RESUMEN
OBJECTIVES/HYPOTHESIS: Immunofluorescence staining methods have been developed to study the distribution of macromolecules in archival formalin-fixed celloidin-embedded human temporal bone tissues. The aim of this study was to investigate the feasibility of utilizing this approach to evaluate the codistribution of more than one molecule of interest in a single tissue section. STUDY DESIGN: Retrospective study of proteoglycan codistribution in archival human temporal bone tissues. METHODS: The chondroitin sulfate and keratan sulfate proteoglycans were selected for evaluating this methodology. Human tissues with known proteoglycan staining patterns were studied as controls. Thirty-one formalin-fixed celloidin-embedded archival human temporal bones were evaluated, and the observations in 11 specimens are described. A dual immunofluorescence staining method was developed using primary antibodies of differing isotypes and secondary antibodies labeled with fluorophores having nonoverlapping emission characteristics. RESULTS: The specificity of the dual immunofluorescence technique for chondroitin sulfate and keratan sulfate proteoglycans was demonstrated in control tissues and confirmed through inhibition studies. The normal human tectorial membrane exhibited intense chondroitin sulfate staining. Cochlear and vestibular hair cells exhibited predominantly keratan sulfate staining. Keratan sulfate staining predominated in spiral ganglion cell bodies and fibers. Alterations in the normal distribution pattern of proteoglycans were observed in cases of presbycusis and otosclerosis. CONCLUSIONS: The dual immunofluorescence staining methodology can be used to study archival formalin-fixed celloidin-embedded human temporal bone tissues. This technique may be applied to the evaluation of other molecules in archival human temporal bone tissues and lead to improvement in our understanding of the function of these molecules and their role in disease processes.
Asunto(s)
Técnica del Anticuerpo Fluorescente , Proteoglicanos/inmunología , Hueso Temporal/inmunología , Cadáver , Proteoglicanos Tipo Condroitín Sulfato/inmunología , Proteoglicanos Tipo Condroitín Sulfato/metabolismo , Estudios de Factibilidad , Humanos , Indicadores y Reactivos , Proteoglicanos/metabolismo , Estudios Retrospectivos , Sensibilidad y Especificidad , Coloración y Etiquetado/métodos , Hueso Temporal/patología , Adhesión del TejidoRESUMEN
Traditional posturographic analysis and four statistical mechanics techniques were applied to center-of-pressure (COP) trajectories of young, older "low-fall-risk" and older "high-fall-risk" individuals. Low-fall-risk older adults were active 3 days per week in a cardiac rehabilitation program, while high-fall-risk older adults were diagnosed with perilymph fistula. Subjects diagnosed with perilymph fistula must have experienced two of the following vestibular findings: constant disequilibrium, positional vertigo and/or a positive fistula test. Non-parametric statistical tests were used to determine whether the posturographic measures could detect differences between the young and older "low-fall-risk" groups (age comparison) and between the older "low-" and "high-risk" groups (risk of falling comparison). The statistical mechanics techniques were more sensitive than the traditional measures: detecting significant differences between the young and older "low-risk" groups, while none of the traditional measures were significantly different. In addition, interpretation of the statistical mechanics techniques may offer more insight into the nature of the process controlling the COP trajectories. However, the methods offered slightly different explanations. For instance, the Hurst rescaled range analysis suggests that the movement of the COP is governed solely by anti-persistent behavior, whereas the stabilogram diffusion analysis suggests a short-term persistence balanced by a long-term anti-persistence. These discrepancies highlight the need for a model that incorporates the biological systems responsible for maintaining balance and experimental methods to directly quantify their status and roles. Until such a model exists, however, the statistical mechanics techniques appear to have some advantages over traditional posturographic measures for studying balance control.