Novel epigenetic target therapy for prostate cancer: a preclinical study.

Epigenetic events are critical contributors to the pathogenesis of cancer, and targeting epigenetic mechanisms represents a novel strategy in anticancer therapy. Classic demethylating agents, such as 5-Aza-29-deoxycytidine (Decitabine), hold the potential for reprograming somatic cancer cells demonstrating high therapeutic efficacy in haematological malignancies. On the other hand, epigenetic treatment of solid tumours often gives rise to undesired cytotoxic side effects. Appropriate delivery systems able to enrich Decitabine at the site of action and improve its bioavailability would reduce the incidence of toxicity on healthy tissues. In this work we provide preclinical evidences of a safe, versatile and efficient targeted epigenetic therapy to treat hormone sensitive (LNCap) and hormone refractory (DU145) prostate cancers. A novel Decitabine formulation, based on the use of engineered erythrocyte (Erythro-Magneto-Hemagglutinin Virosomes, EMHVs) drug delivery system (DDS) carrying this drug, has been refined. Inside the EMHVs, the drug was shielded from the environment and phosphorylated in its active form. The novel magnetic EMHV DDS, endowed with fusogenic protein, improved the stability of the carried drug and exhibited a high efficiency in confining its delivery at the site of action in vivo by applying an external static magnetic field. Here we show that Decitabine loaded into EMHVs induces a significant tumour mass reduction in prostate cancer xenograft models at a concentration, which is seven hundred times lower than the therapeutic dose, suggesting an improved pharmacokinetics/pharmacodynamics of drug. These results are relevant for and discussed in light of developing personalised autologous therapies and innovative clinical approach for the treatment of solid tumours

Potential role of low kV ex vivo micro-CT for 3D morphometry of paraffin embedded coronary vessels before histology

Purpose: Micro CT is an established tool for non destructive 3D inspection of small specimens. Aim of the study was to demonstrate that despite its limitations in differentiation of soft materials, micro CT can reliably display coronary vessel structure and surrounding tissues after paraffin embedding. Data were obtained from coronary specimens of pig, physical phantoms and numerical simulations. Preliminary images with dual-energy techniques are also shown. Methods: A micro CT scanner built by our group was used for the experiments. The x-ray tube was set up in the range of 20-50 kV; the voxel size was set to 21 μm. A phantom composed of formalin fixed fat and myocardium of rat, dehydrated and paraffin embedded, was used to measure the contrast of different tissues with respect to background. Similar acquisitions were simulated numerically. Real samples of pig excised coronary arteries were processed in the same way and acquired with the same settings; resulting images were compared to those obtained by histology. Results: In phantom, the myocardium contrast vs. paraffin varied from 40% at 20 kV to 29% at 50 kV. The fat contrast vs. background was 2% at 20 kV, whereas it was indistinguishable from the background at 50 kV; all the contrasts in phantom appeared lower than those expected from simulations, probably because of tissue shrinkage. In the samples from pig (see Figure), the vessel wall contrast was 25% greater than the myocardium contrast; the pericardium and a balloon induced stenosis were clearly distinguished. All micro CT scans were shorter than 1 hour. Conclusion: Micro CT is a useful complementary tool for the 3D morphometry of coronary vessels after paraffin embedding, and it can help for the preliminary identification of features of interest for subsequent histological analysis

Phenotype Changes of Circulating Monocytes in a Hypercholesterolemic Swine Model of Coronary Artery Disease

Objective: Inflammation and immunity activation play a key role in atherosclerosis (ATS) onset and progression. Aim of this study was to investigate the relationships between phenotype of circulating monocytes and coronary artery disease (CAD) development in a histologically well-characterized swine model of ATS. Methods: Blood samples were obtained from 6 animals at baseline and after 16 weeks high fat cholesterolenriched diet. Flow cytometry monocyte identification was performed (CD172a marker). Adhesion (CD18a, CD11a, CD11R3, CD49d, CD29), differentiation (CD14) and activation receptors (SLA-DR, CD16, CD163) were quantified as percentage of positivity (%) and Relative Fluorescence Intensity (RFI). Lipid parameters (LDL, oxLDL, HDL) and soluble endothelial ICAM-1 were measured and histologic quantitative assessment of coronary ATS was performed. Results: Flow cytometry analysis demonstrated a significant post-diet decrease of CD14 RFI and an increment of % SLA-DR. Pre-diet values of ICAM-1 and % SLA-DR correlated reciprocally (P=0.0191) and with several CAD severity indexes (P≤0.02). Positive correlations between RFI changes of CD29 (P=0.0213) and CD18a (P=0.0341) and morphometric indexes of coronary ATS were found. Post-diet RFI values of CD29, CD18a and CD16 were also closely related to morphometric parameters (P<0.03). A cumulative post-diet tendency to increase of CD14low/ CD163high monocyte fraction (45.07 ± 2.27 vs. 40.14 ± 3.16) and a tight correlation between changes of this monocyte subset and corresponding HDL variations (P=0.0100) were also observed. Conclusions: Blood monocyte orientation towards a macrophage-like phenotype, similar to a HDL-induced maturation, and a close association between markers changes and severity of diet induced coronary ATS could provide new insights into plaque growth and progression in CAD

Ex-vivo micro-CT for the assessment of the structure of paraffin embedded coronary vessels before histology

Purpose: Aim of the study was to demonstrate that absorption-based single low-energy micro-CT can reliably display coronary vessel structure and surrounding tissues after paraffin embedding. Methods and Materials: A micro-CT scanner built by our group was used for the experiments. The x-ray tube was set up in the range of 20-50 kV. A phantom composed of formalin fixed fat and myocardial tissues of rat, dehydrated and paraffin embedded, was used to measure the image contrast of relevant tissues with respect to the background. Similar acquisitions were simulated using standardized attenuation functions and simulated spectra. Real samples of pig excised coronary arteries were also processed in the same way and acquired with similar settings; resulting images were compared to those obtained by histology. The voxel size in the micro-CT images was 21 &#956;m. Results: In phantom, the myocardium contrast vs. paraffin varied from 40% at 20 kV to 29% at 50 kV. The fat contrast was 2% at 20 kV, whereas it was indistinguishable from the background at 50 kV. The pericardium and a balloon induced stenosis were clearly distinguished. A noncalcified fatty streak was also identified. All contrasts in phantom appeared lower than those expected from simulations, probably because of the sample processing. The image quality was sufficient for segmentation purposes, allowing quantitative morphometry within the selected Volume of Interest (VOI). Conclusion: Micro-CT is a useful complementary tool for the assessment of coronary vessels structure after sample embedding in paraffin prior to histological examination. This type of imaging can help for the identification of features for subsequent histological analysis in experimental models of restenosis and atherogenesis

Myocardial interleukin-6 in the setting of left ventricular mechanical assistance: relation with outcome and C-reactive protein

Background: In left ventricular assist device (LVAD) recipients, plasma levels of interleukin (IL)-6 are associated with Interagency Registry for Mechanically Assisted Circulatory Support (INTERMACS) profiles, reflecting postoperative risk. However, it is not clear how the cardiac. Conclusions: Cardiac IL-6 levels do not contribute to improve risk profile of LVAD recipients in relation to clinical inpatient post-implantation. Instead, plasma IL-6 and serum CRP concentrations are more effective in predicting the severity of the clinical course in the early phase of LVAD therapy. level of IL-6, detectable on the tissue samples at the time of implantation, can contribute to predict the post-operative outcome

Non-enhanced micro-CT of paraffin embedded coronary vessels: a tool for experimental atherosclerosis

Purpose. To assess and quantify morphometric parameters of full-length coronary arteries in three dimensions at high resolution by micro-CT (?CT), and to evaluate the possible integration with in vivo information by invasive coronary angiography (ICA) and and intravascular ultrasound (IVUS). Methods and Materials. Excised left coronary artery (LMCA and LAD) and underlying myocardium from pigs on a high-cholesterol diet were fixed in formalin, dehydrated and embedded in paraffin as a whole sample inside a Falcon-type test tube. In vivo information obtained by ICA and IVUS were also available for the same animals. After the ?CT scan, the samples were further processed in order to perform standard histological stains. Results. For each sample, we have obtained a reconstructed volume of 512x512x1400 isotropic voxels, with a voxel size of 57.4^3 ?m^3. The ?CT images provided good discrimination between fat, muscle and calcium, allowing a reliable segmentation of walls and lesions. The profiles of total area, wall area and calcium along the entire coronary artery allowed a comprehensive evaluation of the progress of atherosclerosis in terms of intimal thickening, plaque formation and calcification. Microcalcifications down to 0.2 nl (nanoliters) were localized and displayed in 3D. The integration with IVUS allowed also to recover the lumen area under in vivo physiological state. Conclusion. Micro-CT allows a comprehensive evaluation in 3D and at high resolution of plaque evolution in an experimental model of coronary artery disease in pigs. The integration with ICA and IVUS can lead to the full characterization of disease progression

Prescription appropriateness of anti-diabetes drugs in elderly patients hospitalized in a clinical setting: evidence from the REPOSI Register

Diabetes is an increasing global health burden with the highest prevalence (24.0%) observed in elderly people. Older diabetic adults have a greater risk of hospitalization and several geriatric syndromes than older nondiabetic adults. For these conditions, special care is required in prescribing therapies including anti- diabetes drugs. Aim of this study was to evaluate the appropriateness and the adherence to safety recommendations in the prescriptions of glucose-lowering drugs in hospitalized elderly patients with diabetes. Data for this cross-sectional study were obtained from the REgistro POliterapie-Società Italiana Medicina Interna (REPOSI) that collected clinical information on patients aged ≥ 65 years acutely admitted to Italian internal medicine and geriatric non-intensive care units (ICU) from 2010 up to 2019. Prescription appropriateness was assessed according to the 2019 AGS Beers Criteria and anti-diabetes drug data sheets.Among 5349 patients, 1624 (30.3%) had diagnosis of type 2 diabetes. At admission, 37.7% of diabetic patients received treatment with metformin, 37.3% insulin therapy, 16.4% sulfonylureas, and 11.4% glinides. Surprisingly, only 3.1% of diabetic patients were treated with new classes of anti- diabetes drugs. According to prescription criteria, at admission 15.4% of patients treated with metformin and 2.6% with sulfonylureas received inappropriately these treatments. At discharge, the inappropriateness of metformin therapy decreased (10.2%, P &lt; 0.0001). According to Beers criteria, the inappropriate prescriptions of sulfonylureas raised to 29% both at admission and at discharge. This study shows a poor adherence to current guidelines on diabetes management in hospitalized elderly people with a high prevalence of inappropriate use of sulfonylureas according to the Beers criteria

Clinical features and outcomes of elderly hospitalised patients with chronic obstructive pulmonary disease, heart failure or both

Background and objective: Chronic obstructive pulmonary disease (COPD) and heart failure (HF) mutually increase the risk of being present in the same patient, especially if older. Whether or not this coexistence may be associated with a worse prognosis is debated. Therefore, employing data derived from the REPOSI register, we evaluated the clinical features and outcomes in a population of elderly patients admitted to internal medicine wards and having COPD, HF or COPD + HF. Methods: We measured socio-demographic and anthropometric characteristics, severity and prevalence of comorbidities, clinical and laboratory features during hospitalization, mood disorders, functional independence, drug prescriptions and discharge destination. The primary study outcome was the risk of death. Results: We considered 2,343 elderly hospitalized patients (median age 81 years), of whom 1,154 (49%) had COPD, 813 (35%) HF, and 376 (16%) COPD + HF. Patients with COPD + HF had different characteristics than those with COPD or HF, such as a higher prevalence of previous hospitalizations, comorbidities (especially chronic kidney disease), higher respiratory rate at admission and number of prescribed drugs. Patients with COPD + HF (hazard ratio HR 1.74, 95% confidence intervals CI 1.16-2.61) and patients with dementia (HR 1.75, 95% CI 1.06-2.90) had a higher risk of death at one year. The Kaplan-Meier curves showed a higher mortality risk in the group of patients with COPD + HF for all causes (p = 0.010), respiratory causes (p = 0.006), cardiovascular causes (p = 0.046) and respiratory plus cardiovascular causes (p = 0.009). Conclusion: In this real-life cohort of hospitalized elderly patients, the coexistence of COPD and HF significantly worsened prognosis at one year. This finding may help to better define the care needs of this population

Clinical features and outcomes of elderly hospitalised patients with chronic obstructive pulmonary disease, heart failure or both

Background and objective: Chronic obstructive pulmonary disease (COPD) and heart failure (HF) mutually increase the risk of being present in the same patient, especially if older. Whether or not this coexistence may be associated with a worse prognosis is debated. Therefore, employing data derived from the REPOSI register, we evaluated the clinical features and outcomes in a population of elderly patients admitted to internal medicine wards and having COPD, HF or COPD + HF. Methods: We measured socio-demographic and anthropometric characteristics, severity and prevalence of comorbidities, clinical and laboratory features during hospitalization, mood disorders, functional independence, drug prescriptions and discharge destination. The primary study outcome was the risk of death. Results: We considered 2,343 elderly hospitalized patients (median age 81&nbsp;years), of whom 1,154 (49%) had COPD, 813 (35%) HF, and 376 (16%) COPD + HF. Patients with COPD + HF had different characteristics than those with COPD or HF, such as a higher prevalence of previous hospitalizations, comorbidities (especially chronic kidney disease), higher respiratory rate at admission and number of prescribed drugs. Patients with COPD + HF (hazard ratio HR 1.74, 95% confidence intervals CI 1.16-2.61) and patients with dementia (HR 1.75, 95% CI 1.06-2.90) had a higher risk of death at one year. The Kaplan-Meier curves showed a higher mortality risk in the group of patients with COPD + HF for all causes (p = 0.010), respiratory causes (p = 0.006), cardiovascular causes (p = 0.046) and respiratory plus cardiovascular causes (p = 0.009). Conclusion: In this real-life cohort of hospitalized elderly patients, the coexistence of COPD and HF significantly worsened prognosis at one year. This finding may help to better define the care needs of this population