Circulating microparticles from septic shock patients exert differential tissue expression of enzymes related to inflammation and oxidative stress

Objective: Septic shock is characterized by hypotension and multiple organ failure after infection of microorganisms. Septic shock patients display high levels of circulating microparticles. These are small vesicles released from the plasma membrane of activated or apoptotic cells. Here, we have investigated the effects of in vivo injection of microparticles from nonseptic or septic subjects on protein expression in mouse tissues.Design: Prospective, controlled experiments. Setting: Animal basic science laboratory. Subjects: Male Swiss mice were randomly assigned to one of two groups: 11 animals injected with microparticles isolated from healthy subjects and 15 animals injected with microparticles isolated from septic patients. Interventions: Microparticles were extracted from whole blood of septic and nonseptic subjects and were intravenously injected in mice. After 24 hrs, mice were killed and heart, lungs, liver, and kidneys were isolated for Western blot assays. Organs were also used for direct measurements of nitric oxide and superoxide anion production by electron paramagnetic resonance. Measurements and Main Results: In heart and lungs, microparticles from septic shock patients increased the expression of endothelial and inducible nitric oxide synthases, cyclooxygenase-2, and nuclear factor-κB. However, extracellular superoxide dismutase was only increased in the heart. These effects were associated either with a greater oxidative or nitrative stress in heart and lungs, without affecting nitric oxide production. The liver exhibited an increase in oxidative stress linked to decreased endothelial nitric oxide synthase and manganese superoxide dismutase expression. However, cyclooxygenase-2 expression and IκBα phosphorylation were decreased. Septic microparticles did not change superoxide anion and nitric oxide productions in kidneys. Conclusions: Results suggest that microparticles from septic shock patients exert pleiotropic and differential effects depending on target tissues with regard to the expression of proinflammatory proteins related with nitrative and oxidative stresses. Thus, microparticles might participate in organ dysfunction observed in septic shock patients

Design, fabrication and characterization of piezoelectric cantilever MEMS for underwater application

This work shows a preliminary microfabrication route for a novel directional hydrophone based on a cross-shaped design of piezoelectric cantilevers. A thin layer of aluminum nitride (AlN) using Molybdenum (Mo) thin film as electrodes will be exploited as piezoelectric functional layer for the microfabrication of a cantilever-based ultrasonic micro electro mechanical system (MEMS) hydrophone. A parameterized simulation based on length of these cantilevers between 100 and 1000 μm allowed to set the first resonant mode between 20 kHz and 200 kHz, the desired underwater ultrasonic acoustic range. The microsystem was designed with cantilevers facing each other in a cross configuration in order to have novel MEMS hydrophone with an omnidirectional response. In order to investigate the first resonance frequency mode and displacement measurements, a Laser Doppler Vibrometer was used and good agreement between simulations and experimental results was achieved. Responsivity and directionality measurements of the piezoelectric MEMS cantilevers were performed in water. Maximum sensitivity up to −153 dB with omnidirectional directivity pattern was achieved by fabricated MEMS sensor

Wearable piezoelectric mass sensor based on pH sensitive hydrogels for sweat pH monitoring

Colorimetric and electrochemical (bio)sensors are commonly employed in wearable platforms for sweat monitoring; nevertheless, they suffer from low stability of the sensitive element. In contrast, mass-(bio)sensors are commonly used for analyte detection at laboratory level only, due to their rigidity. To overcome these limitations, a flexible mass-(bio)sensor for sweat pH sensing is proposed. The device exploits the flexibility of piezoelectric AlN membranes fabricated on a polyimide substrate combined to the sensitive properties of a pH responsive hydrogel based on PEG-DA/CEA molecules. A resonant frequency shift is recorded due to the hydrogel swelling/shrinking at several pH. Our device shows a responsivity of about 12 kHz/pH unit when measured in artificial sweat formulation in the pH range 3-8. To the best of our knowledge, this is the first time that hydrogel mass variations are sensed by a flexible resonator, fostering the development of a new class of compliant and wearable devices

Influence of micropaticles harvested from patients affected by obstructive sleep apnea syndrome on endothelial function and vascular reactivity

Obstructive sleep apnea syndrome (OSAS) is a highly prevalent disease characterized by recurrent episodes of partial or complete obstruction of the upper airways during sleep, leading to oxygen desaturation, sleep fragmentation and clinical endothelial dysfunction. Microparticles (MPs) are membrane vesicles released during cell activation and apoptosis. Elevated levels of circulating MPs have been detected in pathologies associated with vascular alterations. We investigated the effects of MPs on endothelial function and vascular reactivity in OSAS. Blood samples were obtained either from 38 OSAS patients without any other cardiovascular comorbidities and 23 healthy subjects. A desaturation index above 10 per hour defined OSAS patients. MPs concentration and origin were assessed using flow cytometer. Male Swiss mice were injected i.v. with MPs from OSAS or healthy subjects, or with saline solution, and sacrified after 24hours. Endothelial function and vascular reactivity were studied on aortic rings and small mesenteric resistance (SMA) arteries by myography and arteriography, respectively. Patients with OSAS did not display increased circulating levels of MPs compared to healthy subjects including those from pro-coagulant, platelet, endothelial, leukocyte and erythrocyte origins. Interestingly, MPs from granulocytes and activated leukocytes were significantly enhanced in OSAS patients. Activated leukocyte MPs positively correlated with oxygen desaturation index. In aorta, MPs from OSAS patients but not those from healthy subjects significantly reduced endothelium-dependent relaxation to acetylcholine. MPs from OSAS increased sensitivity of the aorta in response to serotonin that was greater compared to the effect of MPs from healthy subjects. In SMA, MPs from OSAS but not those from healthy subjects impaired flow-induced dilation without any effect on myogenic tone. Although SMA from mice treated with healthy subjects MPs did not affect flow-induced dilation, these vessels showed a reduced prostacyclin-component that was completely compensated by the NO-component of the response. The endothelial dysfunction induced by MPs from OSAS was caused by the reduction of both NO- and prostacyclin- but not the endothelium-derived hyperpolarizing factor-components of the response in SMA. These data provide evidence that circulating MPs from OSAS patients influence both endothelial function and vascular reactivity

Efficient synthesis of kainic acid analogues

The present paper deals with an improved synthesis of two molecular hybrids of AMPA and KA, compounds CIP-A and CIP-B, and their transformation into CIOP-A and CIOP-B, the corresponding amido derivatives. Exploiting the continuous-flow technology, a significant improvement in the synthesis of the glutamate agonists CIP-A and CIP-B was accomplished, in terms of overall yield, time, and excess of ethyl chlorooximinoacetate. Moreover, we find out the HPLC conditions suitable to separate, at a preparative level, the three intermediates formed in the 1,3-dipolar cycloaddition step

Development and characterization of a novel hybrid tissue engineering-based scaffold for spinal cord injury repair

Spinal cord injury (SCI) represents a significant health and social problem, and therefore it is vital to develop novel strategies that can specifically target it. In this context, the objective of the present work was to develop a new range of three-dimensional (3D) tubular structures aimed at inducing the regeneration within SCI sites. Up to six different 3D tubular structures were initially developed by rapid prototyping: 3D bioplotting–based on a biodegradable blend of starch. These structures were then further complemented by injecting Gellan Gum, a polysaccharide-based hydrogel, in the central area of structures. The mechanical properties of these structures were assessed using dynamic mechanical analysis, under both dry and wet conditions, and their morphologies= porosities were analyzed using micro-computed tomography and scanning electron microscopy. Biological evaluation was carried out to determine their cytotoxicity, using both minimum essential medium (MEM) extraction and MTS tests, as well as by encapsulation of an oligodendrocyte-like cell (M03-13 cell line) within the hydrogel phase. The histomorphometric analysis showed a fully interconnected network of pores with porosity ranging from 70% to 85%. Scaffolds presented compressive modulus ranging from 17.4 to 62.0MPa and 4.42 to 27.4 MPa under dry and wet conditions, respectively. Cytotoxicity assays revealed that the hybrid starch=poly-ecaprolactone= Gellan Gum scaffolds were noncytotoxic, as they did not cause major alterations on cell morphology, proliferation, and metabolic viability. Moreover, preliminary cell encapsulation assays showed that the hybrid scaffolds could support the in vitro culture of oligodendrocyte-like cells. Finally, preliminary in vivo studies conducted in a hemisection rat SCI model revealed that the above-referred structures were well integrated within the injury and did not trigger chronic inflammatory processes. The results herein presented indicate that these 3D systems might be of use in future SCI regeneration approaches.Portuguese Foundation for Science and Technology through funds from Programa Operacional Ciencia, Tecnologia, Inovacao (POCTI) and/or Fundo Europeu de Desenvolvimento Regional (FEDER) programs (funding to ICVS, 3B's Research Group, predoctoral and postdoctoral fellowships to N. A. Silva, J. T. Oliveira, A. J. Salgado, and R. A. Sousa-SFRH/BD/40684/2007; SFRH/BD/17135/2004; SFRH/BPD/17595/2004; SFRH/BPD/17151/2004)

Dysbiosis triggers ACF development in genetically predisposed subjects

Background: Colorectal cancer (CRC) is the third most common cancer worldwide, characterized by a multifactorial etiology including genetics, lifestyle, and environmental factors including microbiota composition. To address the role of microbial modulation in CRC, we used our recently established mouse model (theWinnie-APCMin/+) combining inflammation and genetics. Methods: Gut microbiota profiling was performed on 8-week-old Winnie-APCMin/++ mice and their littermates by 16S rDNA gene amplicon sequencing. Moreover, to study the impact of dysbiosis induced by the mother’s genetics in ACF development, the large intestines of APCMin/++ mice born from wild type mice were investigated by histological analysis at 8 weeks. Results: ACF development in 8-week-old Winnie-APCMin/++ mice was triggered by dysbiosis. Specifically, the onset of ACF in genetically predisposed mice may result from dysbiotic signatures in the gastrointestinal tract of the breeders. Additionally, fecal transplant from Winnie donors to APCMin/++ hosts leads to an increased rate of ACF development. Conclusions: The characterization of microbiota profiling supporting CRC development in genetically predisposed mice could help to design therapeutic strategies to prevent dysbiosis. The application of these strategies in mothers during pregnancy and lactation could also reduce the CRC risk in the offspring

Synthesis of unusual isoxazoline containing β and γ-dipeptides as potential glutamate receptor ligands

New unconventional beta and gamma dipeptides, representing conformationally constrained higher homologues of glutamic acid, have been prepared and tested as new pharmacological tools to investigate the iGluR binding domain, in an attempt to identify potential selective antagonists

Synthesis of L-tricholomic acid analogues and pharmacological characterization at ionotropic glutamate receptors

The synthesis of analogues of the natural compound ltricholomic acid and of its threo diastereoisomer was accomplished in order to explore their affinity for glutamate ionotropic receptors. In this study, fourteen new unnatural amino acids, characterized by a 3-hydroxy-~2 -isoxazoline or 3- hydroxy-~2 -pyrazoline-skeleton, were obtained exploiting, as key reaction, a 1,3-dipolar cycloaddition or an intramolecular cyclization

The use of FDG-PET in the initial staging of 142 patients with follicular lymphoma: a retrospective study from the FOLL05 randomized trial of the Fondazione Italiana Linfomi

BACKGROUND: The role of [(18)F] fluorodeoxyglucose (FDG)-positron emission tomography (PET) in follicular lymphoma (FL) staging is not yet determined. PATIENTS AND METHODS: The aim of the present study was to investigate the role of PET in the initial staging of FL patients enrolled in the FOLL05-phase-III trial that compared first-line regimens (R-CVP, R-CHOP and R-FM). Patients should have undergone conventional staging and have available PET baseline to be included. RESULTS: A total of 142 patients were analysed. PET identified a higher number of nodal areas in 32% (46 of 142) of patients and more extranodal (EN) sites than computed tomography (CT) scan. Also, the Follicular Lymphoma International Prognostic Index (FLIPI) score increased in 18% (26 of 142) and decreased in 6% (9 of 142) of patients. Overall, the impact of PET on modifying the stage was highest in patients with limited stage. Actually, 62% (15 of 24) of cases with limited disease were upstaged with PET. CONCLUSIONS: The inclusion of PET among staging procedures makes the evaluation of patients with FL more accurate and has the potential to modify therapy decision and prognosis in a moderate proportion of patients. Further prospective clinical trials on FL should incorporate PET at different moments, and the therapeutic criteria to start therapy should be re-visited in the views of this new tool