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1.
Adv Protein Chem Struct Biol ; 141: 331-360, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38960479

RESUMO

We recently identified TMEM230 as a master regulator of the endomembrane system of cells. TMEM230 expression is necessary for promoting motor protein dependent intracellular trafficking of metalloproteins for cellular energy production in mitochondria. TMEM230 is also required for transport and secretion of metalloproteinases for autophagy and phagosome dependent clearance of misfolded proteins, defective RNAs and damaged cells, activities that decline with aging. This suggests that aberrant levels of TMEM230 may contribute to aging and regain of proper levels may have therapeutic applications. The components of the endomembrane system include the Golgi complex, other membrane bound organelles, and secreted vesicles and factors. Secreted cellular components modulate immune response and tissue regeneration in aging. Upregulation of intracellular packaging, trafficking and secretion of endosome components while necessary for tissue homeostasis and normal wound healing, also promote secretion of pro-inflammatory and pro-senescence factors. We recently determined that TMEM230 is co-regulated with trafficked cargo of the endomembrane system, including lysosome factors such as RNASET2. Normal tissue regeneration (in aging), repair (following injury) and aberrant destructive tissue remodeling (in cancer or autoimmunity) likely are regulated by TMEM230 activities of the endomembrane system, mitochondria and autophagosomes. The role of TMEM230 in aging is supported by its ability to regulate the pro-inflammatory secretome and senescence-associated secretory phenotype in tissue cells of patients with advanced age and chronic disease. Identifying secreted factors regulated by TMEM230 in young patients and patients of advanced age will facilitate identification of aging associated targets that aberrantly promote, inhibit or reverse aging. Ex situ culture of patient derived cells for identifying secreted factors in tissue regeneration and aging provides opportunities in developing therapeutic and personalized medicine strategies. Identification and validation of human secreted factors in tissue regeneration requires long-term stabile scaffold culture conditions that are different from those previously reported for cell lines used as cell models for aging. We describe a 3 dimensional (3D) platform utilizing non-biogenic and non-labile poly ε-caprolactone scaffolds that supports maintenance of long-term continuous cultures of human stem cells, in vitro generated 3D organoids and patient derived tissue. Combined with animal component free culture media, non-biogenic scaffolds are suitable for proteomic and glycobiological analyses to identify human factors in aging. Applications of electrospun nanofiber technologies in 3D cell culture allow for ex situ screening and the development of patient personalized therapeutic strategies and predicting their effectiveness in mitigating or promoting aging.


Assuntos
Envelhecimento , Organoides , Humanos , Organoides/metabolismo , Envelhecimento/metabolismo , Proteínas de Membrana/metabolismo , Senescência Celular , Feminino , Alicerces Teciduais/química , Glândulas Mamárias Humanas/metabolismo , Glândulas Mamárias Humanas/citologia
2.
Children (Basel) ; 10(9)2023 Aug 28.
Artigo em Inglês | MEDLINE | ID: mdl-37761427

RESUMO

Childbirth education classes represent an antenatal tool for supporting pregnant women and couples in increasing knowledge on pregnancy, delivery, breastfeeding, and newborn care. The aim of this study was to investigate the impact of an additional lesson during the prenatal course regarding the advantage of vaccination to mitigation of maternal anxiety. An observational study was designed that included participants in childbirth education classes and compared courses enhanced by the extra lesson on vaccination during pregnancy versus those who did not receive it. Assessment of the impact of prenatal educational on vaccination was measured by using validated questionnaires (State-Trait Anxiety Inventory, STAI; Perceived Stress Scale, PSS; World Health Organization- Five Well-Being Index, WHO-5). A total of 145 pregnant women participated to the investigation by answering to the online survey. Of them, 33 patients (22.8%) belonged to the course without a lesson on vaccine, while 112 (77.2%) participated to online prenatal education that included an additional meeting on the usefulness of getting vaccinated during pregnancy. No statistical differences were found between study groups in terms of demographics and perinatal outcomes. Participants in the enriched course reported lower basal anxiety levels than those without the vaccine lesson (STAI-State, normal score < 40, 30 vs. 19%, p-value 0.041; STAI-State, mild score 40-50, 78 vs. 67%, p-value 0.037). With reference to the prior two weeks, maternal wellbeing level was improved by the added class (score > 13 as measurement of wellbeing: 62% vs. 80%, p-value < 0.05). Moderate perceived stress assessed by PSS was found in those pregnant women without prenatal education on vaccination (64 vs. 50%, p-value 0.042). The introduction of a lesson regarding vaccination during pregnancy in the program of prenatal education courses improved maternal anxiety levels and wellbeing, in addition to reducing perceived stress.

3.
Protein Sci ; 32(7): e4687, 2023 07.
Artigo em Inglês | MEDLINE | ID: mdl-37243950

RESUMO

The HspB8-BAG3 complex plays an important role in the protein quality control acting alone or within multi-components complexes. To clarify the mechanism underlying its activity, in this work we used biochemical and biophysical approaches to study the tendency of both proteins to auto-assemble and to form the complex. Solubility and Thioflavin T assays, Fourier transform infrared spectroscopy and atomic force microscopy analyses clearly showed the tendency of HspB8 to self-assemble at high concentration and to form oligomers in a "native-like" conformation; otherwise, BAG3 aggregates poorly. Noteworthy, also HspB8 and BAG3 associate in a "native-like" conformation, forming a stable complex. Furthermore, the high difference between dissociation constant values of HspB8-HspB8 interaction with respect to the binding to BAG3 obtained by surface plasmon resonance confirms that HspB8 is an obligated partner of BAG3 in vivo. Lastly, both proteins alone or in the complex are able to bind and affect the aggregation of the Josephin domain, the structured domain that triggers the ataxin-3 fibrillation. In particular, the complex displayed higher activity than HspB8 alone. All this considered, we can assert that the two proteins form a stable assembly with chaperone-like activity that could contribute to the physiological role of the complex in vivo.


Assuntos
Proteínas de Choque Térmico , Proteínas Serina-Treonina Quinases , Proteínas Adaptadoras de Transdução de Sinal/química , Autofagia , Proteínas de Choque Térmico/química , Chaperonas Moleculares/metabolismo , Dobramento de Proteína , Proteínas Serina-Treonina Quinases/química , Humanos , Animais
4.
Sensors (Basel) ; 23(9)2023 May 08.
Artigo em Inglês | MEDLINE | ID: mdl-37177760

RESUMO

By using AFM as a nanografting tool, we grafted micrometer-sized DNA platforms into inert alkanethiol SAMs. Tuning the grafting conditions (surface density of grafting lines and scan rate) allowed us to tailor the molecular density of the DNA platforms. Following the nanografting process, AFM was operated in the low perturbative Quantitative Imaging (QI) mode. The analysis of QI AFM images showed the coexistence of molecular domains of different heights, and thus different densities, within the grafted areas, which were not previously reported using contact AFM imaging. Thinner domains corresponded to low-density DNA regions characterized by loosely packed, randomly oriented DNA strands, while thicker domains corresponded to regions with more densely grafted DNA. Grafting with densely spaced and slow scans increased the size of the high-density domains, resulting in an overall increase in patch height. The structure of the grafted DNA was compared to self-assembled DNA, which was assessed through nanoshaving experiments. Exposing the DNA patches to the target sequence produced an increase in the patch height, indicating that hybridization was accomplished. The relative height increase of the DNA patches upon hybridization was higher in the case of lower density patches due to hybridization leading to a larger molecular reorganization. Low density DNA patches were therefore the most suitable for targeting oligonucleotide sequences.


Assuntos
DNA , Oligonucleotídeos , Microscopia de Força Atômica , DNA/química , Hibridização de Ácido Nucleico
5.
J Colloid Interface Sci ; 632(Pt A): 65-73, 2023 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-36410295

RESUMO

HYPOTHESIS: Spinodal dewetting is one of the basic processes inducing a spontaneous withdrawal of a liquid from a substrate surface. In the accepted theory, thickness fluctuations generated by thermally activated capillary waves are amplified by the competing actions of surface tension and disjoining pressure. Ubiquitous sub-nanometric substrate roughness also produces thickness fluctuations and may play a role analogous but even more efficient in seeding the process. MODELLING: Analytic calculations valid at the early linear stage of the process and simulations extending the study to its whole non-linear development have been performed to compare features and the relative relevance of the two seeding mechanisms. FINDINGS: Calculations and simulations have shown that substrate roughness can replace capillary waves in seeding spinodal dewetting. A typically larger amplitude and a steady nature compared to the transitory one of capillary waves allow us to conclude that, contrary to the common view, substrate roughness is the prevailing seed of the spinodal instability. The consequence of our statement is that spinodal dewetting loses most of its stochastic nature and becomes, in principle, a process that can be tuned by engineering substrate roughness.


Assuntos
Sementes , Tensão Superficial
6.
Eur J Neurosci ; 56(5): 4505-4513, 2022 09.
Artigo em Inglês | MEDLINE | ID: mdl-35848658

RESUMO

The antisecretory factor (AF) is an endogenous protein that counteracts intestinal hypersecretion and various inflammation conditions in vivo. It has been detected in many mammalian tissues and plasma, but its mechanisms of action are largely unknown. To study the pharmacological action of the AF on different GABAA receptor populations in cerebellar granule cells, we took advantage of the two-photon uncaging method as this technique allows to stimulate the cell locally in well-identified plasma membrane parts. We compared the electrophysiological response evoked by releasing a caged GABA compound on the soma, the axon initial segment and neurites before and after administering AF-16, a 16 amino acids long peptide obtained from the amino-terminal end of the AF protein. After the treatment with AF-16, we observed peak current increases of varying magnitude depending on the neuronal region. Thus, studying the effects of furosemide and AF-16 on the electrophysiological behaviour of cerebellar granules, we suggest that GABAA receptors, containing the α6 subunit, may be specifically involved in the increase of the peak current by AF, and different receptor subtype distribution may be responsible for differences in this increase on the cell.


Assuntos
Neuropeptídeos , Receptores de GABA-A , Animais , Cerebelo/fisiologia , Mamíferos/metabolismo , Neurônios/fisiologia , Neuropeptídeos/metabolismo , Ratos , Receptores de GABA-A/metabolismo , Ácido gama-Aminobutírico/metabolismo , Ácido gama-Aminobutírico/farmacologia
7.
Biomol Concepts ; 13(1): 289-297, 2022 Jun 08.
Artigo em Inglês | MEDLINE | ID: mdl-35675587

RESUMO

The study of the GABAA receptor itself and its pharmacology is of paramount importance for shedding light on the role of this receptor in the central nervous system. Caged compounds have emerged as powerful tools to support research in this field, as they allow to control, in space and time, the release of neurotransmitters enabling, for example, to map receptors' distribution and dynamics. Here we focus on γ-aminobutyric acid (GABA)-caged compounds, particularly on a commercial complex called RuBi-GABA, which has high efficiency of uncaging upon irradiation at visible wavelengths. We characterized, by electrophysiological measurements, the effects of RuBi-GABA on GABAA receptors of rat cerebellar granule cells in vitro. In particular, we evaluated the effects of side products obtained after RuBi-GABA photolysis. For this purpose, we developed a procedure to separate the "RuBi-cage" from GABA after uncaging RuBi-GABA with a laser source; then, we compared electrophysiological measurements acquired with and without administering the RuBi-cage in the perfusing bath. In conclusion, to investigate the role of the "cage" molecules both near and far from the cell soma, we compared experiments performed changing the distance of the uncaging point from the cell.


Assuntos
Neurônios , Ácido gama-Aminobutírico , Animais , Neurônios/fisiologia , Ratos , Receptores de GABA-A/fisiologia , Ácido gama-Aminobutírico/farmacologia
8.
Sci Rep ; 10(1): 13380, 2020 08 07.
Artigo em Inglês | MEDLINE | ID: mdl-32770032

RESUMO

Ruthenium-bipyridinetriphenylphosphine-GABA (RuBi-GABA) is a caged compound that allows studying the neuronal transmission in a specific region of a neuron. The inhibitory neurotransmitter γ-aminobutyric acid (GABA) is bound to a caged group that blocks the interaction of the neurotransmitter with its receptor site. Following linear-one-photon (1P)-and non-linear-multi-photon-absorption of light, the covalent bond of the caged molecule is broken, and GABA is released. Such a controlled release in time and space allows investigating the interaction with its receptor in four dimensions (X,Y,Z,t). Taking advantage of this strategy, we succeeded in addressing the modulation of GABAA in rat cerebellar neurons by coupling the photoactivation process, by confocal or two-photon excitation microscopy, with the electrophysiological technique of the patch-clamp in the whole-cell configuration. Key parameters have been comprehensively investigated and correlated in a temporally and spatially confined way, namely: photoactivation laser power, time of exposure, and distance of the uncaging point from the cell of interest along the X, Y, Z spatial coordinates. The goal of studying specific biological events as a function of controlled physical parameters has been achieved.


Assuntos
Neurônios/metabolismo , Neurônios/fisiologia , Neurotransmissores/metabolismo , Receptores de GABA-A/metabolismo , Transmissão Sináptica/fisiologia , Ácido gama-Aminobutírico/metabolismo , Animais , Células Cultivadas , Cerebelo/citologia , Luz , Microscopia Confocal , Imagem Molecular/métodos , Técnicas de Patch-Clamp , Ratos Sprague-Dawley
9.
Pharmacol Res ; 158: 104899, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32407959

RESUMO

SARS-CoV-2 is causing an increasing number of deaths worldwide because no effective treatment is currently available. Remdesivir has shown in vitro activity against coronaviruses and is a possible antiviral treatment for SARS-CoV-2 infection. This prospective (compassionate), open-label study of remdesivir, which was conducted at Luigi Sacco Hospital, Milan, Italy, between February 23 and March 20, 2020, involved patients with SARS-CoV-2 pneumonia aged ≥18 years undergoing mechanical ventilation or with an oxygen saturation level of ≤94 % in air or a National Early Warning Score 2 of ≥4. The primary outcome was the change in clinical status based on a 7-category ordinal scale (1 = not hospitalised, resuming normal daily activities; 7 = deceased). The 35 patients enrolled from February 23 to March 20, 2020, included 18 in intensive care unit (ICU), and 17 in our infectious diseases ward (IDW). The 10-day course of remdesivir was completed by 22 patients (63 %) and discontinued by 13, of whom eight (22.8 %) discontinued because of adverse events. The median follow-up was 39 days (IQR 25-44). At day 28, 14 (82.3 %) patients from IDW were discharged, two were still hospitalized and one died (5.9 %), whereas in ICU 6 (33.3 %) were discharged, 8 (44.4 %) patients died, three (16.7 %) were still mechanically ventilated and one (5.6 %) was improved but still hospitalized. Hypertransaminasemia and acute kidney injury were the most frequent severe adverse events observed (42.8 % and 22.8 % of the cases, respectively). Our data suggest that remdesivir can benefit patients with SARS-CoV-2 pneumonia hospitalised outside ICU where clinical outcome was better and adverse events are less frequently observed. Ongoing randomised controlled trials will clarify its real efficacy and safety, who to treat, and when.


Assuntos
Monofosfato de Adenosina/análogos & derivados , Alanina/análogos & derivados , Betacoronavirus , Ensaios de Uso Compassivo/estatística & dados numéricos , Infecções por Coronavirus/tratamento farmacológico , Hospitalização/estatística & dados numéricos , Unidades de Terapia Intensiva/estatística & dados numéricos , Pneumonia Viral/tratamento farmacológico , Injúria Renal Aguda/induzido quimicamente , Monofosfato de Adenosina/efeitos adversos , Monofosfato de Adenosina/uso terapêutico , Idoso , Alanina/efeitos adversos , Alanina/uso terapêutico , Antivirais/efeitos adversos , Antivirais/uso terapêutico , COVID-19 , Infecções por Coronavirus/sangue , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Pandemias , Pneumonia Viral/sangue , SARS-CoV-2 , Transaminases/sangue , Resultado do Tratamento
11.
Sensors (Basel) ; 20(6)2020 Mar 13.
Artigo em Inglês | MEDLINE | ID: mdl-32183234

RESUMO

The continuous advances of Nanofluidics have been stimulating the development of novel nanostructures and strategies to accumulate very diluted analytes, for implementing a new class of high sensitivity miniaturized polymeric sensors. We take advantage of the electrokinetic properties of these structures, which allow accumulating analytes inside asymmetric microfluidic structures to implement miniaturized sensors able to detect diluted solutions down to nearly 1.2 pg/mL. In particular, exploiting polydimethylsiloxane devices, fabricated by using the junction gap breakdown technique, we concentrate antigens inside a thin microfunnel functionalized with specific antibodies to favor the interaction and, if it is the case, the recognition between antigens in solution and antibodies anchored to the surface. The transduction mechanism consists in detecting the fluorescence signal of labeled avidin when it binds to biotinylated antigens. Here, we demonstrate that exploiting these electrokinetic phenomena, typical of nanofluidic structures, we succeeded in concentrating biomolecules in correspondence of a 1 pL sensing region, a strategy that grants to the device performance comparable to standard immunoassays.


Assuntos
Antígenos/isolamento & purificação , Técnicas Biossensoriais , Imunoensaio/métodos , Dispositivos Lab-On-A-Chip , Anticorpos/química , Antígenos/química , Dimetilpolisiloxanos/química , Humanos , Nanomedicina/tendências
12.
Blood Purif ; 49(6): 685-691, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32200385

RESUMO

BACKGROUND: The current effective delivered dose is a quality indicator for continuous renal replacement therapy. Its periodic assessment might enable physicians to deliver personalised treatments. Yet, its quantification as by extracorporeal urea clearance (Cl) is cumbersome and thus often neglected in routine practice. The aim of this in vitro study is to demonstrate the non-inferior effectiveness of assessing the current effective delivered dose using a simpler, cheaper and faster approach based on measurement of fluoride rather than urea extracorporeal Cl. METHODS: We compared urea and fluoride removal in 3 post-dilution continuous veno-venous haemofiltration (CVVH) and 3 continuous veno-venous haemodialysis (CVVHD) in vitro experimental models. Experiments ran for 180 min, using 3 L of human blood, heparin anticoagulation and a machine dose of 30 mL/kg/h. Urea and fluoride were measured in the inflow, outflow and effluent lines to compare sieving coefficients (SC), saturation coefficients (SA) and transmembrane Cls. RESULTS: In CVVH, the median SC values were 1.06 (1.02-1.07) and 1.02 (1.01-1.04) for fluoride and urea, respectively (discrepancy of 4.3%), while transmembrane convective Cls were 31.28 (30.01-31.31) mL/kg/h and 30.30 (29-31.85) mL/kg/h (discrepancy of 3.13%), respectively. In CVVHD, the median SA values were 1.01 (0.96-1.02) and 1 (0.95-1.01) for fluoride and urea, respectively (discrepancy of 1.6%), while transmembrane dialytic Cls were 30.26 (29.52-31.32) mL/kg/h and 31.16 (30-31.75) mL/kg/h (discrepancy of -2.97%), respectively. CONCLUSION: Fluoride transmembrane removal was close to that observed with urea, in terms of SC, SA and transmembrane Cl. Fluoride seems as much accurate as urea in assessing the current effective delivered dose during both CVVH and CVVHD and might therefore be adopted for dose measurement. Besides accuracy, fluoride bedside assessment could present many advantages over urea, particularly in terms of availability, costs, time requirement and rapidity of assessment.


Assuntos
Terapia de Substituição Renal Contínua/normas , Fluoretos/sangue , Indicadores de Qualidade em Assistência à Saúde , Injúria Renal Aguda/terapia , Nitrogênio da Ureia Sanguínea , Terapia de Substituição Renal Contínua/métodos , Hemofiltração/métodos , Hemofiltração/normas , Humanos , Diálise Renal/métodos , Diálise Renal/normas , Resultado do Tratamento , Ureia/sangue
13.
ACS Biomater Sci Eng ; 6(6): 3649-3663, 2020 06 08.
Artigo em Inglês | MEDLINE | ID: mdl-33463182

RESUMO

Recent studies have suggested that microenvironmental stimuli play a significant role in regulating cellular proliferation and migration, as well as in modulating self-renewal and differentiation processes of mammary cells with stem cell (SCs) properties. Recent advances in micro/nanotechnology and biomaterial synthesis/engineering currently enable the fabrication of innovative tissue culture platforms suitable for maintenance and differentiation of SCs in vitro. Here, we report the design and fabrication of an open microfluidic device (OMD) integrating removable poly(ε-caprolactone) (PCL) based electrospun scaffolds, and we demonstrate that the OMD allows investigation of the behavior of human cells during in vitro culture in real time. Electrospun scaffolds with modified surface topography and chemistry can influence attachment, proliferation, and differentiation of mammary SCs and epigenetic mechanisms that maintain luminal cell identity as a function of specific morphological or biochemical cues imparted by tailor-made fiber post-treatments. Meanwhile, the OMD architecture allows control of cell seeding and culture conditions to collect more accurate and informative in vitro assays. In perspective, integrated systems could be tailor-made to mimic specific physiological conditions of the local microenvironment and then analyze the response from screening specific drugs for more effective diagnostics, long-term prognostics, and disease intervention in personalized medicine.


Assuntos
Engenharia Tecidual , Alicerces Teciduais , Diferenciação Celular , Humanos , Microfluídica , Poliésteres
14.
Nanomaterials (Basel) ; 9(12)2019 Nov 25.
Artigo em Inglês | MEDLINE | ID: mdl-31775220

RESUMO

Nanofluidic structures are often the key element of many lab-on-chips for biomedical and environmental applications. The demand for these devices to be able to perform increasingly complex tasks triggers a request for increasing the performance of the fabrication methods. Soft lithography and poly(dimethylsiloxane) (PDMS) have since long been the basic ingredients for producing low-cost, biocompatible and flexible devices, replicating nanostructured masters. However, when the desired functionalities require the fabrication of shallow channels, the "roof collapse" phenomenon, that can occur when sealing the replica, can impair the device functionalities. In this study, we demonstrate that a "focused drop-casting" of h-PDMS (hard PDMS) on nanostructured regions, provides the necessary stiffness to avoid roof collapse, without increasing the probability of deep cracks formation, a drawback that shows up in the peel-off step, when h-PDMS is used all over the device area. With this new approach, we efficiently fabricate working devices with reproducible sub-100 nm structures. We verify the absence of roof collapse and deep cracks by optical microscopy and, in order to assess the advantages that are introduced by the proposed technique, the acquired images are compared with those of cracked devices, whose top layer, of h-PDMS, and with those of collapsed devices, made of standard PDMS. The geometry of the critical regions is studied by atomic force microscopy of their resin casts. The electrical resistance of the nanochannels is measured and shown to be compatible with the estimates that can be obtained from the geometry. The simplicity of the method and its reliability make it suitable for increasing the fabrication yield and reducing the costs of nanofluidic polymeric lab-on-chips.

15.
Polymers (Basel) ; 11(5)2019 May 20.
Artigo em Inglês | MEDLINE | ID: mdl-31137564

RESUMO

This paper describes a procedure to measure the permeability P, diffusivity D, and rate of adsorption k1, thus determining the solubility S and rate of desorption k2 of He, N2, O2, CH4, and CO2 on a polydimethylsiloxane (PDMS) membrane. The described procedure is able to determine experimentally all the physical quantities that characterize the gas transport process through a thin rubber polymer membrane. The experiments were carried out at room temperature and at a transmembrane pressure of 1 atm. The results are in good agreement with the available data in the literature and offer an evaluation of k1 and k2.

16.
Sci Rep ; 9(1): 585, 2019 01 24.
Artigo em Inglês | MEDLINE | ID: mdl-30679515

RESUMO

We reported the efficacy and safety data for daclatasvir (DCV)-based all-oral antiviral therapy in patients treated in the Italian compassionate-use program. 275 patients were included (202 male-73.5%, mean age: 57.4 years, 62 HIV-coinfected, 94 with recurrence of hepatitis C post-OLT). Forty-nine patients (17.8%) had Child-Pugh B, Genotype(G) distribution was: G1a:72 patients (26.2%), G1b:137 (49.8%); G3:40 (14.5%) and G4:26 (9.5%). Patients received DCV with sofosbuvir(SOF) (n = 221, 129 with ribavirin(RBV) or with simeprevir (SMV) or asunaprevir (ASU) (n = 54, 19 with RBV) for up to 24 weeks. Logistic regression was used to identify baseline characteristics associated with sustained virological response at week 12 post-treatment (SVR12). Liver function changes between baseline and follow up were assessed in 228 patients. 240 patients achieved SVR12 (87.3%), post transplant and HIV co-infected patients were equally distributed among SVR and no SVR (35% vs 34.3%; p = 0.56 and 24.2% vs 11.4%, p = 0.13, respectively). SVR rate was significantly higher with the combination DCV + SOF compared with DCV + SIM or ASU (93.2% vs 63.0%, p < 0.0001). Bilirubin value (OR: 0.69, CI95%: 0.54-0.87, p = 0.002) and regimen containing SOF (OR: 9.99, CI95%: 4.09-24.40; p < 0.001) were independently related with SVR. Mean albumin and bilirubin values significantly improved between baseline and follow-up week 12. DCV-based antiviral therapy was well tolerated and resulted in a high SVR when combined with SOF either in pre-transplant and in OLT patients and in "difficult to treat" HCV genotypes. Regimens containing DCV in combination with NS3 protease inhibitors obtained suboptimal results.


Assuntos
Antivirais/uso terapêutico , Hepatite C Crônica/complicações , Hepatite C Crônica/tratamento farmacológico , Imidazóis/uso terapêutico , Cirrose Hepática/tratamento farmacológico , Cirrose Hepática/patologia , Adulto , Idoso , Antivirais/efeitos adversos , Carbamatos , Quimioterapia Combinada/métodos , Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos , Feminino , Humanos , Imidazóis/efeitos adversos , Isoquinolinas/uso terapêutico , Itália , Testes de Função Hepática , Masculino , Pessoa de Meia-Idade , Pirrolidinas , Ribavirina/uso terapêutico , Sofosbuvir/uso terapêutico , Sulfonamidas/uso terapêutico , Resposta Viral Sustentada , Resultado do Tratamento , Valina/análogos & derivados
18.
EBioMedicine ; 33: 253-268, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30049385

RESUMO

The lack of engineering systems able to faithfully reproduce complex kidney structures in vitro has made it difficult to efficiently model kidney diseases and development. Using polydimethylsiloxane (PDMS) scaffolds and a kidney-derived cell line we developed a system to rapidly engineer custom-made 3D tubules with typical renal epithelial properties. This system was successfully employed to engineer patient-specific tubules, to model polycystic kidney disease (PKD) and test drug efficacy, and to identify a potential new pharmacological treatment. By optimizing our system we constructed functional ureteric bud (UB)-like tubules from human induced pluripotent stem cells (iPSCs), and identified a combination of growth factors that induces budding morphogenesis like embryonic kidneys do. Finally, we applied this assay to investigate budding defects in UB-like tubules derived from a patient with a PAX2 mutation. Our system enables the modeling of human kidney disease and development, drug testing and discovery, and lays the groundwork for engineering anatomically correct kidney tissues in vitro and developing personalized medicine applications.


Assuntos
Células-Tronco Pluripotentes Induzidas/citologia , Túbulos Renais/citologia , Técnicas de Cultura de Órgãos/métodos , Fator de Transcrição PAX2/genética , Doenças Renais Policísticas/patologia , Animais , Diferenciação Celular , Cães , Descoberta de Drogas , Avaliação Pré-Clínica de Medicamentos , Humanos , Células Madin Darby de Rim Canino , Modelos Biológicos , Mutação , Doenças Renais Policísticas/genética , Medicina de Precisão , Alicerces Teciduais
19.
Midwifery ; 64: 53-59, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29929062

RESUMO

OBJECTIVE: To assess current practices of health care providers (HCPs) caring for women experiencing a stillbirth and to explore their needs for training to better support bereaved families. DESIGN: Nationwide cross-sectional survey. The main outcome measures were the evaluation of HCPs cognition, emotions and behaviours with regard to the care of women with a stillbirth care, as well as their compliance with international guidelines. PARTICIPANTS: 750 HCPs, in 11 Italian hospitals, were administered a multiple-choice questionnaire. FINDINGS: The response rate was 89.9%; the majority (94.1%) were female, with a mean age of 37.6 (SD = 10.4) years. Midwives were the most represented (72.8%). Half of the respondents recommended immediate birth; only 55% routinely bathed and dressed stillborn babies for their parents to see, while 44.4% of HCPs immediately took the babies away without allowing parents to properly say goodbye to them. More than half felt inadequate and some even reported having failed to provide support to the family when caring for a woman with stillbirth in the past. The need for professional training courses was expressed by 90.2%, and three-quarters had never previously attended a course on perinatal bereavement care. When answers by Italian HCPs are systematically evaluated with reference to international guidelines, the results were very poor with only 27.9% of respondents reported having created memories of the baby and less than 3% complied with all recommendations in the areas of respect for baby and parents, appropriate birth options, and aftercare. KEY CONCLUSIONS: There is a substantial gap between the standards of care defined by international guidelines and the practices currently in place in Italy. Italian HCPs feel an urgent need to be offered professional training courses to better meet the needs of grieving families. IMPLICATION FOR PRACTICE: Perinatal HCPs should be aware of their pivotal role in helping bereaved parents after stillbirth and perinatal loss, and seek appropriate training.


Assuntos
Competência Clínica/normas , Tocologia/normas , Pais/psicologia , Assistência Perinatal/normas , Natimorto/psicologia , Adulto , Estudos Transversais , Educação/métodos , Feminino , Pesar , Humanos , Itália , Pessoa de Meia-Idade , Tocologia/educação , Assistência Perinatal/métodos , Gravidez , Apoio Social , Inquéritos e Questionários
20.
Sci Rep ; 8(1): 6345, 2018 Apr 20.
Artigo em Inglês | MEDLINE | ID: mdl-29679013

RESUMO

The purpose of this investigation is to fabricate PDMS membranes with reliable surface roughness in order to reduce the surface resistances and to study its impact on the permeation rate. The permeance of CO2 through PDMS membranes with rough surfaces at nanoscale is studied and compared with the one of membranes with flat surfaces. At very low thickness, rough membranes have a permeance greater than that of membranes with flat surfaces. The enhancement occurs in a regime where the gas transport is sorption desorption surface rate limited, and cannot be explained by the increase in surface area due to the corrugation. The analysis, introducing a phenomenological model in analogy with electrical flow, indicates that nano-corrugation reduces the surface resistance. To test the model, the permeance of N2 is also measured in the same experimental conditions and the influence of surface roughness on permeation rate of CO2, He, CH4 and N2 is studied. The comparison among the gases suggests that the Henry's coefficient depends on the surface roughness and allows discussing the role of roughness on membrane selectivity.

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