ANALISI DELL\u2019ESPRESSIONE GENICA DI RUNX-2 E DELLADENSITA\u2019 MINERALE OSSEA IN UNA POPOLAZIONEDI GIOVANI E ADULTI SANI

La densit\ue0 minerale ossea (BMD) e il picco di massa ossea (PBM) sono importanti per la resistenza dello scheletro. Lo sviluppo della densitometria ossea ha permesso di studiare in modo preciso la BMD e le sue variazioni e l\u2019influenza che le componenti genetiche ed ambientali hanno sull\u2019osso. I fattori ereditari influenzano in modo significativo la BMD e Runx-2 \ue8 noto essere un regolatore fondamentale dell\u2019attivit\ue0 osteoblastica e della formazione ossea. Lo scopo del nostro studio \ue8 stato di valutare il comportamento di Runx-2 durante lo sviluppo dello scheletro in una popolazione sana di giovani ed adulti. Abbiamo analizzato mediante densitometria ossea (DXA) la BMD della colonna e del femore di 153 volontari, 98 donne e 55 uomini. In un sottogruppo di questi \ue8 stato prelevato un campione di sangue periferico, al fine di studiare il profilo di espressione di Runx-2, sia a livello delle cellule mesenchimali circolanti (MSCs; 28 soggetti), sia nel totale delle cellule mononucleate del sangue periferico (PBMC; 140 soggetti). Nel nostro lavoro la BMD \ue8 risultata simile nei 2 sessi subito dopo la pubert\ue0, mentre \ue8 diventata progressivamente maggiore negli uomini rispetto alle donne durante la terza e quarta decade di vita. Il PBM \ue8 stato raggiunto nella terza decade nelle donne e nella quarta dagli uomini. Il dato pi\uf9 interessante riguarda il gene Runx-2, la cui espressione \ue8 risultata altamente correlata con la BMD in entrambi i sessi. Inoltre il profilo di espressione di Runx-2 nelle MSCs e nelle PBMC era pressoch\ue9 sovrapponibile. In conclusione, il PBM viene raggiunto prima nelle donne rispetto agli uomini, seppure in questi ultimi la BMD risulti alla fine maggiore, e il PBM e la BMD sono parametri strettamente associati al profilo di espressione di Runx-2. Inoltre le PBMC potrebbero essere prese in considerazione in futuro per lo studio di espressione genica nelle malattie dello scheletro.Bone mineral density (BMD) and peak bone mass (PBM) are important determinants of skeletal resistance. The development of bone densitometry improved the possibility of studying BMD and the influence of genetic and environmental factors on bone. Heredity factors are important for BMD and Runx-2 is accepted as a regulator of osteoblasts and bone formation. The aim of our study was to evaluate the behaviour of Runx-2 during skeletal maturity in the healthy young-adult population. We analyzed spine and hip BMD in 153 volunteers, 98 women and 55 men using dual energy x-ray absorptiometry (DXA). In a subgroup of them, a sample of peripheral blood was taken in order to perform gene expression analysis of Runx-2 both in peripheral mesenchimal stem cells (MSCs; 28 subjects) and peripheral mononuclear cells (PBMC; 140 subjects). In our work BMD was comparable in both genders after puberty, while it became higher in men than women during the third and fourth decade. PBM was achieved in the third decade in women and in the fourth in men. More interestingly Runx-2 gene expression highly correlated with BMD in both genders. MSCs and PBMC showed the same gene expression profile of Runx-2. In conclusion PBM is reached earlier in females, BMD becomes higher in men later and BMD and PBM are strictly associated with Runx-2. In addition, PBMC should be considered an important source for gene expression analysis in bone diseases

Ultrasound guided chest compressions during cardiopulmonary resuscitation

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Safety and tolerability of zoledronic acid and other bisphosphonates in osteoporosis management

Bisphosphonates (BPs) are widely used in the treatment of postmenopausal osteoporosis and other metabolic bone diseases. They bind strongly to bone matrix and reduce bone loss through inhibition of osteoclast activity. They are classified as nitrogen- and non-nitrogen-containing bisphosphonates (NBPs and NNBPs, respectively). The former inhibit farnesyl diphosphate synthase while the latter induce the production of toxic analogs of adenosine triphosphate. These mechanisms of action are associated with different antifracture efficacy, and NBPs show the most powerful action. Moreover, recent evidence indicates that NBPs can also stimulate osteoblast activity and differentiation. Several randomized control trials have demonstrated that NBPs significantly improve bone mineral density, suppress bone turnover, and reduce the incidence of both vertebral and nonvertebral fragility fractures. Although they are generally considered safe, some side effects are reported (esophagitis, acute phase reaction, hypocalcemia, uveitis), and compliance with therapy is often inadequate. In particular, gastrointestinal discomfort is frequent with the older daily oral administrations and is responsible for a high proportion of discontinuation. The most recent weekly and monthly formulations, and in particular the yearly infusion of zoledronate, significantly improve persistence with treatment, and optimize clinical, densitometric, and antifracture outcomes

Role of Ox-PAPCs in the Differentiation of Mesenchymal Stem Cells (MSCs) and Runx2 and PPARγ2 Expression in MSCs-Like of Osteoporotic Patients

BACKGROUND: Mesenchymal stem cells (MSCs) can differentiate into osteoblasts and adipocytes and conditions causing bone loss may induce a switch from the osteoblast to adipocyte lineage. In addition, the expression of Runx2 and the PPARγ2 transcription factor genes is essential for cellular commitment to an osteogenic and adipogenic differentiation, respectively. Modified lipoproteins derived from the oxidation of arachidonate-containing phospholipids (ox-PAPCs: POVPC, PGPC and PEIPC) are considered important factors in atherogenesis. METHODOLOGY: We investigated the effect of ox-PAPCs on osteogenesis and adipogenesis in human mesenchymal stem cells (hMSCs). In particular, we analyzed the transcription factor Runx2 and the PPARγ2 gene expression during osteogenic and adipogenic differentiation in absence and in presence of ox-PAPCs. We also analyzed gene expression level in a panel of osteoblastic and adipogenic differentiation markers. In addition, as circulating blood cells can be used as a "sentinel" that responds to changes in the macro- or micro-environment, we analyzed the Runx2 and the PPARγ2 gene expression in MSCs-like and ox-PAPC levels in serum of osteoporotic patients (OPs). Finally, we examined the effects of sera obtained from OPs in hMSCs comparing the results with age-matched normal donors (NDs). PRINCIPAL FINDINGS: Quantitative RT-PCR demonstrated that ox-PAPCs enhanced PPARγ2 and adipogenic gene expression and reduced Runx2 and osteoblast differentiation marker gene expression in differentiating hMSCs. In OPs, ox-PAPC levels and PPARγ2 expression were higher than in NDs, whereas Runx2 was lower than in ND circulant MSCs-like. CONCLUSIONS: Ox-PAPCs affect the osteogenic differentiation by promoting adipogenic differentiation and this effect may appear involved in bone loss in OPs

Safety and tolerability of zoledronic acid and other bisphosphonates in osteoporosis management

Luca Dalle Carbonare, Mirko Zanatta, Adriano Gasparetto, Maria Teresa ValentiClinic of Internal Medicine D, Department of Medicine, University of Verona, ItalyAbstract: Bisphosphonates (BPs) are widely used in the treatment of postmenopausal ­osteoporosis and other metabolic bone diseases. They bind strongly to bone matrix and reduce bone loss through inhibition of osteoclast activity. They are classified as nitrogen- and non-nitrogen-containing bisphosphonates (NBPs and NNBPs, respectively). The former inhibit farnesyl diphosphate synthase while the latter induce the production of toxic analogs of adenosine triphosphate. These mechanisms of action are associated with different antifracture efficacy, and NBPs show the most powerful action. Moreover, recent evidence indicates that NBPs can also stimulate osteoblast activity and differentiation. Several randomized control trials have demonstrated that NBPs significantly improve bone mineral density, suppress bone turnover, and reduce the incidence of both vertebral and nonvertebral fragility fractures. Although they are generally considered safe, some side effects are reported (esophagitis, acute phase reaction, hypocalcemia, uveitis), and compliance with therapy is often inadequate. In particular, gastrointestinal discomfort is frequent with the older daily oral administrations and is responsible for a high proportion of discontinuation. The most recent weekly and monthly formulations, and in particular the yearly infusion of zoledronate, significantly improve persistence with treatment, and optimize clinical, densitometric, and antifracture outcomes.Keywords: bisphosphonates, osteoporosis, safety, tolerability, zoledronic aci

Point-of-care critical ultrasound in a rural emergency department

Point-of-care critical ultrasound (CCUS) has changed the management of critically ill patients in the emergency department. It is brought to the bed of patient, images are immediately available and therapy can be monitored making real time changes. Although it is difficult to estimate the real efficacy of CCUS, we evaluated the impact of ultrasound in our emergency department. This study is a cross sectional observational study with 241 cases enrolled. All patients were evaluated by the emergency physician and underwent clinical examination and then CCUS. Patients were then independently evaluated by at least one consultant. A final diagnosis was made after an agreement between the emergency physician and the consultant. Percentages of correct final diagnosis were higher after CCUS than after primary survey: 82.5% <em>vs</em> 49.1% of patients with dyspnea (P&lt;0.001), 71.9% <em>vs</em> 40.6 % with thoracic pain (P=0.03), 76.2% <em>vs</em> 45% with abdominal pain (P&lt;0.001), 80.0% <em>vs</em> 43.6% with suspected deep venous thrombosis (P=0.03) and 80.0% <em>vs</em> 20% with shock (P=0.014). Extended fast assessment for trauma was effective for the management of traumatic patients and correctly ruled out complications in 81.1% of patients (P=0.04). A small number of ultrasound guided invasive procedures were safely and successfully performed. In our study the integration of primary survey with CCUS increased diagnostic capability of the emergency physician and improved overall quality of medical assistance

Pre-hospital lung ultrasound for cardiac heart failure and COPD: is it worthwhile?

Abstract Background Pre-hospital ultrasound is a new challenge and lung ultrasound could be an interesting opportunity in the pre-hospital medical service. The aim of our study was to evaluate the efficacy of lung ultrasound in out-of-hospital non-traumatic respiratory insufficiency. Methods We planned a case-controlled study in the ULSS 5 ovest vicentino area (Vicenza—Italy) enrolling subjects with severe dyspnea caused by cardiac heart failure or acute exacerbation of chronic obstructive pulmonary disease. We compared drugs administration, oxygen delivery, and laboratory tests between those patients with ultrasound integrated management and those without ultrasound. Results Pre-hospital lung ultrasound had a high specificity (94.4%) and sensitivity (100%) for the correct identification of alveolar interstitial syndrome using B lines, whereas the percentages obtained with pleural effusion were lower (83.3, 53.3%, respectively). The patients with ultrasound integrated management received a more appropriate pharmacological therapy (p 0.01), as well as non-invasive ventilation (CPAP) was used more frequently in those with an acute exacerbation of chronic obstructive pulmonary disease (p 0.011). Laboratory tests and blood gases analysis were not significant different between the two study groups. In a sub-analysis of the patients with an A profile, we observed a significant lower concentration of PCO2 in those with an ultrasound integrated management (PCO2: 42.62 vs 52.23 p 0.049). According with physicians’ opinion, pre-hospital lung ultrasound gave important information or changed the therapy in the 42.3% of cases, whereas it just confirmed physical examination in the 67.7% of cases. Conclusions Pre-hospital lung ultrasound is easy and feasible, and learning curve is rapid. Our study suggests that cardiac heart failure and acute exacerbation of chronic obstructive pulmonary disease can be considered two indications for pre-hospital ultrasound, and can improve the management of patient with acute respiratory insufficiency

Runx-2 gene expression is associated with age-related changes of bone mineral density in the healthy young-adult population

Bone mineral density (BMD) and peak bone mass (PBM) are important determinants of skeletal resistance. The development of bone densitometry improved the possibility of studying BMD and the influence of genetic and environmental factors on bone. Heredity factors are important for BMD, and Runx-2 is accepted as a regulator of osteoblasts and bone formation. The aim of our study was to evaluate the behavior of Runx-2 during skeletal maturity in the healthy young-adult population. We analyzed spine and hip BMD in 153 volunteers, 98 women and 55 men, using dual-energy X-ray absorptiometry. In a subgroup of these volunteers, a sample of peripheral blood was taken to perform gene expression analysis of Runx-2 both in peripheral mesenchymal stem cells (MSCs; 28 subjects) and in peripheral mononuclear cells (PBMCs; 140 subjects). In our work BMD was comparable in both genders after puberty, then became higher in men than women during the third and fourth decades. PBM was achieved in the third decade in women and in the fourth in men. More interestingly, Runx-2 gene expression highly correlated with BMD in both genders. MSCs and PBMCs showed the same gene expression profile of Runx-2. In conclusion, PBM is reached earlier in females, BMD becomes higher in males later in life, and BMD and PBM are strictly associated with Runx-2. In addition, PBMC should be considered an important source for gene expression analysis in bone diseases