RESUMO
PURPOSE: In clinical practice the assessment of the "vocal cord-arytenoid unit" (VCAU) mobility is crucial in the staging, prognosis, and choice of treatment of laryngeal squamous cell carcinoma (LSCC). The aim of the present study was to measure repeatability and reliability of clinical assessment of VCAU mobility and radiologic analysis of posterior laryngeal extension. METHODS: In this multi-institutional retrospective study, patients with LSCC-induced impairment of VCAU mobility who received curative treatment were included; pre-treatment endoscopy and contrast-enhanced imaging were collected and evaluated by raters. According to their evaluations, concordance, number of assigned categories, and inter- and intra-rater agreement were calculated. RESULTS: Twenty-two otorhinolaryngologists evaluated 366 videolaryngoscopies (total evaluations: 2170) and 6 radiologists evaluated 237 imaging studies (total evaluations: 477). The concordance of clinical rating was excellent in only 22.7% of cases. Overall, inter- and intra-rater agreement was weak. Supraglottic cancers and transoral endoscopy were associated with the lowest inter-observer reliability values. Radiologic inter-rater agreement was low and did not vary with imaging technique. Intra-rater reliability of radiologic evaluation was optimal. CONCLUSIONS: The current methods to assess VCAU mobility and posterior extension of LSCC are flawed by weak inter-observer agreement and reliability. Radiologic evaluation was characterized by very high intra-rater agreement, but weak inter-observer reliability. The relevance of VCAU mobility assessment in laryngeal oncology should be re-weighted. Patients affected by LSCC requiring imaging should be referred to dedicated radiologists with experience in head and neck oncology.
Assuntos
Neoplasias Laríngeas , Prega Vocal , Humanos , Neoplasias Laríngeas/diagnóstico por imagem , Neoplasias Laríngeas/patologia , Masculino , Feminino , Pessoa de Meia-Idade , Idoso , Estudos Retrospectivos , Prega Vocal/diagnóstico por imagem , Prega Vocal/fisiopatologia , Adulto , Reprodutibilidade dos Testes , Idoso de 80 Anos ou mais , Laringoscopia/métodos , Carcinoma de Células Escamosas/diagnóstico por imagem , Carcinoma de Células Escamosas/patologiaRESUMO
Nanostructured SnO2 is a promising material for the scalable production of portable gas sensors. To fully exploit their potential, these gas sensors need a faster recovery rate and higher sensitivity at room temperature than the current state of the art. Here we demonstrate a chemiresistive gas sensor based on vertical SnOx nanopillars, capable of sensing < 5 ppm of H2 at room temperature and 10 ppt at 230 °C. We test the sample both in vacuum and in air and observe an exceptional improvement in the performance compared to commercially available gas sensors. In particular, the recovery time for sensing NH3 at room temperature is more than one order of magnitude faster than a commercial SnO2 sensor. The sensor shows an unique combination of high sensitivity and fast recovery time, matching the requirements on materials expected to foster widespread use of portable and affordable gas sensors.
RESUMO
An easy transfer procedure to obtain graphene-based gas sensing devices operating at room temperature (RT) is presented. Starting from chemical vapor deposition-grown graphene on copper foil, we obtained single layer graphene which could be transferred onto arbitrary substrates. In particular, we placed single layer graphene on top of a SiO2/Si substrate with pre-patterned Pt electrodes to realize a chemiresistor gas sensor able to operate at RT. The responses to ammonia (10, 20, 30 ppm) and nitrogen dioxide (1, 2, 3 ppm) are shown at different values of relative humidity, in dark and under 254 nm UV light. In order to check the sensor selectivity, gas response has also been tested towards hydrogen, ethanol, acetone and carbon oxide. Finally, a model based on linear dispersion relation characteristic of graphene, which take into account humidity and UV light effects, has been proposed.
RESUMO
A low-cost method for carbon nanotubes (CNTs) network production from solutions on flexible polyethylene naphthalate substrates has been adopted to prepare high quality and well characterized SWCNT bundle layers to be used as the active layer in chemiresistor gas sensors. Two types of SWCNTs have been tested: pristine SWCNTs, deposited from a surfactant solution, and covalently functionalized SWCNTs, deposited from a dimethyl-acetamide solution. The humidity effects on the sensitivity of the SWCNTs network to NH3 have been investigated. The results show that relative humidity favors the response to NH3, confirming recent theoretical predictions. The COOH-functionalized sample displays the largest response owing to both its hydrophilic nature, favoring the interaction with H2O molecules, and its largest surface area. Compared to data available in the literature, the present sensors display a remarkable sensitivity well below the ppm range, which makes them quite promising for environmental and medical applications, where NH3 concentrations (mostly of the order of tens of ppb) have to be detected.
RESUMO
A combination of the functionalities of carbon nanotube (CNT)-Si hybrid heterojunctions is presented as a novel method to steer the efficiency of the photovoltaic (PV) cell based on these junctions, and to increase the selectivity and sensitivity of the chemiresistor gas sensor operated with the p-doped CNT layer. The electrical characteristics of the junctions have been tracked by exposing the devices to oxidizing (NO2) and reducing (NH3) molecules. It is shown that when used as PV cells, the cell efficiency can be reversibly steered by gas adsorption, providing a tool to selectively dope the p-type layer through molecular adsorption. Tracking of the current-voltage curve upon gas exposure also allowed to use these cells as gas sensors with an enhanced sensitivity as compared to that provided by a readout of the electrical signal from the CNT layer alone. In turn, the chemiresistive response was improved, both in terms of selectivity and sensitivity, by operating the system under illumination, as the photo-induced charges at the junction increase the p-doping of CNTs making them more sensitive to NH3 and less to NO2.
RESUMO
The gas response of single-wall carbon nanotubes (SWCNT) functionalized with indium tin oxide (ITO) nanoparticles (NP) has been studied at room temperature and an enhanced sensitivity to ammonia and nitrogen dioxide is demonstrated. The higher sensitivity in the functionalized sample is related to the creation of nano-heterojunctions at the interface between SWCNT bundles and ITO NP. Furthermore, the different response of the two devices upon NO2 exposure provides a way to enhance also the selectivity. This behavior is rationalized by considering a gas sensing mechanism based on the build-up of space-charge layers at the junctions. Finally, full recovery of the signal after exposure to NO2 is achieved by UV irradiation for the functionalized sample, where the ITO NP can play a role to hinder the poisoning effects on SWCNT due to NO2 chemisorption.
RESUMO
A photosystem II (PSII) core complex lacking the internal antenna CP43 protein was isolated from the photosystem II of Synechocystis PCC6803, which lacks photosystem I (PSI). CP47-RC and reaction centre (RCII) complexes were also obtained in a single procedure by direct solubilization of whole thylakoid membranes. The CP47-RC subcore complex was characterized by SDS/PAGE, immunoblotting, MALDI MS, visible and fluorescence spectroscopy, and absorption detected magnetic resonance. The purity and functionality of RCII was also assayed. These preparations may be useful for mutational analysis of PSII RC and CP47-RC in studying primary reactions of oxygenic photosynthesis.
Assuntos
Cianobactérias/química , Complexo de Proteínas do Centro de Reação Fotossintética/isolamento & purificação , Western Blotting , Cromatografia Líquida , Eletroforese em Gel de Poliacrilamida , Complexo de Proteínas do Centro de Reação Fotossintética/química , Espectrometria de Fluorescência , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Tilacoides/químicaRESUMO
Photosystem II of higher plants and cyanobacteria is composed of more than 20 polypeptide subunits. The pronounced hydrophobicity of these proteins hinders their purification and subsequent analysis by mass spectrometry. This paper reports the results obtained by application of matrix-assisted laser desorption/ionization mass spectrometry directly to isolated complexes and thylakoid membranes prepared from cyanobacteria and spinach. Changes in protein contents following physiopathological stimuli are also described. Good correlations between expected and measured molecular masses allowed the identification of the main, as well as most of the minor, low molecular weight components of photosystem II. These results open up new perspectives for clarifying the functional role of the various polypeptide components of photosystems and other supramolecular integral membrane complexes.
Assuntos
Cianobactérias/química , Complexo de Proteínas do Centro de Reação Fotossintética/química , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Spinacia oleracea/química , Cromatografia Líquida de Alta Pressão , Cianobactérias/genética , Eletroforese em Gel de Poliacrilamida , Mutação , Peptídeos/química , Complexo de Proteínas do Centro de Reação Fotossintética/genética , Complexo de Proteína do Fotossistema II , Tilacoides/metabolismo , Raios UltravioletaRESUMO
The distinction between passive and active visuo-spatial memory has been useful to interpret various pattern of deficits reported in individual differences studies. However, this interpretation raises the issue of task difficulty, since active tasks could be failed simply because more complex and the corresponding deficit could reflect a reduced capacity of the system. We describe two children with Nonverbal Learning Disability whose performance provides evidence of a dissociation between passive and active memory processes. One of the children showed a selective impairment in passive tasks and performed flawlessly in active tasks, whereas the second child displayed the opposite pattern. These data suggest that a qualitative difference between passive and active processes does exist and that differences in performance do not reflect a lower/higher level of task difficulty. Further, these data underlie the importance of formulating theoretical models of visuo-spatial memory including both material-related (i.e., visual vs spatial) and process-related (i.e., passive vs active) distinctions.
Assuntos
Deficiências da Aprendizagem/diagnóstico , Transtornos da Memória/diagnóstico , Percepção Espacial/fisiologia , Percepção Visual/fisiologia , Adolescente , Criança , Deficiências do Desenvolvimento/diagnóstico , Feminino , Humanos , Deficiências da Aprendizagem/complicações , Masculino , Transtornos da Memória/complicações , Índice de Gravidade de DoençaRESUMO
This study reports the observations gathered from 11 children referred to consulting services because of learning difficulties at school and diagnosed with nonverbal learning disabilities (NVLD). These children had an average verbal IQ, but a WISC-R performance IQ lower than the verbal IQ by at least 15 points and experienced difficulties especially in mathematics and drawing. The children completed a battery of four tasks requiring visuospatial working memory and visual imagery: a memory task composed of pictures and their positions (Pictures task), a task that required them to memorize the positions filled in a matrix (Passive Matrix task), a task that required them to imagine a pathway along a matrix (Active Matrix task) and a task that required them to learn groups made up of three words, using a visual interactive imagery strategy (TV task). In comparison to a control group of 49 children, children with NVLD scored lower in all the tasks, showing deficits in the use of visuospatial working memory and visual imagery. By contrasting subgroups of children of different ages in the control group, it was possible to show that some tasks did not show a clear developmental trend. Thus the deficits shown by the children with NVLD cannot simply be attributed to a developmental delay of these children, but seem to reflect a more severe disability.
Assuntos
Imagens, Psicoterapia , Deficiências da Aprendizagem/diagnóstico , Adolescente , Criança , Deficiências do Desenvolvimento/diagnóstico , Deficiências do Desenvolvimento/psicologia , Diagnóstico Diferencial , Feminino , Humanos , Inteligência , Deficiências da Aprendizagem/psicologia , Masculino , Memória de Curto Prazo , Orientação , Reconhecimento Visual de Modelos , Aprendizagem VerbalRESUMO
Fractionation of photosynthetic membranes from the cyanobacterium Synechocystis 6803 by polyacrylamide gel electrophoresis in the presence of Deriphat-160 allowed the isolation of a number of pigmented bands. Two of them, with molecular masses of 240+/-20 and 110+/-15 kDa respectively, showed peroxidase activity and, by means of polypeptide composition, immunoblotting and N-terminal sequencing, were identified as dimeric and monomeric cytochrome b6/f complexes, containing 1.3+/-0.35 chlorophyll molecules per cytochrome f. Further fractionation of monomeric complexes by mild gel electrophoresis in the presence of sodium dodecyl sulfate indicated that it is the cytochrome b6 polypeptide which provides the actual binding site for the chlorophyll molecule observed in the complex.
Assuntos
Clorofila/metabolismo , Cianobactérias/química , Grupo dos Citocromos b/química , Grupo dos Citocromos b/metabolismo , Complexo III da Cadeia de Transporte de Elétrons , Proteínas de Bactérias/análise , Proteínas de Bactérias/química , Complexo Citocromos b6f , Dimerização , Eletroforese em Gel de Poliacrilamida/métodos , Heme/química , Imidoésteres/química , Immunoblotting , Indicadores e Reagentes/química , Proteínas Ferro-Enxofre/análise , Proteínas Ferro-Enxofre/imunologia , Análise Espectral/métodosRESUMO
The effects of ultraviolet-B radiation (280-320 nm) on photosystem II of Synechocystis sp. PCC 6303 were investigated at the functional and structural levels. Loss of oxygen-evolving and electron-transport activity, measured by various techniques including Clark electrode polarography, fluorescence induction and fluorescence relaxation after a single turnover flash, are discussed in terms of two types of damage caused by ultraviolet-B radiation: (a) depletion of the plastoquinone pool; (b) perturbation and degradation of the D1 protein, with cleavage in the second transmembrane segment. These findings are in full agreement with those obtained, both in vivo and in vitro for higher plants for which a donor-side mechanism involving the water-splitting Mn cluster has been proposed for the main cleavage of the D1 protein. At the structural level, complete disruption of the photosystem II core is documented as a consequence of (or in parallel with) degradation of the D1 protein. From this point of view, ultraviolet-B-induced photoinhibition is unlike the visible-induced type and less susceptible to repair by synthesis and reinsertion of new D1 protein.
Assuntos
Cianobactérias/efeitos da radiação , Complexo de Proteínas do Centro de Reação Fotossintética/efeitos da radiação , Cloroplastos/química , Manganês/metabolismo , Peso Molecular , Oxigênio/metabolismo , Fotossíntese , Complexo de Proteína do Fotossistema II , Plastoquinona/metabolismo , Desnaturação Proteica/efeitos da radiação , Espectrometria de Fluorescência , Raios UltravioletaRESUMO
Photosystem I and II core complexes were resolved in a single step from the thylakoid membrane of Synechocystis sp. PCC 6803 by using a mild solubilization procedure in dodecyl beta-D-maltoside and Deriphat/PAGE. For each photosystem, two green bands were obtained containing oligomeric and monomeric forms of the core complexes of either photosystem. The oligomers are likely to be trimers in the case of photosystem I and dimers for photosystem II. The absorption spectra, polypeptide and pigment composition of green bands corresponding to either photosystem I or photosystem II were identical for monomeric and oligomeric forms. The cytochrome b-559 content of photosystem II was evaluated to be one cytochrome b-559/reaction centre both in the monomeric and dimeric forms. Two new 15-kDa and 22-kDa carotenoid-binding protein were isolated and their polypeptides purified to homogeneity.
Assuntos
Cianobactérias/química , Complexo de Proteínas do Centro de Reação Fotossintética/isolamento & purificação , Sequência de Aminoácidos , Dados de Sequência Molecular , Peso MolecularRESUMO
The in vivo effects of ultraviolet-B radiation (280-320 nm) on photosystem-II activity and degradation of the D1 protein are investigated and compared with the in vitro results on isolated thylakoids and other detergent-extracted photosystem-II preparations. A cleavage site in the second transmembrane segment of the D1 protein, giving rise to a 20-kDa C-terminal and a 13-kDa N-terminal fragment pair, is detected after irradiation of entire leaves as well as in all photosystem-II preparations, irrespective of their actual ability to evolve oxygen but depending on the presence of Mn ions associated with the water-splitting system. Damage to the plastoquinone moiety, observed by other authors, is confirmed and is proposed to be responsible for the impairment of electron-transport activity, but not for the observed cleavage of the D1 protein.
Assuntos
Manganês/química , Complexo de Proteínas do Centro de Reação Fotossintética/química , Complexo de Proteínas do Centro de Reação Fotossintética/efeitos da radiação , Sítios de Ligação , Transporte de Elétrons , Manganês/metabolismo , Peso Molecular , Oxigênio/metabolismo , Fragmentos de Peptídeos/química , Complexo de Proteínas do Centro de Reação Fotossintética/metabolismo , Complexo de Proteína do Fotossistema II , Spinacia oleracea , Raios UltravioletaRESUMO
The effect of photosystem II core phosphorylation on the secondary quinone acceptor of photosystem II (Q(B)) domain environment was analyzed by comparative herbicide-binding studies with photosystem II preparations from spinach (Spinacia oleracea L.). It was found that phosphorylation reduces the binding affinity for most photosynthetic herbicides. The binding of synthetic quinones and of the electron acceptor 2,6-dichlorophenolindophenol is also reduced by photosystem II phosphorylation. Four photosystem II core populations isolated from membranes showed different extents of phosphorylation as well as different degrees of affinity for photosynthetic herbicides. These findings support the idea that heterogeneity of photosystem II observed in vivo could be, in part, due to phosphorylation.
RESUMO
The structural and topological stability of thylakoid components under photoinhibitory conditions (4,500 microE.m-2.s-1 white light) was studied on Mn depleted thylakoids isolated from spinach leaves. After various exposures to photoinhibitory light, the chlorophyll-protein complexes of both photosystems I and II were separated by sucrose gradient centrifugation and analysed by Western blotting, using a set of polyclonals raised against various apoproteins of the photosynthetic apparatus. A series of events occurring during donor side photoinhibition are described for photosystem II, including: (a) lowering of the oligomerization state of the photosystem II core; (b) cleavage of 32-kD protein D1 at specific sites; (c) dissociation of chlorophyll-protein CP43 from the photosystem II core; and (d) migration of damaged photosystem II components from the grana to the stroma lamellae. A tentative scheme for the succession of these events is illustrated. Some effects of photoinhibition on photosystem I are also reported involving dissociation of antenna chlorophyll-proteins LHCI from the photosystem I reaction center.
Assuntos
Luz/efeitos adversos , Complexo de Proteínas do Centro de Reação Fotossintética/metabolismo , Plantas/metabolismo , Apoproteínas/análise , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/efeitos da radiação , Transporte Biológico/efeitos da radiação , Cloroplastos/metabolismo , Cloroplastos/efeitos da radiação , Cloroplastos/ultraestrutura , Membranas Intracelulares/metabolismo , Membranas Intracelulares/efeitos da radiação , Membranas Intracelulares/ultraestrutura , Complexos de Proteínas Captadores de Luz , Manganês/metabolismo , Modelos Biológicos , Oxirredução , Complexo de Proteínas do Centro de Reação Fotossintética/efeitos da radiação , Complexo de Proteínas do Centro de Reação Fotossintética/ultraestrutura , Complexo de Proteína do Fotossistema I , Complexo de Proteína do Fotossistema II , Plantas/efeitos da radiação , Plantas/ultraestrutura , Conformação Proteica/efeitos da radiação , Frações Subcelulares/metabolismo , Frações Subcelulares/efeitos da radiação , Frações Subcelulares/ultraestruturaRESUMO
When isolated photosystem II reaction centers from spinach are exposed to photoinhibitory light in the presence of an electron acceptor, breakdown products of the D2 protein at 28, 25, 23, 18, 9, 5 and 4.5 kDa are detected by immunoblotting with a monospecific anti-D2 polyclonal antibody. In a time-course experiment the 23 and 4.5 kDa fragments show a transient appearance, whilst the others are photoaccumulated. The regions of the D2 protein containing the cleavage sites for the 28 and 18 kDa photoinduced fragments have been identified. Significant degradation of D2 takes place only in the presence of an electron acceptor, and breakdown of the protein is partially prevented by serine-type protease inhibitors.
Assuntos
Fragmentos de Peptídeos/metabolismo , Complexo de Proteínas do Centro de Reação Fotossintética/metabolismo , Plantas/metabolismo , Sequência de Aminoácidos , Western Blotting , Endopeptidases/efeitos dos fármacos , Endopeptidases/metabolismo , Dados de Sequência Molecular , Fragmentos de Peptídeos/isolamento & purificação , Estimulação Luminosa , Complexo de Proteínas do Centro de Reação Fotossintética/isolamento & purificação , Complexo de Proteína do Fotossistema II , Inibidores de Serina Proteinase/farmacologiaRESUMO
When isolated reaction centres of photosystem II are subjected to photoinhibitory illumination, a 41 kDa SDS-PAGE band is observed under all experimental conditions. The same band is also found, together with lower molecular weight fragments of the D1 protein, in whole thylakoids and in all PSII sub-particles investigated up to now. In the case of isolated reaction centres the 41 kDa band is represented by a heterodimer of the D1 polypeptide and the alpha-subunit of cytochrome b559. The cross-linkage between D1 and alpha-cyt b559 involves a region on D1 between the N-terminal residue and Arg-225, and is an early event in photo-induced damage to the D1 protein.
Assuntos
Grupo dos Citocromos b/efeitos da radiação , Complexo de Proteínas do Centro de Reação Fotossintética/efeitos da radiação , Western Blotting , Grupo dos Citocromos b/química , Luz , Fotoquímica , Complexo de Proteínas do Centro de Reação Fotossintética/química , Complexo de Proteína do Fotossistema IIRESUMO
Photoinduced degradation of the photosystem II (PSII) reaction center D1 protein was studied in isolated thylakoids and different PSII subparticles. A 16 kDa fragment corresponding to the C-terminus of the protein is detected in thylakoids when they are inactivated at the donor side before illumination. The same D1 fragment is found in different types of PSII preparations at different integration levels characterized by different polypeptide compositions so long as they have an inactivated donor side and an active electron acceptor for the reduced pheophytin. However, when the PSII particle is equal to or smaller than the 43-less PSII core complex, other fragments are observed which are not found in more integrated systems.