Background: Peripheral artery disease (PAD) is associated to coronary endothelial dysfunction and is an important predictor of future cardiovascular events. MicroRNAs play a critical role in modulating a variety of cellular processes by post-transcriptional regulation of their target genes. They are highly expressed in human endothelial cells (ECs), playing key role in essential biological processes. Thus, the aim of the present study was to evaluate the role, if any, of miRNAs in endothelial dysfunction in a model of PAD. Methods and Results: Male Wistar rats were provided of a standard atherogenic, high cholesterol diet and divided into 4 groups: no treatment (sham); rat subjected to vascular injury by balloon catheter (B.I.); rats underwent common femoral artery ligation (PAD); rats subjected to common femoral artery ligation and, after 21 days, subject to balloon injury (PAD+B.I.). RNA from intima was separately obtained from carotid arteries and evaluated for endothelial specific miRNAs using a stem-loop real-time-PCR. Interestingly, among the most important miRNAs expressed, real time RT-PCR showed an important up-regulation of miR-16 in ECs extracted by injured carotid artery of PAD+BI group respect the ECs of the others groups. A bioinformatics approach revealed highly conserved binding sites for miR-16 in the 3’UTR of SMAD7, an inhibitor of TGF-β1 signaling. The expression level of SMAD7 was measured in ECs treated with antagomiR-16 using immunoblotting analysis. Up-regulation of miR-16 revealed a reduction of SMAD7 levels. Also, we evaluated the effects of miR-16 inhibition in ECs on mRNA and protein levels of eNOS and NF-kB, regulators of endothelial dysfunction. Real-time PCR showed an increased expression of eNOS mRNA in ECs treated with antagomiR-16. Furthermore, immunoblotting revealed increased eNOS protein production as well as its activation in ECs by miR-16 inhibition. Finally, we showed an increased NF-kB activation in ECs after up-regulation of miR-16. Conclusions: Our data demonstrated that in rat with PAD and atherosclerotic diet, miR-16 is up-regulated in endothelial cell of carotid arteries after balloon injury. This findings might of importance to assess future strategies in patients with PAD.
A peptide fraction from the mitochondrial DNA of calf's liver was isolated using Drouin's method (1). This peptide fraction, which was extracted at pH 9.5 from an extensively purified mitochondrial DNA (2), has been shown to exert an in vitro regulatory role on the transcription and duplication activity of DNA (3). The same fraction also binds with mitochondrial DNA with a high affinity constant and stabilizes DNA from calf's thymus against thermal denaturation. The peptides from mitochondrial DNA have been subfractionated by fingerprinting-like techniques and one of them has been sequenced.
The analysis of cardiac damage biomarkers is crucial in the diagnosis and prognosis of acute myocardial infarction. The present work proposes a dynamical model for the analysis of the release in the blood of cardiac troponin T (cTnT). The model, tested against patients data, proves to be an effective tool to extrapolate information of interest to the clinician, like peaks and peak-times of the release curve. Furthermore, the model has been proven efficient to quantitatively differentiate patients with effective and non-effective thrombolysis response, which is a key information to guide revascularization.
Authors tested "in vitro" action of a low molecular weight peptide fraction extracted by mitochondrial DNA from calf liver. The fraction acts upon duplication performed by E. Coli DNA polymerase I on poli dAT dAT template inhibiting chain elongation. The authors hypothesize that a similar role is exerted by this fraction "in vitro" on mitochondrial DNA template.
Abstract Background Short bowel syndrome(SBS)is a severe clinical condition occuring when,following multiple intestinal resections,the remaining small bowel in continuity is ≤200cm.SBS is associated with intestinal failure(IF)when the intestinal absorptive function is compromised,requiring intravenous nutritional or electrolyte supplementation.Crohn’s disease(CD)is one of adults’ most frequent causes of SBS.The aim of this study was to identify clinical predictive factors of SBS in CD pts Methods We performed a prospective study, enrolling consecutive CD outpatients (pts).Statistical univariate comparison for primary outcome was assessed for continuous variables by the Mann-Whitney U test and the Kruskal-Wallis ANOVA test.Categorical variables were compared using the chi-square test.We also assessed a multivariate logistic regression model to identify independent predictors of SBS Results We enrolled 169 CD pts(47.3% male,mean age 43.5 years[±14.1]).45.5% of them were smokers;extraintestinal manifestations(EIMs)were present in 45.5% of pts, and 67.5% of pts had bowel resections(26.6% of them with more than one surgical intervention).Median disease duration and median time between diagnosis and the first resection were 9y(IQR 4-17)and 6.5y(IQR 1-13),respectively.At baseline,101 pts were on anti-TNF therapy,and considering the course of the disease,overall,20.1% of the pts were exposed to ≥3 biologics.10% of pts had SBS and 5.3% IF.In univariate analysis,comparing patients with and without SBS,the following variables were statistically different:disease duration(p=0.000),upper gastrointestinal disease location(p=0.001),ileocolonic location(p=0.008),stenosing behavior(p=0.040),perianal disease(p=0.000)and time elapsing from CD diagnosis to start biological therapy(0.002).At multivariate analysis,disease duration(p=0.000),perianal disease(p=0.05)and time elapsing from CD diagnosis to start biological therapy(p=0.016)resulted independently associated with the development of SBS Conclusion Our data identify the clinical features that would allow the gastroenterologists to predict the risk of developing SBS in a cohort of CD pts.Further studies would be needed to confirm our findings.Funded by:2.1 “Rafforzamento e potenziamento della ricerca biomedica del SSN”,finanziato dall’Unione europea–NextGenerationEU, CUP C53C22001140007. 2022 PNRR Project “Changing the future of intestinal failure in intestinal chronic inflammation:towards innovative predictive factors and therapeutic targets”code:PNRR-MAD-2022-12376791
The diagnosis of an acute myocardial infarction (AMI) is based, among other things, on the measurement of the level of cardiac biomarkers in the plasma. In order to optimize and speed up the analysis of cardiac Troponin T (cTnT), we have previously developed a mathematical model, which can be used to estimate the patient-specific cTnT release curve, based on a discrete number of acquisitions. Here, the effectiveness of our approach is improved by the extending the model to take into account also the release of another biomarker of cardiac damage, namely the creatine kinase MB (CK-MB). Moreover, the description of cTnT release dynamics has also been improved, taking into account a delay mechanism in the release from cytoplasm to plasma. Structural identifiability of the devised model is studied and a procedure for the identification of the parameters is set up and tested on a clinical dataset to assess the effectiveness of the proposed approach.
