RESUMO
OBJECTIVE: To identify relevant aspects of functioning, disability, and contextual factors for adults with hearing loss (HL) from hearing health professional perspective summarized using the ICF classification as reference tool. DESIGN: Internet-based cross-sectional survey using open-ended questions. Responses were analysed using a simplified content analysis approach to link concept to ICF categories according to linking rules. STUDY SAMPLE: Hearing health professionals (experts) recruited through e-mail distribution lists of professional organizations and personal networks of ICF core set for hearing loss steering committee members. Stratified sampling according to profession and world region enhanced the international and professional representation. RESULTS: Sixty-three experts constituted the stratified sample used in the analysis. A total of 1726 meaningful concepts were identified in this study, resulting in 209 distinctive ICF categories, with 106 mentioned by 5% or more of respondents. Most categories in the activities & participation component related to communication, while the most frequent environmental factors related to the physical environment such as hearing aids or noise. Mental functions, such as confidence or emotional functions were also frequently highlighted. CONCLUSIONS: More than half (53.3%) of the entire ICF classification categories were included in the expert survey results. This emphasizes the importance of a multidimensional tool, such as the ICF, for assessing persons with hearing loss.
Assuntos
Perda Auditiva , Classificação Internacional de Funcionalidade, Incapacidade e Saúde , Adulto , Idoso , Estudos Transversais , Coleta de Dados , Feminino , Humanos , Internacionalidade , Masculino , Pessoa de Meia-IdadeRESUMO
K(+) efflux is observed as an early event in the apoptotic process in various cell types. Loss of intracellular K(+) and subsequent reduction in ionic strength are suggested to release the inhibition of proapoptotic caspases. In this work, a new K(+)-specific microelectrode was used to study possible alterations in intracellular K(+) in Xenopus laevis oocytes during chemically induced apoptosis. The accuracy of the microelectrode to detect changes in intracellular K(+) was verified with parallel electrophysiological measurements. In concordance with previous studies on other cell types, apoptotic stimuli reduced the intracellular K(+) concentration in Xenopus oocytes and increased caspase-3 activity. The reduction in intracellular K(+) was prevented by dense expression of voltage-gated K (Kv) channels. Despite this, the caspase-3 activity was increased similarly in Kv channel-expressing oocytes as in oocytes not expressing Kv channels. Thus, in Xenopus oocytes caspase-3 activity is not dependent on the intracellular concentration of K(+).
Assuntos
Apoptose , Caspase 3/metabolismo , Oócitos/metabolismo , Potássio/metabolismo , Animais , Células Cultivadas , Microeletrodos , Oócitos/citologia , Xenopus laevisRESUMO
Ion channels in the plasma membrane are important for the apoptotic process. Different types of voltage-gated ion channels are up-regulated early in the apoptotic process and block of these channels prevents or delays apoptosis. In the present investigation we examined whether ion channels are up-regulated in oocytes from the frog Xenopus laevis during apoptosis. The two-electrode voltage-clamp technique was used to record endogenous ion currents in the oocytes. During staurosporine-induced apoptosis a voltage-dependent Na(+) current increased three-fold. This current was activated at voltages more positive than 0 mV (midpoint of the open-probability curve was +55 mV) and showed almost no sign of inactivation during a 1-s pulse. The current was resistant to the Na(+)-channel blockers tetrodotoxin (1 µM) and amiloride (10 µM), while the Ca(2+)-channel blocker verapamil (50 µM) in the bath solution completely blocked the current. The intracellular Na(+) concentration increased in staurosporine-treated oocytes, but could be prevented by replacing extracellular Na(+) with either K(+) or Choline(+). Prevention of this influx of Na(+) also prevented the STS-induced up-regulation of the caspase-3 activity, suggesting that the intracellular Na(+) increase is required to induce apoptosis. Taken together, we have found that a voltage dependent Na(+) channel is up-regulated during apoptosis and that influx of Na(+) is a crucial step in the apoptotic process in Xenopus oocytes.
Assuntos
Oócitos/efeitos dos fármacos , Oócitos/metabolismo , Amilorida/farmacologia , Animais , Apoptose/efeitos dos fármacos , Técnicas de Patch-Clamp , Bloqueadores dos Canais de Sódio/farmacologia , Tetrodotoxina/farmacologia , Verapamil/farmacologia , Canais de Sódio Disparados por Voltagem/metabolismo , Xenopus laevisRESUMO
Mouse serum alkaline phosphatase (ALP) is frequently measured and interpreted in mammalian bone research. However, little is known about the circulating ALPs in mice and their relation to human ALP isozymes and isoforms. Mouse ALP was extracted from liver, kidney, intestine, and bone from vertebra, femur and calvaria tissues. Serum from mixed strains of wild-type (WT) mice and from individual ALP knockout strains were investigated, i.e., Alpl(-/-) (a.k.a. Akp2 encoding tissue-nonspecific ALP or TNALP), Akp3(-/-) (encoding duodenum-specific intestinal ALP or dIALP), and Alpi(-/-) (a.k.a. Akp6 encoding global intestinal ALP or gIALP). The ALP isozymes and isoforms were identified by various techniques and quantified by high-performance liquid chromatography. Results from the WT and knockout mouse models revealed identical bone-specific ALP isoforms (B/I, B1, and B2) as found in human serum, but in addition mouse serum contains the B1x isoform only detected earlier in patients with chronic kidney disease and in human bone tissue. The two murine intestinal isozymes, dIALP and gIALP, were also identified in mouse serum. All four bone-specific ALP isoforms (B/I, B1x, B1, and B2) were identified in mouse bones, in good correspondence with those found in human bones. All mouse tissues, except liver and colon, contained significant ALP activities. This is a notable difference as human liver contains vast amounts of ALP. Histochemical staining, Northern and Western blot analyses confirmed undetectable ALP expression in liver tissue. ALP activity staining showed some positive staining in the bile canaliculi for BALB/c and FVB/N WT mice, but not in C57Bl/6 and ICR mice. Taken together, while the main source of ALP in human serum originates from bone and liver, and a small fraction from intestine (<5%), mouse serum consists mostly of bone ALP, including all four isoforms, B/I, B1x, B1, and B2, and two intestinal ALP isozymes dIALP and gIALP. We suggest that the genetic nomenclature for the Alpl gene in mice (i.e., ALP liver) should be reconsidered since murine liver has undetectable amounts of ALP activity. These findings should pave the way for the development of user-friendly assays measuring circulating bone-specific ALP in mouse models used in bone and mineral research.
Assuntos
Fosfatase Alcalina/sangue , Isoenzimas/sangue , Fosfatase Alcalina/genética , Fosfatase Alcalina/metabolismo , Animais , Osso e Ossos/enzimologia , Feminino , Intestinos/enzimologia , Fígado/enzimologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos KnockoutRESUMO
This paper presents the growth and structure of ZnO nanorods on a sub-micrometer glass pipette and their application as an intracellular selective ion sensor. Highly oriented, vertical and aligned ZnO nanorods were grown on the tip of a borosilicate glass capillary (0.7 µm in diameter) by the low temperature aqueous chemical growth (ACG) technique. The relatively large surface-to-volume ratio of ZnO nanorods makes them attractive for electrochemical sensing. Transmission electron microscopy studies show that ZnO nanorods are single crystals and grow along the crystal's c-axis. The ZnO nanorods were functionalized with a polymeric membrane for selective intracellular measurements of Naâº. The membrane-coated ZnO nanorods exhibited a Naâº-dependent electrochemical potential difference versus an Ag/AgCl reference micro-electrode within a wide concentration range from 0.5 mM to 100 mM. The fabrication of functionalized ZnO nanorods paves the way to sense a wide range of biochemical species at the intracellular level.
RESUMO
ZnO nanorods were grown on a silver-coated tip of a borosilicate glass capillary (0.7 µm in tip diameter) and used as selective potentiometric sensor of intracellular free Mg(2+). To functionalize the ZnO nanorods for selectivity of Mg(2+), a polymeric membrane with Mg(2+)-selective ionophores were coated on the surface of the ZnO nanorods. These functionalized ZnO nanorods exhibited a Mg(2+)-dependent electrochemical potential difference versus an Ag/AgCl reference microelectrode within the concentration range from 500 nM to 100 mM. Two types of cells, human adipocytes and frog oocytes, were used for the intracellular Mg(2+) measurements. The intracellular concentration of free Mg(2+) in human adipocytes and frog oocytes were 0.4-0.5 and 0.8-0.9 mM, respectively. Such type of nanoelectrode device paves the way to enable analytical measurements in single living cells and to sense other bio-chemical species at the intracellular level.
Assuntos
Técnicas Biossensoriais/métodos , Líquido Intracelular/química , Magnésio/análise , Nanotubos/química , Óxido de Zinco/química , Adipócitos/química , Animais , Técnicas Biossensoriais/instrumentação , Cátions Bivalentes/análise , Técnicas Eletroquímicas , Feminino , Humanos , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Nanotubos/ultraestrutura , Oócitos/química , Xenopus laevisRESUMO
In this article, we report a functionalised ZnO-nanorod-based selective electrochemical sensor for intracellular glucose. To adjust the sensor for intracellular glucose measurements, we grew hexagonal ZnO nanorods on the tip of a silver-covered borosilicate glass capillary (0.7 microm diameter) and coated them with the enzyme glucose oxidase. The enzyme-coated ZnO nanorods exhibited a glucose-dependent electrochemical potential difference versus an Ag/AgCl reference microelectrode. The potential difference was linear over the concentration range of interest (0.5-1000 microM). The measured glucose concentration in human adipocytes or frog oocytes using our ZnO-nanorod sensor was consistent with values of glucose concentration reported in the literature; furthermore, the sensor was able to show that insulin increased the intracellular glucose concentration. This nanoelectrode device demonstrates a simple technique to measure intracellular glucose concentration.