Guiding stem cell self-organization and differentiation through material nanopatterning

Biophysical stimuli in the local microenviroment proved to be effective in influencing various aspects of cell behavior such as adhesion, spreading, migration and differentiation [1]. In particular, many studies focused on biomaterials with nanometric scale surface topography able to dictate specific cell shapes, which in turn control cell fate and functions through mechanotransduction pathways [2]. While the role of biophysical signals and more specifically of topographic signals on single cell behavior is well established, there are comparatively less studies on their role on the collective cell behavior. In particular it is not clear how biophysical signals may affect cells self-organization into a three-dimensional tissue. Here, we used nanopatterned substrates able to control adhesion and contractility processes to generate ordered tissues in vitro. In particular we show that by changing the combination of the initial conditions for cell adhesion we obtained different cell behaviours in terms of self-organization and subsequent tissue development. Bone-marrow-derived hMSCs were cultured on polydimethylsiloxane (PDMS) substrates containing parallel and straight channels having ridge to groove width ratio of 1:1, previously treated with the oxygen plasma, that increases the hydrophilicity, improving cell adhesion. Pattern features used, were 350 nm width and depth of 100 nm. Focal adhesions, cytoskeleton and matrix production were visualized with confocal, scanning electron and transmission electron microscopy. Expression of relevant genes was assessed by RT-PCR analysis. In this culturing setup, hMSCs proliferated and organized into dense cells sheets displaying a multi-level order. Molecular analysis suggests that these conditions create a microenvironment that allows the maintenance of stemness and the enhance of the expression of the pluripotency genes into hMSC. By modifying the initial conditions, dramatically changes the cellular behavior were observed. In fact, by increasing the size of the topographic patterns (channels of 700 nm width and 250 nm depth), without surface chemical treatments, the hMSC were stimulated to arrange themselves into self-organized structures that shared similarities with the tendon tissue, in terms of macroscopic morphology, internal cellular organization and molecular profile. Thses data suggest that the pattern provides an initial guidance for FAs and subsequent cell alignment. Aligned cell exert a polarized contractility that leads to the formation of ordered structures. In conclusion, our results demonstrate that the chemical-physical characteristics of the substrate, were able to strongly affect cell behavior in terms of cell fate and in vitro generated tissue functions. In conclusion, nanoengineered material surfaces can be in principle employed to set off the hMSC program toward tissue genesis in a deterministic manner by using the correct combination of initial biophysical signals. References 1. Ventre et al. J R Soc Interface, 9, 2017-2032, 2012 2. McNamara et al. J Tissue Eng, 120623, 201

Neuro-hormonal effects of physical activity in the elderly.

Thanks to diagnostic and therapeutic advances, the elderly population is continuously increasing in the western countries. Accordingly, the prevalence of most chronic age-related diseases will increase considerably in the next decades, thus it will be necessary to implement effective preventive measures to face this epidemiological challenge. Among those, physical activity exerts a crucial role, since it has been proven to reduce the risk of cardiovascular diseases, diabetes, obesity, cognitive impairment and cancer. The favorable effects of exercise on cardiovascular homeostasis can be at least in part ascribed to the modulation of the neuro-hormonal systems implicated in cardiovascular pathophysiology. In the elderly, exercise has been shown to affect catecholamine secretion and biosynthesis, to positively modulate the renin-angiotensin-aldosterone system and to reduce the levels of plasma brain natriuretic peptides. Moreover, drugs modulating the neuro-hormonal systems may favorably affect physical capacity in the elderly. Thus, efforts should be made to actually make physical activity become part of the therapeutic tools in the elderly. © 2013 Femminella, de Lucia, Iacotucci, Formisano, Petraglia, Allocca, Ratto, DAmico, Rengo, Pagano, Bonaduce, Rengo and Ferrara

Zoonotic Risk of Encephalitozoon cuniculi in Animal-Assisted Interventions: Laboratory Strategies for the Diagnosis of Infections in Humans and Animals

The involvement of animals for therapeutic purposes has very ancient roots. To date, it is clear that animal-assisted interventions (AAIs), in addition to ensuring the replacement of missing or deficient affects, improves psychophysiological parameters connected to human health. However, AAI could potentially present risks related to the transmission of infectious agents from animals to humans. Among these microorganisms, E. cuniculi is a microspore which induces pathological effects (fever, headache, nausea, vomiting, diarrhea, breathlessness, respiratory symptoms, and weakness) in both humans and animals. Consequently, an accurate and fast diagnosis of E. cuniculi infection, as well as the identification of new diagnostic approaches, is of fundamental importance. This literature review was carried out to provide an extensive and comprehensive analysis of the most recent diagnostic techniques to prevent and care for E. cuniculi-associated risks in the AAI field

Changes of Oxytocin and Serotonin Values in Dialysis Patients after Animal Assisted Activities (AAAs) with a Dog-A Preliminary Study

Simple Summary This study aimed to improve the moment of dialysis because the emotional management of a person during treatment can help to reduce stress, anxiety and depression. This process positively affects the acceptance and progress of treatment and improves the self-management of the disease, a very important achievement in chronic kidney disease. Serotonin and oxytocin are important neuromodulators of different human behaviours, such as affectivity and socialization, and are involved in the control of stress, anxiety and social cooperation. The relationship between humans and domestic animals provides psychophysical well-being and can facilitate interpersonal bonds by favouring mechanisms involved in social relations. Dogs due to their ethological characteristics, allow the establishment of an active relationship through play, communication and interaction. Animal-assisted activities (AAAs) are structured interventions aimed at improving the psychophysical conditions of people in stressful conditions. Our study was aimed at determining the circulating levels of serotonin and oxytocin in patients who participated in an AAAs program with a dog during dialysis treatment. Our study aimed to measure the levels of serotonin and oxytocin in patients affected by end-stage renal disease (ESRD), undergoing dialysis and participating in a program of animal-assisted activities (AAAs) with a dog. Ten patients with comparable levels of ESRD were enrolled. A blood sample was taken before the start of the study in order to establish basal levels. Eleven meetings were held once a week for 3 months during the last hour of dialysis, and blood samples were collected before and after AAAs. Two more meetings, one month apart from each other, were held two months later without the dog but with the same veterinarian zootherapist. Blood was drawn at the beginning and at the end of each meeting. The samples were then processed for the measurement of serotonin and oxytocin, and data obtained were analysed using analysis of variance with mixed effect models. The results show an increasing level of both serotonin and oxytocin between subsequent meetings with the dog and an increasing trend of inter-intervention levels. Overall, the results suggest that AAAs lead to modifications of serotonin and oxytocin levels, which are also accompanied by behavioural changes of patients

CDK 4/6 inhibitors as single agent in advanced solid tumors

Cyclin-dependent kinases (CDK) 4/6 inhibitors, namely abemaciclib, palbociclib, and ribociclib, interfere with cell cycle progression, induce cell senescence and might promote cancer cell disruption by a cytotoxic T cells-mediated effect. Phase III randomized clinical trials have proven that CDK4/6 inhibitors (CDK4/6i) in combination with several endocrine agents improve treatment efficacy over endocrine agents alone for hormone receptor positive (HR+) HER2 negative (HER2-) metastatic breast cancer (MBC). Based on such results, these combinations have been approved for clinical use. Preclinical studies in cell cultures and mouse models proved that CDK4/6i are active against a broad spectrum of solid tumors other than breast cancer, including liposarcoma, rhabdomyosarcoma, non-small cell lung cancer, glioblastoma multiforme, esophageal cancer, and melanoma. The role of CDK4/6i in monotherapy in several solid tumors is currently under evaluation in phase I, II, and III trials. Nowadays, abemaciclib is the only of the three inhibitors that has received approval as single agent therapy for pretreated HR+ HER2- MBC. Here we review biological, preclinical and clinical data on the role of CDK4/6 inhibitors as single agents in advanced solid tumors

Predictive Value of Left Ventricular Myocardial Deformation for Left Ventricular Remodeling in Patients With Classical Low-Flow, Low-Gradient Aortic Stenosis Undergoing Transcatheter Aortic Valve Replacement

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Omi/HtrA2 promotes cell death by binding and degrading the anti-apoptotic protein ped/pea-15

ped/pea-15 is a ubiquitously expressed 15-kDa protein featuring a broad anti-apoptotic function. In a yeast two-hybrid screen, the pro-apoptotic Omi/HtrA2 mitochondrial serine protease was identified as a specific interactor of the ped/pea-15 death effector domain. Omi/HtrA2 also bound recombinant ped/pea-15 in vitro and co-precipitated with ped/pea-15 in 293 and HeLa cell extracts. In these cells, the binding of Omi/HtrA2 to ped/pea-15 was induced by UVC exposure and followed the mitochondrial release of Omi/HtrA2 into the cytoplasm. Upon UVC exposure, cellular ped/pea-15 protein expression levels decreased. This effect was prevented by the ucf-101 specific inhibitor of the Omi/HtrA2 proteolytic activity, in a dose-dependent fashion. In vitro incubation of ped/pea-15 with Omi/HtrA2 resulted in ped/pea-15 degradation. In intact cells, the inhibitory action of ped/pea-15 on UVC-induced apoptosis progressively declined at increasing Omi/HtrA2 expression. This further effect of Omi/HtrA2 was also inhibited by ucf-101. In addition, ped/pea-15 expression blocked Omi/HtrA2 co-precipitation with the caspase inhibitor protein XIAP and caspase 3 activation. Thus, in part, apoptosis following Omi/HtrA2 mitochondrial release is mediated by reduction in ped/pea-15 cellular levels. The ability of Omi/HtrA2 to relieve XIAP inhibition on caspases is modulated by the relative levels of Omi/HtrA2 and ped/pea-15

Omi/HtrA2 promotes cell death by binding and degrading the anti-apoptotic protein ped/pea-15

ped/pea-15 is a ubiquitously expressed 15-kDa protein featuring a broad anti-apoptotic function. In a yeast two-hybrid screen, the pro-apoptotic Omi/HtrA2 mitochondrial serine protease was identified as a specific interactor of the ped/pea-15 death effector domain. Omi/HtrA2 also bound recombinant ped/pea-15 in vitro and co-precipitated with ped/pea-15 in 293 and HeLa cell extracts. In these cells, the binding of Omi/HtrA2 to ped/pea-15 was induced by UVC exposure and followed the mitochondrial release of Omi/HtrA2 into the cytoplasm. Upon UVC exposure, cellular ped/pea-15 protein expression levels decreased. This effect was prevented by the ucf-101 specific inhibitor of the Omi/HtrA2 proteolytic activity, in a dose-dependent fashion. In vitro incubation of ped/pea-15 with Omi/HtrA2 resulted in ped/pea-15 degradation. In intact cells, the inhibitory action of ped/pea-15 on UVC-induced apoptosis progressively declined at increasing Omi/HtrA2 expression. This further effect of Omi/HtrA2 was also inhibited by ucf-101. In addition, ped/pea-15 expression blocked Omi/HtrA2 co-precipitation with the caspase inhibitor protein XIAP and caspase 3 activation. Thus, in part, apoptosis following Omi/HtrA2 mitochondrial release is mediated by reduction in ped/pea-15 cellular levels. The ability of Omi/HtrA2 to relieve XIAP inhibition on caspases is modulated by the relative levels of Omi/HtrA2 and ped/pea-15

Extended Non-destructive Testing for Surface Quality Assessment

AbstractThis chapter introduces various extended non-destructive testing (ENDT) techniques for surface quality assessment, which are first characterized, then enhanced, and finally applied to assess the level of pre-bond contaminations intentionally applied to carbon fiber reinforced plastic (CFRP) adherends following the procedures described in the previous chapter. Based on two user cases comprising different scenarios that are characteristic of either aeronautical production or repair, the detailed tests conducted on two types of sample geometry, namely flat coupons and scarfed pilot samples with a more complex shape, form the basis for applying the advanced ENDT procedures for the monitoring of realistic and real aircraft parts, as will be described in Chap. 10.1007/978-3-319-92810-4_5. Specifically, the reported investigations were performed to assess the surface quality of first ground and then intentionally contaminated CFRP surfaces using the following ENDT tools: the aerosol wetting test (AWT), optically stimulated electron emission (OSEE), two differently implemented approaches based on electronic noses, laser-induced breakdown spectroscopy (LIBS), Fourier-transform infrared (FTIR) spectroscopy, laser-induced fluorescence (LIF), and laser vibrometry