Molecular effects of cardiac contractility modulation in patients with heart failure of ischemic aetiology uncovered by transcriptome analysis.

Cardiac contractility modulation (CCM) is based on electrical stimulation of the heart without alteration of action potential and mechanical activation, the data on its fundamental molecular mechanisms are limited. Here we demonstrate clinical and physiological effect of 12 months CCM in 29 patients along with transcriptomic molecular data. Based on the CCM effect the patients were divided into two groups: responders (n = 13) and non-responders (n = 16). RNA-seq data were collected for 6 patients before and after CCM including 3 responders and 3 non-responders. The overall effect of CCM on gene expression was mainly provided by samples from the responder group and included the upregulation of the genes involved in the maintenance of proteostasis and mitochondrial structure and function. Using pathway enrichment analysis, we found that baseline myocardial tissue samples from responder group were characterized by upregulation of mitochondrial matrix-related genes, Z disc-protein encoding genes and muscle contraction-related genes. In summary, twelve months of ССM led to changes in signaling pathways associated with cellular respiration, apoptosis, and autophagy. The pattern of myocardial remodeling after CCM is associated with initial expression level of myocardial contractile proteins, adaptation reserves associated with mitochondria and low expression level of inflammatory molecules.

Desmin mutations impact the autophagy flux in C2C12 cell in mutation-specific manner.

Desmin is the main intermediate filament of striated and smooth muscle cells and plays a crucial role in maintaining the stability of muscle fiber during contraction and relaxation cycles. Being a component of Z-disk area, desmin integrates autophagic pathways, and the disturbance of Z-disk proteins' structure negatively affects chaperone-assisted selective autophagy (CASA). In the present study, we focused on alteration of autophagy flux in myoblasts expressing various Des mutations. We applied Western blotting, immunocytochemistry, RNA sequencing, and shRNA approach to demonstrate that DesS12F, DesA357P, DesL345P, DesL370P, and DesD399Y mutations. Mutation-specific effect on autophagy flux being most severe in aggregate-prone Des mutations such as DesL345P, DesL370P, and DesD399Y. RNA sequencing data confirmed the most prominent effect of these mutations on expression profile and, in particular, on autophagy-related genes. To verify CASA contribution to desmin aggregate formation, we suppressed CASA by knocking down Bag3 and demonstrated that it promoted aggregate formation and lead to downregulation of Vdac2 and Vps4a and upregulation of Lamp, Pink1, and Prkn. In conclusion, Des mutations showed a mutation-specific effect on autophagy flux in C2C12 cells with either a predominant impact on autophagosome maturation or on degradation and recycling processes. Aggregate-prone desmin mutations lead to the activation of basal autophagy level while suppressing the CASA pathway by knocking down Bag3 can promote desmin aggregate formation.

Multifunctional mats by antimicrobial nanoparticles decoration for bioinspired smart wound dressing solutions.

Developing advanced materials for wound dressings is a very challenging, yet unaddressed task. These systems are supposed to act as temporary skin substitutes, performing multiple functions, including fluid absorption and antimicrobial action, supporting cell proliferation and migration in order to promote the skin regeneration process. Following a global bioinspired approach, in this study, we developed a multifunctional textile for wound dressing applications. Biodegradable polyhydroxybutyrate/poly-3-caprolactone (PHB/PCL) mats were fabricated by electrospinning to mimic the extracellular matrix (ECM), thus providing structural and biochemical support to tissue regeneration. Furthermore, inspired by nature's strategy which exploits melanin as an effective weapon against pathogens infection, PHB/PCL mats were modified with hybrid Melanin-TiO2 nanostructures. These were combined to PHB/PCL mats following two different strategies: in-situ incorporation during electrospinning process, alternately ex-post coating by electrospraying onto obtained mats. All samples revealed huge water uptake and poor cytotoxicity towards HaCat eukaryotic cells. Melanin-TiO2 coating conferred PHB/PCL mats significant antimicrobial activity towards both Gram(+) and Gram(-) strains, marked hydrophilic properties as well as bioactivity which is expected to promote materials-cells interaction. This study is going to provide a novel paradigm for the design of active wound dressings for regenerative medicine.

Syngonanthus nitens (Bong.) Ruhland Derivatives Loaded into a Lipid Nanoemulsion for Enhanced Antifungal Activity Against Candida parapsilosis.

BACKGROUND: Vaginal infections caused by non-albicans species have become common in women of all age groups. The resistance of species such as Candida parapsilosis to the various antifungal agents is a risk factor attributed to these types of infections, which instigates the search for new sources of active compounds in vulvovaginal candidiasis (VCC) therapy. OBJECTIVE: This study evaluated the antifungal activity of Syngonanthus nitens Bong. (Ruhland) derivatives and employed a lipid nanoemulsion as a delivery system.' METHODS: In this study, a lipid nanoemulsion was employed as a delivery system composed of Cholesterol (10%), soybean phosphatidylcholine: Brij 58 (1: 2) and PBS (pH 7.4) with the addition of 0.5% of a chitosan dispersion (80%), and evaluated the antifungal activity of S. nitens Bong. (Ruhland) derivatives against planktonic cells and biofilms of Candida parapsilosis. By a biomonitoring fractionation, the crude extract (EXT) and one fraction (F2) were selected and incorporated into a lipid nanoemulsion (NL) composed of cholesterol (10%), a 1:2 mixture of soybean phosphatidylcholine:polyoxyethylene -20- cetyl ether (10%), and phosphate buffer solution (pH 7.4) with a 0.5% chitosan dispersion (80%). The NL presented a diameter size between 50-200 nm, pseudoplastic behavior, and positive charge. The EXT and five fractions were active against planktonic cells. RESULTS AND DISCUSSION: The incorporation of EXT and F2 into the NL increased antifungal activity and enhanced the anti-biofilm potential. This study classified the use of an NL as an important tool for the administration of S. nitens derivatives in cases of infections caused by this C. parapsisilosis. CONCLUSION: This work concluded that S. nitens derivatives were important sources of active molecules against C. parapsilosis and the use of a lipid nanoemulsion was an important tool to promote more effective F2 release and to improve the antifungal activity aiming the control of C. parapsilosis infections.

Mass-spring models for the simulation of mitral valve function: Looking for a trade-off between reliability and time-efficiency.

Patient-specific finite element (FE) models can assess the impact of mitral valve (MV) repair on the complex MV anatomy and function. However, FE excessive time requirements hamper their use for surgical planning; mass-spring models (MSMs) represent a more approximate approach but can provide almost real-time simulations. On this basis, we implemented MSMs of three healthy MVs from cardiac magnetic resonance (cMR) imaging to simulate the systolic MV closure, including the in vivo papillary muscles and annular kinematics, and the anisotropic and non-linear mechanical response of MV tissues. To test MSM reliability we compared the systolic peak configurations computed by MSMs and FE: mismatches by less than twice the in-plane cMR image resolution were detected over 75% of the leaflets' surface, independently of the MSM mesh refinement and of the specific MV anatomy. Data on MSMs time-efficiency and data from the comparison of MSMs vs. FE models suggest that MSM could represent a suitable trade-off between almost real-time simulations and reliability when computing MV systolic configuration, with the potential to be used in a clinical setting either as a support to the decisional process or as a virtual training tool.

Influence of the aortic valve leaflets on the fluid-dynamics in aorta in presence of a normally functioning bicuspid valve.

In this work, we consider the blood fluid-dynamics in the ascending aorta in presence of a normally functioning bicuspid aortic valve (BAV). In particular, we perform an unsteady finite element study in real geometries with physiological velocity boundary conditions at the inlet to assess the effect of the inclusion of the leaflets on the fluid-dynamic abnormalities characterizing BAV cases. To this aim, we perform a comparison in two geometries (a dilated and a non-dilated ones) among three scenarios which are built up for each geometry: BAV without leaflets, BAV with leaflets, and tricuspid case with leaflets. For each case, we compute four indices quantifying flow asymmetry, reversal flows, helical patterns, and wall shear stresses. Our results show that the inclusion of the leaflets increases the fluid-dynamics abnormalities, especially for the non-dilated configuration, which presents a greater increment of the indices. In particular, we observe that the values of the time-averaged wall shear stress and of the systolic jet asymmetry increase by approximatively 100 and 40%, respectively, when considering the leaflets.

Fluorescent (rhodamine), folate decorated and doxorubicin charged, PEGylated nanoparticles synthesis.

PEGylated silica nanoparticles, giving very stable aqueous sols, were successfully functionalised with rhodamine, one of the more stable fluorophore; they were also decorated with the targeting agent folic acid (FA) and charged with the well known drug doxorubicin. Rhodamine functionalization required a modification of the synthesis route of the nanoparticles (NP). Functionalization with FA required activation with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride. Folate decorated NP were easily charged with doxorubicin. The experimental results proved the successfulness of the functionalization. The bond to the NP does not reduce the therapeutic efficacy of the drug. The calculated encapsulation efficiency (32 %) was only a little lower than the value (47 %) reported for the very popular PEGylated PLGA NP.

Results with expanded donor acceptance criteria in heart transplantation.

OBJECTIVE: Over the past years both donor and recipient profiles have changed in heart transplantation. Satisfactory clinical outcomes of marginal donors in candidates >60 years of age have led us to allocate suboptimal donors to younger recipients as well. Therefore, we retrospectively reviewed our experience. METHODS: Among 199 patients undergoing heart transplantation from January 2000 to February 2010, there were 83 (41%) aged 61-72 years. The other 116 (59%) ranged in age between 18 and 60 years. According to their clinical conditions as heart transplantation candidates, They were classified into 4 groups: younger recipients (n=116) of either optimal donors (n=72; group 1 [G1]) or marginal donors (n=44; group 2 [G2]) and older recipients (n=83) of either marginal grafts (n=70, group 3 [G3]) or optimal grafts (n=13; group 4 [G4]). The gender distribution, cause of end-stage heart failure, preoperative pulmonary hypertension incidence, pretransplantation clinical status, and mean follow-up were not significantly different among the 4 groups. RESULTS: Overall 30-day survival was 90 ± 1% and 10-year rate was 78 ± 9%. Among the groups, 30-day and 10-year actuarial survival rates were, respectively: 94 ± 4% and 87 ± 1% for G1; 86 ± 5% and 84 ± 7% for G2; 88 ± 4% and 71 ± 7% for G3 and were 100% and 82 ± 7% for G4 (P=.7). In comparison among the 4 groups, there was no significant difference regarding freedom from graft failure (P=.3), right ventricular failure (P=.3), acute rejection episodes (P = .2), chronic rejection (P=.2), neoplasia (P=.5), or chronic renal failure (P=.1). Older recipients of marginal donors [G3] had a 4% (n=3) prevalence of permanent pacemaker implant, versus G2: 3% (n=2) among (P=.1). CONCLUSION: Our results suggest that extended donor and recipient criteria do not compromise clinical outcomes after transplantation.

Relationship between leptin and all–cause and cardiovascular mortality in chronic hemodialysis patients

Background: We aimed to evaluate the relationship between serum leptin and the leptin/body mass index (BMI) ratio with prevalent cardiovascular disease (CVD), and their influence on all–cause and CVD–related mortality in patients on hemodialysis (HD). Methods: 118 stable HD patients (50 women, median [interquartile range] age, 65.1 [54.7–72.2] years) were studied. All patients had baseline measurement of serum leptin concentrations. Relationships between leptin and all–cause and CVD mortality were studied by means of survival analysis and Cox regression analysis. Results: The leptin/BMI ratio was similar in patients with and without CVD at baseline (0.65 [0.29–2.23] vs. 0.68 [0.29–1.49] ng·m2/ml·kg, respectively, NS). Multiple logistic regression analysis showed that there was not an independent association between leptin/BMI ratio and prevalent CVD. During the follow–up time, 52 (44.1%) patients died. CVD was the cause of death in 27 out of 52 (51.9%) deceased patients. Survival analysis and Cox proportional multivariate regression analysis showed that there were no significant relationships between leptin levels or the leptin/BMI ratio and all–cause and CVD–related mortality. Conclusion: These results do not support that, in stable HD patients, serum leptin concentrations and the leptin/BMI ratio are related with prevalent CVD. Leptin/BMI ratio seems not to be a risk factor for mortality in these patients (AU)

Introducción: El objetivo del presente estudio ha sido evaluar la relación entre la leptina sérica y el cociente leptina/índice de masa corporal (IMC) con la enfermedad cardiovascular (ECV) prevalente y su influencia en la mortalidad global y en la mortalidad por ECV en pacientes en hemodiálisis (HD). Métodos: Se estudiaron 118 pacientes estables en HD (50 mujeres, edad mediana [recorrido intercuartílico], 65,1 [54,7–72,2] años). En todos los pacientes se cuantificó la concentración basal de leptina. La relación entre leptina y la mortalidad se evaluó mediante análisis de supervivencia y análisis de regresión de Cox. Resultados: El cociente leptina/IMC fue similar en pacientes con y sin ECV prevalente (0,65 [0,29–2,23] frente a 0,68 [0,29–1,49] ng·m2/ml·kg, respectivamente, NS). El análisis de regresión logística mostró que no existía una asociación independiente entre el cociente leptina/IMC y la enfermedad cardiovascular prevalente. Durante el seguimiento 52 pacientes fallecieron (44,1%). La ECV fue causa de muerte en 27 de 52 pacientes fallecidos (51,9%). El análisis de supervivencia y el análisis multivariante de Cox mostraron que no hubo relación significativa entre los niveles de leptina o el cociente leptina/IMC y la mortalidad global o por causa de ECV. Conclusión: Estos resultados no apoyan la hipótesis de que, en pacientes estables en HD, las concentraciones de leptina y el cociente leptina/IMC estén relacionados con la ECV prevalente. Más aún, el cociente leptina/IMC no parece ser un factor de riesgo de mortalidad en estos pacientes (AU)

[Kidney and sports]. / Rene e sport.

Several renal function modifications can be related to sports activity, as pre-existing renal dysfunction can influence the assessment of sports practice capability. The multiplicity of aetiologies requires an accurate diagnosis to correctly define the treatment approach and feasible activities. Previous nephrectomy or chronic renal failure are conditions that complicate the sport exercise fitness assessment and every patient should be assessed individually by a multidisciplinary medical team.

Negative regulation of UCP2 by TGFß signaling characterizes low and intermediate-grade primary breast cancer.

The histological manifestation of growth-regulating and differentiation-inducing signals in cancer cells is considered as a key component for clinical outcome prediction and commonly defined as tumor differentiation grade. However, the molecular and functional framework underlying this clinical parameter remains poorly understood. Our correlative data display a significant association (P>0.001) between mitochondrial uncoupling protein 2 (UCP2) and tumor grade in primary breast cancer (n=234). Through mechanistic analyses, we show a synergistic link between UCP2 and established cellular pathways in conferring grade-associated functional phenotypes. Here, the application of well to moderately differentiated primary tumor cell lines has enabled direct observation of SMAD recruitment to the UCP2 promoter underlying repression of gene transcription. In contrast, poorly differentiated tumor cells, known to be TGFß resistant, displayed aberrant UCP2 regulation, and consequently, gene overexpression, which reduced mitochondrial calcium and facilitated the maintenance of mitochondrial membrane potential, thereby significantly decreasing oxidative stress and inhibiting cell death. Conversely, UCP2 silencing in such cells rapidly led to the induction of apoptosis and cell differentiation, concurrent with reduced cell survival and proliferation, confirming gene-specific effects. Demonstration of a biologically driven role for UCP2 dysregulation in promoting multiple characteristics of tumor aggressiveness strongly endorses assessment of gene expression at clinical presentation to augment therapeutic decision-making and improve patient outcome through personalized targeting approaches.

Iron, zinc and aluminium ferritin content of hemodialysis hyperferritinemic patients: comparison with other hyperferritinemic clinical conditions and normoferritinemic blood donors.

The present study describes the specific content of ferritin iron, zinc and aluminium in four different groups: 1) hemodialysis hyperferritinemic patients; 2) septic patients; 3) iron overloaded patients with hematologic diseases; and 4) blood donors. In all four groups high levels of aluminium and zinc were found in addition to those of iron. However, the sum of the ferritin ions of the control group is significantly higher than that of the other three groups. Furthermore, while ferritin of hemodialysis patients has the same molecular ratio of metal ions as control group (high Al content vs. Fe and Zn), a lower Al/Fe ratio is found both in septic and hematological patients. The results of the present paper might help to explain the high percentage of hyperferritinemia found in hemodialysis patients also in presence of low transferrin saturation and in absence of inflammatory markers. Moreover, the high content of ions other than iron in the ferritin core leads us to believe that ferritin is not only an iron storage protein but rather a regulator of redox active ions.

A neonate with cyanosis and tortuous great arteries.

This report describes an unusual case with tortuosity of the great vessels in a neonate who presented at birth with cyanosis. The diagnosis was made with magnetic resonance imaging (MRI), then confirmed by genetic analysis.

Sphingosine 1-phosphate induces differentiation of adipose tissue-derived mesenchymal stem cells towards smooth muscle cells.

Sphingosine 1-phosphate (S1P) is a bioactive sphingolipid which regulates multiple biological parameters in a number of cell types, including stem cells. Here we report, for the first time, that S1P dose-dependently stimulates differentiation of adipose tissue-derived mesenchymal stem cells (ASMC) towards smooth muscle cells. Indeed, S1P not only induced the expression of smooth muscle cell-specific proteins such as alpha-smooth muscle actin (alpha SMA) and transgelin, but also profoundly affected ASMC morphology by enhancing cytoskeletal F-actin assembly, which incorporated alpha SMA. More importantly, S1P challenge was responsible for the functional appearance of Ca(2+) currents, characteristic of differentiated excitable cells such as smooth muscle cells. By employing various agonists and antagonists to inhibit S1P receptor subtypes, S1P(2) turned out to be critical for the pro-differentiating effect of S1P, while S1P(3) appeared to play a secondary role. This study individuates an important role of S1P in AMSC which can be exploited to favour vascular regeneration.

[Malnutrition in patients on chronic hemodialysis: prevalence, pathogenesis, and treatment]. / La malnutrizione nel paziente in emodialisi cronica: prevalenza, patogenesi, terapia.

Malnutrition is common in patients on hemodialysis and is a strong predicor of morbidity and mortality. Much progress has been made in recent years in identifying the causes and pathogenesis of malnutrition in hemodialysis patients as well as in recognizing the link between malnutrition and morbidity and mortality. Nevertheless, there is no consensus concerning its management. Conventional interventions such as nutritional counseling, oral nutritional supplements and intradialytic parenteral nutrition and novel preventive and therapeutic strategies such as appetite stimulants, growth hormone, androgenic anabolic steroids, and antiinflammatory drugs have been tested with contradictory and inconclusive results. Malnutrition still remains an important challenge for the nephrologist in the third millennium.

In-situ sol-gel synthesis and characterization of bioactive pHEMA/SiO2 blend hybrids.

A novel procedure to synthesize poly(2-hydroxyethylmethacrylate)-silica blend hybrids is presented. Methacrylate monomers bearing an alkoxysilyl unit, prepared by Michael addition of 2-hydroxyethylmethacrylate (HEMA) to 3-Aminopropyltriethoxysilane (APTS) were employed. By (13)C NMR and mass analysis it was possible to establish the formation of coupling hybrid species. Hybrid materials, with final concentration ranging from 10% to 30% w/w of silica gel to the mass of polymer, were obtained through basic catalyzed sol-gel process of tetraethoxysilane (TEOS) and the alkoxysilyl unit of the hybrid monomer, followed by in-situ free-radical polymerization. The hybrids were characterized as far as concerns their thermal properties (glass transition temperature, decomposition temperature), their sorption behavior in water, and in-vitro bioactivity. Optical transparency, higher glass transition temperature, and higher decomposition temperature than pHEMA suggest an increase in either density or intensity of cross-links between the organic and the inorganic phases. The swelling ratio of the 30% hybrids is comparable to pHEMA, whereas it is lower for the other compositions. In-vitro bioactivity of the hybrids, due to the inorganic phase, was ascertained. Soaking time required for apatite deposition on the samples surface decreases as the content of silica gel increases. Therefore, the obtained bioactive hybrids can be used to make bioactive scaffolds for bone engineering.

Bioactive poly(2-hydroxyethylmethacrylate)/silica gel hybrid nanocomposites prepared by sol-gel process.

A hybrid of poly(2-hydroxyethylmethacrylate) (pHEMA), a polymer widely employed for biomedical applications, and silica gel, exhibiting a well-known bioactivity, was produced by sol-gel. The amount of the inorganic precursor, tetraethoxysilane (TEOS), was mixed to the organic monomer, so as to have a final concentration of 30% (w/w) of silica gel to the mass of polymer. The nanocomposite was characterized for its composition by thermogravimetric (TG) analysis, swelling behavior, glass transition temperature using differential thermal analysis (DTA), morphology through scanning electron microscopy (SEM), and bioactivity using FT-IR spectroscopy, SEM, and energy dispersive system (EDS). The nanocomposite showed phase separation between the polymer and the silica gel, improved thermal stability and swelling properties and higher glass transition temperature than pHEMA. Moreover, bioactive SiO(2) gel nanoparticles promoted apatite formation on the surface of the modified hydrogel, when it was soaked in SBF. Therefore, the obtained bioactive nanocomposite can be used to make bioactive scaffold for bone engineering.

Effect of the solution flow rate on the in vitro bioactivity of 2.5CaO x 2SiO(2) glass.

A comparative study of in vitro bioactivity of 2.5CaO x 2SiO(2) glass has been carried out by soaking it in a simulated body fluid, with continuously and periodic exchange of this solution (dynamic and differential protocols). Dynamic assays were carried out at different solution flow rates, 3 mL/h, 6 mL/h, 12 mL/h, to study the influence of flow rate on glass reactivity. Glass surface was studied by Fourier transform infrared spectroscopy, scanning electron microscopy, and energy dispersive spectroscopy so as to compare the behavior in the two procedures, revealing that in both cases an apatite layer is formed on the glass surface, although there are differences on deposition rate and morphology, which are also influenced by the solution flow rate.