On the functional compartmentalization of the normal middle ear. Morpho-histological modelling parameters of its mucosa.

BACKGROUND: Middle ear physiology includes both sound pressure transmission and homeostasis of its static air pressure. Pressure gradients are continuously created by gas exchange over the middle ear mucosa as well as by ambient pressure variations. Gas exchange models require actual values for regional mucosa thickness, blood vessel density, and diffusion distance. Such quantitative data have been scarce and limited to few histological samples from the tympanic cavity (TC) and the antrum. However, a detailed regional description of the morphological differences of the TC and mastoid air cell system (MACS) mucosa has not been available. The aim of the present study was to provide such parameters. METHODS: The study included sets of three histological H&E-slides from 15 archived healthy temporal bones. We performed a comparison of the mucosa morphology among the following regions: (1) anterior TC; (2) inferior TC; (3) posterior TC; (4) superior TC; (5) MACS antrum; (6) superior MACS; (7) central MACS; (8) inferior MACS. RESULTS: Regions (1)-(3), situated below the inter-attico-tympanic diaphragm, had the largest proportion of high respiratory epithelium, cilia and loose lamina propria within the mucosa, as well as the thickest mucosa and the largest diffusion distance. Regions (6)-(8), situated above the diaphragm, had the thinnest mucosa, the shortest distance to the blood vessels, together with the largest proportion of flat epithelium and very few cilia. Regions (4)-(5), still supradiaphragmatic, had intermediary values for these parameters, but generally closer to regions (6)-(8). The blood vessel density and the proportion of active mucosa were not significantly different among the regions. CONCLUSION: Mucosa of regions (1), (2) and (3) represented a predominantly clearance-specific morphology, whereas in regions (4)-(8) it seemed adapted to gas exchange. However, the lack of statistically significant differences in blood vessel density and proportion of active mucosa indicated that all regions could be involved in gas exchange with the highest adaptation in the superior MACS. This pattern divides the middle ear functionally along the inter-attico-tympanic diaphragm rather than the anatomical division into TC and MACS.

Proteome stability analysis of snap frozen, RNAlater preserved, and formalin-fixed paraffin-embedded human colon mucosal biopsies.

Large repositories of well characterized RNAlater preserved samples and formalin-fixed, paraffin-embedded samples have been generated worldwide. However, the impact on the proteome of the preservation methods remain poorly described. Therefore, we analyzed the impact on the proteome of preserving samples in RNAlater, and by formalin-fixation, paraffin-embedding on human soft tissue, using directly frozen samples as a control ("Comparing the proteome of snap frozen, RNAlater preserved, and formalin-fixed paraffin-embedded human tissue samples" [1]). We here report the data from the analysis. The comparative analysis was performed on 24 colon mucosa biopsies, extracted from the sigmoideum of two gastroenterologically healthy participants for the purpose of this study. A set of biopsies were additionally stored for 30 min at room temperature prior to formalin-fixation. The samples were analyzed by high throughput gel free quantitative proteomics. The MS proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PRIDE: PXD002029.

Pressure buffering by the tympanic membrane. In vivo measurements of middle ear pressure fluctuations during elevator motion.

OBJECTIVES: The tympanic membrane (TM) represents a pressure buffer, which contributes to the overall pressure regulation of the middle ear (ME). This buffer capacity is based on its viscoelastic properties combined with those of the attached ossicular chain, muscles and ligaments. The current work presents a set of in vivo recordings of the ME pressure variations normally occurring in common life: elevator motion. This is defined as a situation of smooth ambient pressure increase or decrease on a limited range and at a low rate of pressure change. Based on these recordings, the purpose was a quantitative analysis of the TM buffer capacity including the TM compliance. METHODS: The pressure changes in seven normal adult ME's with intact TM's were continuously recorded directly inside the ME cavity during four different elevator trips using a high precision instrument. The TM buffer capacity was determined by the ratio between the changes in ME and the ambient pressure. Further, the ME volumes were calculated by Boyle's Law from pressure recordings during inflation-deflation tests; subsequently the TM compliance could also be calculated. Finally, the correlation between the ME volume and buffer function was determined. RESULTS: Twenty-one elevator trips could be used for the analysis. The overall mean TM pressure buffering capacity was 23.3% (SEM = 3.4), whereas the mean overall compliance was 28.9 × 10-3 µL/Pa (SEM = 4.8). A strong negative linear correlation was found between the TM buffer capacity and the ME volumes (R2 = 0.92). CONCLUSIONS: These results were in fair agreement with the literature obtained in clinical as well as temporal bone experiments, and they provide an in vivo reference for the normal ME function as well as for ME modeling. The TM buffer capacity was found more efficient in smaller mastoids. Possible clinical implications are discussed.

â¿¿Comparing the proteome of snap frozen, RNAlater preserved, and formalin-fixed paraffin-embedded human tissue samples.

Large biobanks exist worldwide containing formalin-fixed, paraffin-embedded samples and samples stored in RNAlater. However, the impact of tissue preservation on the result of a quantative proteome analysis remains poorly described. Human colon mucosal biopsies were extracted from the sigmoideum and either immediately frozen, stabilized in RNAlater, or stabilized by formalin-fixation. In one set of biopsies, formalin stabilization was delayed for 30 min. The protein content of the samples was characterized by high throughput quantitative proteomics. We were able to identify a similar high number of proteins in the samples regardless of preservation method, with only minor differences in protein quantitation.

Eustachian tube pressure equilibration. Temporal analysis of pressure changes based on direct physiological recordings with an intact tympanic membrane.

Eustachian tube function is important in pressure regulation of the middle ear. The efficacy or magnitude of pressure equilibration by tube openings should be determined by the gradient between middle ear and ambient pressure, but in theory also the duration of the tube opening may play a role. This study employed direct measurements of middle ear pressure in patients, who after parotidectomy had a catheter inserted into the mastoid with a pressure transducer connected. Thus, monitoring of the middle ear pressure in response to experimentally induced pressure changes could be performed under physiological conditions with an intact tympanic membrane. A set of six experiments was performed in four healthy subjects with different pressure deviations, where the counter-regulation was recorded over 10 min's time frames; a total of 75 events of tube openings were recorded. The transducer had a high accuracy of ±0.1 daPa, and data were sampled at 10 Hz, so that detailed parameters for each tube opening event could be obtained: the pressure change, the pressure gradient, and the duration of the opening were determined. The pressure changes in response to Eustachian tube openings showed significant positive correlation to the pressure gradient and ambient pressure (p < 0.001). However, the duration of the opening time was not related to the pressure gradient (p = 0.16), as well as the pressure change was also not related to the duration of the opening time (p = 0.34). This meant that the magnitude of a pressure equilibration during tube openings was only determined by the pressure gradient and not variations in the duration of the opening time. Additional correlations were investigated including the pressure change rate. In conclusion, under physiological conditions the opening of the Eustachian tube behaves similarly to a reflex mechanism with relative constant duration. Therefore, in order to equilibrate higher pressure gradients, series of Eustachian tube openings are needed, rather than the tube will open during a longer period of time. This article is part of a special issue entitled "MEMRO 2012".