Johan Aerts

Viscoelastic properties of the human tympanic membrane studied with stroboscopic holography and finite element modeling

A new anatomically-accurate Finite Element (FE) model of the tympanic membrane (TM) and malleus was combined with measurements of the sound-induced motion of the TM surface and the bony manubrium, in an isolated TM-malleus preparation. Using the results, we were able to address two issues related to how sound is coupled to the ossicular chain: (i) Estimate the viscous damping within the tympanic membrane itself, the presence of which may help smooth the broadband response of a potentially highly resonant TM, and (ii) Investigate the function of a peculiar feature of human middle-ear anatomy, the thin mucosal epithelial fold that couples the mid part of the human manubrium to the TM. Sound induced motions of the surface of ex vivo human eardrums and mallei were measured with stroboscopic holography, which yields maps of the amplitude and phase of the displacement of the entire membrane surface at selected frequencies. The results of these measurements were similar, but not identical to measurements made in intact ears. The holography measurements were complemented by laser-Doppler vibrometer measurements of sound-induced umbo velocity, which were made with fine-frequency resolution. Comparisons of these measurements to predictions from a new anatomically accurate FE model with varied membrane characteristics suggest the TM contains viscous elements, which provide relatively low damping, and that the epithelial fold that connects the central section of the human manubrium to the TM only loosely couples the TM to the manubrium. The laser-Doppler measurements in two preparations also suggested the presence of significant variation in the complex modulus of the TM between specimens. Some animations illustrating the model results are available at our website (www.uantwerp.be/en/rg/bimef/downloads/tympanic-membrane-motion).

A reliable liquid chromatography–tandem mass spectrometry method for simultaneous determination of multiple mycotoxins in fresh fish and dried seafoods

A reliable liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for simultaneous determination of nine mycotoxins, i.e., aflatoxin B1 (AFB1), aflatoxin B2 (AFB2), aflatoxin G1 (AFG1), aflatoxin G2 (AFG2), ochratoxin A (OTA), zearalenone (ZEN), T-2 toxin, HT2 toxin and deoxynivalenol (DON), in fresh fish (muscle and entrails) as well as dried seafoods. Special focus was given to sample pretreatment which is crucial for an accurate and reliable analytical method. With regards to the high complexity of the matrices, extraction solvent, time, and temperature as well as clean-up cartridges were optimized to improve extraction efficiency and reduce matrix effects. The optimum procedure included ultrasound-assisted extraction with acetonitrile/water/acetic acid (79/20/1, v/v/v) at 40 °C for 30 min, defatting with n-hexane and purification by Oasis HLB cartridges. The method was further validated by determining the linearity (R(2) ≥ 0.9989), sensitivity (limit of detection ≤ 2 μg/kg, limit of quantitation ≤ 3 μg/kg), recovery (72.2-119.9%) and precision (≤ 18.3%) in muscle and entrails of fresh crucian carp (Carassius carassius) as well as dried fish products. The method was proven to be suitable for its intended purposes. Mycotoxins of OTA, ZEN and AFB2 have been found in fresh fish and dried seafoods for the first time.

Nonlinearity in eardrum vibration as a function of frequency and sound pressure.

It is generally accepted that the middle ear acts mainly as a linear system for sound pressures up to 130 dB SPL in the auditory frequency range. However, at quasi-static pressure loads a strong nonlinear response has been demonstrated. Consequently, small nonlinear distortions may also be present in the middle ear response in the auditory frequency range. A new measurement method was developed to quickly determine vibration response, nonlinear distortions and noise level of acoustically driven biomechanical systems. Specially designed multisines are used for the excitation of the test system. The method is applied on a gerbil eardrum for sound pressures ranging from 90 to 120 dB SPL and for frequencies ranging from 125 Hz to 16 kHz. The experiments show that nonlinear distortions rise above noise level at a sound pressure of 96 dB SPL, and they grow as sound pressure increases. Post-mortem changes in the middle ear influence the nonlinear distortions rapidly until a stabilization occurs after approximately 3h.

Open access high-resolution 3D morphology models of cat, gerbil, rabbit, rat and human ossicular chains.

High-resolution 3D morphology models of cat, gerbil, rabbit, rat and human ossicular chains are presented. The models are based on high-resolution CT measurements. The resolution of the CT images, from which the models are segmented, varies from 5.6 to 33.5 μm. Models are freely available in different formats at our website (http://www.ua.ac.be/bimef/models) for research and educational purposes.

Scales tell a story on the stress history of fish

Fish faced with stressful stimuli launch an endocrine stress response through activation of the hypothalamic-pituitary-interrenal (HPI-) axis to release cortisol into the blood. Scientifically validated biomarkers to capture systemic cortisol exposure over longer periods of time are of utmost importance to assess chronic stress in governmental, wildlife, aquaculture and scientific settings. Here we demonstrate that cortisol in scales of common carp (Cyprinus carpio L.) is the long-sought biomarker for chronic stress. Undisturbed (CTR) and daily stressed (STRESS) carp were compared. Dexamethasone (DEX) or cortisol (CORT) fed fish served as negative and positive controls, respectively. Scale cortisol was quantified with a validated ultra-performance liquid chromatography tandem mass spectrometry method. An increase in scale cortisol content was found in STRESS and CORT but not in CTR and DEX fish. Scale cortisol content reflects its accumulation in a stressor and time dependent manner and validates the scale cortisol content as biomarker for chronic stress. Plasma analyses confirmed that (i) CTR, DEX and CORT treatments were effective, (ii) plasma cortisol of STRESS fish showed no signs of chronic HPI-axis activation, and (iii) plasma cortisol is a poor predictor for chronic stress. The expression of HPI key genes crf, pomc, and star were up-regulated in STRESS fish in the absence of a plasma cortisol response, as was the target gene of cortisol encoding subunit α1 of the Na+/K+-ATPase in gills. When lost, scales of fish regenerate fast. Regenerated scales corroborate our findings, offering (i) unsurpassed time resolution for cortisol incorporation and as such for stressful events, and (ii) the possibility to investigate stress in a well defined and controlled environment and time frame creating novel opportunities for bone physiological research. We conclude that the cortisol content in ontogenetic and regenerated scales is an innovative biomarker for chronic stress offering ample applications in science and industry.

Effect of the ratio of dietary n-3 fatty acids eicosapentaenoic acid and docosahexaenoic acid on broiler breeder performance, egg quality, and yolk fatty acid composition at different breeder ages.

When added to the feed of broiler breeder hens, dietary polyunsaturated fatty acids (FA) can be incorporated into the yolk and therefore become available to the progeny during their early development. The mechanism involved in lipid metabolism and deposition in the egg may be influenced by breeder age. Before the effect of an elevated concentration of certain polyunsaturated FA on the embryo can be investigated, the effect at breeder level and egg quality must be further assessed. The aim of the present experiment was to evaluate the effects of dietary n-6/n-3 ratios and dietary eicosapentaenoic acid (EPA, 20:5 n-3) and docosahexaenoic acid (DHA, 22:6 n-3) ratios, provided to broiler breeder hens, in terms of their zoo technical performance, egg quality, and yolk FA composition. Starting at 6 wk of age, 640 Ross-308 broiler breeder hens were fed 1 of 4 different diets. The control diet was a basal diet, rich in n-6 FA. The 3 other diets were enriched in n-3 FA, formulated to obtain a different EPA/DHA ratio of 1/1 (EPA = DHA), 1/2 (DHA), or 2/1 (EPA). In fact, after analysis the EPA/DHA ratio was 0.8, 0.4, or 2.1, respectively. Dietary EPA and DHA addition did not affect the performance of the breeder hens, except for egg weight. Egg weight was lower (P < 0.001) for all n-3 treatments. Dietary EPA improved number of eggs laid in the first 2 wk of the production cycle (P = 0.029). The absolute and relative yolk weight of eggs laid by EPA = DHA fed hens was lowest (P = 0.004 and P = 0.025, respectively). The EPA and DHA concentrations in the yolk were highly dependent on dietary EPA and DHA concentrations with a regression coefficient equal to 0.89. It can be concluded that dietary EPA and DHA can be incorporated in the breeder egg yolk to become available for the developing embryo, without compromising the performance and egg quality of the flock.

Details of human middle ear morphology based on micro-CT imaging of phosphotungstic acid stained samples

A multitude of morphological aspects of the human middle ear (ME) were studied qualitatively and/or quantitatively through the postprocessing and interpretation of micro‐CT (micro X‐ray computed tomography) data of six human temporal bones. The samples were scanned after phosphotungstic acid staining to enhance soft‐tissue contrast. The influence of this staining on ME ossicle configuration was shown to be insignificant. Through postprocessing, the image data were converted into surface models, after which the approaches diverged depending on the topics of interest. The studied topics were: the ME ligaments; morphometric and mechanical parameters of the ossicles relating to inertia and the ossicular lever arm ratio; the morphology of the distal incus; the contact surface areas of the tympanic membrane (TM) and of the stapes footplate; and the thickness of the TM, round window of the cochlea, ossicle joint spaces, and stapedial annular ligament. Some of the resulting insights are relevant in ongoing discussions concerning ME morphology and mechanical functions, while other results provide quantitative data to add to existing data. All findings are discussed in the light of other published data and many are relevant for the construction of mechanical finite element simulations of the ME. J. Morphol. 276:1025–1046, 2015.

Cortisol directly impacts Flavobacterium columnare in vitro growth characteristics.

Teleost fish faced with stressful stimuli launch an endocrine stress response through activation of the hypothalamic-pituitary-interrenal axis to release glucocorticoids, in particular cortisol, into the blood. For the majority of bacterial fish pathogens, stress is considered a key factor in disease outbreaks. Based upon studies in mammals, there is considerable evidence to suggest that, besides impairing the immune system, cortisol can have a direct effect on bacterial cells. Hitherto, this intriguing field of microbial endocrinology has remained largely unexplored in aquatic diseases. The present study investigated in vitro the impact of cortisol on phenotypic traits of the fresh water fish pathogen Flavobacterium columnare. Colonies obtained from the highly virulent (HV) isolates resulted in significantly larger and more spreading colonies compared to those from the low virulent (LV) isolates. High cortisol doses added displayed a direct effect on the bacterial cells and induced a significant decrease in colony size. An additional intriguing finding was the inverse relationship between cortisol concentrations added to the broth and the spreading character of colonies retrieved, with higher cortisol doses resulting in less rhizoid to rough and even smooth colony formation (the latter only in the LV trout isolate), suggesting a dose–response effect. The loss of the rhizoid appearance of the F. columnare colonies upon administration of cortisol, and hence the loss of motility, might indicate a phenotypic change to the biofilm state. These findings form the basis for further research on the impact of glucocorticoids on other virulence factors and biofilm formation of F. columnare.

Do wild-caught urban house sparrows show desensitized stress responses to a novel stressor?

ABSTRACT While urbanization exposes individuals to novel challenges, urban areas may also constitute stable environments in which seasonal fluctuations are buffered. Baseline and stress-induced plasma corticosterone (cort) levels are often found to be similar in urban and rural populations. Here we aimed to disentangle two possible mechanisms underlying such pattern: (i) urban environments are no more stressful or urban birds have a better ability to habituate to stressors; or (ii) urban birds developed desensitized stress responses. We exposed wild-caught urban and rural house sparrows (Passer domesticus) to combined captivity and diet treatments (urban versus rural diet) and measured corticosterone levels both in natural tail feathers and in regrown homologous ones (cortf). Urban and rural house sparrows showed similar cortf levels in the wild and in response to novel stressors caused by the experiment, supporting the growing notion that urban environments are no more stressful during the non-breeding season than are rural ones. Still, juveniles and males originating from urban populations showed the highest cortf levels in regrown feathers. We did not find evidence that cortf was consistent within individuals across moults. Our study stresses the need for incorporating both intrinsic and environmental factors for the interpretation of variation in cortf between populations. Summary: Corticosterone in natural and regrown feathers was similar between urban and rural birds. Sex and age related to corticosterone in regrown feathers. Feather corticosterone was not consistent across different moults.

High resolution gas volume change sensor

Changes of gas quantity in a system can be measured either by measuring pressure changes or by measuring volume changes. As sensitive pressure sensors are readily available, pressure change is the commonly used technique. In many physiologic systems, however, buildup of pressure influences the gas exchange mechanisms, thus changing the gas quantity change rate. If one wants to study the gas flow in or out of a biological gas pocket, measurements need to be done at constant pressure. In this article we present a highly sensitive sensor for quantitative measurements of gas volume change at constant pressure. The sensor is based on optical detection of the movement of a droplet of fluid enclosed in a capillary. The device is easy to use and delivers gas volume data at a rate of more than 15 measurements/s and a resolution better than 0.06 microl. At the onset of a gas quantity change the sensor shows a small pressure artifact of less than 15 Pa, and at constant change rates the pressure artifact is smaller than 10 Pa or 0.01% of ambient pressure.