MiRNA 126 as a New Predictor Biomarker in Venous Thromboembolism of Persistent Residual Vein Obstruction: A Review of the Literature Plus a Pilot Study.

Venous thromboembolism (VTE) is the third most common cardiovascular disease. Interleukins (ILs) and micro-ribonucleic acids (miRNAs) have been proposed as molecules able to modulate endothelial inflammation and platelet hyperactivity. At present, no early biomarkers are available to predict the outcome of VTE. We investigated in a pilot study a selected number of miRNAs and ILs as prognostic VTE biomarkers and reviewed literature in this setting. Twenty-three patients (aged 18-65) with a new diagnosis of non-oncological VTE and free from chronic inflammatory diseases were enrolled. Twenty-three age- and sex-matched healthy blood donors were evaluated as control subjects. Serum miRNAs (MiRNA 126, 155, 17.92, 195), inflammatory cytokines (IL-6, tumor necrosis factor-α, IL-8), and lymphocyte subsets were evaluated in patients at enrolment (T0) and in controls. In VTE patients, clinical and instrumental follow-up were performed assessing residual vein obstruction, miRNA and ILs evaluation at 3 months' follow-up (T1). At T0, IL-8, activated T lymphocytes, Treg lymphocytes, and monocytes were higher in patients compared with healthy controls, as were miRNA 126 levels. Moreover, miRNA 126 and IL-6 were significantly increased at T0 compared with T1 evaluation in VTE patients. Higher levels of MiR126 at T0 correlated with a significant overall thrombotic residual at follow-up. In recent years an increasing number of studies (case-control studies, in vivo studies in animal models, in vitro studies) have suggested the potential role of miRNAs in modulating the cellular and biohumoral responses involved in VTE. In the frame of epidemiological evidence, this pilot study with a novel observational approach supports the notion that miRNA can be diagnostic biomarkers of VTE and first identifies miRNA 126 as a predictor of outcome, being associated with poor early recanalization.

Rectus femoris myotendinous lesion treated with PRP: a case report.

BACKGROUND AND AIM OF WORK: Musculoskeletal injuries are the most common cause of severe, chronic pain and physical disability for the majority of all sport-related injuries. Platelet-rich plasma is being used more frequently to promote healing of muscle injuries. We report a case of 39 years old non professional soccer player who came to our attention for a quadriceps muscle pain onset after kicking the ball during a match. METHODS: Clinical and instrumental evaluation revealed a myotendinous junction rupture of the rectus femoris with retraction of 1.5 cm from the anterior inferior iliac spine. We decided to treat the patient with PRP ultrasound guided injections and a specific rehabilitation protocol. RESULTS: Clinical evaluation 45 days following the end of the treatment showed the resolution of the pain and the full recovery of strength and range of motion. Muscle healing was documented by magnetic resonance imaging. CONCLUSIONS: Even if the role of PRP in muscle injury is not still clear, the result observed confirms that it could be used in the treatment of muscle lesions.

Human macrophage differentiation induces OCTN2-mediated L-carnitine transport through stimulation of mTOR-STAT3 axis.

l-Carnitine, in addition to playing a fundamental role in the ß-oxidation of fatty acids, has been recently identified as a modulator of immune function, although the mechanisms that underlie this role remain to be clarified. In this study, we addressed the modulation of l-carnitine transport and expression of related transporters during differentiation of human monocytes to macrophages. Whereas monocytes display a modest uptake of l-carnitine, GM-CSF-induced differentiation massively increased the saturable Na+-dependent uptake of l-carnitine. Kinetic and inhibition analyses demonstrate that in macrophage l-carnitine transport is mediated by a high-affinity component (Km ∼4 µM) that is identifiable with the operation of OCTN2 transporter and a low-affinity component (Km > 10 mM) that is identifiable with system A for neutral amino acids. Consistently, both SLC22A5/OCTN2 and SLC38A2/SNAT2 are induced during the differentiation of monocytes to macrophages at gene and protein levels. Elucidation of GM-CSF signaling demonstrates that the cytokine causes the activation of mTOR kinase, leading to the phosphorylation and activation of STAT3, which, in turn, is responsible for OCTN2 transcription. SLC22A5/OCTN2 therefore emerges as a novel member of the set of genes markers of macrophage differentiation.

Adipose-derived Stem Cells Added to Platelet-rich Plasma for Chronic Skin Ulcer Therapy.

INTRODUCTION: Adipose-derived stem cells (ASCs) hold great promise for regenerative medicine applications due to their ability to promote the healing process through in situ differentiation and secretion of paracrine factor. The aim of this paper is to present a clinical adjunct for chronic skin wound therapy based on ASCs added to platelet-rich plasma (PRP), to obtain an enhanced PRP (e-PRP). MATERIALS AND METHODS: For 18 months, 24 control-group patients with 31 chronic skin ulcers were treated with standard wound care, while 16 experimental-group patients with 21 chronic skin ulcers were treated with standard wound care and 1 e-PRP injection. The patients were randomly assigned to the control or experimental group. Outpatients had weekly follow-up visits where they were subjected to standard treatment and the wound healing process was assessed. RESULTS: At the end of the study, the control and experimental groups had similar healing rates but wound closure rates were significantly different (P = 0.0257): 0.0890 cm(2) x day and 0.2287 cm(2) x day respectively, resulting in a faster recovery for the group treated with e-PRP. No side effects were reported. CONCLUSION: In the authors' experience, e-PRP significantly enhanced wound closure rates when compared to standard wound care, without causing any serious complications. This finding highlights e-PRP as a valuable resource for chronic wound treatment.

Use of platelet-rich plasma in the care of sports injuries: our experience with ultrasound-guided injection.

BACKGROUND: Platelet-rich plasma is being used more frequently to promote healing of muscle injuries. The growth factors contained in platelet-rich plasma accelerate physiological healing processes and the use of these factors is simple and minimally invasive. The aim of this study was to demonstrate the efficacy of ultrasound-guided injection of platelet-rich plasma in muscle strains and the absence of side effects. MATERIALS AND METHODS: Fifty-three recreational athletes were enrolled in the study. The patients were recruited from the Emergency Room in the University Hospital at Parma according to a pre-defined protocol. Every patient was assessed by ultrasound imaging to evaluate the extent and degree of muscle injuries. Only grade II lesions were treated with three ultrasound-guided injections of autologous platelet-rich plasma every 7 days. Platelet concentrate was produced according to standard methods, with a 10% variability in platelet count. The platelet gel for clinical use was obtained by adding thrombin to the concentrates under standardised conditions. Outcomes assessed were: pain reduction, muscle function recovery and return to sports activity, ultrasound-imaging tissue healing, relapses, local infections, and any side effect during the treatment. RESULTS: In all cases muscle lesions healed fully on ultrasound-imaging, the pain disappeared, and muscle function recovery was documented with a return to sports activity. A single patient had a relapse 1 year after treatment. DISCUSSION: Platelet-rich plasma injected into the injury site is one of the most important factors rendering the treatment effective. To maximise its efficacy the preliminary ultrasound must be done accurately to localise the lesion and guide the needle into the corresponding lesion. According to the current results, which document full muscle recovery and no relapse except for one case, platelet-rich plasma ultrasound-guided injection represents a valid mini-invasive treatment for muscle injuries.

Platelet gel in the treatment of cutaneous ulcers: the experience of the Immunohaematology and Transfusion Centre of Parma.

BACKGROUND: Platelet gel is being ever more frequently used to promote healing of cutaneous ulcers. However, the factors that determine the often variable clinical outcome of this procedure are still incompletely understood. AIMS: The aims of this study were to demonstrate that platelet gel, even when obtained under strictly controlled conditions, produces highly variable outcomes in patients with cutaneous ulcers and to propose a method for in vitro standardisation of the biological properties of platelet gel. MATERIAL AND METHODS.: Patients were enrolled on the basis of a pre-defined protocol. Platelet concentrate was produced with standard methods, with a variability in platelet count among the different samples of less than 10%. The platelet gel for clinical use was obtained, under strictly standardized conditions, by adding thrombin and calcium gluconate to the concentrates. For in vitro studies, platelet gel, obtained from platelet-rich plasma from four donors, was frozen and thawed twice so as to increase gel contraction. The supernatant was used to modify cell proliferation, protein synthesis, and the expression of selected genes in cultures of human diploid fibroblasts. RESULTS: Seventeen patients (aged 44-78 years) with ulcers (4 diabetic, 11 vascular, 1 post-traumatic, 1 decubitus) were treated with platelet gel (4 autologous, 13 homologous). Complete re-epithelialisation of four ulcers (1 diabetic, 1 post-traumatic, 2 vascular) was obtained after applications of platelet gel (2 autologous, 2 homologous); in 11 other cases there was a greater than 50% reduction in the size of the ulcer. Two patients had no benefit. The supernatant of the platelet gel was able to promote dose-dependent proliferation and changes in gene expression as well as in metabolic activities related to protein synthesis. CONCLUSIONS: Although the use of platelet gel in the treatment of cutaneous ulcers is increasing, and conditions for its production are better standardised, very considerable variability of clinical outcomes is still observed, even within single centres, suggesting that there are differences in biological properties of platelet concentrates from individual patients which cannot be readily controlled with current techniques. The biological effects of the platelet gel supernatant described in this article may provide the basis for a simple biological validation of platelet preparations before their clinical use, so as to reduce this potentially important source of variability.

Evaluation of cardiovascular risk in blood donors: results of the CARDIORISK study in the Parma Transfusion Service.

BACKGROUND: Cardiovascular disease, which is one of the main causes of mortality in industrialised countries, is ever increasingly the focus of prevention. In this study, called "Cardiorisk", we evaluated cardiovascular risk in the population of blood donors at the Service of Immunohaematology and Transfusion Medicine in Parma. PATIENTS AND METHODS: Between January 2007 and December 2008, 6,172 consecutive blood donors (aged 35-65 years) were enrolled in this project which entailed calculating each subject's cardiovascular risk score, based on an evaluation of both unalterable risk factors (age and gender) and modifiable risk factors (total cholesterol, HDL, LDL, triglycerides, glycaemia, smoking, hypertension) as well as anti-hypertensive and/or cholesterol-lowering therapy. RESULTS: Of the 6,172 donors enrolled in the study, 5,039 (81.7%) had a low cardiovascular risk (score from 0-10), 774 (12.5%) had a moderate cardiovascular risk (score from 11-19) and 359 (5.8%) donors had a high cardiovascular risk (score from 20-28). CONCLUSIONS: In our opinion, the calculation of cardiovascular risk is an important instrument for preventive medicine in blood donors.

Treatment of chronic venous leg ulcers by platelet gel.

Chronic venous leg ulcers (CVLU) are chronic wounds, associated with long-standing venous hypertension, which have a poor prognosis for healing. In the process of wound healing the first step is represented by platelet aggregation and subsequent release of growth factors and other mediators, which play a key role in the repair response. Platelet gel (PG), a hemocomponent obtained by mixing platelets, thrombin, and calcium, is able, when applied topically, to release platelet mediators that likely favor CVLU healing. However, unstandardized protocols have been described in studies utilizing PG for the regeneration of a number of tissues, including CVLU; the relative clinical outcomes were hence highly variable. In our experience the topical use of PG, together with the strict adherence to the principles of good wound care, quickly promoted increased granulation tissue, followed by a complete CVLU epithelization. Although further studies and trials are needed to establish the major outcome affecting rules for optimal indications, preparation, and use of PG for CVLU treatment, PG can be undoubtedly considered a useful tool, able to improve the management of CVLU.