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Lung cancer is one of the most aggressive malignancies, classified into two major histological subtypes: non-small cell lung cancer (NSCLC), that accounts for about 85% of new diagnosis, and small cell lung cancer (SCLC), the other 15%. In the case of NSCLC, comprehensive genome sequencing has allowed the identification of an increasing number of actionable targets, which have become the cornerstone of treatment in the advanced setting. On the other hand, the concept of oncogene-addiction is lacking in SCLC, and the only innovation of the last 30 years has been the introduction of immune checkpoint inhibitors in extensive stage disease. Dysregulation of cell cycle is a fundamental step in carcinogenesis, and Aurora kinases (AURKs) are a family of serine/threonine kinases that play a crucial role in the correct advance through the steps of the cycle. Hyperexpression of Aurora kinases is a common protumorigenic pathway in many cancer types, including NSCLC and SCLC; in addition, different mechanisms of resistance to anticancer drugs rely on AURK expression. Hence, small molecule inhibitors of AURKs have been developed in recent years and tested in several malignancies, with different results. The aim of this review is to analyze the current evidences of AURK inhibition in lung cancer, starting from preclinical rationale to finish with clinical trials available up to now.

Background Duodenal duplication cyst is an uncommon foregut malformation usually diagnosed at birth or during infancy. Differently from elsewhere, sited small bowel duplications cannot be removed with simple bowel resection, because of the proximity of the biliary and pancreatic ducts also possibly with abnormal course. Case presentation We report a duodenal duplication cyst in a newborn female requiring early surgery because of nutritional difficulties. The cyst was located adjoined to the second portion of the duodenum sharing part of its muscle wall with the bowel. It was treated by removal of all the esophytic cyst while the remaining mucosa on the common wall with the duodenum was ablated with argon plasma coagulation, preserving the bowel integrity. Early postoperative period was uneventful, and the child could be fed per os on the second day. Yearly follow-up was maintained until 16 years for the risk of recurrence and cancer change due to the incomplete excision. Clinic and echographic controls had always been stayed free from any sequelae. Conclusions Foregut duplications should be removed totally to prevent complications and the long-term risk of cancer, but a duodenal resection can be a harmful surgery in neonatal age. Duplication cysts that are impossible to remove totally can be treated by unroofing and argon plasma coagulation of mucosal surface remnants, avoiding the risks of major procedures also in newborns.

Hereditary and sporadic renal cell carcinomas (RCCs) are often associated with Von Hippel-Lindau (VHL)-gene inactivation. Patients with VHL disease have an increased risk of RCC, leading to bilateral nephrectomy and dialysis. In patients with advanced RCC, no standard second-lines are available after progression to immune checkpoint inhibitors (ICIs), and new agents are required to manage progression. HIFs have emerged as a promising target for metastatic RCC patients who have progressed to ICI-based combinations, as well as for those with RCC and VHL syndrome where the goal is to delay surgery and/or and preserve kidney function and avoid dialysis. This review describes the available evidence supporting the use of the small-molecule HIF-2 alpha inhibitor, belzutifan (MK-6482), as well as other new anti-HIF molecules that have demonstrated significant efficacy in VHL disease-related RCCs as well as for sporadic RCC that has progressed after the use of ICI-based combinations.

Stability studies performed by the pharmaceutical industry are principally designed to fulfill licensing requirements. Thus, post-dilution or post-reconstitution stability data are frequently limited to 24 h only for bacteriological reasons, regardless of the true physicochemical stability which could, in many cases, be longer. In practice, the pharmacy-based centralized preparation may require preparation in advance for administration, for example, on weekends, holidays, or in general when pharmacies may be closed. We report an innovative strategy for storing resuspended solutions of azacitidine, a well-known chemotherapic agent, for which the manufacturer lists maximum stability of 22 h. By placing the syringe with the azacitidine reconstituted suspension between two refrigerant gel packs and storing it at 4 °C, we found that the concentration of azacitidine remained above 98% of the initial concentration for 48 h, and no change in color nor the physicochemical properties of the suspension were observed throughout the study period. The physicochemical and microbiological properties were evaluated by HPLC–UV and UHPLC-HRMS analysis, FTIR spectroscopy, pH determination, visual and subvisual examination, and sterility assay. The HPLC-UV method used for evaluating the chemical stability of azacitidine was validated according to ICH. Precise control of storage temperature was obtained by a digital data logger. Our study indicates that by changing the storage procedure of azacitidine reconstituted suspension, the usage window of the drug can be significantly extended to a time frame that better copes with its use in the clinical environment.

Mesenchymal stem cells (MSCs) have been recently introduced in veterinary medicine as a potential therapeutic tool for several pathologies. The large-scale in vitro expansion needed to ensure the preparation of a suitable number of MSCs for clinical application usually requires the use of xenogeneic supplements like the fetal bovine serum (FBS). The substitution of FBS with species-specific supplements would improve the safety of implanted cells, reducing the risk of undesired immune responses following cell therapy. We have evaluated the effectiveness of canine adipose tissue-derived stromal vascular fraction (SVF) and MSCs (ADMSCs) expansion in the presence of canine blood-derived supplements. Cells were cultured on traditional plastic surface and inside a 3D environment derived from the jellification of different blood-derived products, i.e., platelet-poor plasma (PPP), platelet-rich plasma (PRP), or platelet lysate (PL). PPP, PRP, and PL can contribute to canine ADMSCs in vitro expansion. Both allogeneic and autologous PPP and PL can replace FBS for ADMSCs culture on a plastic surface, exhibiting either a similar (PPP) or a more effective (PL) stimulus to cell replication. Furthermore, the 3D environment based on homospecific blood-derived products polymerization provides a strong stimulus to ADMSCs replication, producing a higher number of cells in comparison to the plastic surface environment. Allogeneic or autologous blood products behave similarly. The work suggests that canine ADMSCs can be expanded in the absence of xenogeneic supplements, thus increasing the safety of cellular preparations. Furthermore, the 3D fibrin-based matrices could represent a simple, readily available environments for effective in vitro expansion of ADMSCs using allogeneic or autologous blood-products.

349 Background: 25-30% of renal cell carcinoma presents with metastases (mRCC) at diagnosis. The activity of immune checkpoint inhibitor (ICI)-combinations on the primary tumor (PT) is debated. Patients and Methods: mRCC patients (pts) with PT who received first-line nivolumab plus ipilimumab (N/I) or pembrolizumab plus axitinib (P/A) were included. We investigated the early primary tumor response (EPTR) at the first radiological assessment. Results: 73 pts were included. The median early reduction of the PT longest diameter was 12.4% with P/A versus 6.2% with N/I (p = 0.42). We evaluated if the type of EPTR could affect the metastases response. Among pts with PT stable disease (SD), 8.3% had metastatic disease progression (PD) with P/A and 34.8% with N/I. Early PT partial response (PR) was associated with no metastatic PD with both N/I and P/A. The 2 pts with PT PD had also metastatic PD to P/A. Of the 3 PT with PD to N/I, 1 had metastatic SD and 2 PD. In the overall population, of the 94.1% without PT progression (PR+SD), 47.5% had metastatic PR, 35.6% SD, 16.9% PD. Conclusions: ICIs-combinations achieved an early PT PR in about 10-20%, without any complete responses. Only a small percentage of PT had an early PD, mainly associated with metastatic PD. However, among those PT without an early progression, metastatic PR can be achieved in approximately 50% of cases.[Table: see text]

To assess the predictive power of the comet assay in the context of occupational exposure to pesticides.The recruited subjects completed a structured questionnaire and gave a blood sample. Exposure to pesticides was measured by means of an algorithm based on Dosemeci's work (Agricultural Health Study). Approximately 50 images were analyzed for each sample via fluorescence microscopy. The extent of DNA damage was estimated by tail moment (TM) and is the product of tail DNA (%) and tail Length.Crude significant risks (odds ratios, ORs) for values higher than the 75th percentile of TM were observed among the exposed subjects (score 1). The frequency of some confounding factors (sex, age and smoking) was significantly higher among the exposed workers. A significant dose-effect relationship was observed between TM and exposure score. Significant high-risk estimates (ORs), adjusted by the studied confounding factors, among exposure to pesticides and TM, % tail DNA and tail length were confirmed using unconditional logistic regression models.The adjusted associations (ORs) between the comet parameters and exposure to pesticides were significant. The sensitivity of the comet test was low (41%), the specificity (89%) and the predictive positive value (0.77) were found acceptable.

Background Occupational exposure is known to play a role in the aetiology of lymphomas. The aim of the present work was to explore the occupational risk of the major B-cell lymphoma subtypes using a case–control study design. Methods From 2009 to 2014, we recruited 158 lymphoma cases and 76 controls in the provinces of Bari and Taranto (Apulia, Southern Italy). A retrospective assessment of occupational exposure based on complete work histories and the Carcinogen Exposure (CAREX) job-exposure matrix was performed. Results After adjusting for major confounding factors, farmers showed an increased risk of diffuse large B-cell lymphoma (DLBCL) [odds ratio (OR) = 10.9 (2.3–51.6)] and multiple myeloma (MM) [OR = 16.5 (1.4–195.7)]; exposure to the fungicide Captafol was significantly associated with risk of non-Hodgkin lymphoma (NHL) [OR = 2.6 (1.1–8.2)], particularly with the risk of DLBCL [OR = 5.3 (1.6–17.3)]. Conclusions Agricultural activity seems to be a risk factor for developing lymphoma subtypes, particularly DLBCL, in the provinces of Bari and Taranto (Apulia Region, Southern Italy). Exposure to the pesticides Captafol, Paraquat and Radon might be implicated. Trial registration Protocol number UNIBA 2207WEJLZB_004 registered 22/09/2008.

A 3D cellular model that mimics the potential clinical application of a biomaterial is here applied for the first time to a bioactive glass, in order to assess its biological potential. A recently developed bioactive glass (BGMS10), whose composition contained strontium and magnesium, was produced in the form of granules and fully investigated in terms of biocompatibility in vitro. Apart from standard biological characterization (Simulated Body Fluid (SBF) testing and biocompatibility as per ISO10993), human bone marrow mesenchymal stromal/stem cells (BM-MSCs) were used to investigate the performance of the bioactive glass granules in an innovative 3D cellular model. The results showed that BGMS10 supported human BM-MSCs adhesion, colonization, and bone differentiation. Thus, bioactive glass granules seem to drive osteogenic differentiation and thus look particularly promising for orthopedic applications, bone tissue engineering and regenerative medicine.

Background Occupational exposure is known to play a role in the aetiology of lymphomas. The aim of the present work was to explore the occupational risk of the major B-cell lymphoma subtypes using a case–control study design. Methods From 2009 to 2014, we recruited 158 lymphoma cases and 76 controls in the provinces of Bari and Taranto (Apulia, Southern Italy). A retrospective assessment of occupational exposure based on complete work histories and the Carcinogen Exposure (CAREX) job-exposure matrix was performed. Results After adjusting for major confounding factors, farmers showed an increased risk of diffuse large B-cell lymphoma (DLBCL) [odds ratio (OR) = 10.9 (2.3–51.6)] and multiple myeloma (MM) [OR = 16.5 (1.4–195.7)]; exposure to the fungicide Captafol was significantly associated with risk of non-Hodgkin lymphoma (NHL) [OR = 2.6 (1.1–8.2)], particularly with the risk of DLBCL [OR = 5.3 (1.6–17.3)]. Conclusions Agricultural activity seems to be a risk factor for developing lymphoma subtypes, particularly DLBCL, in the provinces of Bari and Taranto (Apulia Region, Southern Italy). Exposure to the pesticides Captafol, Paraquat and Radon might be implicated. Trial registration Protocol number UNIBA 2207WEJLZB_004 registered 22/09/2008.

A 3D cellular model that mimics the potential clinical application of a biomaterial is here applied for the first time to a bioactive glass, in order to assess its biological potential. A recently developed bioactive glass (BGMS10), whose composition contained strontium and magnesium, was produced in the form of granules and fully investigated in terms of biocompatibility in vitro. Apart from standard biological characterization (Simulated Body Fluid (SBF) testing and biocompatibility as per ISO10993), human bone marrow mesenchymal stromal/stem cells (BM-MSCs) were used to investigate the performance of the bioactive glass granules in an innovative 3D cellular model. The results showed that BGMS10 supported human BM-MSCs adhesion, colonization, and bone differentiation. Thus, bioactive glass granules seem to drive osteogenic differentiation and thus look particularly promising for orthopedic applications, bone tissue engineering and regenerative medicine.

Introduction: The study of oxidative stress induced in peripheral lymphocytes by aneuploidizzanti substances may be useful to identify biological markers that predict malignancy and evaluate repair...

IntroductionWhite matter hyperintensities (WMHs) are one the most common neuroimaging findings in patients with bipolar disorder (BD). It has been suggested that WMHs are associated with impaired insight in schizophrenia and schizoaffective patients; however, the relationship between insight and WMHs in BD type I has not been directly investigated.MethodsPatients with BD-I (148) were recruited and underwent brain magnetic resonance imaging (MRI). Affective symptoms were assessed using Young Mania Rating Scale (YMRS) and Hamilton Depression Rating Scale (HDRS17); the presence of impaired insight was based on the corresponding items of YMRS and HDRS17.ResultsMultiple punctate periventricular WMHs (PWMHs) and deep WMHs (DWMHs) were observed in 49.3% and 39.9% of the cases, respectively. Subjects with lower insight for mania had significantly more PWMHs (54.6% vs 22.2%; p < 0.05) when compared to BD-I patients with higher insight for mania. The presence of PWMHs was independently associated with lower insight for mania: patients who denied illness according to the YMRS were 4 times more likely to have PWMHs (95% CI: 1.21/13.42) than other patients.ConclusionsImpaired insight in BD-I is associated with periventricular WMHs. The early identification of BD-I subjects with PWMHs and impaired insight may be crucial for clinicians.

Ricostruire in laboratorio, ossia ex situ, ghiandole endocrine bioartificiali e una prospettiva innovativa della ricerca traslazionale applicata alla medicina rigenerativa dei disturbi endocrino-metabolici. Utilizzando cellule staminali o primarie e possibile ingegnerizzare organoidi funzionali mediante ricellularizzazione sia di matrici tridimensionali (3D) acellulari allogeniche o xenogeniche, derivate da una ghiandola endocrina come quella che si vuole riprodurre, sia di supporti reticolari 3D biocompatibili amorfi o che mimano la morfologia originaria delle ghiandola, cioe organomorfi. Con questo metodo sono stati ricostruiti in modelli animali insule pancreatiche, follicoli ovarici e tiroidei, tubuli seminiferi del testicolo e parte della corteccia surrenale. Utilizzando cellule umane e stato riprodotto un abbozzo di ovaio e microaggregati di paratiroide. Le ghiandole endocrine bioartificiali promettono di fornire un’alternativa alla terapia sostitutiva con ormoni di sintesi, che richiede compliance da parte del paziente, integrandosi nei circuiti naturali di feed-back. Inoltre, quando generate con cellule autologhe e supporti organomorfi individualizzati, e plausibile riproducano una condizione secretiva simile a quella originale del soggetto donatore (terapia personalizzata). Infine, il loro uso potrebbe ridurre i costi sostenuti dal Servizio Sanitario Nazionale per le terapie croniche, superando i limiti etici e farmacologici connessi al trapianto cellulare/tissutale da cadavere e donatore vivente allogenico o xenogenico e promuovendo il mercato della salute attraverso lo sviluppo delle biotecnologie mediche, analogamente a quanto accadde, oltre un trentennio addietro, per quello delle tecnologie informatiche, con l’avvento dei semiconduttori e dei calcolatori elettronici.

We describe an innovative methodology combining Additive Layer Manufacturing (ALM) and indirect replication to reconstruct reticular-like, three-dimensional (3D) structures mimicking the vascular network of soft tissue and endocrine organs. Using a fractal-like algorithm capable of modelling the intraparenchymal vascular distribution of these viscera, single intraglandular branches of the human thyroid arteries were prototyped with synthetic resin, based on the algorithmic standard to layer (STL) output and ALM techniques. Satisfactory dimensional accuracy was obtained for these models, which were used as masters to evaluate protocols for their indirect replication, through both single and double procedures. Additional studies were conducted using casts of the human kidney arteries, obtained by injection / corrosion of the isolated organ. Satisfactory 3D reproduction of the external morphology of the kidney vessels was achieved. We conclude that our approach has the potential to develop up to the reconstr...

The high-throughput, label-free Corning Epic assay has applications in drug discovery, pharmacogenomics, cell receptor signaling, cell migration, and viral titration. The utility of Epic technology for biocompatibility testing has not been well established. In manufacturing of medical devices, in vitro and in vivo biocompatibility assessments are mandatory, according to ISO 10993. The new medical device regulation MDR 745/2017 specifies that ex vivo assays that can closely recapitulate in vivo scenarios are needed to better evaluate biomedical devices. We propose herein that Epic technology-which enables detection of variations in cell mass distribution-is suitable for biocompatibility screening of compounds. In this study, we challenged primary human osteoblasts, endothelial cells, and neurons derived from induced pluripotent stem cells with specific concentrations of methyl methacrylate (MMA). Polymeric MMA has long been applied in cranioplasty, where it makes contact with multiple cell types. Application of Epic technology yielded real-time cytotoxicity profiles for all considered cell types. The results were compared with those from microscopic observation of the same culture plate used in the Epic analyses. The Epic assay should be further examined for its utility for cell biology, genomics, and proteomics companion assays. Our results suggest that Epic technology can be applied to biocompatibility evaluation of human cells in medical device development.

This paper studies the Riboflavin release from compressed disc modules of Dome Matrix® technology using tapioca starch–ethylmethacrylate (TSEMA) and tapioca hydroxypropylstarch–ethylmethacrylate (THSEMA), graft copolymers produced by two different drying methods. The comparison with the release behaviour of similar HPMC modules was performed. Two different shape modules have been made, identified as female and male modules, in order to obtain their assemblage by interlocking the disc bases. HPMC matrices showed quasi-linear Riboflavin release in case of both female and male modules, with faster drug release than TSEMA modules. In the case of THSEMA modules, a faster release was observed compared to HPMC modules. Furthermore, matrices obtained with TSEMA copolymers remained nearly intact after dissolution process, while matrices containing HPMC experimented a complete dissolution of the modules. Combining these results with the release curve analysis using the Korsmeyer and Peppas exponential equation, HPMC modules controlled the drug release by polymer relaxation or erosion. For TSEMA and THSEMA, the drug release mechanism was controlled mainly by drug diffusion. The pronounced faster releases for the matrices containing THSEMA copolymers compared with the ones with TSEMA were due to a more important erosive support; however, the main structure of the matrix remains coherent. Porosity and tortuosity values and the shape of the modules explained the drug release observed.

Biofilms are assemblages of bacterial cells irreversibly associated with a surface where moisture is present. In particular, they retain a relevant impact on public health since through biofilms bacteria are able to survive and populate biomedical devices causing severe nosocomial infections that are generally resistant to antimicrobial agents. Therefore, controlling biofilm formation is a mandatory feature during medical device manufacturing and during their use. In this study, combining a crystal violet staining together with advanced stereomicroscopy, we report an alternative rapid protocol for both qualitative and semi-quantitative biofilm determination having high specificity, high repeatability, and low variability. The suggested approach represents a reliable and versatile method to detect, monitor, and measure biofilm colonization by an easy, more affordable, and reproducible method.

Patients with cryptorchidism can have such short spermatic vessels that it is impossible to place the testicle in a satisfactory scrotal position using conventional orchiopexy. In these cases the most commonly used operation is 1 to 2-stage Fowler-Stephens orchiopexy. We present our surgical experience using staged inguinal orchiopexy without section of the spermatic vessels in patients with short spermatic vessels.We used 2-stage inguinal orchiopexy in 38 children with intra-abdominal testis or testis peeping through the internal ring and short spermatic vessels (7 bilateral). Spermatic vessels were not sectioned, but were lengthened through progressive traction of the spermatic cord wrapped in polytetrafluoroethylene pericardial membrane (Preclude). In the first stage we mobilized the spermatic cord in the retroperitoneal space and then wrapped it in the polytetrafluoroethylene membrane. We subsequently attached the testis to the invaginated scrotal bottom. At 9 to 12 months we performed the second stage, which involved removing the polytetrafluoroethylene membrane.From the first to the second stage we observed progressive descent of the testicle toward the scrotum. At 1 to 8-year followup after the second stage all 45 testicles were palpable in a satisfactory scrotal position with stable or increased testicular volume.This technique represents an alternative to Fowler-Stephens orchiopexy, which can be associated with a greater risk of testicular ischemia.

Novel 3,4-diaryl β-lactams were prepared with high stereoselectivity in an efficient manner by a palladium-catalyzed [2+2] carbonylative cycloaddition of benzyl halides with heteroarylidene amines. The type of alkyl group linked to the nitrogen atom influences the reaction’s stereoselectivity. Moreover, using chiral imines, separable diastereomeric mixtures of chiral 3,4-diaryl-β-lactams were isolated with good yields and high trans diastereoselections.

The aim of this research was to study, in vitro by resultant-weight measurement and in vivo by γ-scintigraphy experiments in humans, the floatation behavior of systems obtained by modules assembled in void configuration. The assembled system technology allowed the manufacturing of a system characterized by the presence of an internal void space that provided an apparent density lower than water. The gastro-retention times of floating assembled systems were determined in comparison with non-floating systems having the same mass and composition. In vitro the floatation of the system started immediately after immersion in water and lasted for more than 5 h. The in vivo studies confirmed that the in vitro floating ability of void configuration was maintained also in the human stomach where the system stayed for periods of time ranging from 2.5 to 5.0 h, depending on the food regimen and the sex of the subject. Reiterate eating and drinking further prolonged the stomach residence time. © 2008 Elsevier B.V. All rights reserved.

Ricostruire in laboratorio, ossia ex situ, ghiandole endocrine bioartificiali è una prospettiva innovativa della ricerca traslazionale applicata alla medicina rigenerativa dei disturbi endocrino-metabolici. Utilizzando cellule staminali o primarie è possibile ingegnerizzare organoidi funzionali mediante ricellularizzazione sia di matrici tridimensionali (3D) acellulari allogeniche o xenogeniche, derivate da una ghiandola endocrina come quella che si vuole riprodurre, sia supporti reticolari 3D biocompatibili amorfi o che mimano la morfologia originaria delle ghiandola, cioè organomorfi. Con questo metodo sono stati ricostruiti in modelli animali insule pancreatiche, follicoli ovarici e tiroidei, tubuli seminiferi del testicolo e parte della corteccia surrenale. Utilizzando cellule umane è stato riprodotto un abbozzo di ovaio e microaggregati di paratiroide. Le ghiandole endocrine bioartificiali promet- tono di fornire un’alternativa alla terapia sostitutiva con ormoni di sintesi, che richiede compliance da parte del paziente, integrandosi nei circuiti naturali di feed-back. Inoltre, quando generate con cellule autologhe e sup- porti organomorfi individualizzati, è plausibile riproducano una condizione secretiva simile a quella originale del soggetto donatore (terapia personaliz- zata). Infine, il loro uso potrebbe ridurre i costi sostenuti dal Servizio Sanitario Nazionale per le terapie croniche, superando i limiti etici e farma- cologici connessi al trapianto cellulare/tissutale da cadavere e donatore viven- te allogenico o xenogenico e promuovendo il mercato della salute attraverso lo sviluppo delle biotecnologie mediche, analogamente a quanto accadde, oltre un trentennio addietro, per quello delle tecnologie informatiche, con l’avvento dei semiconduttori e dei calcolatori elettronici.

Regenerative medicine of endocrine glands is one of the newest fields in biomedical research, and promises to become a primary strategy to treat a number of endocrine disorders. Using computational bioengineering we have recently suggested that the three-dimensional (3D) geometry of the thyroid, stromal / vascular matrix may act as an epigenetic guidance for growth and differentiation of developing thyrocytes (1-3). To test this hypothesis, we have bioengineered ex situ (i.e. on the laboratory bench) a bioartificial rat thyroid lobe using its decellularized stromal/vascular matrix as a natural 3D scaffold, to be eventually recellularized with thyroid stem/precursor cells. Sprague-Dawley male rats (125-225 g) were used as thyroid donors, and lobe matrixes were prepared using a N2 freezing/Trypsin-EDTA / Triton-deoxycholate processing. Test matrixes were made electrondense, and analyzed by microTC (Skyscan 1172) to define their architecture. Primary thyroid cells were isolated after 72 h in primary monolayer culture, whereas presence of ABCG2-positive stem/precursor elements was determined using Western blotting. Following trypsinization, 250 - 450 x 103 cells were harvested and dropped onto the empty follicular cavities of the inner matrix of single lobe halves for homing . Cultures were kept in static conditions up to 14 days. The recellularized matrixes were either fixed in aldheydes and analyzed with light, transmission, and scanning electron microscopy, or denaturated and processed for ABCG2 immunoblotting (polyclonal rabbit anti-human 1:2000, Cell Signalling). Culture supernatants were collected every 48 h for assessment of FT3 and FT4 by immuno-chemiluminescence (Beckman-Coulter). Complete decellularization and maintenance of the 3D native architecture of the lobe matrix were achieved. Thyroid-derived cells including thyrocytes, epithelial- mesenchymal elements, and stem/precursors were found to migrate inside matrix septa, self-assemble like follicule, and recellularize the decellularized native follicular spaces. Thyroid hormone secretion occurred for at least 7 days. These results show that the 3D matrix of the rat thyroid may guide both differentiated and stem-like elements to self-assemble into functional follicular units, up to the lobar recellularization. This raises the possibility that a bioartificial, immuno-tolerant thyroid gland be bioengineered ex situ using autologous stem cells, and eventually transplanted.

We describe an innovative methodology combining Additive Layer Manufacturing (ALM) and indirect replication to reconstruct reticular-like, three-dimensional (3D) structures mimicking the vascular network of soft tissue and endocrine organs. Using a fractal-like algorithm capable of modelling the intraparenchymal vascular distribution of these viscera, single intraglandular branches of the human thyroid arteries were prototyped with synthetic resin, based on the algorithmic standard to layer (STL) output and ALM techniques. Satisfactory dimensional accuracy was obtained for these models, which were used as masters to evaluate protocols for their indirect replication, through both single and double procedures. Additional studies were conducted using casts of the human kidney arteries, obtained by injection / corrosion of the isolated organ. Satisfactory 3D reproduction of the external morphology of the kidney vessels was achieved. We conclude that our approach has the potential to develop up to the reconstruction with biomaterials of an entire, intraparenchymal vascular tree of soft tissue and endocrine organs.

We are currently developing innovative methodologies for rapid prototyping of threedimensional (3D) replicas of the vascular matrix of soft and hard tissue organs, primarily the thyroid and bones, with the intent of obtaining organomorphic scaffolds for their ex situ reconstruction (1, 2). To identify biocompatible materials suitable for this application, we have studied the effect of poly-L-lactic acid (PLLA) and poly-ε-caprolactone (PCL) on survival, adhesion and hormonal secretory activity of primary rat thyroid cells. Sprague- Dawley male rats (225-250 g) were used as thyroid donors. After penthobarbital anesthesia rats were thyroidectomised, thyroids surgically excised, and primary cells prepared using enzymatic breaking. After 72 hs in standard monolayer culture, cells were trypsinized and seeded (20 x103 / cm2) onto microporous scaffold sheets. Biomaterial scaffolds were prepared using either 3.5-5% PLLA (Lacea H100E, Mistui Chemicals) in anydrous dichloromethane / amylene (Sigma) or 3% PCL (Aldrich) in anydrous tetrahydrofurane / BHT (Aldrich). Solutions were dropped onto a wet glass support, and let polymerize as a thin layer at 25°C in the absence of forced air flow. Pattern of polymerization was assessed with thermogravimetry. Cells were let grow on biomaterials up to 8 days. Single biomaterial sheets without cells, and standard multiwell cultures were used as controls. Every 2 days, percentage of viable cells was assessed using a count chamber and Trypan blue exclusion, whereas morphology of growing cells was analyzed using a scanning electron microscopy (SEM) technique without critical point drying. Culture supernatants were collected every 48 hs, and free forms of thyroid hormone (FT3 and FT4) assessed with chemiluminescent immunoassays. PLLA and PCL scaffold sheets exhibited a quite homogeneous thickness (10-12 μm), a repetitive and regular pore geometry, and a crystalline pattern of polymerization. Both biomaterials promoted survival, adhesion and proliferation of primary thyroid elements. Secretion of FT3 and FT4 was variably maintained during the culture period, and resulted statistically higher with respect to monolayer cultures on standard multiwells. Our results indicate that PLLA and PCL are suitable for growth and differentiation of adult rat thyroid cells in culture, and suggests that they might be exploited as bioerodible materials for rapid prototyping of vascular-like, organomorphic scaffolds., Italian Journal of Anatomy and Embryology, Vol 117, No 2 (Supplement) 2012

Recently, we designed a bioreactor system for bioengineering ex situ (i.e. on the laboratory bench) a bioartificial thyroid gland suitable for transplantation. It is based on the organomorphic principle, i.e. the bioreactor mimics the macro-microscopic architecture of the thyroid stromal-vascular scaffold (SVS). To prove the reliability of this approach, we have initiated a pilot study using as a model the rat thyroid, and its natural decellularized 3D matrix to be eventually recellularized up to formation of a viable 3D thyroid lobe ex situ. Sprague-Dawley male rats (220-240 g) were used as thyroid donors. After penthobarbital anesthesia, rats were thyroidectomised and thyroid matrixes obtained by decellularization of the native SVS. Initially, we applied a sequence of liquid N2 freezing at - 80°C / thawing at 4°C for a total of 72 h, various washings with 0.02% trypsin / 0.05% EDTA for 1 h at 37°C, 3% Triton X-100 for 72 h at 4°C, and 4% deoxycholic acid for 24 h at 4°C, followed by sterilization with 0.1% peracetic acid, and 1% penicillin / streptomycin / fungizone for 24 h. Test matrixes were made electrondense with uranium / bismuth / lead counterstaining, and analyzed by microtomography (microTC). Primary thyroid cultures were prepared using enzymatic breaking of the native thyroid tissue. Cells were seeded at 19.000 / cm 2, and grown 72 h in low-glucose DMEM supplemented with 10% FBS / 5% FCS. Following trypsinization, 450.000 cells were harvested to coat the inner surface of the matrix. After 7 and 14 days, colonized matrixes were fixed in aldheydes and processed for light (LM), transmission (TEM) and scanning electron (SEM) microscopy. Culture supernatants were collected every 48 h, and thyroid hormones assessed with chemiluminescent immunoassays. Complete decellularization and maintenance of the 3D architecture of the thyroid SVS were achieved. Thyroid-derived cells were found to aggregate, link and give rise to intracytoplasmic cavities up to follicular coating, whereas secretory de-differentiation occurred. These results show that the 3D matrix of the rat thyroid can be used as a natural scaffold to recellularize the thyroid lobe with progenitor-like elements, supporting the validity of the organomorphic principle for ex situ bioengineering of a bioartificial thyroid gland. Grants FIL09, PRIN082008ZCCJX4, FIRB2010RBAP10MLK7

Ex situ bioengineering is one of the most promising perspectives in the field of regenerative medicine allowing for organ reconstruction outside the living body; i.e. on the laboratory bench. A number of hollow viscera of the cardiovascular, respiratory, genitourinary, and digestive systems have been successfully bioengineered ex situ, exploiting biocompatible scaffolds with a 3D morphology that recapitulates that of the native organ (organomorphic scaffold). In contrast, bioengineering of entire soft tissue organs and, in particular endocrine glands still remains a substantial challenge. Primary reasons are that no organomorphic scaffolding for endocrine viscera have as yet been entirely assembled using biocompatible materials, nor is there a bioreactor performance capable of supporting growth within the thickness range of the regenerating cell mass which has proven to be reliable enough to ensure formation of a complete macroscopic gland ex situ. Current technical options for reconstruction of endocrine viscera include either biocompatible 3D reticular scaffolds lacking any organomorphic geometry, or allogenic/xenogenic acellular 3D matrices derived from a gland similar to that to be bioengineered, eventually recellularized by autologous/heterologous cells. In 2007, our group designed, using biocompatible material, an organomorphic scaffold-bioreactor unit for bioengineering ex situ the human thyroid gland, chosen as a model for its simple anatomical organization (repetitive follicular cavities). This unit reproduces both the 3D native geometry of the human thyroid stromal/vascular scaffold, and the natural thyrocyte/vascular interface. It is now under intense investigation as an experimental tool to test cellular 3D auto-assembly of thyroid tissue and its related vascular system up to the ex situ generation of a 3D macroscopic thyroid gland. We believe that these studies will lay the groundwork for a new concept in regenerative medicine of soft tissue and endocrine organs: i.e. that the organomorphism of a biocompatible scaffold-bioreactor complex is essential to both the 3D organization of seeded stem cells/precursor cells and their phenotypic fate as glandular/parenchymal/vascular elements, eventually leading to a physiologically competent and immuno-tolerant bioconstruct, macroscopically suitable for transplantation and clinical applications. (C) 2011 Elsevier GmbH. All rights reserved.

The aim of this case report was to evaluate the usefulness of combined biliary and duodenal stenting in the palliation of pancreatic cancer. We report a series of 4 consecutive patients (2 men and 2 women, mean age 58.5 years, range 38-77 years) who underwent combined biliary and duodenal stenting in our department between March 2000 and April 2001. All patients had cancer of the head of the pancreas causing stricture of the common bile duct and second portion of the duodenum. Biliary and duodenal stents were successfully positioned, with relief of symptoms in all cases. No early complications were observed, except for a transient increase in serum lipase and amylase in one case. Mean follow-up was 7.5 months (range 5-14 months). One patient presenting recurrence of vomiting after 4 months because of tumour overgrowth at the distal edge of the prosthesis was successfully treated by insertion of a partially overlapping second coaxial stent. Combined biliary and duodenal stenting for the palliation of pancreatic cancer was performed safely and successfully. Stents allowed effective re-canalization of the biliary tract and duodenum, relieving both jaundice and vomiting. This procedure should be considered as an alternative to palliative surgery, especially in critically ill patients.

The aim of this open study was to compare the outcomes and side effects of plasmapheresis (PP) in patients with proliferative lupus nephritis treated with cyclophosphamide (Cyc) boluses. The study involved 28 consecutive patients. All of the patients met the ACR modified criteria for SLE and underwent a qualifying renal biopsy. In group I, patients were treated with synchronised therapy (PP, 50 ml/kg, followed by pulse Cyc, 750 mg/m(2), repeated monthly for 6 months), whereas in group II, they were given only intermittent Cyc boluses (at the same dosage). The data were collected in the patients' records according to a standardised protocol. Patients were followed-up for a mean of 4 years. The disease-free survival was analysed using Kaplan-Meier estimated survival curves ([S(t)]). At the end of the 6-month treatment period, a statistically significant number of patients in group I (75%) was in complete remission in comparison to group II (31%) (P < 0.02), whereas at long-term follow-up, these percentages were similar (41% vs. 50%, P = n.s.). The main functional and immunological parameters showed a normalisation in both groups. The risk of a poor renal outcome significantly correlated with high serum creatinine levels at the onset of nephritis (P < 0.05). We documented a higher rate of infectious complications in group I. This study reports that synchronised therapy is useful in inducing a faster remission in patients with proliferative lupus nephritis. However, it is not superior to conventional therapy at long term follow-up analysis. Positive results should be reinforced by a long-term maintenance therapy.

Novel 3,4-diaryl beta-lactams were prepared with high stereoselectivity in an efficient manner by a palladium- catalyzed [2+2] carbonylative cycloaddition of benzyl halides with heteroarylidene amines. The type of alkyl group linked to the nitrogen atom influences the reaction’s stereoselectivity. Moreover, using chiral imines, separable diastereomeric mixtures of chiral 3,4-diaryl-beta-lactams were isolated with good yields and high trans diastereoselections.

During my PhD,I have been develop an innovative technique to reproduce in vitro the 3D thymic microenvironment, to be used for growth and differentiation of thymocytes, and possible transplantation replacement in conditions of depressed thymic immune regulation. The work has been developed in the laboratory of Tissue Engineering at the University Hospital in Basel, Switzerland, under the tutorship of Prof.Ivan Martin. Since a number of studies have suggested that the 3D structure of the thymic microenvironment might play a key role in regulating the survival and functional competence of thymocytes, I’ve focused my effort on the isolation and purification of the extracellular matrix of the mouse thymus. Specifically, based on the assumption that TEC can favour the differentiation of pre-T lymphocytes, I’ve developed a specific decellularization protocol to obtain the intact, DNA-free extracellular matrix of the adult mouse thymus. Two different protocols satisfied the main characteristics of a decellularized matrix, according to qualitative and quantitative assays. In particular, the quantity of DNA was less than 10% in absolute value, no positive staining for cells was found and the 3D structure and composition of the ECM were maintained. In addition, I was able to prove that the decellularized matrixes were not cytotoxic for the cells themselves, and were able to increase expression of MHC II antigens compared to control cells grown in standard conditions. I was able to prove that TECs grow and proliferate up to ten days on top the decellularized matrix. After a complete characterization of the culture system, these innovative natural scaffolds could be used to improve the standard culture conditions of TEC, to study in vitro the action of different factors on their differentiation genes, and to test the ability of TECs to induce in vitro maturation of seeded T lymphocytes.

Adult stem cells have been recently isolated from the human and mouse thyroid. Identification has been possible by their capacity to form floating cell spheroids or thyrospheres when primary cells are cultured in the absence of serum but presence of epidermal growth factor and basic fibroblast growth factor, as well as by the presence of stem cell markers like the breast cancer-resistant protein 1 (Bcrp1)/ATP-binding cassette subfamily G member 2 (ABCG2). Using this strategy, and an innovative in vitro growing system, we have attempted identification of stem / progenitor elements from the adult rat thyroid. Sprague-Dawley male rats (50-75 gr) were used as thyroid donors. After penthobarbital anesthesia rats were thyroidectomised, thyroids surgically excised, and primary cells prepared using enzymatic breaking. After 72 hs in standard monolayer culture, cells were trypsinized and either seeded (20 x103/cm2) and grown for 8 days in a 3D Matrigel (12.5-50%) system using low-glucose DMEM and serum, or immediately cytospinned (1200 RPM x 5 min) for immunocytochemistry, or harvested and frozen with lysis buffer for Western blotting (WB). Bcrp1/ ABCG2-immunoreactivity (IR) was detected using a rabbit anti-human, polyclonal antibody (1:500, Cell Signalling), and visualized either with the ABC technique and DAB as a chromogen, or with a chemiluminescence-based staining. The human plasmocytoma cell line, RPMI 8226 (B lymphocytes) and the acute lymphoblastic leukemia cell line CCRF-CEM (T lymphocytes) were used as positive and negative controls, respectively. Thyrosphere-like aggregates were transiently observed after initial monolayer expansion and, more consistently, at day 3 in 50% Matrigel culture, followed by rapid cell differentiation (days 4-8), including epithelial-mesenchymal transitions, formation of follicles and pavment layering. Similar differentiation changes were also detected after seeding of primary thyroid cells onto decellularized rat thyroid matrixes, as previously reported [1]. Less than 0.4% of cytospinned thyroid cells exhibited cytoplasmic Bcrp1/ABCG2-IR, and a band of around 72kD was detected by WB in cell lysates. We conclude that the thyroid of the adult rat contains a small population of stem / progenitor-like elements, likely contributing to the regenerative processes that occur during ex situ recellularization of acellular thyroid matrixes [1, 2]., Italian Journal of Anatomy and Embryology, Vol 117, No 2 (Supplement) 2012

Recently, we designed a bioreactor system for bioengineering ex situ (i.e. on the laboratory bench) a bioartificial thyroid gland suitable for transplantation. It is based on the organomorphic principle, i.e. the bioreactor mimics the macro-microscopic architecture of the thyroid stromal-vascular scaffold (SVS). To prove the reliability of this approach, we have initiated a pilot study using as a model the rat thyroid, and its natural decellularized 3D matrix to be eventually recellularized up to formation of a viable 3D thyroid lobe ex situ. Sprague-Dawley male rats (220-240 g) were used as thyroid donors. After penthobarbital anesthesia, rats were thyroidectomised and thyroid matrixes obtained by decellularization of the native SVS. Initially, we applied a sequence of liquid N2 freezing at - 80°C / thawing at 4°C for a total of 72 h, various washings with 0.02% trypsin / 0.05% EDTA for 1 h at 37°C, 3% Triton X-100 for 72 h at 4°C, and 4% deoxycholic acid for 24 h at 4°C, followed by sterilization with 0.1% peracetic acid, and 1% penicillin / streptomycin / fungizone for 24 h. Test matrixes were made electrondense with uranium / bismuth / lead counterstaining, and analyzed by microtomography (microTC). Primary thyroid cultures were prepared using enzymatic breaking of the native thyroid tissue. Cells were seeded at 19.000 / cm 2, and grown 72 h in low-glucose DMEM supplemented with 10% FBS / 5% FCS. Following trypsinization, 450.000 cells were harvested to coat the inner surface of the matrix. After 7 and 14 days, colonized matrixes were fixed in aldheydes and processed for light (LM), transmission (TEM) and scanning electron (SEM) microscopy. Culture supernatants were collected every 48 h, and thyroid hormones assessed with chemiluminescent immunoassays. Complete decellularization and maintenance of the 3D architecture of the thyroid SVS were achieved. Thyroid-derived cells were found to aggregate, link and give rise to intracytoplasmic cavities up to follicular coating, whereas secretory de-differentiation occurred. These results show that the 3D matrix of the rat thyroid can be used as a natural scaffold to recellularize the thyroid lobe with progenitor-like elements, supporting the validity of the organomorphic principle for ex situ bioengineering of a bioartificial thyroid gland. Grants FIL09, PRIN082008ZCCJX4, FIRB2010RBAP10MLK7, Italian Journal of Anatomy and Embryology, Vol 116, No 1 (Supplement) 2011

Texto completo: acesso restrito. p.47–55. Submitted by Ana Valéria de Jesus Moura (anavaleria_131@hotmail.com) on 2012-02-29T17:21:05Z No. of bitstreams: 1 Affective temperamental profiles are associated with white matter.pdf: 236313 bytes, checksum: 437ecb29de9d4e5412d6bfe66d2dc6dd (MD5) Made available in DSpace on 2012-02-29T17:21:05Z (GMT). No. of bitstreams: 1 Affective temperamental profiles are associated with white matter.pdf: 236313 bytes, checksum: 437ecb29de9d4e5412d6bfe66d2dc6dd (MD5) Previous issue date: 2011 Background: Patients with white matter hyperintensities (WMH) may be at higher risk for affective disorders and suicide. Affective temperaments may play a significant role in mood disorders. This study aimed to evaluate the eventual association between WMH, affective temperaments and suicidal behaviour in major affective disorder. Methods: A total of 318 patients with major affective disorders were consecutively admitted as psychiatric inpatient. A total of 247 were included and given, brain magnetic resonance imaging (MRI) and assessed with the Mini International Neuropsychiatric Interview (MINI), the Beck Hopelessness Scale (BHS), the Hamilton Depression Rating Scale (HDRS17), the Young Mania Rating Scale (YMRS) and the Temperament Evaluation of Memphis, Pisa, Paris and San Diego (TEMPS-A). Results: A total of 48% of patients had periventricular WMH (PWMH) and 39% of them had deep WMH (DWMH). Patients with higher dysthymia and lower hyperthymia (H-DCIA group) were more likely to have higherBHS scores (BHS 9=77% vs. 52%; pN0.001),moreWMH(46% vs. 29%; 2 n=3=9.90; pb0.05), higherMINI suicidal risk (54% vs. 42%; pb0.05), and more recent suicide attempts (24% vs. 14%; pb0.05), than patients with higher hyperthymia and lower dysthymia (H-H group). Limitations: The small sample size did not allow the generalization of the present findings. Conclusions: Differences among temperament groupsmeasured by the TEMPS-Aare associatedwith differences in their MRIs, indicating that different temperament profiles are associated with differences in the subcortical structures of the brain. The implications of the resultswere discussed.

High-dose non ablative chemotherapy followed by growth factors efficiently mobilizes and amplifies Pheripheral Blood stem Cells (PBSC). Cytofluorimetric PBSC monitoring reduces the number of leukapheresis needed to collect sufficient amounts of progenitors to restore hemopoiesis after myeloablative therapy. Twenty-eight patients, affected by lymphoproliferative disorders, were primed with non myeloablative chemotherapy followed by G-CSF 5 μg/kg/die subcutaneously, until leukapheresis. A total number of 90 leukaphereses was performed (median: 3 per patient) using blood cell separator CS 3000 Plus Baxter; we collected 1±0.8 x 108/kg mononuclear cells (MNC), 6±9 x 104/kg CFU-GM and 4±5 x 106 CD34+ cells for each procedure. The statistical analysis showed that the number of progenitors collected was dependent on the age, number and type of previous chemotherapies and interval between the last chemotherapy and the priming; the type of priming, type and status of disease, sex, and bone marrow involvement were not significant. Duration of neutropenia after megachemotherapy was very short; in two cases platelet support was necessary and only two patients needed hospitalization. Our experience shows that high-dose non ablative chemotherapy followed by G-CSF is safe and yields large amounts of PBSC; several factors influence the quality of collections mainly regarding age and the previous treatment.

The aim of this research was to design and study norfloxacin (NFX) release in floating conditions from compressed hydrophilic matrices of hydroxypropylmethylcellulose (HPMC) or poly(ethylene oxide) (PEO). Module assembling technology for drug delivery system manufacturing was used. Two differently cylindrical base curved matrix/modules, identified as female and male, were assembled in void configuration by friction interlocking their concave bases obtaining a floating release system. Drug release and floatation behavior of this assembly was investigated. Due to the higher surface area exposed to the release medium, faster release was observed for individual modules compared to their assembled configuration, independently on the polymer used and concentration. The release curves analyzed using the Korsmeyer exponential equation and Peppas & Sahlin binomial equation showed that the drug release was controlled both by drug diffusion and polymer relaxation or erosion mechanisms. However, convective transport was predominant with PEO and at low content of polymers. NFX release from PEO polymeric matrix was more erosion dependent than HPMC. The assembled systems were able to float in vitro for up to 240 min, indicating that this drug delivery system of norfloxacin could provide gastro-retentive site-specific release for increasing norfloxacin bioavailability. © 2010 Elsevier B.V. All rights reserved.

This paper studies the Riboflavin release from systems made of assembled modules of Dome Matrix® technology using tapioca starch-ethylmethacrylate (TSEMA) and tapioca hydroxypropylstarch-ethylmethacrylate (THSEMA) graft copolymers produced by two different drying methods. Two different shape modules were manufactured for this study, i.e., female and male modules, in order to facilitate their assemblage in "void configuration", a system with an internal void space. Drug release studies on void configurations based on THSEMA show faster releases than TSEMA; HPMC systems used as a comparative reference showed intermediate release. Moreover, using void configurations made with one module of TSEMA and the other of THSEMA is possible to average the drug release, without difference between the drying methods used for the polymers. With respect to the floatation characteristics, all the void configurations floated immediately and, due to the mass center of the system, the floatation position of the system was always axial with the female module up and the male down. The drug release studies performed with a sinker to force the immersion of the systems in the medium did not show differences with respect to the dissolution test without a sinker. The combination of floatation capability of the assembled modules and the prolonged drug release provided with the graft copolymers make these assembled modules candidates as controlled release gastro-retentive dosage forms.

The aim of this work was to study the clindamycin release kinetics from floating delivery systems consisting of two modules assembled in void configuration, according to the modified release technology platform known as Dome Matrix®. Two modules differently shaped, i.e., female and male, formulated as swellable matrices and containing clindamycin, were assembled by friction interlocking. Then, by stacking additional female modules without drug on the assembled two-module floating system, modulation of clindamycin release rate and kinetics was attained. The additional modules stacked on the assembled system acted as a transient barrier to clindamycin release from the void configuration. Inertness, dissolution/erosion or swelling behavior characterized their performance as matrices in simulated gastric fluid. In particular, we found that stacking additional barrier modules on the bases of void configuration, the drug release rate and kinetics of the assembled system were modified in dependence on the composition of module added. In particular, the quickly soluble module exerted an influence on the release rate in the late time of delivery. The swellable module produced a significant reduction in release rate of void assembly, but the release mechanism remained the same. Finally, the inert module led to a substantial linearization of the release profile with a minimal reduction in release rate.

Patients with white matter hyperintensities (WMH) may be at higher risk for affective disorders and suicide. Affective temperaments may play a significant role in mood disorders. This study aimed to evaluate the eventual association between WMH, affective temperaments and suicidal behaviour in major affective disorder.A total of 318 patients with major affective disorders were consecutively admitted as psychiatric inpatient. A total of 247 were included and given, brain magnetic resonance imaging (MRI) and assessed with the Mini International Neuropsychiatric Interview (MINI), the Beck Hopelessness Scale (BHS), the Hamilton Depression Rating Scale (HDRS(17)), the Young Mania Rating Scale (YMRS) and the Temperament Evaluation of Memphis, Pisa, Paris and San Diego (TEMPS-A).A total of 48% of patients had periventricular WMH (PWMH) and 39% of them had deep WMH (DWMH). Patients with higher dysthymia and lower hyperthymia (H-DCIA group) were more likely to have higher BHS scores (BHS≥9=77% vs. 52%; p0.001), more WMH (46% vs. 29%; χ(2)(n=3)=9.90; p0.05), higher MINI suicidal risk (54% vs. 42%; p0.05), and more recent suicide attempts (24% vs. 14%; p0.05), than patients with higher hyperthymia and lower dysthymia (H-H group).The small sample size did not allow the generalization of the present findings.Differences among temperament groups measured by the TEMPS-A are associated with differences in their MRIs, indicating that different temperament profiles are associated with differences in the subcortical structures of the brain. The implications of the results were discussed.

The aim of this work was to study a multi-kinetics and site-specific oral antimalaria drug delivery system (MKS_DDS), containing artesunate and clindamycin, based on the Dome Matrix® module assembly technology. The MKS_DDS assembled system comprises of four modules, i.e., two controlled release (CR) modules for delivery of 160. mg of clindamycin phosphate, one immediate release module containing 50. mg of artesunate and one immediate release module containing 80. mg of clindamycin phosphate. These modules have been assembled in stacked and void configurations. The void configuration is able to float and showed gastro-retentive behavior. The MKS_DDS was investigated for its mechanical characteristics, system behavior during release, drug release rate and mechanism.A bioavailability study (dogs) showed that the clindamycin plasma curve of the MKS_DDS system exhibited a quasi constant release rate, up to 8. h.The MKS_DDS system containing clindamycin and artesunate allows the use of one tablet containing one immediate release dose of artesunate and of clindamycin and a portion of clindamycin released over a prolonged time, by exploiting the gastro-retentive properties of a floating system. © 2010 Elsevier B.V.

Novel 3,4-diaryl β-lactams were prepared with high stereoselectivity in an efficient manner by a palladium-catalyzed [2+2] carbonylative cycloaddition of benzyl halides with heteroarylidene amines. The type of alkyl group linked to the nitrogen atom influences the reaction’s stereoselectivity. Moreover, using chiral imines, separable diastereomeric mixtures of chiral 3,4-diaryl-β-lactams were isolated with good yields and high trans diastereoselections.

La tecnologia di assemblaggio dei moduli di rilascio, denominata Dome Matrix®, è un’alternativa alle esistenti tecnologie per la somministrazione di farmaci in modalità controllata. Il modulo o unità di rilascio (che semplificando può essere assimilata ad una compressa) esibente una propria cinetica di liberazione del farmaco. Il modulo Dome Matrix è una compressa avente forma di disco con una base concava e una convessa. La particolare forma è funzionale all’assemblaggio. Esistono due tipi di moduli: maschi e femmina. I moduli maschi presentavano una protrusione anulare sulla faccia concava, assente sul modulo femmina. I moduli di rilascio possono essere assemblati in due configurazioni definite a camera vuota e stacked. Assemblando due o più moduli è possibile ottenere un rilascio nel tempo e nello spazio che dipende fortemente da come i moduli sono stati messi insieme. Lo scopo che in questa tesi è quello di produrre un sistema di rilascio di farmaco multi cinetico e sito specifico per l’associazione di farmaci per cura della malaria utilizzando la tecnologia modulare Dome Matrix®. Nella prima parte della ricerca è stata studiata la fattibilità della produzione per compressione di moduli Dome Matrix® maschio e femmina a partire da granulati ottenuti con una tecnica di granulazione in high shear mixer. Lo studio di compressione di granulati di alginato ha mostrato come la diversa forma dei moduli influenzi la durezza e la friabilità dei compatti ottenuti. Nel caso dei moduli maschio era possibile nella maggior parte dei casi ottenere compresse dure e poco friabili; di contro dei moduli femmina questo avveniva solo per granulati aventi dimensioni inferiori a 500µm con bassa friabilità e quando suddetti granuli venivano compressi con una forza di 30KN. Nella seconda parte della ricerca è stato studiato il processo di rivestimento in letto fluido dei moduli Dome Matrix®. La ricopertura dei moduli non è uguale su tutta la superficie: a causa della sua forma a cupola entrambi in moduli mostrano una differente ricopertura nella parte concava e convessa. Lo strato di rivestimento presente sui moduli Dome Matrix® influenza la forza con cui i moduli sono assemblati in configurazione a camera vuota mediante incastro delle reciproche facce concave di un modulo Dome Matrix® maschio e uno femmina. Nella terza parte della ricerca è stato affrontato lo studio delle proprietà di galleggiamento in vivo e in vitro di una formulazione placebo costruita assemblando i moduli Dome Matrix® in configurazione a camera vuota. In vitro il sistema si dimostra capace di galleggiare immediatamente mantenendo inalterata questa caratteristica per più di 9 ore. In vivo nell'ambiente gastrico il sistema rimane nello stomaco per un periodo di tempo compreso fra le 2.5 e le 5 ore, la lunghezza di questo intervallo dipende sia dal regime dietetico sia sesso del soggetto. Nella quarta ed ultima parte della ricerca è stato progettata e realizzata una forma farmaceutica a rilascio modificato contenente Clindamicina e Artesunato per la cura dell’infezione malarica utilizzando la tecnologia Dome Matrix®. Tale assemblato in vitro si comporta nella seguente maniera: il sistema affonda rapidamente e il modulo a rilascio immediato di artesunato disgrega in meno di un minuto rilasciando in 30 minuti circa 80% della dose. Il modulo di clindamicina, anch’esso a rilascio immediato, disgrega in 3 minuti liberando coi i due moduli assemblati in configurazione a camera vuota e rilasciando in 10 minuti un terzo della dose totale somministrata. A seguito della disgregazione dei moduli esterni l’assemblato in configurazione a camera vuota viene distaccato dal sistema a quattro moduli raggiungendo una densità che gli consente di riemergere e galleggiare. Il resto della dose di clindamicina viene quindi rilasciata in maniera graduale per più di 10 ore. I test preliminari in vivo condotti su tre cani di razza beagle ha mostrato che il sistema terapeutico è in grado di mantener concentrazioni plasmatiche di farmaco più alte rispetto alla M.I.C.

A veiculação de fármacos para o cólon tem atraído a atenção de muitos investigadores interessados no tratamento de patologias locais e no seu potencial para a veiculação de proteínas e peptídeos. para desenvolver estes sistemas é fundamental compreender o cólon enquanto local de veiculação de fármacos, em particular a sua capacidade de absorção e o tempo necessário para o atingir. Dois tipos de abordagens podem ser utilizados, libertação retardada ou libertação específica no cólon. o primeiro caso é menos específico e consiste em formulações preparadas de forma a retardar a libertação de fármaco até o sistema farmacêutico atingir o cólon. o segundo caso explora as propriedades de pH, actividade enzimática e pressão intraluminal do cólon para promover a libertação colónica de fármacos. colon targeting has been attracting interest for those interested in treating local pathologies and administrating proteins and peptides. in order to develop these systems a perfect understanding of the colonic region is compulsory. two main strategies are used: delayed release and specific release. the first one, less specific, consists in postponing release from the system until it reaches de colon. the second one uses colon properties as pH, enzymatic activity and intraluminal pressure to trigger colonic drug release.