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Polysialic acid (polySia) is a posttranslational modification found on only a handful of proteins in the central nervous and immune systems. The addition of polySia to therapeutic proteins improves pharmacokinetics and reduces immunogenicity. To date, polysialylation of therapeutic proteins has only been achieved in vitro by chemical or chemoenzymatic strategies. In this work, we develop a biosynthetic pathway for site-specific polysialylation of recombinant proteins in the cytoplasm of Escherichia coli. The pathway takes advantage of a bacterial cytoplasmic polypeptide-glycosyltransferase to establish a site-specific primer on the target protein. The glucose primer is extended by glycosyltransferases derived from lipooligosaccharide, lipopolysaccharide and capsular polysaccharide biosynthesis from different bacterial species to synthesize long chain polySia. We demonstrate the new biosynthetic route by modifying green fluorescent proteins and a therapeutic DARPin (designed ankyrin repeat protein).
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Escherichia coli , Modificação Traducional de Proteínas/genética , Ácidos Siálicos , Escherichia coli/genética , Escherichia coli/metabolismo , Glicosilação , Proteínas de Fluorescência Verde/biossíntese , Proteínas de Fluorescência Verde/genética , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/genética , Ácidos Siálicos/genética , Ácidos Siálicos/metabolismoRESUMO
This work concerns the verification of the self-healing ability of PP-co-HUPy copolymers dispersed in epoxy systems. PP is the acronym for the Poly-PEGMA polymer, and HUPy refers to the HEMA-UPy copolymers based on ureidopyrimidinone (UPy) moieties. In particular, this work aims to verify whether this elastomer characterized by an intrinsic self-healing ability can activate supramolecular interactions among polymer chains of an epoxy resin, as in the elastomer alone. The elastomer includes a class of polyethylene glycol monomethyl ether methacrylate-based copolymers, with different percentages of urea-N-2-amino-4-hydroxy-6-methyl pyrimidine-N'-(hexamethylene-n-carboxyethyl methacrylate) (HEMA-UPy) co-monomers. The self-healing capability of these copolymers based on possible quadruple hydrogen bond interactions between polymer chains has been verified. The formulated epoxy samples did not show self-healing efficiency. This can be attributed to the formation of phase segregation that originates during the curing process of the samples, although the PP-co-HUPy copolymers are completely soluble in the liquid epoxy matrix EP. The morphological investigation highlighted the presence of crystals of PP-co-HUPy copolymers, which are in greater quantity in the sample containing the highest weight percentage (7.8 wt%) of HUPy units. Furthermore, the crystals act as promotors for increasing the curing degree (DC) of the epoxy systems containing HUPy units. DC goes from 91.6% for EP to 96.1% and 95.4% for the samples containing weight percentages of 2.5 and 7.8 wt% of HUPy units, respectively. Dynamic mechanical analysis (DMA) shows storage modulus values for epoxy systems containing PP-co-HUPy units lower than that of the unfilled resin EP. The values of maximum in Tan δ (Tg), representing the temperature at which the glass transition occurs, are 220 for the unfilled resin EP, 228 for the sample containing 2.5 wt% of HEMA-UPy units, and 211 for the sample containing 7.8 wt% of HEMA-UPy units.
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Radioligand therapy with [177Lu]Lu-PSMA is a theranostic approach for heavily treated mCRPC patients with positive PSMA PET in the absence of relevant PSMA-negative metastases assessed through CT, MRI, bone scan or FDG PET. In this case, we described a mCRPC patient treated with RLT with discordant PSA values and PSMA images, in which Choline PET confirmed a biochemically suspected disease progression (PD), showing metastatic lesions not revealed by PSMA imaging.
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An effective non-covalent compatibilization method for graphite and low-density polyethylene is reported. To obtain this result, pyren-1-yl-stearate (P1S) was synthesized, characterized and mixed with graphite to provide a better dispersion in polyethylene matrix. The P1S improves the dispersion of carbon filler in polyethylene through non-covalent compatibilization: the pyrenyl group gives π-π stacking interactions with graphite and the stearyl chain provides van der Waals interaction with the polymer chain (specifically London dispersion forces). In this study, different P1S/graphite fillers were prepared with a ratio by weight of 90/10 and 50/50, respectively, by using manual and ball-milling mixing. Their stability, interaction and morphology were evaluated through TGA, RX, and SEM. Thermogravimetric analyses showed that ball-milling mixing is more effective than manual mixing in promoting π-π stacking interactions of molecules such as P1S ester containing an alkyl chain and aromatic rings. The role of ball milling is confirmed by X-ray diffraction measurements since it was possible to observe both exfoliation and intercalation phenomena when this technique was used to mix the P1S ester with graphite. SEM analyses of polyethylene containing 1% of the carbon fillers again highlighted the importance of ball milling to promote the interaction of the ester with graphite and, simultaneously, the importance of the alkyl chain in order to achieve polyethylene-graphite compatibilization.
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Background: In differentiated thyroid cancer (DTC) patients, the response to initial treatments is evaluated 6-12 months after radioiodine therapy (RIT) according to the 2015 American Thyroid Association (2015 ATA) criteria. In selected patients, diagnostic 131-radioiodine whole-body scintigraphy (Dx-WBS) is recommended. We evaluated the diagnostic performance of 123I-Dx-WBS-SPECT/CT imaging in detecting incomplete structural responses in the early follow-up of DTC patients and, additionally, derived optimized basal-Tg value as a yardstick for scintigraphic imaging. Methods: We reviewed the records of 124 low or intermediate-risk DTC patients with negative anti-thyroglobulin antibody. All patients had undergone (near)-total-thyroidectomy followed by RIT. The response to initial treatments was evaluated 6-12 months after RIT. Results: According to the 2015 ATA criteria, 87, 19 and 18 DTC patients were classified to have excellent response (ER), indeterminate/incomplete biochemical response (BIndR/BIR) or structural incomplete response (SIR), respectively. Among patients with less than ER, 18 had a positive 123I-Dx-WBS-SPECT/CT. Metastatic disease at 123I-Dx-WBS-SPECT/CT mainly involved lymph nodes within the central compartment, and corresponding neck ultrasound examinations were negative. The ROC curve analysis was performed to define the best basal-Tg cut-off (i.e., 0.39 ng/mL; AUC = 0.852) able to discriminate patients with and without positive 123I-Dx-WBS-SPECT/CT, respectively. The overall sensitivity, specificity, accuracy, PPV and NPV were 77.8%, 89.6%, 87.9%, 56.0% and 95.9%, respectively. Basal-Tg cut-off was an independent risk factor for having a positive 123I-Dx-WBS-SPECT/CT. Conclusion:123I-Dx-WBS-SPECT/CT identified lymph node metastases in 14/37 patients with less than ER and a negative neck ultrasound, thus modifying the management of such patients. The diagnostic performance of 123I-Dx-WBS-SPECT/CT significantly increased in patients with basal-Tg values ≥ 0.39 ng/mL.
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This prospective clinical trial aimed at evaluating the clinical performance of three-unit posterior zirconia fixed dental prostheses (FDPs) after 5 years of clinical function. Thirty-seven patients received 48 three-unit zirconia-based FDPs. The restorations replaced either a premolar or a molar. Specific inclusion criteria were needed. Tooth preparation was standardized. Computer-aided design/computer-assisted manufacturing frameworks with a 9-mm(2) cross section of the connector and a 0.6-mm minimum thickness of the retainer were made. The restorations were luted with resin cement. The patients were recalled after 1, 6, 12, 24, 36, 48, and 60 months. The survival and success of the ceramics and zirconia were evaluated. The technical and aesthetic outcomes were examined using the United States Public Health Service criteria. The biologic outcomes were analyzed at abutment and contralateral teeth. Descriptive statistics were performed. All FDPs completed the study, resulting in 100% cumulative survival rate and 91.9% and 95.4% cumulative success rates for patients wearing one and two FDPs, respectively. No losses of retention were recorded. Forty-two restorations were rated alpha in all measured parameters. A minor chipping of the ceramics was detected in three restorations. No significant differences between the periodontal parameters of the test and control teeth were observed. Five-year clinical results proved that three-unit posterior zirconia-based FDPs were successful in the medium term for both function and aesthetic. Zirconia can be considered a promising substitute of metal frameworks for the fabrication of short-span posterior prostheses.
Assuntos
Desenho Assistido por Computador , Porcelana Dentária , Planejamento de Prótese Dentária , Prótese Parcial Fixa , Zircônio , Adulto , Idoso , Dente Pré-Molar , Cimentação , Retenção em Prótese Dentária , Adesivos Dentinários , Feminino , Humanos , Estimativa de Kaplan-Meier , Masculino , Pessoa de Meia-Idade , Dente Molar , Estudos Prospectivos , Cimentos de Resina , Estatísticas não Paramétricas , Adulto JovemRESUMO
For prostate cancer (PCa) biochemical recurrence (BCR), the primarily suggested imaging technique by the European Association of Urology (EAU) guidelines is prostate-specific membrane antigen (PSMA) positron emission tomography/computer tomography (PET/CT). Indeed, the increased detection rate of PSMA PET/CT for early BCR has led to a fast and wide acceptance of this novel technology. However, PCa is a very heterogeneous disease, not always easily assessable with the highly specific PSMA PET with around 10% of cases occuring without PSMA expression. In this paper, we present the case of a patient with PCa BCR that resulted negative on [68Ga]Ga-PSMA-11 PET/CT, but positive on [18F]Fluoromethylcholine (Choline) PET/CT.
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Radioisótopos de Gálio , Neoplasias da Próstata , Masculino , Humanos , Tomografia por Emissão de Pósitrons combinada à Tomografia Computadorizada/métodos , Neoplasias da Próstata/diagnóstico por imagem , Colina , Tomografia por Emissão de Pósitrons , ComputadoresRESUMO
The great concern over the environmental impact of wastewaters has led to the designing of advanced treatment processes to upgrade conventional treatment plants and achieve a significant reduction of contaminants in receiving waters. In the present study we combined chemical and ecotoxicological analyses, aiming to evaluate the reduction of toxicity effects associated with the removal of micropollutants and to define the contribution of the detected compounds to the overall toxicity of the mixtures in a series of wastewater effluents collected from a secondary treatment (OUT 2) and from a tertiary activated carbon treatment (OUT 3) plant. The target compounds were selected after a screening procedure among pharmaceuticals, musk fragrances, and trace metals. The classical algal growth inhibition test was conducted on the original effluent samples and on different fractions obtained by solid-phase extraction (SPE) treatment. A good accordance was found between the removal of toxicity (30%-80%) and organic compounds (70%-80%) after the tertiary treatment, suggesting its high efficiency to improve the wastewater quality. The discrepancy between the contribution to the overall toxicity of the nonadsorbable compounds (i.e., inorganic or very polar organic compounds) as experimentally measured by the SPE bioassays (18%-76%) and calculated by the concentration addition approach (>97%) could be mitigated by including the bioavailability correction in metal-toxicity modeling of wastewater mixtures. For the organic compounds, the toxic equivalency method enabled us to quantify the portion of toxicity explained by the detected chemicals in both OUT 2 (82%-104%) and OUT 3 (5%-57%), validating the selection of the target molecules. The applied integrating approach could be implemented by the inclusion of both additional target chemicals and toxicity endpoints. Environ Toxicol Chem 2022;41:2404-2419. © 2022 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.
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Águas Residuárias , Poluentes Químicos da Água , Carvão Vegetal , Ecotoxicologia , Compostos Orgânicos , Preparações Farmacêuticas , Eliminação de Resíduos Líquidos/métodos , Águas Residuárias/química , Poluentes Químicos da Água/análise , Poluentes Químicos da Água/toxicidadeRESUMO
The analytical scenario for determining contaminants in the food and feed sector is constantly prompted by the progress and improvement of knowledge and expertise of researchers and by the technical innovation of the instrumentation available. Mycotoxins are agricultural contaminants of fungal origin occurring at all latitudes worldwide and being characterized by acute and chronic effects on human health and animal wellness, depending on the species sensitivity. The major mycotoxins of food concern are aflatoxin B1 and ochratoxin A, the first for its toxicity, and the second for its recurrent occurrence. However, the European legislation sets maximum limits for mycotoxins, such as aflatoxin B1, ochratoxin A, deoxynivalenol, fumonisins, and zearalenone, and indicative limits for T-2 and HT-2 toxins. Due to the actual probability that co-occurring mycotoxins are present in a food or feed product, nowadays, the availability of reliable, sensitive, and versatile multi-mycotoxin methods is assuming a relevant importance. Due to the wide range of matrices susceptible to mycotoxin contamination and the possible co-occurrence, a multi-mycotoxin and multi-matrix method was validated in liquid chromatography-tandem mass spectrometry (LC-MS/MS) with the purpose to overcome specific matrix effects and analyze complex cereal-based samples within the Italian Total Diet Study project.
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Grão Comestível/química , Contaminação de Alimentos/análise , Micotoxinas/análise , Pão/análise , Cromatografia Líquida , Farinha/análise , Oryza/química , Espectrometria de Massas em Tandem , Triticum/químicaRESUMO
BACKGROUND: Recent data suggest that donor intraislet endothelial cells may survive islet transplantation and participate to the events that influence islet engraftment. However, the mechanisms that regulate islet endothelial behavior in this setting are poorly known. METHODS: We obtained immortalized human (hIECs) and mouse (mIECs) islet endothelial cells by transfection with SV40-T-large antigen and studied the synthesis and response to Platelet-activating factor (PAF), a multipotent phospholipid that acts as endothelial mediator of both inflammation and angiogenesis. RESULTS: HIECs showed typical endothelial markers such as expression of vWF, CD31, and CD105, uptake of acetylated-LDL and binding to ULE-A lectin. Moreover, they expressed nestin, the PAF-receptor and possess surface fenestrations and in vitro angiogenic ability of forming tubular structures on Matrigel. Likewise, mIECs showed expression of vWF, CD31, nestin, PAF-receptor and CD105, and uptake of acetylated-LDL. HIECs and mIECs rapidly produced PAF under stimulation with thrombin in a dose-dependent way. Exogenous PAF or thrombin-induced PAF synthesis increased leukocyte adhesion to hIECS and mIECs and cell motility of both endothelial cell lines. Moreover, PAF or thrombin-induced PAF synthesis accelerated in vitro formation of vessel-like tubular structures when hIECs are seeded on Matrigel. Notably, gene-microarray analysis detected up-regulation of beta3 integrin gene on hIECs stimulated with PAF, that was confirmed at the protein level. CONCLUSIONS: Based on the novel development of immortalized islet endothelium, these results suggest that PAF may have a dual role that links inflammation to angiogenesis in the early events of islet transplantation.
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Endotélio Vascular/fisiologia , Transplante das Ilhotas Pancreáticas/fisiologia , Ilhotas Pancreáticas/fisiologia , Fator de Ativação de Plaquetas/biossíntese , Animais , Antígenos Transformantes de Poliomavirus/genética , Linhagem Celular , Movimento Celular , Células Cultivadas , Humanos , Ilhotas Pancreáticas/irrigação sanguínea , Camundongos , Análise de Sequência com Séries de Oligonucleotídeos , Fator de Ativação de Plaquetas/genética , TransfecçãoRESUMO
We report a case of an ectopic/hypoplastic kidney removed by radio-guided surgery. A 7-year-old girl, with a history of vaginal drainage of urine, underwent renal scintigraphy with Tc-DMSA. SPECT/CT revealed a focal uptake in the pelvis, corresponding to hypoplastic kidney as confirmed by MRI. Based on SPECT/CT findings, the patient underwent laparoscopic surgery, using Tc-DMSA scan to help the surgeon to detect the small ectopic kidney. Intraoperatory histological report confirmed the renal origin of the specimen.
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Nefropatias/cirurgia , Rim/anormalidades , Compostos Radiofarmacêuticos , Cirurgia Assistida por Computador , Ácido Dimercaptossuccínico Tecnécio Tc 99m , Tomografia Computadorizada de Emissão de Fóton Único , Criança , Feminino , Humanos , Rim/diagnóstico por imagem , Rim/cirurgia , Nefropatias/diagnóstico por imagem , LaparoscopiaRESUMO
Knowledge on the functional properties of tumor-derived endothelial cells (TEC) can be relevant for the development of antiangiogenic therapeutic strategies. In the present study, we obtained and characterized endothelial cell lines from human renal carcinomas. TEC did not undergo senescence and showed constant expression of markers of endothelial activation and angiogenesis. In vitro, TEC, in contrast to normal endothelial cells, were resistant to apoptosis, proadhesive for renal carcinoma cells, and able to grow and organize in the absence of serum in persistent capillary-like structures. In vivo, TEC were able to grow in immunodeficient mice and to form vascular structures connected with the circulation. At a molecular level, gene array analysis showed an increased expression of genes involved in survival and cell adhesion compared with expression in normal microvascular endothelial cells. Moreover, expression of angiopoietin-1 and vascular endothelial growth factor (VEGF)-D and the Akt survival pathway were up-regulated. Inhibition of interaction of VEGFR-2 or VEGFR-3 with VEGF-D but not of Tie-2-angiopoietin-1 interaction with soluble receptors abrogated Akt activation and survival of TEC. These results indicate that at least some of the TEC within a tumor display abnormal characteristics in terms of survival and angiogenic properties and also indicate the presence of a functional autocrine pathway related to VEGF-D.
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Carcinoma de Células Renais/irrigação sanguínea , Neoplasias Renais/irrigação sanguínea , Neovascularização Patológica , Proteínas Serina-Treonina Quinases , Indutores da Angiogênese/biossíntese , Angiopoietina-1 , Animais , Apoptose , Carcinoma de Células Renais/metabolismo , Carcinoma de Células Renais/patologia , Adesão Celular , Sobrevivência Celular , Senescência Celular , Fatores de Crescimento Endotelial/biossíntese , Endotélio Vascular/metabolismo , Perfilação da Expressão Gênica , Humanos , Neoplasias Renais/metabolismo , Neoplasias Renais/patologia , Glicoproteínas de Membrana/biossíntese , Camundongos , Camundongos SCID , Modelos Biológicos , PTEN Fosfo-Hidrolase , Monoéster Fosfórico Hidrolases/metabolismo , Proteínas Proto-Oncogênicas/metabolismo , Proteínas Proto-Oncogênicas c-akt , Células Tumorais Cultivadas , Proteínas Supressoras de Tumor/metabolismo , Fator D de Crescimento do Endotélio VascularRESUMO
PURPOSE: Platelet-activating factor (PAF), a phospholipid mediator of inflammation, has been recently detected on tumor cells but its effect in tumor development is largely undefined. EXPERIMENTAL DESIGN: To address its potential role in tumor biology, we inhibited intratumor PAF activity by engineering tumor cell lines to express plasma PAF-acetylhydrolase (PAF-AH), the major PAF-inactivating enzyme, and studied their behavior in vitro and in vivo. RESULTS: When transfected with PAF-AH, KS-Imm human Kaposi's sarcoma cells implanted in SCID mice and B16F10 mouse melanoma cells implanted in syngenic C57Bl/6J mice showed significantly reduced vascularization and growth allowing longer survival compared with control tumors. The amounts of bioactive PAF extracted from PAF-AH-transfected tumors were significantly reduced. In vitro, expression of PAF-AH did not influence cell proliferation, whereas it inhibited PAF-dependent cell motility in Kaposi's sarcoma cells that express PAF-receptor but not in melanoma cells that did not express it. On the other hand, PAF-induced endothelial tubulogenesis in Matrigel was inhibited by incubation with supernatant from PAF-AH-transfected melanoma cells, indicating that PAF-AH inhibits in vitro neoangiogenesis. CONCLUSIONS: We demonstrated that in situ PAF inactivation affects tumor vascularization and growth through inhibition of neoangiogenesis and, in the case of cells expressing PAF receptor, also tumor cell motility.
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1-Alquil-2-acetilglicerofosfocolina Esterase/metabolismo , Divisão Celular/fisiologia , Melanoma Experimental/irrigação sanguínea , Neovascularização Patológica/prevenção & controle , Fator de Ativação de Plaquetas/antagonistas & inibidores , 1-Alquil-2-acetilglicerofosfocolina Esterase/genética , Animais , Sequência de Bases , Linhagem Celular Tumoral , Movimento Celular/fisiologia , Quimiotaxia/fisiologia , Primers do DNA , Humanos , Melanoma Experimental/patologia , Camundongos , RNA Mensageiro/genética , Proteínas Recombinantes/metabolismo , TransfecçãoRESUMO
BACKGROUND: The Procera AllCeram (Nobel Biocare AB, Göteborg, Sweden, and Procera Sandvik AB, Stockholm, Sweden) technique is one alternative to metal-ceramic restorations. However, few long-term evaluations of its use for single crowns on natural and implant-supported abutments are available. PURPOSE: The aim of the present study was to assess the clinical performance of Procera AllCeram single crowns when placed in aesthetic sites supported by either natural teeth or implants over a period of 48 months. MATERIALS AND METHODS: Eighty-six single crowns were fabricated and used in 51 patients. The restorations were examined according to the California Dental Association's quality assessment system. RESULTS: One crown was lost after 20 months of follow-up. Of the 85 restorations that completed the 48-month follow-up, only one crown (1.2%) showed a veneering porcelain chip. All crowns were ranked as either excellent or acceptable. The success rates of single crowns supported by natural tooth and implant-supported abutments were 100% and 98.3%, respectively; the total crown success rate was 98.8%. CONCLUSION: Within the limitations of the present study, Procera AllCeram crowns proved to be a reliable therapeutic choice for the restoration of anterior teeth on both natural and implant-supported abutments. The hybrid glass-ionomer cement used in the present study appeared to be a reliable luting agent.
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Coroas , Porcelana Dentária , Prótese Dentária Fixada por Implante , Adolescente , Adulto , Óxido de Alumínio , Cimentação , Resinas Compostas , Dente Canino , Adaptação Marginal Dentária , Falha de Restauração Dentária , Feminino , Cimentos de Ionômeros de Vidro , Humanos , Incisivo , Masculino , Pessoa de Meia-Idade , Estudos RetrospectivosRESUMO
Met, the receptor for hepatocyte growth factor (HGF), is overexpressed in approximately 90% papillary thyroid carcinomas. To investigate the role of the HGF-Met system in these tumors, we examined HGF and Met expression in a variety of primary cultures, normal or malignant thyroid cells, and tissue specimens and analyzed the different HGF effects (promotion of mitogenesis, branching morphogenesis, and cell motility and invasion). In cancer specimens, HGF was produced at high levels by tumor stromal cells, and Met was constitutively phosphorylated in malignant cells. No HGF production was found in a panel of malignant thyroid cancer cells. Biological effects of HGF were examined in papillary cancer cell cultures with either high or low Met expression. High-Met cells were more sensitive to the growth effect of HGF (ED50 = 3-5 ng/ml and 10-15 ng/ml in high- or low-Met cells, respectively). Moreover, only high-Met cells underwent branching morphogenesis in response to HGF. In contrast, high-Met cells showed a reduced migration. Met down-regulation by small interfering RNAs resulted in enhanced cell migration and abrogation of branching morphogenesis in response to HGF. Conversely, Met overexpression impaired cell migration while favoring branching morphogenesis and cell adherence to substrate. These data suggest that both Met and HGF are overexpressed in papillary thyroid carcinomas, that Met is frequently activated in these carcinomas and may favor tumor growth, and that the abundance of Met expression may differently regulate cell growth, morphogenesis, and migration in response to HGF.
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Carcinoma Papilar/fisiopatologia , Fator de Crescimento de Hepatócito/metabolismo , Proteínas Proto-Oncogênicas c-met/metabolismo , Neoplasias da Glândula Tireoide/fisiopatologia , Linhagem Celular Tumoral , Regulação para Baixo , Quinase 1 de Adesão Focal , Proteína-Tirosina Quinases de Adesão Focal , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/fisiologia , Fator de Crescimento de Hepatócito/genética , Fator de Crescimento de Hepatócito/farmacologia , Humanos , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Fosforilação , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas Tirosina Quinases/metabolismo , Proteínas Proto-Oncogênicas/metabolismo , Proteínas Proto-Oncogênicas c-akt , Proteínas Proto-Oncogênicas c-met/genética , Transfecção , Células Tumorais CultivadasRESUMO
Nitric oxide (NO) and platelet-activating factor (PAF) can modulate the interaction between endothelial lining and circulating leukocytes. Several studies implicated the production of PAF and NO in the pathogenesis of microcirculatory alterations occurring in septic shock. However, the reciprocal interaction between PAF and NO has not been fully elucidated. In the present study, we evaluated whether the basal synthesis of NO could modulate the production of PAF by neutrophils (PMN), monocytes (MO), and endothelial cells (EC) unstimulated or stimulated with lipopolysaccharides (LPS) or tumor necrosis factor (TNF). PMN, MO, and EC, when incubated with N(omega)-nitro-L-arginine methyl ester (L-NAME) spontaneously synthesized PAF, with an early peak at 30 min. The effective inhibition of NO production was visualized on MO cells as generation of fluorescence reactivity by cell-permeable NO reactive dye DAF-2 DA. Also, monomethyl-L-arginine (L-NMMA) induced PAF synthesis by PMN, whereas the biologically inactive D-enantiomers of NAME (D-NAME) and of NMMA (D-NMMA) did not. Stimulation of PMN with L-NAME in presence of the exogenous NO donor nitroprusside, of the NO secondary mediator cGMP, or of the NO synthase substrate L-arginine reduced PAF synthesis, suggesting the involvement of an NO-dependent pathway on the modulation of PAF synthesis. The synthesis of PAF was enhanced by combined treatment with L-NAME and TNF or LPS. These results indicate an inhibitor effect of NO on the spontaneous and TNF or LPS-induced synthesis of PAF by human PMN, MO, and EC.
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Endotélio Vascular/efeitos dos fármacos , Monócitos/efeitos dos fármacos , Neutrófilos/efeitos dos fármacos , Óxido Nítrico/fisiologia , Fator de Ativação de Plaquetas/biossíntese , Adulto , Arginina/farmacologia , Células Cultivadas/efeitos dos fármacos , Células Cultivadas/metabolismo , GMP Cíclico/farmacologia , Sinergismo Farmacológico , Endotélio Vascular/citologia , Endotélio Vascular/metabolismo , Inibidores Enzimáticos/farmacologia , Humanos , Lipopolissacarídeos/farmacologia , Monócitos/metabolismo , NG-Nitroarginina Metil Éster/farmacologia , Neutrófilos/metabolismo , Doadores de Óxido Nítrico/farmacologia , Óxido Nítrico Sintase/antagonistas & inibidores , Óxido Nítrico Sintase/metabolismo , Nitroprussiato/farmacologia , Fator de Ativação de Plaquetas/genética , Estereoisomerismo , Fator de Necrose Tumoral alfa/farmacologia , ômega-N-Metilarginina/farmacologiaRESUMO
PURPOSE: To compare the efficacy and safety of 0.1% dexamethasone/0.3% netilmicin (Netildex), with that of 0.1% dexamethasone/0.3% tobramycin (Tobradex) in the treatment of external ocular inflammation requiring antibiotic therapy. METHODS: In this randomized, double-blind study, 139 subjects with conjunctival inflammation associated with signs of ocular infection were treated with Netildex (n = 71) or Tobradex (n = 68) four times daily for 6 days. The primary efficacy analysis was based on the percentage of patients with at least a 50% decrease in conjunctival hyperemia at the endpoint visit (Day 6 [± 1]) with respect to baseline (responder rate). An equivalence margin of 20% was set for this study. A follow-up visit was performed at Day 14 (± 1). Other efficacy parameters were: conjunctival edema, conjunctival discharge, lid hyperemia, lid edema, presence of ocular infection, and symptoms of ocular discomfort. Safety evaluations included intraocular pressure, visual acuity, and adverse events. RESULTS: At Day 6, a decrease of conjunctival hyperemia was observed in 87.3% and 90.9% of the patients treated with Netildex and Tobradex, respectively. The 95% confidence interval for the difference between groups (-15.3 ÷ 8.0) satisfied the equivalence hypothesis. Subjects treated with Netildex had a better control of lid hyperemia (P = 0.016), tearing (P = 0.001), burning (P = 0.007), and stinging (P = 0.004). No adverse reactions were observed during the study except one case of keratitis in the Tobradex group. CONCLUSION: Netildex was as effective and safe as Tobradex in reducing signs and symptoms in patients with conjunctival inflammation when ocular infection was suspected.
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PURPOSE: The aim of this study was to investigate the effects of an ophthalmic gel containing sodium hyaluronate and xanthan gum in addition to the antibiotic netilmicin in the management of traumatic corneal abrasions. PATIENTS AND METHODS: PATIENTS WITH TRAUMATIC CORNEAL ABRASIONS WERE RANDOMLY TREATED AS FOLLOWS: Group A (n = 20) with an occlusive patching for 12 hours plus one drop of an eye gel containing 0.15% sodium hyaluronate, 1% xanthan gum and 0.3% netilmicin qid for 5 days; and Group B (n = 20) with an occlusive patching for 2-3 days plus one application of 0.3% netilmicin ophthalmic ointment qid for 5 days. All patients were evaluated after the third and seventh day by slit-lamp examination, fluorescein staining, and corneal defect photograph in order to assess corneal re-epithelialization. Conjunctival hyperaemia, lid oedema, subjective symptoms of discomfort, and conjunctival swabs were also evaluated. RESULTS: No statistically significant difference was observed between the groups in terms of the extent of corneal healing after 3 days of treatment. Both treatments were also highly effective in decreasing the erosion score and the conjunctival hyperemia (P < 0.0001, P < 0.005, respectively) without any significant difference between the two types of treatment. Subjective symptoms of discomfort and conjunctival swabs were also evaluated. CONCLUSION: In the management of traumatic corneal abrasions, the administration of an eye gel containing sodium hyaluronate and xanthan gum is able to reduce the length of occlusive patching. In addition, the presence of netilmicin guarantees good antibiotic prophylaxis during the wound repair process.
RESUMO
OBJECTIVE: The interest of dental research in metal-free restorations has been rising in the last 20 years following the introduction of innovative all-ceramic materials in the daily practice. In particular, high strength ceramics and related CAD/CAM techniques have widely increased the clinical indications of metal-free prostheses, showing more favourable mechanical characteristics compared to the early ceramic materials. The purpose of the present paper is providing a brief review on the all-ceramic dental materials, evaluating pros and cons in the light of the most recent scientific results and of the authors' clinical experience. MATERIALS: A structured review of the literature was given on the basis of medical and engineering papers published in the last decades on the use of dental ceramics and zirconia in particular. The experimental and clinical findings of the most relevant researches were reported. RESULTS: Zirconia is one of the most promising restorative materials, because it yields very favourable mechanical properties and reasonable esthetic. Several in vitro and in vivo investigations reported suitable strength and mechanical performances of zirconia, compatible with clinical serviceability as a framework material for both single crowns and short-span fixed partial dentures. However, clinical results are not comparable, at the moment, with conventional metal-ceramic restorations, neither is there sufficient long-term data for validating the clinical potential of zirconia in the long run. SIGNIFICANCE: The use of zirconia frameworks for long-span fixed partial dentures or for implant-supported restorations is currently under evaluation and further in vivo, long-term clinical studies will be needed to provide scientific evidence for drawing solid guidelines.