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1.
J Endocrinol Invest ; 44(11): 2367-2374, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-33683662

RESUMO

PURPOSE: Brain tumors are the most common solid tumor in children. The prevalence of survivors from these cancers has been increasing, presenting endocrine sequelae in more than 40% of the cases. Our aim was to characterize the endocrinopathies diagnosed in this population, exploring the outcomes of growth hormone treatment. METHODS: We have performed a retrospective analysis of the survivors that were followed-up through a close protocol at our endocrine late-effects clinic. RESULTS: 242 survivors, followed during 6.4 (0-23.4) years, were considered. The median age at tumor diagnosis was 6.7 (0-18) years and pilocytic astrocytoma was the most frequent neoplasm (33.5%). The prevalence of endocrinopathies was of 71.5%, with growth hormone deficiency being the most frequent (52.9%). An indirect correlation between the age at the beginning of somatropin and growth velocity in the first year of treatment was observed. Those treated with craniospinal radiotherapy presented a smaller final upper/lower segments ratio comparing with those that only received cranial radiotherapy. However, their final height was not compromised when compared to their family height target. We found pubertal delay in 12%; accelerated/precocious puberty in 13.2%; central and primary hypogonadism in 21.9% and 3.3%, respectively; primary and central hypothyroidism in 23.6% and 14.5%, respectively; thyroid nodules in 7.4%; ACTH deficiency in 10.3% and diabetes insipidus in 12%. CONCLUSION: This study reveals a higher prevalence of endocrinopathies in brain tumors survivors and explores the influence of craniospinal irradiation in the adult body proportions. It reinforces the importance of routine follow-up among survivors.


Assuntos
Neoplasias Encefálicas , Sobreviventes de Câncer/estatística & dados numéricos , Doenças do Sistema Endócrino , Transtornos do Crescimento , Hormônio do Crescimento , Radioterapia , Adolescente , Fatores Etários , Neoplasias Encefálicas/epidemiologia , Neoplasias Encefálicas/patologia , Neoplasias Encefálicas/terapia , Criança , Diabetes Insípido/diagnóstico , Diabetes Insípido/etiologia , Doenças do Sistema Endócrino/diagnóstico , Doenças do Sistema Endócrino/epidemiologia , Doenças do Sistema Endócrino/etiologia , Feminino , Seguimentos , Transtornos do Crescimento/epidemiologia , Transtornos do Crescimento/etiologia , Transtornos do Crescimento/metabolismo , Transtornos do Crescimento/terapia , Hormônio do Crescimento/análise , Hormônio do Crescimento/deficiência , Hormônio do Crescimento/uso terapêutico , Hormônio do Crescimento Humano/uso terapêutico , Humanos , Hipotireoidismo/diagnóstico , Hipotireoidismo/epidemiologia , Hipotireoidismo/etiologia , Masculino , Portugal/epidemiologia , Prevalência , Puberdade Tardia/diagnóstico , Puberdade Tardia/epidemiologia , Puberdade Tardia/etiologia , Radioterapia/efeitos adversos , Radioterapia/métodos , Radioterapia/estatística & dados numéricos
2.
Pharm Res ; 37(7): 139, 2020 Jul 13.
Artigo em Inglês | MEDLINE | ID: mdl-32661727

RESUMO

PURPOSE: Lipid nanoparticles (SLN and NLC) were functionalized with the RVG29 peptide in order to target the brain and increase the neuronal uptake through the nicotinic acetylcholine receptors. These nanosystems were loaded with quercetin to take advantage of its neuroprotective properties mainly for Alzheimer's disease. METHODS: The functionalization of nanoparticles with RVG29 peptide was confirmed by NMR and FTIR. Their morphology was assessed by transmission electron microscopy and nanoparticles size, polydispersity and zeta potential were determined by dynamic light scattering. The in vitro validation tests were conducted in hCMEC/D3 cells, a human blood-brain barrier model and thioflavin T binding assay was conducted to assess the process of amyloid-beta peptide fibrillation typical of Alzheimer's disease. RESULTS: RVG29-nanoparticles displayed spherical morphology and size below 250 nm, which is compatible with brain applications. Zeta potential values were between -20 and -25 mV. Quercetin entrapment efficiency was generally higher than 80% and NLC nanoparticles were able to encapsulate up to 90%. The LDH assay showed that there is no cytotoxicity in hCMEC/D3 cell line and RVG29-nanoparticles clearly increased in 1.5-fold the permeability across the in vitro model of blood-brain barrier after 4 h of incubation compared with non-functionalized nanoparticles. Finally, this nanosystem was capable of inhibiting amyloid-beta aggregation in thioflavin T binding assay, suggesting its great potential for neuroprotection. CONCLUSIONS: RVG29-nanoparticles that simultaneously target the blood-brain barrier and induce neurons protection against amyloid-beta fibrillation proved to be an efficient way of quercetin delivery and a promising strategy for future approaches in Alzheimer's disease. Graphical Abstract.


Assuntos
Doença de Alzheimer/tratamento farmacológico , Antígenos Virais/metabolismo , Encéfalo/metabolismo , Lipídeos/química , Fármacos Neuroprotetores/metabolismo , Fragmentos de Peptídeos/metabolismo , Quercetina/metabolismo , Proteínas do Envelope Viral/metabolismo , Doença de Alzheimer/metabolismo , Doença de Alzheimer/patologia , Peptídeos beta-Amiloides/metabolismo , Antígenos Virais/química , Barreira Hematoencefálica/metabolismo , Encéfalo/efeitos dos fármacos , Encéfalo/patologia , Permeabilidade Capilar , Linhagem Celular , Composição de Medicamentos , Humanos , Lipossomos , Nanopartículas , Fármacos Neuroprotetores/administração & dosagem , Fármacos Neuroprotetores/química , Fragmentos de Peptídeos/química , Agregados Proteicos , Agregação Patológica de Proteínas , Quercetina/administração & dosagem , Quercetina/química , Receptores Nicotínicos/metabolismo , Distribuição Tecidual , Proteínas do Envelope Viral/química
3.
J Chem Phys ; 147(4): 044908, 2017 Jul 28.
Artigo em Inglês | MEDLINE | ID: mdl-28764377

RESUMO

We study the compression and extension dynamics of a DNA-like polymer interacting with non-DNA binding and DNA-binding proteins, by means of computer simulations. The geometry we consider is inspired by recent experiments probing the compressional elasticity of the bacterial nucleoid (DNA plus associated proteins), where DNA is confined into a cylindrical container and subjected to the action of a "piston"-a spherical bead to which an external force is applied. We quantify the effect of steric interactions (excluded volume) on the force-extension curves as the polymer is compressed. We find that non-DNA-binding proteins, even at low densities, exert an osmotic force which can be a lot larger than the entropic force exerted by the compressed DNA. The trends we observe are qualitatively robust with respect to changes in protein sizes and are similar for neutral and charged proteins (and DNA). We also quantify the dynamics of DNA expansion following removal of the "piston": while the expansion is well fitted by power laws, the apparent exponent depends on protein concentration and protein-DNA interaction in a significant way. We further highlight an interesting kinetic process which we observe during the expansion of DNA interacting with DNA-binding proteins when the interaction strength is intermediate: the proteins bind while the DNA is packaged by the compression force, but they "pop-off" one-by-one as the force is removed, leading to a slow unzipping kinetics. Finally, we quantify the importance of supercoiling, which is an important feature of bacterial DNA in vivo.


Assuntos
Cromossomos Bacterianos/química , DNA Bacteriano/química , Proteínas de Ligação a DNA/química , Elasticidade , Entropia , Simulação de Dinâmica Molecular , Cinética
4.
Indoor Air ; 25(5): 557-68, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25263282

RESUMO

This work aims to characterize levels and phase distribution of polycyclic aromatic hydrocarbons (PAHs) in indoor air of preschool environment and to assess the impact of outdoor PAH emissions to indoor environment. Gaseous and particulate (PM1 and PM(2.5)) PAHs (16 USEPA priority pollutants, plus dibenzo[a,l]pyrene, and benzo[j]fluoranthene) were concurrently sampled indoors and outdoors in one urban preschool located in north of Portugal for 35 days. The total concentration of 18 PAHs (ΣPAHs) in indoor air ranged from 19.5 to 82.0 ng/m(3) ; gaseous compounds (range of 14.1-66.1 ng/m(3)) accounted for 85% ΣPAHs. Particulate PAHs (range 0.7-15.9 ng/m(3)) were predominantly associated with PM1 (76% particulate ΣPAHs) with 5-ring PAHs being the most abundant. Mean indoor/outdoor ratios (I/O) of individual PAHs indicated that outdoor emissions significantly contributed to PAH indoors; emissions from motor vehicles and fuel burning were the major sources.


Assuntos
Poluição do Ar em Ambientes Fechados/análise , Material Particulado/análise , Hidrocarbonetos Policíclicos Aromáticos/análise , Escolas Maternais/estatística & dados numéricos
5.
Indoor Air ; 24(6): 618-28, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-24689947

RESUMO

UNLABELLED: The aim of this work was to assess ultrafine particles (UFP) number concentrations in different microenvironments of Portuguese preschools and to estimate the respective exposure doses of UFP for 3-5-year-old children (in comparison with adults). UFP were sampled both indoors and outdoors in two urban (US1, US2) and one rural (RS1) preschool located in north of Portugal for 31 days. Total levels of indoor UFP were significantly higher at the urban preschools (mean of 1.82 × 10(4) and 1.32 × 10(4) particles/cm(3) at US1 an US2, respectively) than at the rural one (1.15 × 10(4) particles/cm(3) ). Canteens were the indoor microenvironment with the highest UFP (mean of 5.17 × 10(4) , 3.28 × 10(4) , and 4.09 × 10(4) particles/cm(3) at US1, US2, and RS1), whereas the lowest concentrations were observed in classrooms (9.31 × 10(3) , 11.3 × 10(3) , and 7.14 × 10(3) particles/cm(3) at US1, US2, and RS1). Mean indoor/outdoor ratios (I/O) of UFP at three preschools were lower than 1 (0.54-0.93), indicating that outdoor emissions significantly contributed to UFP indoors. Significant correlations were obtained between temperature, wind speed, relative humidity, solar radiation, and ambient UFP number concentrations. The estimated exposure doses were higher in children attending urban preschools; 3-5-year-old children were exposed to 4-6 times higher UFP doses than adults with similar daily schedules. PRACTICAL IMPLICATIONS: This study reports information on ultrafine particles (UFPs) in various indoor and outdoor microenvironments (canteens, classrooms, gymnasiums, and outdoor) of urban and rural preschools. It identifies the potential sources and origins, characterizes the influence of meteorological parameters on UFP levels, and performs a comparison with other existing international studies. To this date, relatively few studies have investigated UFP in preschools (none in Portugal) and none assessed exposure dose for different age-groups. The obtained findings showed that levels of UFP in various microenvironments of schools differed significantly. Therefore, to obtain an accurate representation of child's overall preschool exposure profiles, the exposures occurring in these different microenvironments should be always accounted for.


Assuntos
Poluição do Ar em Ambientes Fechados/análise , Instituições Acadêmicas , Adulto , Pré-Escolar , Monitoramento Ambiental , Feminino , Humanos , Exposição por Inalação , Masculino , Tamanho da Partícula , Material Particulado/análise , Portugal , Tempo (Meteorologia)
6.
Int Endod J ; 47(5): 416-24, 2014 May.
Artigo em Inglês | MEDLINE | ID: mdl-23992346

RESUMO

AIM: To synthesize and characterize zinc oxide (ZnO) nanocrystals and assess their biological properties for applications in dentistry, particularly in endodontics, by means of intraosseous implants. METHODOLOGY: ZnO nanocrystals were synthesized and characterized by micro-Raman spectroscopy and X-ray Diffraction. Ten guinea pigs were divided into two groups representing experimental periods of 4 and 12 weeks and received implants on both sides of the mandible in the region of the symphysis. The connective tissue response along the lateral wall outside the cup served as the negative control. The animals were euthanized at the end of each observation period and prepared for routine histological examinations which evaluated inflammatory response and material biocompatibility. RESULTS: ZnO nanocrystals were highly pure, crystalline, and averaged 21 nm in size. After 12 weeks, tissue analysis revealed the presence of a thin layer of connective tissue with some giant cells between the implanted material and underlying bone tissue. While signs of mild inflammation could be seen, bone resorption and remodeling were not apparent. CONCLUSION: ZnO nanocrystals were biocompatible, well tolerated and allowed new bone formation and bone remodelling.


Assuntos
Materiais Biocompatíveis , Implantação Dentária Endóssea , Nanopartículas/química , Óxido de Zinco/química , Cristalografia por Raios X , Análise Espectral Raman
7.
Genet Mol Res ; 13(4): 10035-48, 2014 Nov 28.
Artigo em Inglês | MEDLINE | ID: mdl-25501215

RESUMO

The cuticular chemical composition plays a significant role in the recognition of nest mates in social insects, thus functioning as a chemical signature of the colony. The structure of cuticular chemicals is subject to interference from genetic and exogenous factors, including diet. In this study, various colonies of the Ectatomma brunneum ant were removed from their natural environment and housed in a laboratory to monitor the response of the cuticular chemical composition to dietary changes. Analyses were performed using gas chromatography and Fourier transform infrared photoacoustic spectroscopy, which has not been previously used for this type of analysis. The results indicate that this method is useful for analyzing biological and natural systems. We observed changes in the chemical signature with food traces in the first 30 days under feed control. Therefore, genetic information may not be the only criterion that can be used to describe the chemical signature of a species; environmental variations also influence recognition signals. Furthermore, these results reinforce the reliability of the Fourier transform infrared photoacoustic spectroscopy method.


Assuntos
Formigas/química , Dieta , Tegumento Comum/fisiologia , Técnicas Fotoacústicas , Animais , Cromatografia Gasosa , Análise Discriminante , Ionização de Chama , Hidrocarbonetos/análise , Espectroscopia de Infravermelho com Transformada de Fourier , Vibração
8.
Parasitology ; 140(2): 171-80, 2013 02.
Artigo em Inglês | MEDLINE | ID: mdl-22975090

RESUMO

Heparin-binding proteins (HBPs) play a key role in Trypanosoma cruzi-host cell interactions. HBPs recognize heparan sulfate (HS) at the host cell surface and are able to induce the cytoadherence and invasion of this parasite. Herein, we analysed the biochemical properties of the HBPs and also evaluated the expression and subcellular localization of HBPs in T. cruzi trypomastigotes. A flow cytometry analysis revealed that HBPs are highly expressed at the surface of trypomastigotes, and their peculiar localization mainly at the flagellar membrane, which is known as an important signalling domain, may enhance their binding to HS and elicit the parasite invasion. The plasmon surface resonance results demonstrated the stability of HBPs and their affinity to HS and heparin. Additionally, gelatinolytic activities of 70 kDa, 65·8 kDa and 59 kDa HBPs over a broad pH range (5·5-8·0) were revealed using a zymography assay. These proteolytic activities were sensitive to serine proteinase inhibitors, such as aprotinin and phenylmethylsulfonyl fluoride, suggesting that HBPs have the properties of trypsin-like proteinases.


Assuntos
Membrana Celular/metabolismo , Flagelos/enzimologia , Proteínas de Protozoários/metabolismo , Serina Proteases/metabolismo , Trypanosoma cruzi/fisiologia , Membrana Celular/enzimologia , Ativação Enzimática/efeitos dos fármacos , Citometria de Fluxo , Gelatina/metabolismo , Regulação da Expressão Gênica , Heparina/metabolismo , Interações Hospedeiro-Parasita , Concentração de Íons de Hidrogênio , Ligação Proteica , Inibidores de Serina Proteinase/farmacologia , Trypanosoma cruzi/enzimologia
9.
Animal ; 17(4): 100734, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-36871322

RESUMO

DM intake (DMI) for individual pens of cattle is recorded daily or averaged across each week by most commercial feedlots as an index of performance. Numerous factors impact DMI by feedlot cattle. Some are available at the start of the feedlot period (initial BW, sex), and others become available early in the feeding period (daily DMI during adaptation) or more continuously (daily DMI from the previous week). To evaluate the relative impact of these factors on daily DMI during individual weeks within the feedlot period, we employed a dataset compiled from 2009 to 2014 from one commercial feedlot, including 4 132 pens (485 458 cattle), which were split into two fractions: 80% were used to calculate DMI regressions on these factors to develop a prediction equation for mean DMI for each week of the feeding period, and 20% were reserved to test the adequacy of these prediction equations. Correlations were used to determine the relationship between all available variables with observed DMI. These variables were then included in the generalized least squares regression models. A veracity test of the model was performed against the reserved data. Daily DMI from previous week was the factor most highly correlated with daily DMI (P < 0.10) during from week 6 to week 31, accounting for approximately 70% of the variation, followed by mean daily DMI during adaptation period (weeks 1-4), including in the prediction model from weeks 5 to 12. Initial shrunk BW (ISBW) was the third most correlated factor, which was included in prediction equations from week 5 to week 20. Sex entered the prediction model only after week 8. Daily DMI for each test week within the feeding period was predicted closely (r2 = 0.98) by these four factors (RMSE = 0.155 kg). In conclusion, the mean daily DMI during each week of the finishing period for a pen of cattle could be predicted closely based on mean daily DMI intake during the previous week plus other variables available early in a feedlot period (daily DMI during adaptation period, ISBW and sex).


Assuntos
Ração Animal , Dieta , Bovinos , Animais , Dieta/veterinária , Ração Animal/análise
10.
J Periodontal Res ; 47(2): 149-58, 2012 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-21883230

RESUMO

BACKGROUND AND OBJECTIVE: Ciclosporin A (CsA)-induced gingival overgrowth is attributed to an exaggerated accumulation of extracellular matrix, which is mainly due to an increased expression of transforming growth factor-ß1 (TGF-ß1). Herein, the in vitro investigation of effects of overexpression of Smad7, a TGF-ß1 signaling inhibitor, in the events associated with CsA-induced extracellular matrix accumulation was performed. MATERIAL AND METHODS: The effects of Smad7 were assessed by stable overexpression of Smad7 in fibroblasts from normal gingiva. Smad7-overexpressing cells and control cells were incubated with CsA, and synthesis of type I collagen, production and activity of MMP-2 and cellular proliferation were evaluated by ELISA, zymography, growth curve, bromodeoxyuridine incorporation assay and cell cycle analysis. The effects of CsA on cell viability and apoptosis of fibroblasts from normal gingiva were also evaluated. Western blot and immunofluorescence for phospho-Smad2 were performed to measure the activation of TGF-ß1 signaling. RESULTS: Although the treatment with CsA stimulated TGF-ß1 production in both control and Smad7-overexpressing fibroblasts, its signaling was markedly inhibited in Smad7-overexpressing cells, as revealed by low levels of phospho-Smad2. In Smad7-overexpressing cells, the effects of CsA on proliferation, synthesis of type I collagen and the production and activity of MMP-2 were significantly blocked. Smad7 overexpression blocked CsA-induced fibroblast proliferation via p27 regulation. Neither CsA nor Smad7 overexpression induced cell death. CONCLUSION: The data presented here confirm that TGF-ß1 expression is related to the molecular events associated with CsA-induced gingival overgrowth and suggest that Smad7 overexpression is effective in blocking these events, including proliferation, type I collagen synthesis and MMP-2 activity.


Assuntos
Ciclosporina/efeitos adversos , Fibroblastos/efeitos dos fármacos , Gengiva/efeitos dos fármacos , Crescimento Excessivo da Gengiva/induzido quimicamente , Proteína Smad7/farmacologia , Antimetabólitos , Apoptose/efeitos dos fármacos , Bromodesoxiuridina , Técnicas de Cultura de Células , Ciclo Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Colágeno Tipo I/efeitos dos fármacos , Inibidor de Quinase Dependente de Ciclina p27/metabolismo , Ciclosporina/antagonistas & inibidores , Matriz Extracelular/efeitos dos fármacos , Matriz Extracelular/metabolismo , Fibroblastos/metabolismo , Regulação da Expressão Gênica/genética , Gengiva/citologia , Gengiva/metabolismo , Humanos , Masculino , Metaloproteinase 2 da Matriz/efeitos dos fármacos , Fosforilação , Inibidores de Proteínas Quinases/metabolismo , Transdução de Sinais/efeitos dos fármacos , Proteína Smad2/efeitos dos fármacos , Proteína Smad7/genética , Transfecção , Fator de Crescimento Transformador beta1/antagonistas & inibidores , Adulto Jovem
11.
Parasitology ; 139(6): 735-43, 2012 May.
Artigo em Inglês | MEDLINE | ID: mdl-22310218

RESUMO

Heparin-binding proteins (HBPs) have been demonstrated in both infective forms of Trypanosoma cruzi and are involved in the recognition and invasion of mammalian cells. In this study, we evaluated the potential biological function of these proteins during the parasite-vector interaction. HBPs, with molecular masses of 65·8 kDa and 59 kDa, were isolated from epimastigotes by heparin affinity chromatography and identified by biotin-conjugated sulfated glycosaminoglycans (GAGs). Surface plasmon resonance biosensor analysis demonstrated stable receptor-ligand binding based on the association and dissociation values. Pre-incubation of epimastigotes with GAGs led to an inhibition of parasite binding to immobilized heparin. Competition assays were performed to evaluate the role of the HBP-GAG interaction in the recognition and adhesion of epimastigotes to midgut epithelial cells of Rhodnius prolixus. Epithelial cells pre-incubated with HBPs yielded a 3·8-fold inhibition in the adhesion of epimastigotes. The pre-treatment of epimastigotes with heparin, heparan sulfate and chondroitin sulfate significantly inhibited parasite adhesion to midgut epithelial cells, which was confirmed by scanning electron microscopy. We provide evidence that heparin-binding proteins are found on the surface of T. cruzi epimastigotes and demonstrate their key role in the recognition of sulfated GAGs on the surface of midgut epithelial cells of the insect vector.


Assuntos
Células Epiteliais/parasitologia , Heparina/metabolismo , Interações Hospedeiro-Parasita , Proteínas de Protozoários/farmacologia , Rhodnius/parasitologia , Trypanosoma cruzi/fisiologia , Animais , Adesão Celular/efeitos dos fármacos , Adesão Celular/fisiologia , Trato Gastrointestinal/citologia , Trato Gastrointestinal/parasitologia , Proteínas de Protozoários/metabolismo , Trypanosoma cruzi/crescimento & desenvolvimento
12.
Water Sci Technol ; 66(4): 704-11, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22766856

RESUMO

One of the most important causes for poor water quality in urban rivers in Brazil is the low collection efficiency of the sewer system due to unforeseen interconnections with the stormwater drainage system. Since the beginning of the 20th century, Brazilian cities have adopted separate systems for sanitary sewers and stormwater runoff. Gradually these two systems became interconnected. A major challenge faced today by water managers in Brazil is to find efficient and low cost solutions to deal with this mixed system. The current situation poses an important threat to the improvement of the water quality in urban rivers and lakes. This article presents an evaluation of the water quality parameters and the diffuse pollution loads during rain events in the Pinheiros River, a tributary of the Tietê River in São Paulo. It also presents different types of integrated solutions for reducing the pollution impact of combined systems, based on the European experience in urban water management. An evaluation of their performance and a comparison with the separate system used in most Brazilian cities is also presented. The study is based on an extensive water quality monitoring program that was developed for a special investigation in the Pinheiros River and lasted 2.5 years. Samples were collected on a daily basis and water quality variables were analyzed on a daily, weekly or monthly basis. Two hundred water quality variables were monitored at 53 sampling points. During rain events, additional monitoring was carried out using an automated sampler. Pinheiros River is one of the most important rivers in the São Paulo Metropolitan Region and it is also a heavily polluted one.


Assuntos
Rios/química , Poluentes da Água/análise , Poluição da Água/prevenção & controle , Amônia/análise , Análise da Demanda Biológica de Oxigênio , Brasil , Cidades , Drenagem Sanitária , Monitoramento Ambiental , Escherichia coli/isolamento & purificação , Nitratos/análise , Nitritos/análise , Fósforo/análise , Esgotos , Eliminação de Resíduos Líquidos , Movimentos da Água
13.
J Antimicrob Chemother ; 66(5): 1036-43, 2011 May.
Artigo em Inglês | MEDLINE | ID: mdl-21393196

RESUMO

OBJECTIVES: Antimicrobial resistance is a major public health concern, particularly in hospitals and other healthcare settings. For the rational design of disinfection strategies, it is of utmost importance to understand the mechanisms of action of antimicrobials. In this study, the mechanism of action of benzyldimethyldodecylammonium chloride (BDMDAC) was assessed against Pseudomonas fluorescens. METHODS: The targets of antimicrobial action were studied using different bacterial physiological indices. The MIC, MBC, membrane permeabilization, intracellular potassium release, physico-chemical surface properties, surface charge, outer membrane protein (OMP) expression and morphological changes were assessed after BDMDAC exposure. RESULTS: The MIC was found to be 20 mg/L and the MBC was 10 mg/L. BDMDAC led to a significant change in cell surface hydrophobicity and induced propidium iodide uptake. Such results suggest cytoplasmic membrane damage, corroborated by the release of intracellular potassium. The results obtained from the zeta potential measurement demonstrate a -31.2 mV value for untreated cells and -21.0 mV for cells at the MIC. Scanning electron microscopy revealed that cells treated with 20 mg/L were less bulky, and their membrane seemed to be rougher, wrinkled and deformed when compared with untreated cells. The overall bactericidal events occurred without detectable changes in OMP expression. CONCLUSIONS: BDMDAC is an effective biocide against P. fluorescens. It binds by ionic and hydrophobic interactions to the cell membrane, causing changes in membrane properties and function, as manifested by phenomena such as cellular disruption and loss of membrane integrity with consequent leakage of essential intracellular constituents.


Assuntos
Antibacterianos/farmacologia , Compostos de Benzalcônio/farmacologia , Pseudomonas fluorescens/efeitos dos fármacos , Compostos de Amônio Quaternário/farmacologia , Proteínas da Membrana Bacteriana Externa/metabolismo , Permeabilidade da Membrana Celular/efeitos dos fármacos , Humanos , Testes de Sensibilidade Microbiana , Viabilidade Microbiana/efeitos dos fármacos , Eletricidade Estática , Propriedades de Superfície
14.
Parasitology ; 138(5): 593-601, 2011 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-21269549

RESUMO

Cell surface glycosaminoglycans (GAGs) play an important role in the attachment and invasion process of a variety of intracellular pathogens. We have previously demonstrated that heparan sulfate proteoglycans (HSPG) mediate the invasion of trypomastigote forms of Trypanosoma cruzi in cardiomyocytes. Herein, we analysed whether GAGs are also implicated in amastigote invasion. Competition assays with soluble GAGs revealed that treatment of T. cruzi amastigotes with heparin and heparan sulfate leads to a reduction in the infection ratio, achieving 82% and 65% inhibition of invasion, respectively. Other sulfated GAGs, such as chondroitin sulfate, dermatan sulfate and keratan sulfate, had no effect on the invasion process. In addition, a significant decrease in infection occurred after interaction of amastigotes with GAG-deficient Chinese Hamster Ovary (CHO) cells, decreasing from 20% and 28% in wild-type CHO cells to 5% and 9% in the mutant cells after 2 h and 4 h of infection, respectively. These findings suggest that amastigote invasion also involves host cell surface heparan sulfate proteoglycans. The knowledge of the mechanism triggered by heparan sulfate-binding T. cruzi proteins may provide new potential candidates for Chagas disease therapy.


Assuntos
Doença de Chagas/parasitologia , Proteoglicanas de Heparan Sulfato/metabolismo , Heparina/farmacologia , Heparitina Sulfato/farmacologia , Trypanosoma cruzi/fisiologia , Animais , Células CHO , Adesão Celular/efeitos dos fármacos , Membrana Celular/metabolismo , Células Cultivadas , Cricetinae , Cricetulus , Citometria de Fluxo , Interações Hospedeiro-Parasita/efeitos dos fármacos , Camundongos , Microscopia Eletrônica de Transmissão , Mutação , Miócitos Cardíacos/parasitologia , Fatores de Tempo , Trypanosoma cruzi/efeitos dos fármacos , Trypanosoma cruzi/patogenicidade
15.
Hum Exp Toxicol ; 40(11): 1962-1973, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34002645

RESUMO

The application of gold nanoparticle-peptide conjugates as theranostic agents for colorectal cancer shows much promise. This study aimed at determining the neurotoxic impact of 14 nm gold nanoparticles (AuNPs) functionalized with colorectal cancer-targeting peptides (namely p.C, p.L or p.14) in a rat model. Brain tissue samples, obtained from Wistar rats that received a single injection of citrate-capped AuNPs, polyethylene glycol-coated (PEG) AuNPs, p.C-PEG-AuNPs, p.L-PEG-AuNPs or p.14-PEG-AuNPs, and sacrificed after 2- and 12-weeks, respectively, were analysed. Inflammation marker (tumour necrosis factor-α, interleukin-6, interleukin-1ß), oxidative stress (superoxide dismutase, catalase, glutathione peroxidase) and apoptotic biomarker (cytochrome c, caspase-3) levels were measured. Gold nanoparticle-treated groups sacrificed after 2-weeks did not exhibit any significant inflammatory, oxidative stress or apoptotic effects in brain tissue compared to the untreated control group. In brain tissue from rats that were exposed to citrate-capped AuNPs for 12-weeks, tumour necrosis factor-α and interleukin-6 levels were significantly increased compared to the untreated control. Exposure to PEG-AuNP, p.C-PEG-AuNP, p.L-PEG-AuNP and p.14-PEG-AuNP did not elicit significant toxic effects compared to the control after 12-weeks, as evidenced by the absence of inflammatory, oxidative stress and apoptotic effects in brain tissue. We thus report on the safety of PEG-coated AuNP-peptide conjugates for potential application in the diagnosis of colorectal cancer; however, exposure to citrate-capped AuNPs could induce delayed neuro-inflammation, and as such, warrants further investigation.


Assuntos
Neoplasias Colorretais/tratamento farmacológico , Ouro/toxicidade , Ouro/uso terapêutico , Nanopartículas Metálicas/toxicidade , Nanopartículas Metálicas/uso terapêutico , Síndromes Neurotóxicas/etiologia , Síndromes Neurotóxicas/fisiopatologia , Animais , Masculino , Terapia de Alvo Molecular/efeitos adversos , Ratos , Ratos Wistar
16.
Nat Commun ; 12(1): 5756, 2021 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-34599163

RESUMO

The discovery that overexpressing one or a few critical transcription factors can switch cell state suggests that gene regulatory networks are relatively simple. In contrast, genome-wide association studies (GWAS) point to complex phenotypes being determined by hundreds of loci that rarely encode transcription factors and which individually have small effects. Here, we use computer simulations and a simple fitting-free polymer model of chromosomes to show that spatial correlations arising from 3D genome organisation naturally lead to stochastic and bursty transcription as well as complex small-world regulatory networks (where the transcriptional activity of each genomic region subtly affects almost all others). These effects require factors to be present at sub-saturating levels; increasing levels dramatically simplifies networks as more transcription units are pressed into use. Consequently, results from GWAS can be reconciled with those involving overexpression. We apply this pan-genomic model to predict patterns of transcriptional activity in whole human chromosomes, and, as an example, the effects of the deletion causing the diGeorge syndrome.


Assuntos
Redes Reguladoras de Genes , Genoma Humano , Modelos Genéticos , Fatores de Transcrição/metabolismo , Cromatina/química , Cromatina/metabolismo , Cromossomos Humanos/química , Cromossomos Humanos/metabolismo , Estudo de Associação Genômica Ampla , Humanos , Polímeros/química , Polímeros/metabolismo , Locos de Características Quantitativas , Transcrição Gênica
17.
Horm Metab Res ; 42(1): 50-5, 2010 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-19798623

RESUMO

Biochemical markers for remission on acromegaly activity are controversial. We studied a subset of treated acromegalic patients with discordant nadir GH levels after oral glucose tolerance test (oGTT) and IGF-I values to refine the current consensus on acromegaly remission. We also compared GH results by two GH immunoassays. From a cohort of 75 treated acromegalic patients, we studied 13 patients who presented an elevated IGF-I despite post-oGTT nadir GH of < or =1 microg/l. The 12-h daytime GH profile (GH-12 h), nadir GH after oGTT, and basal IGF-I levels were studied in patients and controls. Bland-Altman method showed high concordance between GH assays. Acromegalic patients showed higher mean GH-12 h values (0.71+/-0.36 vs. 0.31+/-0.28 microg/l; p<0.05) and nadir GH after oGTT (0.48+/-0.32 vs. 0.097+/-0.002 microg/l; p<0.05) as compared to controls. Nadir GH correlated with mean GH-12 h (r=0.92, p<0.05). The mean GH-12 h value from upper 95% CI of controls (0.54 microg/l) would correspond to a theoretical normal nadir GH of < or =0.27 microg/l. Patients with GH nadir < or =0.3 microg/l had IGF-I between 100-130% ULNR (percentage of upper limit of normal range) and mean GH-12 h of 0.35+/-0.15, and patients with GH nadir >0.3 and < or =1 microg/l had IGF-I >130% ULNR and mean GH-12 h of 0.93+/-0.24 microg/l. Our data integrate daytime GH secretion, nadir GH after oGTT, and plasma IGF-I concentrations showing a continuum of mild residual activity in a subgroup of treated acromegaly with nadir GH values < or =1 microg/l. The degree of increased IGF-I levels and nadir GH after oGTT are correlated with the subtle abnormalities of daytime GH secretion.


Assuntos
Acromegalia/metabolismo , Hormônio do Crescimento Humano/sangue , Fator de Crescimento Insulin-Like I/metabolismo , Acromegalia/radioterapia , Acromegalia/cirurgia , Adulto , Idoso , Biomarcadores/sangue , Estudos de Casos e Controles , Estudos de Coortes , Feminino , Glucose/metabolismo , Teste de Tolerância a Glucose , Humanos , Masculino , Pessoa de Meia-Idade
18.
Lung ; 188(3): 263-8, 2010 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-20049470

RESUMO

This study was designed to use volumetric capnography to evaluate the breathing pattern and ventilation inhomogeneities in patients with chronic sputum production and bronchiectasis and to correlate the phase 3 slope of the capnographic curve to spirometric measurements. Twenty-four patients with cystic fibrosis (CF) and 21 patients with noncystic fibrosis idiopathic bronchiectasis (BC) were serially enrolled. The diagnosis of cystic fibrosis was based on the finding of at least two abnormal sweat chloride concentrations (iontophoresis sweat test). The diagnosis of bronchiectasis was made when the patient had a complaint of chronic sputum production and compatible findings at high-resolution computed tomography (HRCT) scan of the thorax. Spirometric tests and volumetric capnography were performed. The 114 subjects of the control group for capnographic variables were nonsmoker volunteers, who had no respiratory symptoms whatsoever and no past or present history of lung disease. Compared with controls, patients in CF group had lower SpO(2) (P < 0.0001), higher respiratory rates (RR) (P < 0.0001), smaller expiratory volumes normalized for weight (V(E)/kg) (P < 0.028), smaller expiratory times (Te) (P < 0.0001), and greater phase 3 Slopes normalized for tidal volume (P3Slp/V(E)) (P < 0.0001). Compared with controls, patients in the BC group had lower SpO(2) (P < 0.0001), higher RR (P < 0.004), smaller V(E)/kg (P < 0.04), smaller Te (P < 0.007), greater P3Slp/V(E) (P < 0.0001), and smaller VCO(2) (P < 0.0002). The pooled data from the two patient groups compared with controls showed that the patients had lower SpO(2) (P < 0.0001), higher RR (P < 0.0001), smaller V(E)/kg (P < 0.05), smaller Te (P < 0.0001), greater P3Slp/V(E) (P < 0.0001), and smaller VCO(2) (P < 0.0003). All of the capnographic and spirometric variables evaluated showed no significant differences between CF and BC patients. Spirometric data in this study reveals that the patients had obstructive defects with concomitant low vital capacities and both groups had very similar abnormalities. The capnographic variables in the patient group suggest a restrictive respiratory pattern (greater respiratory rates, smaller expiratory times and expiratory volumes, normal peak expiratory flows). Both groups of patients showed increased phase III slopes compared with controls, which probably indicates the presence of diffuse disease of small airways in both conditions leading to inhomogeneities of ventilation.


Assuntos
Bronquiectasia/fisiopatologia , Capnografia/métodos , Fibrose Cística/fisiopatologia , Adulto , Testes Respiratórios , Feminino , Volume Expiratório Forçado , Humanos , Masculino , Pessoa de Meia-Idade , Ventilação Pulmonar , Taxa Respiratória , Espirometria , Escarro/metabolismo , Capacidade Vital
19.
Biofouling ; 26(2): 205-12, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-19937490

RESUMO

This study presents a new technological approach to minimize the use of antimicrobial (AMB) agents and their deleterious effects, based on the principle of drug-delivery systems whereby the AMB chemicals are transported on microparticles. The efficacy of microparticles carrying the quaternary ammonium compound (QAC), benzyldimethyldodecyl ammonium chloride (BDMDAC), was assessed against Pseudomonas fluorescens in both the planktonic and the biofilm state. The microparticles were prepared using a layer-by-layer (LBL) self-assembly technique. Oppositely charged molecules of polyethyleneimine (PEI), sodium polystyrene sulfonate (PSS), and BDMDAC were assembled on polystyrene (PS) cores. BDMDAC-coated particles were observed by CryoSEM and their composition analyzed by X-ray microanalysis. Zeta potential measurements indicated that changes in surface charge were compatible with a BDMDAC/particle interaction. This biocidal carrier structure had significant stability, verified by the release of only 15% of the BDMDAC when immersed in water for 18 months. Biocidal carrier activity was evaluated by determining the survival ratio of P. fluorescens planktonic and biofilm cells after different exposure periods to BDMDAC-coated particles. Tests with biofilm cells were also performed with the free QAC. An efficient AMB effect (minimum bactericidal concentration) against suspended cells was found for a concentration of 9.2 mg l(-1) of BDMDAC on coated particles after incubation for 30 min and 6.5 mg l(-1) of BDMDAC on coated particles after 60 min. Exposure of biofilms to PS-PEI/PSS/BDMDAC (0.87 mg l(-1)) resulted in a decrease in viability of 60.5% and 66.5% of the total biofilm population for 30 and 60 min exposure times, respectively. Exposure for 60 min to 6.33 mg l(-1) and 11.75 mg l(-1) of BDMDAC in PS-PEI/PSS/BDMDAC particles promoted inactivation of 80.6% and 87.2% of the total population, respectively. The AMB effects obtained with the application of free BDMDAC were statistically similar to those promoted by the application of BDMDAC coated particles. The overall results indicate that this novel AMB strategy has potential for the control of microbial growth of planktonic cells and biofouling. Moreover, the technique allows the reuse of AMB molecules and consequently reduces the environmental risks associated with excessive use of AMB agents, thereby providing real benefits to public health.


Assuntos
Compostos de Benzalcônio/administração & dosagem , Incrustação Biológica/prevenção & controle , Desinfetantes/administração & dosagem , Pseudomonas fluorescens , Biofilmes , Microesferas , Plâncton
20.
Int J Sports Med ; 31(12): 866-9, 2010 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-21072739

RESUMO

The purpose of this study was to examine the effect of two between set rest intervals (RI) on isokinetic knee extension peak torque (PT) produced by sedentary women and men. Seventeen young women (27.18 ± 4.05 yrs) and 16 young men (26.75 ± 4.73 yrs) performed 3 sets of 10 unilateral isokinetic knee extension at 60° and 180°/s. The RI between sets was 60 and 120 s, counterbalanced across 2 testing days. Statistical evaluation of the data was performed using a 3-way mixed factor ANOVA (gender x rest interval x sets). Males and females exhibited decreases (p<0.05) in PT with 120 s RI at 60°/s. There was no significant decline in PT in the female group during both RI at 180°/s. Men showed a significant decrease in PT only with 60 s RI. Young women and men require more than 120 s of RI to recover full PT at 60°/s. However, full quadriceps's muscle strength recovery can be attained with a 60 and 120 s at 180°/s in women, but in men only with a 120 s at the same velocity.


Assuntos
Articulação do Joelho/fisiologia , Contração Muscular/fisiologia , Fadiga Muscular/fisiologia , Músculo Esquelético/fisiologia , Adulto , Análise de Variância , Feminino , Humanos , Contração Isométrica/fisiologia , Masculino , Fatores Sexuais , Fatores de Tempo , Torque , Adulto Jovem
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