Multiscale modeling of mechanotransduction in the utricle.

We review recent progress in using numerical models to relate utricular hair bundle and otoconial membrane (OM) structure to the functional requirements imposed by natural behavior in turtles. The head movements section reviews the evolution of experimental attempts to understand vestibular system function with emphasis on turtles, including data showing that accelerations occurring during natural head movements achieve higher magnitudes and frequencies than previously assumed. The structure section reviews quantitative anatomical data documenting topographical variation in the structures underlying macromechanical and micromechanical responses of the turtle utricle to head movement: hair bundles, OM, and bundle-OM coupling. The macromechanics section reviews macromechanical models that incorporate realistic anatomical and mechanical parameters and reveal that the system is significantly underdamped, contrary to previous assumptions. The micromechanics: hair bundle motion and met currents section reviews work based on micromechanical models, which demonstrates that topographical variation in the structure of hair bundles and OM, and their mode of coupling, result in regional specializations for signaling of low frequency (or static) head position and high frequency head accelerations. We conclude that computational models based on empirical data are especially promising for investigating mechanotransduction in this challenging sensorimotor system.

On the high frequency transfer of mechanical stimuli from the surface of the head to the macular neuroepithelium of the mouse.

Vestibular macular sensors are activated by a shearing motion between the otoconial membrane and underlying receptor epithelium. Shearing motion and sensory activation in response to an externally induced head motion do not occur instantaneously. The mechanically reactive elastic and inertial properties of the intervening tissue introduce temporal constraints on the transfer of the stimulus to sensors. Treating the otoconial sensory apparatus as an overdamped second-order mechanical system, we measured the governing long time constant (Τ(L)) for stimulus transfer from the head surface to epithelium. This provided the basis to estimate the corresponding upper cutoff for the frequency response curve for mouse otoconial organs. A velocity step excitation was used as the forcing function. Hypothetically, the onset of the mechanical response to a step excitation follows an exponential rise having the form Vel(shear) = U(1-e(-t/TL)), where U is the applied shearing velocity step amplitude. The response time of the otoconial apparatus was estimated based on the activation threshold of macular neural responses to step stimuli having durations between 0.1 and 2.0 ms. Twenty adult C57BL/6 J mice were evaluated. Animals were anesthetized. The head was secured to a shaker platform using a non-invasive head clip or implanted skull screws. The shaker was driven to produce a theoretical forcing step velocity excitation at the otoconial organ. Vestibular sensory evoked potentials (VsEPs) were recorded to measure the threshold for macular neural activation. The duration of the applied step motion was reduced systematically from 2 to 0.1 ms and response threshold determined for each duration (nine durations). Hypothetically, the threshold of activation will increase according to the decrease in velocity transfer occurring at shorter step durations. The relationship between neural threshold and stimulus step duration was characterized. Activation threshold increased exponentially as velocity step duration decreased below 1.0 ms. The time constants associated with the exponential curve were Τ(L) = 0.50 ms for the head clip coupling and T(L) = 0.79 ms for skull screw preparation. These corresponded to upper -3 dB frequency cutoff points of approximately 318 and 201 Hz, respectively. T(L) ranged from 224 to 379 across individual animals using the head clip coupling. The findings were consistent with a second-order mass-spring mechanical system. Threshold data were also fitted to underdamped models post hoc. The underdamped fits suggested natural resonance frequencies on the order of 278 to 448 Hz as well as the idea that macular systems in mammals are less damped than generally acknowledged. Although estimated indirectly, it is argued that these time constants reflect largely if not entirely the mechanics of transfer to the sensory apparatus. The estimated governing time constant of 0.50 ms for composite data predicts high frequency cutoffs of at least 318 Hz for the intact otoconial apparatus of the mouse.

How does high-frequency sound or vibration activate vestibular receptors?

The mechanism by which vestibular neural phase locking occurs and how it relates to classical otolith mechanics is unclear. Here, we put forward the hypothesis that sound and vibration both cause fluid pressure waves in the inner ear and that it is these pressure waves which displace the hair bundles on vestibular receptor hair cells and result in activation of type I receptor hair cells and phase locking of the action potentials in the irregular vestibular afferents, which synapse on type I receptors. This idea has been suggested since the early neural recordings and recent results give it greater credibility.

Turtle utricle dynamic behavior using a combined anatomically accurate model and experimentally measured hair bundle stiffness.

Anatomically correct turtle utricle geometry was incorporated into two finite element models. The geometrically accurate model included appropriately shaped macular surface and otoconial layer, compact gel and column filament (or shear) layer thicknesses and thickness distributions. The first model included a shear layer where the effects of hair bundle stiffness was included as part of the shear layer modulus. This solid model's undamped natural frequency was matched to an experimentally measured value. This frequency match established a realistic value of the effective shear layer Young's modulus of 16 Pa. We feel this is the most accurate prediction of this shear layer modulus and fits with other estimates (Kondrachuk, 2001b). The second model incorporated only beam elements in the shear layer to represent hair cell bundle stiffness. The beam element stiffness's were further distributed to represent their location on the neuroepithelial surface. Experimentally measured striola hair cell bundles mean stiffness values were used in the striolar region and the mean extrastriola hair cell bundles stiffness values were used in this region. The results from this second model indicated that hair cell bundle stiffness contributes approximately 40% to the overall stiffness of the shear layer-hair cell bundle complex. This analysis shows that high mass saccules, in general, achieve high gain at the sacrifice of frequency bandwidth. We propose the mechanism by which this can be achieved is through increase the otoconial layer mass. The theoretical difference in gain (deflection per acceleration) is shown for saccules with large otoconial layer mass relative to saccules and utricles with small otoconial layer mass. Also discussed is the necessity of these high mass saccules to increase their overall system shear layer stiffness. Undamped natural frequencies and mode shapes for these sensors are shown.

Experimental measurement of utricle system dynamic response to inertial stimulus.

The membranous utricle sac of the red-eared turtle was mounted in a piezoelectric actuated platform mounted on the stage of a light microscope. The piezoelectric actuator oscillated the base of the neuroepithelium along a linear axis. Displacements were in the plane of the utricle and consisted of a linear sinusoidal-sweep signal starting at 0 and increasing to 500 Hz over 5 s. This inertial stimulus caused measurable shear displacement of the otoconial layer's dorsal surface, resulting in shear deformation of the gelatinous and column filament layers. Displacements of the otoconial layer and a reference point on the neuroepithelium were filmed at 2,000 frames/s with a high-speed video camera during oscillations. Image registration was performed on the video to track displacements with a resolution better than 15 nm. The displacement waveforms were then matched to a linear second-order model of the dynamic system. The model match identified two system mechanical parameters-the natural circular frequency ω n and the damping ratio ζ-that characterized the utricle dynamic response. The median values found for the medial-lateral axis on 20 utricles with 95 % confidence intervals in parenthesis were as follows: ω n = 374 (353, 396) Hz and ζ = 0.50 (0.47, 0.53). The anterior-posterior axis values were not significantly different: ω n = 409 (390, 430) Hz and ζ = 0.53 (0.48, 0.57). The results have two relevant and significant dynamic system findings: (1) a higher than expected natural frequency and (2) significant under damping. Previous to this study, utricular systems were treated as overdamped and with natural frequencies much lower that measured here. Both of these system performance findings result in excellent utricle time response to acceleration stimuli and a broad frequency bandwidth up to 100 Hz. This study is the first to establish the upper end of this mechanical system frequency response of the utricle in any animal.

Diel activity patterns of the fish community in a temperate stream.

The diel activity patterns of fishes in a temperate New Brunswick stream were studied during the summer over 5 years. Young-of-the year Atlantic salmon Salmo salar and blacknose dace Rhinichthys atratulus were more active during the day than at night, whereas lake chub Couesius plumbeus, brook trout Salvelinus fontinalis and adult white suckers Catostomus commersonii were more active at night than during the day. Because fishes were as likely to be nocturnal as diurnal, the data suggest that more night-time sampling is needed to provide an unbiased view of fish community structure in temperate streams.

Experimental measurement of utricle dynamic response.

The utricle of the red-eared turtle was subjected to forced sinusoidal oscillations across various frequencies (10-125 Hz) and amplitudes (5-9 µm) to determine dynamic characteristics of the utricle under natural inertial stimulation. The utricle was maintained in physiologic solution during the entire experiment. Utricular specimens were prepared so that the Otoconial Layer (OL) crystals were exposed yet undisturbed, and the neuroepithelium was secured to a glass slide with dental floss strands. A piezoelectric-actuated platform, fitted to the stage of the microscope, created controlled sinusoidal displacement along the utricle's medial-lateral direction. The OL surface displacement was measured through the microscope with high-speed video at 1500 fps. A sub-pixel image registration algorithm was used to achieve displacement resolution ⩽ 15 nm. The Membranous Shelf (MS), that overlies the macula, was recorded with high-speed video under identical amplitude and frequency inputs and was used as a reference point. Maximum displacement amplitudes of the OL and MS were used to determine the Amplitude Ratio (AR) of the OL relative to the MS. ARs at various frequencies were fit to a single degree of freedom model of the utricle to determine the utricle's natural frequency of 363 Hz (95% confidence intervals: 328, 397) with a damping ratio of 0.96 (0.8, 1.12).

Steady-state stiffness of utricular hair cells depends on macular location and hair bundle structure.

Spatial and temporal properties of head movement are encoded by vestibular hair cells in the inner ear. One of the most striking features of these receptors is the orderly structural variation in their mechanoreceptive hair bundles, but the functional significance of this diversity is poorly understood. We tested the hypothesis that hair bundle structure is a significant contributor to hair bundle mechanics by comparing structure and steady-state stiffness of 73 hair bundles at varying locations on the utricular macula. Our first major finding is that stiffness of utricular hair bundles varies systematically with macular locus. Stiffness values are highest in the striola, near the line of hair bundle polarity reversal, and decline exponentially toward the medial extrastriola. Striolar bundles are significantly more stiff than those in medial (median: 8.9 µN/m) and lateral (2.0 µN/m) extrastriolae. Within the striola, bundle stiffness is greatest in zone 2 (106.4 µN/m), a band of type II hair cells, and significantly less in zone 3 (30.6 µN/m), which contains the only type I hair cells in the macula. Bathing bundles in media that break interciliary links produced changes in bundle stiffness with predictable time course and magnitude, suggesting that links were intact in our standard media and contributed normally to bundle stiffness during measurements. Our second major finding is that bundle structure is a significant predictor of steady-state stiffness: the heights of kinocilia and the tallest stereocilia are the most important determinants of bundle stiffness. Our results suggest 1) a functional interpretation of bundle height variability in vertebrate vestibular organs, 2) a role for the striola in detecting onset of head movement, and 3) the hypothesis that differences in bundle stiffness contribute to diversity in afferent response dynamics.

Pseudoepitheliomatous, keratotic and micaceous balanitis after penile squamous cell carcinoma.

Pseudoepitheliomatous keratotic and micaceous balanitis (PKMB) is a rare form of balanitis, with only a handful of cases reported since the disease was first described. Although the condition has been described as benign, there is increasing evidence of its premalignant potential, with several of the reported cases progressing to verrucous or squamous cell carcinoma (SCC). We report a case of PKMB following penile SCC and discuss the literature on this rare condition.

Combined effects of chemical and visual information in eliciting antipredator behaviour in juvenile Atlantic salmon Salmo salar.

Under natural conditions, both young-of-the-year (YOY; 0+ year) and parr (1+ year) Atlantic salmon Salmo salar exhibited strong antipredator behaviour (e.g. increase in latency to resume foraging) following the exposure to damage-released chemical alarm cues relative to a stream water control. Subsequent exposure to a novel visual stimulus had contrasting results. Parr increased their reactive distance to the visual stimulus if they had been previously exposed to a chemical alarm cue, whereas YOY did not. On the other hand, both YOY and parr took significantly longer to resume foraging when exposed to a visual stimulus if they had been previously exposed to a chemical alarm cue than control groups. While YOY and parr differed in the type and intensity of antipredator responses to both chemical and visual stimuli, perhaps due to differential costs and benefits associated with age, both used the chemical and the visual information in a combined manner.

Layer thickness and curvature effects on otoconial membrane deformation in the utricle of the red-ear slider turtle: static and modal analysis.

Finite element models of otoconial membrane (OM) were developed to investigate the effects of three geometric variables on static and modal response of the OM: (1) curvature of the macular surface, (2) spatial variation in thicknesses of three OM layers, and (3) shape of the macular perimeter. A geometrically accurate model of a turtle utricle was constructed from confocal images. Modifying values for each variable formed variants of this model: modeling the macula surface as flat, OM layer thicknesses as spatially invariant, and the macular perimeter as a rectangle. Static tests were performed on each modified OM model, and the results were compared to determine the effects of each geometric variable on static mechanical gain (deflection per unit acceleration). Results indicate that all three geometric variables affect the magnitude and directional properties of OM static mechanical gain. In addition, through modal analysis, we determined the natural frequencies and displacement modes of each model, which illustrate the effects of the three geometric variables on OM dynamics. This study indicates the importance of considering three-dimensional OM geometry when attempting to understand responses of the OM and, therefore, the modulation of hair cell signals to accelerations during head movements.

Cutaneous metastases from a hypopharyngeal malignancy.

We present an unusual case of carcinoma of the piriform fossa, presenting with cutaneous metastases. These metastases are very rare, present in the late stages of disease, and are usually associated with distant metastases and poor prognosis. They often suggest aggressive disease. We believe that cutaneous metastases from a malignancy in the piriform fossa are more unusual than those from other sites of the head and neck.

Primary central nervous system lymphoma: a single-centre experience of 55 unselected cases.

AIMS: Current treatment for primary central nervous system lymphoma (PCNSL) involves high-dose methotrexate (HDMTX) with or without radiotherapy. Many published studies describing this approach include a highly selected group of patients. We report a single-centre experience of unselected cases of PCNSL. MATERIALS AND METHODS: We retrospectively reviewed the case notes of 55 consecutive patients diagnosed with biopsy-proven PCNSL between 1995 and 2003 at Addenbrooke's Hospital Cambridge, UK. We describe the treatment and outcome, including survival, treatment-related toxicity and long-term functional disability. RESULTS: At diagnosis, 45% of patients were considered unfit to receive treatment with HDMTX, owing to poor performance status or comorbidity. These patients had a median survival of 46 days and may not have been included in other published studies. The remaining patients were treated with a chemotherapy regimen, which included HDMTX. Patients who received at least one cycle of a chemotherapy containing HDMTX had a median survival of 31 months. Forty per cent did not complete planned chemotherapy owing to toxicity, disease progression or death. The median survival of patients treated with HDMTX aged 60 years compared with patients aged under 60 years was 26 months vs 41 months (P = 0.07), respectively. Younger patients treated with HDMTX, who achieved complete remission with chemotherapy, had a median survival of 56 months. We identified a high incidence of functional disability among survivors, resulting from a combination of the tumour itself, the neurosurgical procedure required for diagnosis and the late neurotoxicity of combined chemoradiotherapy. CONCLUSION: The treatment of PCNSL is associated with significant early and late toxicity. Further attempts to improve treatment should address mechanisms to reduce this toxicity. In particular, the benefit of radiotherapy in patients who achieve complete remission with HDMTX will remain uncertain until it is addressed in a multicentre, randomised trial.

Effect of fluid forcing on vestibular hair bundles.

A dynamic 3-D hair bundle model including inertia and viscous fluid drag effects based on the finite element method is presented. Six structural components are used to construct the hair bundle--kinocilium, stereocilia, upper lateral links, shaft links, tip links, and kinocilial links. Fluid drag is distributed on the surface of cilia columns. Bundle mechanics are analyzed under two distinct loading conditions: (1) drag caused by the shear flow of the surrounding endolymph fluid (fluid-forced), (2) a single force applied to the tip of the kinocilium (point-forced). A striolar and a medial extrastriolar vestibular hair cell from the utricle of a turtle are simulated. The striolar cell bundle shows a clear difference in tip link tension profile between fluid-forced and point-forced cases. When the striolar cell is fluid forced, it shows more evenly distributed tip link tensions and is far more sensitive, responding like an on/off switch. The extrastriolar cell does not show noticeable differences between the forcing types. For both forcing conditions, the extrastriolar cell responds serially--the nearest tip links to the kinocilium get tensed first, then the tension propagates to the farther tip links.

The effect of food abundance on territory size and population density of juvenile steelhead trout (Oncorhynchus mykiss).

Optimal territory size models predict a decrease in territory size with increasing food abundance. However, most of these models may not be applicable to juvenile salmonids in streams, because they defend contiguous territories at high densities. The optimal size of a contiguous territory is predicted to (1) be independent of food abundance when food is rare and (2) decrease only when food abundance is high enough to induce a reduction in territory size below the contiguous optimum. To test these predictions, we raised equal densities of juvenile steelhead trout in outdoor stream channels over a 32-fold range of food abundance in the absence of emigration for 25 days. Increasing competition for scarce food resulted in increasing mortality, higher willingness to emigrate, higher variance in body mass, lower growth, lower population density and lower biomass. The size of territories decreased with increasing local population density, and increased with increasing body size. However, territory size did not change with food abundance, a result consistent with the prediction of a contiguous territory size model. On average, total salmonid biomass increased 5.7 times in response to the 32-fold increase in food abundance. Our data provide strong support for an earlier quantitative relationship between the abundance of stream salmonids and their food.

Malignant eccrine poroma in breast cancer-related lymphoedema.

We describe the first case of malignant eccrine poroma arising in a lymphoedematous site. The patient had long-standing lymphoedema of the upper limb following breast cancer treatment. Lymphoedema is a recognised complication of breast cancer treatment and a risk factor for the subsequent development of malignancy. Possible mechanisms for this are discussed.

Aberrant expression of minichromosome maintenance protein-2 and Ki67 in laryngeal squamous epithelial lesions.

Histological classification of laryngeal epithelial lesions is highly subjective, and methods of cytological detection are not well developed. Improved determination of aberrant cell cycle entry may allow increased objectivity in histological assessment and enable the development of less invasive diagnostic cytology tests. Sections of normal larynx (n=10), laryngeal dysplasia (n=20) and laryngeal squamous cell carcinoma (SCC) (n=10) were classified according to the Ljubljana classification and stained for markers of cell cycle entry, minichromosome maintenance protein-2 (Mcm-2) and Ki67. Expression patterns were compared using double labelling confocal microscopy. There was a correlation between Mcm-2 and Ki67 labelling indices (rho=0.93; 95% CI [0.84, 0.97]) and both markers showed increased expression from normal epithelium to SCC (Mcm-2, P=0.001; Ki67, P=0.0002). Importantly, there was minimal expression of Mcm-2 or Ki67 in the most superficial layers of normal larynx and abnormal or atypical hyperplasia, in contrast to carcinoma in situ and SCC. Clusters of Mcm-2/5-positive cells were present in cytological preparations from SCC, but not from those showing atypical hyperplasia or inflammation in non-neoplastic tissue. Minichromosome maintenance protein-2 staining may increase the objectivity and reliability of histological grading of laryngeal epithelial lesions. Laryngeal brushings, combined with immuno-enhanced liquid-based cytology, could be useful, as a less invasive approach, to the detection of laryngeal malignant and premalignant lesions.

Influence of an interdental full pin on stability of an acrylic external fixator for rostral mandibular fractures in dogs.

OBJECTIVE: To determine total stiffness and gap stiffness of an external fixation system in a canine mandibular fracture gap model incorporating a full interdental pin as the only point of rostral fixation in a bilateral type-I external fixator. SAMPLE POPULATION: 10 canine mandibles. PROCEDURE: Bilateral mandibular ostectomies were performed between premolars 3 and 4. A type-I external fixator incorporating a full interdental pin was placed to stabilize a 0.5-cm fracture gap. Four pin configurations (intact mandibular bodies with fixator; ostectomized mandibular bodies and complete fixator; ostectomized mandibular bodies with caudal pins of rostral fragment cut; ostectomized mandibular bodies with all pins of rostral fragment cut) were tested in dorsoventral bending 5 times on each mandible. The full interdental pin remained intact in all configurations. Total stiffness and gap stiffness were determined for each configuration on a materials testing machine. RESULTS: Total stiffness of intact mandibles was significantly greater than that of ostectomized mandibles, regardless of external fixator configuration. However, total stiffness and gap stiffness were not significantly different among different external fixator configurations applied to ostectomized mandibles. CONCLUSION AND CLINICAL RELEVANCE: External fixator configurations with only the full interdental pin engaging the rostral fragment were as stiff as configurations that had 2 or 4 additional pins in the rostral fragment for the applied loads. External fixators for rostral mandibular fractures may be rigidly secured with rostral fragment implants applied extracortically, avoiding iatrogenic trauma to teeth and tooth roots.

Spread of a recurrent lentigo maligna into a graft: a case for conservative treatment.

Lentigo maligna is an in situ malignant melanoma for which the treatment of choice is surgical excision. The recurrence rate in lentigo maligna is high and hence other treatments, such as cryotherapy, laser therapy, radiotherapy and Mohs' chemosurgery, have been described. Despite the high recurrence rate, spread of a lentigo maligna into a skin graft is rare. We describe a case of a recurrent lentigo maligna spreading into a skin graft, which, along with the cases described in the literature, highlights the presence of a group of low-grade malignant lentiginous lesions that may be managed by careful follow-up and observation.