The effect of bone allografts combined with bone marrow stromal cells on the healing of segmental bone defects in a sheep model

Background: The repair of large bone defects is a major orthopedic challenge because autologous bone grafts are not available in large amounts and because harvesting is often associated with donor-site morbidity. Considering that bone marrow stromal cells (BMSC) are responsible for the maintenance of bone turnover throughout life, we investigated bone repair at a site of a critically sized segmental defect in sheep tibia treated with BMSCs loaded onto allografts. The defect was created in the mid-portion of the tibial diaphysis of eight adult sheep, and the sheep were treated with ex-vivo expanded autologous BMSCs isolated from marrow aspirates and loaded onto cortical allografts (n = 4). The treated sheep were compared with control sheep that had been treated with cell-free allografts (n = 4) obtained from donors of the same breed as the receptor sheep. Results: The healing response was monitored by radiographs monthly and by computed tomography and histology at six, ten, fourteen, and eighteen weeks after surgery. For the cell-loaded allografts, union was established more rapidly at the interface between the host bone and the allograft, and the healing process was more conspicuous. Remodeling of the allograft was complete at 18 weeks in the cell-treated animals. Histologically, the marrow cavity was reestablished, with intertrabecular spaces being filled with adipose marrow and with evidence of focal hematopoiesis. Conclusions: Allografts cellularized with AOCs (allografts of osteoprogenitor cells) can generate great clinical outcomes to noncellularized allografts to consolidate, reshape, structurally and morphologically reconstruct bone and bone marrow in a relatively short period of time. These features make this strategy very attractive for clinical use in orthopedic bioengineering

Application-oriented Mixed Integer Non-Linear Programming

In the most recent years there is a renovate interest for Mixed Integer Non-Linear Programming (MINLP) problems. This can be explained for different reasons: (i) the performance of solvers handling non-linear constraints was largely improved; (ii) the awareness that most of the applications from the real-world can be modeled as an MINLP problem; (iii) the challenging nature of this very general class of problems. It is well-known that MINLP problems are NP-hard because they are the generalization of MILP problems, which are NP-hard themselves. However, MINLPs are, in general, also hard to solve in practice. We address to non-convex MINLPs, i.e. having non-convex continuous relaxations: the presence of non-convexities in the model makes these problems usually even harder to solve. The aim of this Ph.D. thesis is to give a flavor of different possible approaches that one can study to attack MINLP problems with non-convexities, with a special attention to real-world problems. In Part 1 of the thesis we introduce the problem and present three special cases of general MINLPs and the most common methods used to solve them. These techniques play a fundamental role in the resolution of general MINLP problems. Then we describe algorithms addressing general MINLPs. Parts 2 and 3 contain the main contributions of the Ph.D. thesis. In particular, in Part 2 four different methods aimed at solving different classes of MINLP problems are presented. Part 3 of the thesis is devoted to real-world applications: two different problems and approaches to MINLPs are presented, namely Scheduling and Unit Commitment for Hydro-Plants and Water Network Design problems. The results show that each of these different methods has advantages and disadvantages. Thus, typically the method to be adopted to solve a real-world problem should be tailored on the characteristics, structure and size of the problem. Part 4 of the thesis consists of a brief review on tools commonly used for general MINLP problems, constituted an integral part of the development of this Ph.D. thesis (especially the use and development of open-source software). We present the main characteristics of solvers for each special case of MINLP.

Carlos Martí Arís e i suoi eteronimi. Vocazione all'anonimo

Questa ricerca indaga la prospettiva investigativa di Carlos Martí Arís. È stato assunto il campo d’azione da lui prediletto, ovvero l’articolato rapporto che in architettura si instaura tra teoria e pratica, comprensivo delle svariate ricadute nel mondo dell’arte e della produzione umana in genere, che fanno del progetto architettonico un campo disciplinare complesso. La sua figura è però assunta in modo strumentale, come grimaldello per addentrarsi in un articolato ambito culturale, che se da un lato coincide con la sua città, Barcellona, dall’altro la trascende grazie a quei “ponti della conoscenza” che CMA interrottamente ha teso al suo intorno. Ci riferiamo alla sua costruzione teorica destinata a consolidare la storica reciprocità tra Italia e Spagna, le cui tematiche urbane e tipologiche ne sono la base, Milano e Barcellona ne sono gli estremi. Ci riferiamo al suo sguardo sull’esperienza del Movimento Moderno e il relativo tema della residenza. Ci riferiamo alla sua naturale vocazione al silenzio, che si oppone al fragoroso rumore della contemporaneità e affianca la discreta parola del mestiere: un modo per porsi all’ascolto. All’ascolto dell’altro e del mondo. Ci riferiamo, insomma, alla sua idea di architettura intesa come «territorio dissodato da tempi remoti»; come trama di corrispondenze sincroniche tra terre, tempi, fatti, uomini, vicini e lontani: condizione ideale per chi voglia disciogliere il proprio lavoro nei labirintici sentieri del mondo, indifferente al rischio di perdersi nell’oblio. Oggi, in cui il progetto architettonico risulta sempre più spesso veicolo di soggettive e arbitrarie sperimentazioni formali, la lezione di CMA indica una via d’uscita: un mo(n)do condiviso che all’arroganza opponga la discrezione, che persuada a celarsi nella tradizione e a porsi umilmente all’ombra dei maestri. Tradizione e Maestri, Eteronimi e Nomi, complementarità a cui CMA affida il suo progetto di anonimato, sovrapersonale e ostinatamente teso a rilevarne le relazioni inedite.

Software Defined Networks: alcuni casi di studio

In questo elaborato si descrive l'emergente approccio alle reti, il Software Defined Network, ed i suoi benefici. Successivamente viene preso in considerazione un importante componente di questa nuova architettura: il protocollo OpenFlow; si spiega che cos'è e si elencano i benefici che può apportare ad un'architettura SDN a sostegno di questi vengono mostrati quattro differenti casi d'uso di OF, comparati poi ad altri scenari equivalenti che non usano questo protocollo. Infine si è pensato ad alcuni possibili studi e sviluppi circa quest'architettura.

Effetto dei farmaci presenti in ambiente sulle larve di orata (Sparus aurata)

La presenza di residui dei farmaci ad uso umano e veterinario nelle acque superficiali è in costante aumento a causa del loro elevato consumo. L’impatto ambientale dei prodotti farmaceutici è riconosciuto in tutto il mondo ma attualmente ancora non sono presenti degli Standard di qualità ambientale per queste sostanze in ambiente acquatico. L’agenzia europea per i farmaci (EMEA) ha introdotto delle linee guida per la valutazione del rischio ambientale per tutti i nuovi farmaci prima di provvedere alla registrazione, ma in nessun caso la loro autorizzazione in commercio è vietata. Una volta assunti, i farmaci sono escreti dagli organismi in forma nativa o come metaboliti, e attraverso gli scarichi urbani raggiungono i depuratori che li rimuovono solo in parte. Di conseguenza, i residui dei farmaci vengono ritrovati nei fiumi, nei laghi, fino alle acque marine costiere. Anche se presenti a basse concentrazioni (ng-μg/L) nelle acque superficiali, i farmaci possono provocare effetti avversi negli organismi acquatici. Queste specie rappresentano involontari bersagli. Tuttavia molti di essi possiedono molecole target simili a quelle dell’uomo, con i quali i farmaci interagiscono per indurre gli effetti terapeutici; in questo caso i farmaci ambientali possono causare effetti specifici ma indesiderati sulla fisiologia degli animali acquatici. Le interazioni possono essere anche non specifiche perché dovute agli effetti collaterali dei farmaci, ad esempio effetti ossidativi, con potenziali conseguenze negative su vertebrati ed invertebrati. In questo lavoro sono stati valutati i potenziali effetti indotti nelle larve di orata da quattro classi di farmaci ovvero: carbamazepina (antiepilettico), ibuprofene (antinfiammatorio non steroideo), caffeina (stimolante) e novobiocina (antibiotico). In particolare, in questo lavoro si è valutato inizialmente il tasso di sopravvivenza delle larve di orata esposte ai farmaci, per verificare se l’esposizione determinasse effetti di tossicità acuta; successivamente si è passati alla valutazione di due biomarker : il danno al DNA e la perossidazione lipidica per verificare la presenza di effetti tossici sub-letali. Le larve sono state esposte per 96 ore alle concentrazioni di 0.1, 1 (MEC), 10 e 50 µg/L (>MEC) di carbamazepina e novobiocina, a 0.1, 5 (MEC),10 e 50 µg/L (> MEC) di ibuprofene ed a 0.1, 5 (MEC),15 e 50 µg/L (> MEC) di caffeina, rappresentative delle concentrazioni riscontrate in ambiente acquatico e al di sopra di quest’ultimo. L’analisi dei dati sulla sopravvivenza ha dimostrato che la carbamazepina, l’ibuprofene, la novobiocina e la caffeina non hanno effetti significativi alle concentrazioni testate. La valutazione dei biomarker ha evidenziato un generale decremento significativo dei livelli di danno primario al DNA e per la perossidazione lipidica è stato generalmente osservato un decremento alle dosi dei farmaci più basse, seguito da un aumento a quelle più elevate. Nell’insieme i dati indicano che alle concentrazioni testate, i farmaci carbamazepina, caffeina, ibuprofene e novobiocina non hanno prodotto alterazioni attribuibili alla comparsa di effetti avversi nelle larve di S. aurata dopo 96 ore di esposizione.

Comparison of the binding and internalization properties of 12 DOTA-coupled and ¹¹¹In-labelled CCK2/gastrin receptor binding peptides: a collaborative project under COST Action BM0607

Specific overexpression of cholecystokinin 2 (CCK2)/gastrin receptors has been demonstrated in several tumours of neuroendocrine origin. In some of these cancer types, such as medullary thyroid cancer (MTC), a sensitive diagnostic modality is still unavailable and therapeutic options for inoperable lesions are needed. Peptide receptor radionuclide therapy (PRRT) may be a viable therapeutic strategy in the management of these patients. Several CCK2R-targeted radiopharmaceuticals have been described in recent years. As part of the European Union COST Action BM0607 we studied the in vitro and in vivo characteristics of 12 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA)-conjugated CCK2R binding peptides. In the present study, we analysed binding and internalization characteristics. Stability, biodistribution and imaging studies have been performed in parallel by other centres involved in the project.

MicroRNA-21 suppression impedes medulloblastoma cell migration

Medulloblastoma (MB), the most common malignant brain tumour in children, is characterised by a high risk of leptomeningeal dissemination. But little is known about the molecular mechanisms that promote cancer cell migration in MB. Aberrant expression of miR-21 is recognised to be causatively linked to metastasis in a variety of human neoplasms including brain tumours; however its function in MB is still unknown. In this study we investigated the expression level and the role of miR-21 in MB cell migration. miR-21 was found to be up-regulated, compared to normal cerebellum, in 29/29 MB primary samples and 6/6 MB-derived cell lines. Inverse correlation was observed between miR-21 expression and the metastasis suppressor PDCD4, while miR-21 repression increased the release of PDCD4 protein, suggesting negative regulation of PDCD4 by miR-21 in MB cells. Anti-miR-21 decreased protein expression of the tumour cell invasion mediators MAP4K1 and JNK, which are also known to be negatively regulated by PDCD4, and down-regulated integrin protein that is essential for MB leptomeningeal dissemination. Moreover miR-21 knockdown in MB cells increased the expression of two eminent negative modulators of cancer cell migration, E-Cadherin and TIMP2 proteins that are known to be positively regulated by PDCD4. Finally and importantly, suppression of miR-21 decreased the motility of MB cells and reduced their migration across basement membranes in vitro. Together, these compelling data propose miR-21 pathway as a novel mechanism impacting MB cell dissemination and raises the possibility that curability of selected MB may be improved by pharmaceutical strategies directed towards microRNA-21.

Rumen-protected choline supplementation in periparturient dairy goats: effects on liver and mammary gland

The current study investigated the effects of supplementing rumen-protected choline (RPC) on metabolic profile, selected liver constituents and transcript levels of selected enzymes, transcription factors and nuclear receptors involved in mammary lipid metabolism in dairy goats. Eight healthy lactating goats were studied: four received no choline supplementation (CTR group) and four received 4g RPC chloride/day (RPC group). The treatment was administered individually starting 4 weeks before expected kidding and continuing for 4 weeks after parturition. In the first month of lactation, milk yield and composition were measured weekly. On days 7, 14, 21 and 27 of lactation, blood samples were collected and analysed for glucose, beta-hydroxybutyrate, non-esterified fatty acids and cholesterol. On day 28 of lactation, samples of liver and mammary gland tissue were obtained. Liver tissue was analysed for total lipid and DNA content; mammary tissue was analysed for transcripts of lipoprotein lipase (LPL), fatty acid synthase (FAS), sterol regulatory binding proteins 1 and 2, peroxisome proliferator-activated receptor gamma and liver X receptor alpha. Milk yield was very similar in the two groups, but R PC goats had lower (P < 0.05) plasma beta-hydroxybutyrate. The total lipid content of liver was unaffected (P = 0.890), but the total lipid/DNA ratio was lower (both P < 0.05) in RPC than CTR animals. Choline had no effect on the expression of the mammary gland transcripts involved in lipid metabolism. The current plasma and liver data indicate that choline has a positive effect on liver lipid metabolism, whereas it appears to have little effect on transcript levels in mammary gland of various proteins involved in lipid metabolism. Nevertheless, the current results were obtained from a limited number of animals, and choline requirement and function in lactating dairy ruminants deserve further investigation.