Il recettpre Kit nella regolazione delle MSCs: analisi della proteina durante la differenziazione adipogenica.

Il recettore Kit nella regolazione delle MSCs : analisi della proteina durante la differenziazione adipogenica. Le Mesenchymal Stem Cells o Marrow Stromal Cells (MSCs) sono cellule stromali multipotenti che possono differenziare, sia in vivo che in vitro, verso lineages diversi (osteogenico, condrogenico, adipogenico, miogenico), hanno capacità migratorie e secernono un ampio spettro di molecole trofiche e/o immunoregolatorie. La loro fonte principale è il midollo osseo, dove rappresentano approssimativamente lo 0.01%-0.001% delle cellule nucleate, ma sono presenti anche in molti altri tessuti adulti. Queste caratteristiche, associate alla facile reperibilità e maneggiabilità, le hanno poste al centro dell’attenzione per il loro potenziale impiego terapeutico. Un’applicazione clinica ottimale richiede però una completa conoscenza, ancora non raggiunta, dei meccanismi biologici alla base del self-renewal e dei processi differenziativi delle MSCs. Dati recenti sottolineano l’importanza di geni chiave coinvolti nella regolazione di tessuti anche molto diversi fra loro. Uno di questi è Kit, che codifica il recettore tirosina-chinasico dello Stem Cell Factor (SCF) e che ha un ruolo fondamentale in diversi tipi di cellule staminali. Kit è, infatti, attivo nelle Primordial Germ Cells (PGCs), nelle cellule staminali cardiache, nelle cellule staminali e nei precursori ematopoietici, nelle cellule staminali neurali, nei melanoblasti e nei mesangioblasti, mentre risulta ancora controverso il suo coinvolgimento nelle MSCs . Nel laboratorio presso il quale ho svolto il mio lavoro di tesi, sono in corso studi volti a chiarire il possibile ruolo di Kit nelle MSCs. In primo luogo, è stata analizzata l’attività trascrizionale di Kit sia in MSCs umane che murine, riscontrando un’espressione a bassi livelli nelle MSCs indifferenziate e una modulazione dell’espressione durante la loro differenziazione. In particolare, durante le prime fasi della differenziazione adipogenica l’mRNA di Kit è maggiormente espresso, mentre durante quella osteogenica il gene è down-regolato. Scopo del mio lavoro di tesi è stato analizzare l’espressione della proteina Kit, per confermarne il coinvolgimento anche nella regolazione delle MSCs, focalizzandomi soprattutto sulla differenziazione adipogenica di MSCs murine. A tale scopo ho effettuato saggi di immunocitochimica e analisi al FACS (Fluorescence Activated Cell Sorting), usando anticorpi diretti contro il dominio extra-citoplasmatico di Kit. I risultati hanno evidenziato un’espressione bassa in MSCs allo stato indifferenziato e un aumento del prodotto genico nelle prime fasi della differenziazione adipogenica, seguito da un ritorno ai livelli basali nelle fasi terminali. Nel nostro laboratorio è stata caratterizzata una linea di topi transgenici Kit-GFP, in cui il gene codificante per la Green Fluorescent Protein (GFP) si trova sotto il controllo trascrizionale di alcuni elementi regolatori del gene Kit. Esperimenti precedenti hanno evidenziato che il transgene è attivo nei diversi tipi cellulari in cui è attivo Kit endogeno e ne ricapitola in maniera fedele l’espressione. Questo topo transgenico si rivela quindi un ottimo modello per studiare Kit in diversi tessuti. Pertanto ho analizzato al FACS MSCs di topi Kit-GFP indotte a differenziare in senso adipogenico, evidenziando una corrispondenza fra l’aumento della fluorescenza dovuta al transgene e il comportamento del Kit endogeno. Inoltre ho monitorato la differenziazione adipogenica e osteogenica di MSCs di topi Kit-GFP attraverso microscopio a fluorescenza, riscontrando un’analogia con i dati ottenuti tramite l’analisi al FACS. In conclusione, i risultati suggeriscono che Kit è attivo anche nelle MSCs , in particolare nelle fasi precoci della differenziazione adipogenica. Inoltre i dati dimostrano che nel transgene Kit-GFP sono contenuti gli elementi regolatori necessari per la sua corretta espressione anche nelle MSCs. Questo nuovo ruolo di Kit può avere interessanti implicazioni anche nelle patologie del tessuto adiposo

Plasma actuation for lifted flame stabilization in coaxial methane-air flow

The flame stabilization represents a relevant issue in aero-engine design. In fact, the growing demand of pollutant emissions reduction without significant losses of the combustion efficiency has driven the efforts of the scientific community towards lean flames. Lean fuel mixtures, characterized by low temperature flames, could manifest an unstable behaviour which can easily lead to the flame extinction due to the establishment of the blowout condition. This requires the implementation of control systems to avoid flame instability occurrence. The present work shows an investigation of the impact of dielectric barrier discharge (DBD) plasma actuation on lifted flame stabilization in a methane CH4-air Bunsen burner at ambient conditions. Two different plasma actuator configurations, powered with a high voltage (HV)/high frequency sinusoidal signal, have been investigated. Once the best actuator configuration was selected, the efficiency of the plasma actuation has been evaluated in terms of the flame lift-off distance, length and shape. In particular, in order to change the actuator power dissipation, different peak-to-peak voltages Vpp were tested, while the actuation frequency was kept equal to 20 kHz. The application of plasma discharges to flame stabilization leads to plasma-attached flames or plasma-enhanced lifted flames, depending on the air and fuel flow rates. At air flow rate of 1.54 g/s, plasma actuation allowed to decrease the lift-off height until the fuel jet velocity was below about 0.05 m/s thanks to the extension of the flame region upstream, toward the burner exit section. Beyond this value, it had no significant impact on the flame lift-off height, even though the amplitude of the lift-off height oscillations reduced coupled with a more stable behaviour of the lifting flame. The benefit of the plasma actuation increased by reducing the air flow rate to 1.35 g/s. In this condition, plasma-assisted flame reattachment was evident at each fuel velocity, in combination with an increasing flame height proportionally to the fuel jet velocity

Multiple cues produced by a robotic fish modulate aggressive behaviour in Siamese fighting fishes

The use of robotics to establish social interactions between animals and robots, represents an elegant and innovative method to investigate animal behaviour. However, robots are still underused to investigate high complex and flexible behaviours, such as aggression. Here, Betta splendens was tested as model system to shed light on the effect of a robotic fish eliciting aggression. We evaluated how multiple signal systems, including a light stimulus, affect aggressive responses in B. splendens. Furthermore, we conducted experiments to estimate if aggressive responses were triggered by the biomimetic shape of fish replica, or whether any intruder object was effective as well. Male fishes showed longer and higher aggressive displays as puzzled stimuli from the fish replica increased. When the fish replica emitted its full sequence of cues, the intensity of aggression exceeded even that produced by real fish opponents. Fish replica shape was necessary for conspecific opponent perception, evoking significant aggressive responses. Overall, this study highlights that the efficacy of an artificial opponent eliciting aggressive behaviour in fish can be boosted by exposure to multiple signals. Optimizing the cue combination delivered by the robotic fish replica may be helpful to predict escalating levels of aggression

Dynamic titanium prosthesis based on 3D-printed replica for chest wall resection and reconstruction

3D-printing technologies can assist the surgical planning and prosthesis engineering for the management of extended chest wall resection. Different types of prosthesis have been utilized over time, but some concerns remain about their impact on the respiratory function. Here we present a new kind of 3D-printed titanium prosthesis designed to be either strong and flexible. The prosthesis was created on a 1:1 3D-printed anatomic replica of the chest, used to delineate surgical margins and to define the reconstructive requirements

New Challenges for Sustainable Organizations in Light of Agenda 2030 for Sustainability

Sustainability is one of humanity’s most daunting issues at present. Increasing popula- tion, escalation of anthropogenic activities, industrialization, modern agricultural practices that pollute water, air, and soil around the world, and ever-increasing greenhouse gas emissions mean that sustainability is now in doubt [1]. In response to these critical concerns, the world has come up with several initiatives including Agenda 2030. Agenda 2030 is a commitment to eradicate poverty and achieve sustainable development worldwide, ensuring that no one is left behind by 2030. Its adoption was a landmark achievement, providing a shared vision towards sustainable development for all. Its 17 Sustainable Development Goals (SDGs) and 169 targets aim to end the plethora of development problems and deliver a better univers

Can Public-Private Partnerships Foster Investment Sustainability in Smart Hospitals?

This article addresses the relationship between Public-Private Partnerships (PPP) and the sustainability of public spending in smart hospitals. Smart (technological) hospitals represent long-termed investments where public and private players interact with banking institutions and eventually patients, to satisfy a core welfare need. Characteristics of smart hospitals are critically examined, together with private actors\u2019 involvement and flexible forms of remuneration. Technology-driven smart hospitals are so complicated that they may require sophisticated PPP. Public players lack innovative skills, whereas private actors seek additional compensation for their non-routine efforts and higher risk. PPP represents a feasible framework, especially if linked to Project Financing (PF) investment patterns. Whereas the social impact of healthcare investments seems evident, their financial coverage raises growing concern in a capital rationing context where shrinking public resources must cope with the growing needs of chronic elder patients. Results-Based Financing (RBF) is a pay-by-result methodology that softens traditional PPP criticalities as availability payment sustainability or risk transfer compensation. Waste of public money can consequently be reduced, and private bankability improved. In this study, we examine why and how advanced Information Technology (IT) solutions implemented in \u201cSmart Hospitals\u201d should produce a positive social impact by increasing at the same time health sustainability and quality of care. Patient-centered smart hospitals realized through PPP schemes, reshape traditional healthcare supply chains with savings and efficiency gains that improve timeliness and execution of care

Starost i rast ranih razvojnih stadija srdele (Sardina pilchardus) u zapadnom Jadranu

Age and growth of early life stages of sardine, Sardina pilchardus, were investigated by microincrement counts on sagittal otoliths. Postlarval and juvenile specimens were collected in the coastal waters off Ortona (central Adriatic Sea) from December 1996 to May 1997. Otolith microstructure analysis was conducted on 286 specimens ranging from 30 to 68 mm TL. Age estimates ranged from 65 to 151 days. The precision of age estimates was tested by applying both the average percent error (APE) and the coefficient of variation (CV). The hatch date distribution, back-calculated from the date of capture, was spread over a long period lasting from September to March, with two main cohorts hatched in autumn (October) and winter (February), consistently with a long spawning season reported for this species. Applying the Gompertz growth model to the age-length data set, a growth curve was obtained for each cohort. The mean absolute daily growth rate was 0.33 mm day-1and 0.22 mm day-1 for the autumn and winter cohorts, respectively. Otolith growth increment patterns was rather similar in the two cohorts, but they differed for the amplitude and timing of increment width deposition, being otoliths accurate proxies of larval fish growth rate. Strong seasonality of environmental features, such as water temperature and food resources, and endogenous factors linked to the larval ontogeny, largely contributed to affect the early life history stages of sardine.Istraživani su starost i rast ranih razvojnih stadija srdele Sardina pilchardus, te mikro prirast na sagitalnim otolitima. Ličinke i juvenilni uzorci prikupljeni su u obalnim vodama Ortone (srednji Jadran) od prosinca 1996. do svibnja 1997. godine. Analiza mikrostrukture otolita provedena je na 286 jedinki u rasponu od 30 do 68 mm ukupne dužine (TL). Procijenjena starost kolebala je od 65 do 151-og dana. Preciznost procjene starosti testirana je primjenom prosječnog postotka pogreške (APE) i koeficijenta varijacije (CV). Datum izvaljivanja ličinki bio je naknadno izračunat prema datumu ulova koji traje od rujna do ožujka, s dvije glavne skupine koje su se izvalile u jesen (listopad) i tijekom zime (veljača), konzistentno s ustanovljenom dugom sezonom mrijesta za ove vrste. Primjenom Gompertz modela rasta na skup podataka o starosno-duljinskom odnosu dobivena je krivulja rasta za svaku skupinu. Apsolutna srednja vrijednost dnevnog rasta iznosila je 0.33 mm dan-1 i 0.22 mm dan-1 za jesenske i zimske skupine, respektivno. Prirast otolita bio je prilično sličan u dvije skupine, mada se razlikovao s obzirom na vrijeme formiranja i učestalost istih. Jak utjecaj sezonskih ekoloških značajki, kao što su temperatura vode i izvor hrane, te endogeni čimbenici povezani s ontogenezom ličinki, u velikoj mjeri utječu na rane razvone stadije srdele

The role of geological origin of smectites and of their physico-chemical properties on aflatoxin adsorption

Since 2013, bentonite in the form of dioctahedral smectite is an additive authorised in the EU as a substance for the reduction of the contamination of feed by aflatoxins. Several studies indicate a big difference in the effectiveness of smectites in sequestering aflatoxins. A clear correlation between mineralogical and physico-chemical properties of smectites and aflatoxin adsorption has not been well established. In the effort to identify the most critical mineralogical, chemical, and physical properties that affect aflatoxin adsorption by smectites, 29 samples of bentonites obtained from different sources around the world were evaluated. “As received” samples were divided into two main groups, i.e. hydrothermal (n=14) and sedimentary (n=15) bentonites depending on their geological origin. The characterization studies showed that all samples contained dioctahedral smectite as major mineral; a moderate CEC value (60-116 cmol/kg); the presence of iron; a small organic matter content; a near-neutral pH; and a fine and uniform particle size (<45μm). They differed substantially in their sodium, calcium and magnesium contents, and in the swelling properties depending on the geological origin. Several in vitro adsorption studies showed that they also differed in a significant manner in adsorbing aflatoxin B1 (AFB1). A correlation between geological origin and AFB1 adsorption capacity was found (p<0.001), being sedimentary smectites significantly more effective than hydrothermal ones in adsorbing the toxin at different pH values. The extent of AFB1 adsorption by all samples was negatively and linearly correlated to the extent of desorption, and sedimentary smectites were significantly more effective than hydrothermal smectites in keeping bound the adsorbed fraction of the toxin (p < 0.001). In addition, correlation studies using the Pearson statistical method showed a significant relationship among some physico-chemical properties of smectites and the amounts of adsorbed toxin. In particular, AFB1 adsorption by smectites correlated positively with sodium content and swell index, but negatively with d001-value, magnesium and calcium contents. In conclusion, it seems that the geological origin of smectite is a useful guide for the selection of bentonites for AFB1 detoxification. Sedimentary bentonites containing sodium/swelling-smectite should be preferred to hydrothermal samples as potential aflatoxin binders. Taking into account the geographical origin of our samples, this approach should be applicable to bentonites worldwide

Linking neuroanatomical abnormalities in autism spectrum disorder with gene expression of candidate ASD genes: A meta-analytic and network-oriented approach

BACKGROUND: Autism spectrum disorder (ASD) is a set of developmental conditions with widespread neuroanatomical abnormalities and a strong genetic basis. Although neuroimaging studies have indicated anatomical changes in grey matter (GM) morphometry, their associations with gene expression remain elusive. METHODS: Here, we aim to understand how gene expression correlates with neuroanatomical atypicalities in ASD. To do so, we performed a coordinate-based meta-analysis to determine the common GM variation pattern in the autistic brain. From the Allen Human Brain Atlas, we selected eight genes from the SHANK, NRXN, NLGN family and MECP2, which have been implicated with ASD, particularly in regards to altered synaptic transmission and plasticity. The gene expression maps for each gene were built. We then assessed the correlation between the gene expression maps and the GM alteration maps. Lastly, we projected the obtained clusters of GM alteration-gene correlations on top of the canonical resting state networks, in order to provide a functional characterization of the structural evidence. RESULTS: We found that gene expression of most genes correlated with GM alteration (both increase and decrease) in regions located in the default mode network. Decreased GM was also correlated with gene expression of some ASD genes in areas associated with the dorsal attention and cerebellar network. Lastly, single genes were found to be significantly correlated with increased GM in areas located in the somatomotor, limbic and ganglia/thalamus networks. CONCLUSIONS: This approach allowed us to combine the well beaten path of genetic and brain imaging in a novel way, to specifically investigate the relation between gene expression and brain with structural damage, and individuate genes of potential interest for further investigation in the functional domain

Addressing charge‐transfer and locally‐excited states in a twisted biphenyl push‐pull chromophore

We present the synthesis and spectroscopic characterization of a twisted push‐pull biphenyl molecule undergoing photoinduced electron transfer. Steady‐state and transient absorption spectra suggest, in this rigid molecular structure, a subtle interplay between locally‐excited and charge‐transfer states, whose equilibrium and dynamics is only driven by solvation. A theoretical model is presented for the solvation dynamics and, with the support of quantum chemical calculations, we demonstrate the existence of two sets of states, having either local or charge‐transfer character, that only “communicate” thanks to solvation, which is the sole driving force for the charge‐separation process