth factor (IGF) – 1 isoforms on muscle homeostasis, regeneration, differentiation, and diseases has increased significantly. Inclusion or exclusion of exon 5 into the IGF-1 mRNA gives rise to three transcripts, IGF-1Ea, IGF-1Eb and IGF-1Ec, which yield three different C-terminal extensions called Ea, Eb and Ec peptides. Protein-coding sequences of exon 5 showed low rate of synonymous mutations and contain disorder-promoting amino acids, suggesting a regulatory role for these domains (Annibalini et al. 2016). To setup the analysis, the supernatants of HEK293 cells transfected with the specific IGF-1 isoform constructs as described in (De Santi et al. 2016) were studied by limited proteolysis combined with mass spectrometry (MS) using a Q-TOF microTM MS/MS (Micromass, Manchester, UK). Preliminary data showed that the C-terminal region of IGF-1Ea has lower resistance to trypsin digestion compared to the mature IGF-1 demonstrating IDRs in the Ea peptide. MS analyses to the detection of IGF-1E isoforms allowed us to identify both mature IGF-1 and IGF-1Ea isoform in transfected HEK293 cell culture supernatant. Analytical methods to correctly detect and quantify the IGF-1 isoforms are not currently available. In fact, the current existing methods rely on the use of antibodies that primarily recognize the mature IGF-1 peptide, thereby underestimate the isoforms. This finding could provide evidence allowing the detection and identification of the “E-tails” of IGF-1 and targeting these regulatory elements may represent a new strategy to control IGF-1 bioavailability in physio-pathological conditions.
1. Annibalini G, et al. MIR retroposon exonization promotes evolutionary variability and generates species-specific expression of IGF-1 splice variants BBAGRM 2016;1859:757-68.
2. De Santi M, et al. Cell Oncol (Dordr) 2016;39:149-59
The isoprenoid pathway in the ectomycorrhizal fungus Tuber borchii Vittad.: cloning and characterisation of the tbhmgr, tbfpps and tbsqs genes
No full textDownregulation of inflammatory genes expression in cultured human endothelial cells exposed to hydrogen peroxide
No full textTHE ISOPRENOID BIOSYNTHESIS DURING TUBER BORCHII FRUIT BODY MATURATION
No full textThe first insight into T. borchii isoprenoid pathway, also known as mevalonic acids pathway (MVA) during ascoma maturation were here discussed . Three genes coding for the most important enzymes of the isoprenoid biosynthesis were characterised, and the activity of the major regulatory enzyme of the MVA was determined in particular during the ascomata maturation. Furthermore several isoprenoids were identified among the volatile compounds produced by ripe T. borchii ascomat
Can non-invasive motor unit analysis reveal distinct neural strategies of force production in young with uncomplicated type 1 diabetes?
No full textPurposeto investigate the early consequences of type 1 diabetes (T1D) on the neural strategies of muscle force production.Methodsmotor unit (MU) activity was recorded from the vastus lateralis muscle with High-Density surface Electromyography during isometric knee extension at 20 and 40% of maximum voluntary contraction (MVC) in 8 T1D (4 males, 4 females, 30.5 +/- 3.6 years) and 8 matched control (4 males, 4 females, 27.3 +/- 5.9 years) participants. Muscle biopsies were also collected from vastus lateralis for fiber type analysis, including myosin heavy chain (MyHC) isoform content via protein and mRNA expression.ResultsMVC was comparable between groups as well as MU conduction velocity, action potentials' amplitude and proportions of MyHC protein isoforms. Nonetheless, MU discharge rate, relative derecruitment thresholds and mRNA expression of MyHC isoform I were lower in T1D.Conclusionsyoung people with uncomplicated T1D present a different neural control of muscle force production. Furthermore, differences are detectable non-invasively in absence of any functional manifestation (i.e., force production and fiber type distribution). These novel findings suggest that T1D has early consequences on the neuromuscular system and highlights the necessity of a better characterization of neural control in this population
The isoprenoid pathway in the ectomycorrhizal fungus Tuber borchii Vittad.: cloning and characterisation of the tbhmgr, tbfpps and tbsqs genes
No full textThe isoprenoid pathway of the ectomycorrhizal fungus Tuber borchii Vittad is investigated to better understand the molecular mechanisms at work, in particular during the maturation of the complex ascomata (the so-called "truffles"). Three T. borchii genes coding for the most important regulatory enzymes of the isoprenoid biosynthesis, 3-hydroxy-3-methylglutaryl-CoA reductase, farnesyl-diphosphate synthase (FPPS) and squalene synthase (SQS), were cloned and characterised. The analyses of their nucleotide and deduced amino acid sequences led us to identify the typical domains shown in homologous proteins. By using a quantitative real-time PCR the expression pattern of the three genes was analysed in the vegetative phase and during the complex ascoma maturation process, revealing an over-expression in the mature ascomata. The enzymatic activity of the T. borchii 3-hydroxy-3-methylglutaril-CoA reductase (HMGR) was investigated with a HPLC method, confirming that the significant isoprenoid biosynthesis in ripe ascomata proceeds not only via a transcriptional activation, but also via an enzyme activity control. These findings imply that isoprenoids play a fundamental role in Tuber ascomata, particularly in the last phases of their maturation, when they could be involved in antifungal or/and antimicrobial processes and contribute to the famous flavour of the truffle ascomata
Association between metabolic syndrome, insulin resistance, and IGF-1 in breast cancer survivors of DIANA-5 study
No full textPurposeCirculating insulin-like growth factor-1 (IGF-1) is positively associated with the risk of BC recurrence, and is more frequently dysregulated in older people, especially in those with metabolic syndrome (MetS) and obesity. This study aimed to analyze the association between IGF-1 levels and indices of MetS and insulin resistance in BC survivors.MethodsBaseline data of 563 BC survivors enrolled in the DIet and ANdrogen-5 (DIANA-5; NCT05019989) study were analyzed.ResultsLower circulating IGF-1 levels in subjects with MetS than in those without MetS were found. After stratification of the patients according to the diagnosis of MetS, we highlighted that the insulin was the main predictor of elevated IGF-1 levels only in subjects without MetS. Moreover, we found an interaction between high-density lipoprotein cholesterol (HDL-C), glycemia, and IGF-1 levels, showing a positive correlation between HDL-C and IGF-1, especially in subjects with higher values of glycemia and without a diagnosis of MetS.ConclusionsWhile IGF-1 levels appear to be much more impaired in subjects diagnosed with MetS, in non-MetS subjects, IGF-1 levels may respond better to metabolic parameters and lifestyle changes. Further studies are needed to analyze the role of physical activity and/or dietary intervention in modulating IGF-1 concentrations in BC survivors.Implications for cancer survivorsThese results could have important clinical implications for planning customized strategies aimed at modulating IGF-1 levels in BC survivors. In fact, while the IGF-1 system seems to be much more compromised in subjects with a diagnosis of MetS, in noMetS subjects, IGF-1 levels could better respond to lifestyle changes
Effect of hyaluronic acid local injections on Achilles tendinitis: an observational study on tendon viscoelastic properties in recreational runners
No full textPurpose: Achilles tendinitis (AT) causes 5% of professional athletes
to end their careers; in Europe AT affects ca. 10 millions recreational
runners. The aim of this study was to evaluate the clinical and
functional efficacy of a three-local injections regimen of hyaluronic
acid (HA) in ameliorating pain score and function in middle aged
recreational runners with AT, whose functional and clinical param-
eters were recorded integrating multiple techniques.
Methods: 8 recreational runners previously diagnosed for monolat-
eral AT were enrolled. AT was confirmed before the first local HA
injection (T0) by clinical examination, MRI and thermography. At T0
patients were assessed for maximal voluntary isometric contraction
(MVI) involving Achilles tendon (both injured and healthy), and pain
level with a Likert scale; Achilles tendon viscoelastic state, i.e. tone
and stiffness, were then measured at relaxed state and at 10% of MVI
with MyotonPro (Myoton Ltd, UK). Finally patients received the first
HA injection (RegenFlex T&M, a blend of 2–1000 kDa HA,
Regenyal, IT). All the measurements were repeated at T1 (15 days
after the first injections and immediately prior the second), at T2
(15 days after the second injection and prior the third) and at T3
(15 days after the third injection), i.e. over a total of 45 days in which
clinical visits were also performed. Furthermore, before each injec-
tion, injured tendon exudates were collected by needle aspiration.
Results: At T0, tone and stiffness values were significantly different
between injured and contralateral tendons, especially when measured
at the relaxed state. Pain score and MVI, coherently with clinical
outcomes, were significantly higher and lower in injured tendons,
respectively. Interestingly, the above differences gradually disap-
peared at T1, 2 and 3. In keeping with these results, tendon exudates
volumes also decreased
Differential effect of creatine on oxidatively-injured mitochondrial and nuclear DNA
No full textCreatine is a naturally occurring compound obtained in humans from endogenous production and consumption through the diet. It is used as an ergogenic aid to improve exercise performance and increase fat-free mass. Lately, creatine’s positive therapeutic benefits in various oxidative stress-associated diseases have been reported in literature and, more recently, creatine has also been shown to exert direct antioxidant effects. Oxidatively-challenged DNA was analysed to show possible protective effects of creatine. Acellular and cellular studies were carried out. Acellular assays, performed using molecular approaches, showed that creatine protects circular and linear DNA from oxidative attacks. Nuclear and mitochondrial DNAs from oxidatively-injured human umbilical vein endothelial cells were analyzed. Creatine supplementation showed significant genoprotective activity on mitochondrial DNA. This evidence suggests that creatine may play an important role in mitochondrial genome stability in that it could normalize mitochondrial mutagenesis and its functional consequences. Thus, creatine supplementation could be used to prevent or ameliorate diseases related to mitochondrial DNA mutations, and possibly to delay aging
Effects of acute aerobic, resistance and combined exercises on 24-h glucose variability and skeletal muscle signalling responses in type 1 diabetics
No full textPurpose: To compare the effect of high-intensity aerobic (AER), resistance (RES), and combined (COMB: RES + AER) exercise, on interstitial glucose (IG) variability and skeletal muscle signalling pathways in type 1 diabetes (T1D). Methods: T1D participants (6 M/6F) wore a flash glucose monitoring system in four randomized sessions: one control (CONT), and one AER, RES and COMB (40 min each). Mean amplitude of glycemic excursions (MAGE), standard deviation (SD) and coefficient variation (CV) of IG were used to compare the 24 h post-exercise IG variability. Blood and muscle samples were collected to compare exercise-induced systemic and muscle signalling responses related to metabolic, growth and inflammatory adaptations. Results: Both RES and COMB decreased the 24 h MAGE compared to CONT; additionally, COMB decreased the 24 h SD and CV. In the 6–12 h post-exercise, all exercise modalities reduced the IG CV while SD decreased only after COMB. Both AER and COMB stimulated the PGC-1α mRNA expression and promoted the splicing of IGF-1Ea variant, while Akt and p38MAPK phosphorylation increased only after RES and COMB. Additionally, COMB enhanced eEF2 activation and RES increased myogenin and MRF4 mRNA expression. Blood lactate and glycerol levels and muscle IL-6, TNF-α, and MCP-1 mRNAs increased after all exercise sessions, while serum CK and LDH level did not change. Conclusion: COMB is more effective in reducing IG fluctuations compared to single-mode AER or RES exercise. Moreover, COMB simultaneously activates muscle signalling pathways involved in substrate metabolism and anabolic adaptations, which can help to improve glycaemic control and maintain muscle health in T1D
