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PubMed2004

Providing QoS through machine-learning-driven adaptive multimedia applications.

We investigate the optimization of the quality of service (QoS) offered by real-time multimedia adaptive applications through machine learning algorithms. These applications are able to adapt in real time their internal settings (i.e., video sizes, audio and video codecs, among others) to the unpredictably changing capacity of the network. Traditional adaptive applications just select a set of settings to consume less than the available bandwidth. We propose a novel approach in which the selected set of settings is the one which offers a better user-perceived QoS among all those combinations which satisfy the bandwidth restrictions. We use a genetic algorithm to decide when to trigger the adaptation process depending on the network conditions (i.e., loss-rate, jitter, etc.). Additionally, the selection of the new set of settings is done according to a set of rules which model the user-perceived QoS. These rules are learned using the SLIPPER rule induction algorithm over a set of examples extracted from scores provided by real users. We will demonstrate that the proposed approach guarantees a good user-perceived QoS even when the network conditions are constantly changing.

Algorithms↗

PubMed2003

Three-dimensional method for comparing in vivo interventional MR images of thermally ablated tissue with tissue response.

PURPOSE: To investigate the ability of magnetic resonance (MR) to monitor radio-frequency (RF) ablation treatments by comparing MR images of thermal lesions to histologically assayed cellular damage. We developed a new methodology using three-dimensional registration for making spatial correlations. MATERIALS AND METHODS: A low-field, open MRI system was used to guide an ablation probe into rabbit thigh muscle and acquire MR volumes after ablation. After fixation, we sliced and photographed the tissue at 3-mm intervals, using a specially designed apparatus, to obtain a volume of tissue images. Histologic samples were digitized using a video microscopy system. For our three-dimensional registration method, we used the tissue images as the reference, and registered histology and MR images to them using two different computer alignment steps. First, the MR volume was aligned to the volume of tissue images by registering needle fiducials placed near the tissue of interest. Second, we registered the histology images with the tissue images using a two-dimensional warping technique that aligned internal features and the outside boundary of histology and tissue images. RESULTS: The MR and histology images were very well aligned, and registration accuracy, determined from displacement of needle fiducials, was 1.32 +/- 0.39 mm (mean +/- SD), which compared favorably to the MR voxel dimensions (0.70 mm in-plane and 3.0 mm thick). A preliminary comparison of MR and tissue response showed that the region inside the elliptical hyperintense rim in MR closely corresponds to the region of necrosis as established by histology, with a mean absolute distance between MR and histology boundaries of 1.17 mm, slightly smaller than the mean registration error. The MR region slightly overestimated the region of necrosis, with a mean signed distance between boundaries of 0.85 mm. CONCLUSION: Our results suggest that our methodology can be used to achieve three-dimensional registration of histology and in vivo MR images. In MR lesion images, the inner border of the hyperintense region corresponds to the border of irreversible cell damage. This is good evidence that during RF ablation treatments, iMRI lesion images can be used for real-time feedback.

Animals↗

PubMedDate not supplied

Real-time markerless tracking for augmented reality: the virtual visual servoing framework.

Tracking is a very important research subject in a real-time augmented reality context. The main requirements for trackers are high accuracy and little latency at a reasonable cost. In order to address these issues, a real-time, robust, and efficient 3D model-based tracking algorithm is proposed for a "video see through" monocular vision system. The tracking of objects in the scene amounts to calculating the pose between the camera and the objects. Virtual objects can then be projected into the scene using the pose. Here, nonlinear pose estimation is formulated by means of a virtual visual servoing approach. In this context, the derivation of point-to-curves interaction matrices are given for different 3D geometrical primitives including straight lines, circles, cylinders, and spheres. A local moving edges tracker is used in order to provide real-time tracking of points normal to the object contours. Robustness is obtained by integrating an M-estimator into the visual control law via an iteratively reweighted least squares implementation. This approach is then extended to address the 3D model-free augmented reality problem. The method presented in this paper has been validated on several complex image sequences including outdoor environments. Results show the method to be robust to occlusion, changes in illumination, and mistracking.

Algorithms↗

PubMed1992

Potentiation of inositol trisphosphate-induced Ca2+ mobilization in Xenopus oocytes by cytosolic Ca2+.

1. The ability of cytosolic Ca2+ ions to modulate inositol 1,4,5-trisphosphate (Insp3)-induced Ca2+ liberation from intracellular stores was studied in Xenopus oocytes using light flash photolysis of caged InsP3. Changes in cytosolic free Ca2+ level were effected by inducing Ca2+ entry through ionophore and voltage-gated plasma membrane channels and by injection of Ca2+ through a micropipette. Their effects on Ca2+ liberation were monitored by video imaging of Fluo-3 fluorescence and by voltage clamp recording of Ca(2+)-activated membrane Cl- currents. 2. Treatment of oocytes with the Ca2+ ionophores A23187 and ionomycin caused a transient elevation of cytosolic Ca2+ level when cells were bathed in Ca(2+)-free solution, which probably arose because of release of Ca2+ from intracellular stores. 3. Membrane current and Fluo-3 Ca2+ signals evoked by photoreleased InsP3 in ionophore-treated oocytes were potentiated when the intracellular Ca2+ level was elevated by raising the Ca2+ level in the bathing solution. 4. Responses to photoreleased InsP3 were similarly potentiated following activation of Ca2+ entry through voltage-gated Ca2+ channels expressed in the plasma membrane. 5. Ca(2+)-activated membrane currents evoked by depolarization developed a delayed 'hump' component during sustained photorelease of InsP3, probably because Ca2+ ions entering through the membrane channels triggered liberation of Ca2+ from intracellular stores. 6. Ba2+ and Sr2+ ions were able to substitute for Ca2+ in potentiating InsP3-mediated Ca2+ liberation. 7. Gradual photorelease of InsP3 by weak photolysis light evoked Ca2+ liberation that began at particular foci and then propagated throughout, but not beyond that area of the oocyte exposed to the light. Local elevations of intracellular Ca2+ produced by microinjection of Ca2+ acted as new foci for the initiation of Ca2+ liberation by InsP3. 8. In resting oocytes, intracellular injections of Ca2+ resulted only in localized elevation of intracellular Ca2+, and did not evoke propagating waves. 9. The results show that cytosolic Ca2+ ions potentiate the ability of InsP3 to liberate Ca2+ from intracellular stores. This process may be important for the positive feedback mechanism underlying the generation of Ca2+ spikes and waves, and for interactions between the InsP3 pathway and Ca2+ ions entering cells through voltage- and ligand-gated channels.

Animals↗

PubMed2003

Oscillatory nucleocytoplasmic shuttling of the general stress response transcriptional activators Msn2 and Msn4 in Saccharomyces cerevisiae.

Msn2 and Msn4 are two related transcriptional activators that mediate a general response to stress in yeast Saccharomyces cerevisiae by eliciting the expression of specific sets of genes. In response to stress or nutritional limitation, Msn2 and Msn4 migrate from the cytoplasm to the nucleus. Using GFP-tagged constructs and high-resolution time-lapse video microscopy on single cells, we show that light emitted by the microscope also triggers this migration. Unexpectedly, the population of Msn2 or Msn4 molecules shuttles repetitively into and out of the nucleus with a periodicity of a few minutes. A large heterogeneity in the oscillatory response to stress is observed between individual cells. This periodic behavior, which can be induced by various types of stress, at intermediate stress levels, is not dependent upon protein synthesis and persists when the DNA-binding domain of Msn2 is removed. The cAMP-PKA pathway controls the sensitivity of the oscillatory nucleocytoplasmic shuttling. In the absence of PKA, Msn4 continues to oscillate while Msn2 is maintained in the nucleus. We show that a computational model based on the possibility that Msn2 and Msn4 participate in autoregulatory loops controlling their subcellular localization can account for the oscillatory behavior of the two transcription factors.

Binding Sites↗

PubMed2003

Lateral pelvic displacement during gait: abnormalities after stroke and changes during the first month of rehabilitation.

OBJECTIVES: To measure the amplitude and symmetry of lateral pelvic displacement (LPD) in patients with acute hemiparetic stroke; to assess the relationship between LPD and walking speed; and to quantify changes in LPD during 1- and 4-week intervals in the early stages of gait rehabilitation. DESIGN: LPD amplitude and symmetry were measured in stroke patients on admission to acute rehabilitation, 1 week later, and at 4-week follow-up. Performance was compared with sex-, height-, and age-matched control subjects. SETTING: Urban geriatric inpatient rehabilitation facility in Australia. PARTICIPANTS: Fifteen patients with a single-hemisphere stroke, confirmed by computed tomography, were compared with the data from 12 control subjects. Patients' FIM trade mark instrument scores ranged from 54 to 124. INTERVENTION: Gait rehabilitation involved twice-daily individual physical therapy sessions of 45 to 60 minutes, 5 days a week, incorporating whole and part practice, mental rehearsal, verbal feedback on performance, manual guidance, and strengthening techniques. MAIN OUTCOME MEASURES: Three-dimensional motion analysis of LPD amplitude and symmetry; and preferred walking speed over 10m. RESULTS: Compared with controls, stroke patients initially showed increased amplitude of LPD, with no difference in LPD symmetry. A statistically significant linear relationship existed between walking speed and amplitude of LPD (r=-.53; P=.04), yet not between walking speed and symmetry (r=-.41, P=.13). Amplitude and symmetry values remained consistent during the 4-week period of rehabilitation. CONCLUSIONS: These results provide baseline LPD values for patients with acute hemiparetic stroke and demonstrate the relationship between LPD and walking speed. Change in LPD during inpatient rehabilitation was not uniform or predictable, particularly during longer periods. This highlights the need for therapists to regularly reassess each patient during the early rehabilitation phase after stroke, especially given that individual differences can be marked.

Aged↗

PubMed1992

Capillary as a communicating medium in the microvasculature.

The preceding study (Dietrich and Tyml, 1992. Microvasc. Res. 43) demonstrated that a local application of norepinephrine (NE) on a capillary in a skeletal muscle produces a temporary reduction in blood flow within this capillary. The reduction is mediated via constriction of the supplying arteriole. The objective of the present study was to address the mechanism by which the local NE stimulus is propagated from the capillary to the arteriole. Using intravital video microscopy we measured red blood cell velocity in capillaries, and diameter of supplying arterioles, in the sartorius muscle in anesthetized frogs. Velocity responses were measured following iontophoretic application of NE (3 mM in the pipette) on the capillary, with or without pretreatment with 0.9 mM tetrodotoxin (nerve-specific sodium channel blocker), 30 mM lidocaine (nonspecific sodium channel blocker), and 30 mM yohimbine (alpha 2-receptor blocker). Diameter responses were measured before and after capillary damage introduced by microcautery. Tetrodotoxin did not block the NE-induced velocity reduction (i.e., from 0.2 to 0.07 mm/sec), while lidocaine attenuated it. Yohimbine blocked it only when applied on the same site as NE. Capillary damage abolished the NE-induced arteriolar constriction (i.e., from 27.8 to 21.5 microns). We conclude that the observed responses were not due to (1) direct diffusion of NE from the capillary to the arteriole, (2) conduction along adrenergic nerves, or (3) venous-arteriolar diffusional cross-talk. We interpret our data to indicate that the capillary itself could function as a communicating medium.

Animals↗

PubMed2001

Reduced skilfulness of arm motor behaviour among motor stroke patients with good clinical recovery: does it indicate reduced automaticity? Can it be improved by unilateral or bilateral training? A kinematic motion analysis study.

Functional cortical reorganisation had been demonstrated to accompany recovery from motor stroke. In agreement with a previous study, quantitative kinematic analysis of aimed movements in 14 almost completely recovered hemiparetic stroke patients and 14 healthy control subjects indicated a reduced skilfulness of both the more ballistic initial movement phase and the more feedback-guided late homing-in phase of aimed movements. By means of two dual motor tasks it was further investigated whether the reduced skilfulness of patients was due to an increased attentional demand and thus a reduced automaticity of motor control. Interference effects by dual tasks, however, were similar for patients and control subjects. Thus, the notion of reduced automaticity could not be supported empirically, and reduced skilfulness seemed rather related to residual pyramidal motor deficits. By means of a repetitive daily training for 1 week based on the Arm Ability Training approach, patients were able to reduce their performance deficits in both movement phases significantly. Whether patients practised with the affected arm or simultaneously with both the affected and non-affected arm only marginally modified outcome, unilateral training being slightly more efficacious. These findings demonstrate that even clinically minor residual motor deficits can be improved by a structured training for the affected arm.

Adult↗

PubMed1994

Instantaneous helical axis estimation from 3-D video data in neck kinematics for whiplash diagnostics.

To date, the diagnosis of whiplash injuries has been very difficult and largely based on subjective, clinical assessment. The work by Winters and Peles Multiple Muscle Systems--Biomechanics and Movement Organization. Springer, New York (1990) suggests that the use of finite helical axes (FHAs) in the neck may provide an objective assessment tool for neck mobility. Thus, the position of the FHA describing head-trunk motion may allow discrimination between normal and pathological cases such as decreased mobility in particular cervical joints. For noisy, unsmoothed data, the FHAs must be taken over rather large angular intervals if the FHAs are to be reconstructed with sufficient accuracy; in the Winters and Peles study, these intervals were approximately 10 degrees. in order to study the movements' microstructure, the present investigation uses instantaneous helical axes (IHAs) estimated from low-pass smoothed video data. Here, the small-step noise sensitivity of the FHA no longer applies, and proper low-pass filtering allows estimation of the IHA even for small rotation velocity omega of the moving neck. For marker clusters mounted on the head and trunk, technical system validation showed that the IHAs direction dispersions were on the order of one degree, while their position dispersions were on the order of 1 mm, for low-pass cut-off frequencies of a few Hz (the dispersions were calculated from omega-weighted errors, in order to account for the adverse effects of vanishing omega). Various simple, planar models relating the instantaneous, 2-D centre of rotation with the geometry and kinematics of a multi-joint neck model are derived, in order to gauge the utility of the FHA and IHA approaches. Some preliminary results on asymptomatic and pathological subjects are provided, in terms of the 'ruled surface' formed by sampled IHAs and of their piercing points through the mid-sagittal plane during a prescribed flexion-extension movement of the neck.

Adult↗

PubMed2004

Mechanics and control of the flat versus normal foot during the stance phase of walking.

OBJECTIVE: To compare walking stance phase rearfoot and forefoot motion, ankle joint moments and extrinsic foot muscle EMG profiles between normal and pes planus feet. DESIGN: A cross-sectional comparative study. BACKGROUND: Musculoskeletal conditions are often attributed to pes planus, in which motion is assumed to be excessive and the muscle control inadequate. However, many of the speculated differences in mechanics and control between the normal and the pes planus foot have not been quantified. METHODS: Kinematic and kinetic data were obtained from video recordings of skin surface markers and a force plate, and EMG was recorded with surface electrodes. Analysis of variance was carried out to compare the group profiles. RESULTS: In the pes planus group: the forefoot was less adducted (P < .05) at toe-off, and total transverse plane range of motion, at 8 degrees versus 10 degrees, was less (P < .01); the peak plantarflexor ankle moment at push-off was greater (P < .05); the invertor moment was greater at foot flat (P < .05); for the EMG profiles, activity early in stance, relative to the mean stance phase activity was higher (P < .01) in tibialis anterior and lower (P < .05) in the peronei, soleus and medial and lateral gastrocnemius. CONCLUSIONS: Despite reaching statistical significance, the group differences were small for the task of laboratory walking at a natural pace. The main implications of the differences were for restraint of motion. The expectations of excessive motion and muscle effort in the pes planus group were therefore not substantiated. RELEVANCE: Symptomatic pes planus subjects did not reveal the expected biomechanical differences from normal subjects. The underlying causes of symptoms were not identifiable.

Adaptation, Physiological↗

PubMed2001

Tyrosine kinase involvement in renal arteriolar constrictor responses to angiotensin II.

Experiments were performed to test the hypothesis that tyrosine kinase activity contributes to renal arteriolar contractile responses to angiotensin (Ang) II. Rats were subjected to short-term enalaprilat treatment to decrease endogenous Ang II formation before tissue was harvested for experiments with the in vitro blood-perfused juxtamedullary nephron technique. Acute surgical papillectomy was used to avoid the indirect afferent arteriolar effect of Ang II that arises through increased tubuloglomerular feedback sensitivity. Arteriolar lumen diameter responses to 1 and 10 nmol/L Ang II were monitored by videomicroscopic methods before and during treatment with various tyrphostin compounds: 100 micromol/L AG18 (broad-spectrum tyrosine kinase inhibitor), 100 nmol/L AG1478 (selective epidermal growth factor receptor tyrosine kinase inhibitor), or 100 micromol/L AG9 (inactive analog). Baseline afferent arteriolar lumen diameter averaged 23.5+/-1.2 micrometer and was not influenced by any tyrphostin. Ang II (10 nmol/L) decreased afferent diameter by 11.1+/-1.0 micrometer under untreated conditions, a response that was not altered by AG9 but significantly blunted by AG18 (34+/-9% inhibition) or AG1478 (52+/-8% inhibition). AG18 did not suppress afferent arteriolar contractile responses to membrane depolarization (20 to 55 mmol/L K(+ )bath). Efferent arteriolar baseline diameter averaged 24.1+/-0.8 micrometer and was unaltered by AG18 or AG1478; however, efferent diameter responses to 10 nmol/L Ang II were diminished 52+/-10% by AG18 and 51+/-13% by AG1478. These observations indicate that Ang II signaling in renal afferent and efferent arteriolar vascular smooth muscle is either mediated or modulated by tyrosine kinase activity, including that of the epidermal growth factor receptor tyrosine kinase.

Angiotensin II↗

PubMed1991

Atrial natriuretic factor decreases cell volume of rabbit atrial and ventricular myocytes.

The effect of atrial natriuretic factor (ANF) on cell volume was studied using video microscopy of rabbit atrial and ventricular myocytes. Each cell served as its own control, and relative cell volumes were determined. ANF (1 microM) significantly decreased relative cell volume to 0.929 +/- 0.006 (n = 7) in atrial and 0.930 +/- 0.013 (n = 5) in ventricular myocytes (normalized to volume without ANF). Reduction of volume was detectable at greater than or equal to 0.01 microM ANF, and the ED50 was 0.072 +/- 0.007 microM (n = 15). The effect of ANF also was examined under hypotonic (0.55T, 168 mosmol/l) and hypertonic (1.82T, 560 mosmol/l) conditions; osmolarity was adjusted using mannitol with NaCl fixed at 65 mM. In 0.55T, 1 microM ANF decreased cell volume to 0.941 +/- 0.014 (n = 5) in atrial and 0.942 +/- 0.017 (n = 7) in ventricular cells (normalized to 0.55T without ANF). In contrast, 1 microM ANF had no effect on atrial (n = 13) or ventricular (n = 11) cell volume in 1.82T. The hypothesis that ANF decreases cell volume by inhibiting Na(+)-K(+)-2Cl- cotransport was tested by blocking the cotransporter with bumetanide (10 microM). After inhibition of Na(+)-K(+)-2Cl- cotransport, 1 microM ANF failed to reduce cell volume in either atrial (n = 6) or ventricular myocytes (n = 6). Block of ANF-induced cell shrinkage by bumetanide was not due to changes in cell volume, since similar results were obtained using atrial (n = 7) and ventricular (n = 7) cells swollen in hypotonic (0.80T, 244 mosmol/l) solution. Replacement of Na+ with N-methyl-D-glucamine or Cl- with methanesulfonate abolished the ability of both ANF and bumetanide to decrease volume of atrial and ventricular cells in 1T and 0.8T solution. These data suggest that ANF can decrease the volume of atrial and ventricular cells under isotonic and hypotonic conditions by a mechanism that may involve Na(+)-K(+)-2Cl- cotransport. An ANF-induced decrease in cell volume may act as negative feedback and inhibit the stretch-induced release of ANF from atrial and ventricular cells. Furthermore, it may contribute to cell volume maintenance in myocytes in the setting of congestive heart failure or myocardial hypoxia when ANF release is elevated.

Animals↗

PubMed2003

Exploring the use of computer games and virtual reality in exposure therapy for fear of driving following a motor vehicle accident.

Specific phobia, situational type-driving, induced by accident (accident phobia) occurs in 18-38% of those involved in a vehicular accident of sufficient severity to warrant referral to the emergency departments of a general hospital. The objective is to investigate, in an open study, the effectiveness of the combined use of computer generated environments involving driving games (game reality [GR]) and a virtual reality (VR) driving environment in exposure therapy for the treatment of driving phobia following a motor vehicle accident (MVA) program. Fourteen subjects who met DSM-IV criteria for Simple Phobia/Accident Phobia and were referred from the emergency department of a general hospital were exposed to a Virtual Driving Environment (Hanyang University Driving Phobia Environment) and computer driving games (London Racer/Midtown Madness/Rally Championship). Patients who experienced "immersion" (i.e., a sense of presence with heightened anxiety) in one of the driving simulations (defined as an increase in SUD ratings of 3 and/or an increase of heart rate > 15 BPM in a 1-h trial session of computer simulation driving) were exposed to a cognitive behavioral program of up to 12 1-h sessions involving graded driving simulation tasks with self-monitoring, physiological feedback, diaphragmatic breathing and cognitive reappraisal. Subjects were assessed at the beginning and end of therapy with measurements of: physiological responsivity (heart rate), subjective ratings of distress (SUD), rating scales for severity of fear of driving (FDI), Posttraumatic Stress Disorder (CAPS) and depression (HAM-D) and achievement of target behaviors. Of all patients 7/14 (50%) became immersed in the driving environments. This immersed group (n = 7) completed the exposure program. Pre- and post-treatment comparisons showed significant post treatment reductions on all measures SUDS (p = 0.008), FDI (p = 0.008), CAPS (p = 0.008), HR (p = 0.008), CAPS (p = 0.008), HAM-D (p = 0.031). Further analysis of the FDI showed significant reductions in all three subscales: travel distress (p = 0.008), travel avoidance (p = 0.008), and maladaptive driving strategies (p = 0.016). The findings of this study suggest that VR and GR may have a useful role in the treatment of driving phobia post-accident even when co-morbid conditions such as post-traumatic stress disorder and depression are present.

Accidents, Traffic↗

PubMed1999

Radiometric surface temperature measurements during dye-assisted laser skin closure: in vitro and in vivo results.

BACKGROUND AND OBJECTIVE: A thermal camera was used to measure surface temperatures during laser skin welding to provide feedback for optimization of the laser parameters. STUDY DESIGN/MATERIALS AND METHODS: Two-centimeter-long, full-thickness incisions were made in guinea pig skin in vitro and in vivo. India ink was applied to the incision edges, which were then mechanically apposed. Continuous-wave, 1.06-microm Nd:YAG laser radiation was scanned over the incisions, producing an effective pulse duration of approximately 100 msec. Cooling durations between scans of 1.6, 4.0, and 8.0 sec were studied in vitro. A 5-mm-diameter laser spot was used with the power kept constant at 10 W. Thermal images were obtained at 30 frames per second with a thermal camera detecting 3-5 microm radiation. Surface temperatures were recorded at 0, 1, and 6 mm from the center line of the incision. RESULTS/CONCLUSIONS: Cooling durations of 1.6 and 4.0 seconds in vitro resulted in temperatures at the weld site that remained above approximately 65 degrees C for prolonged periods of time. Cooling durations of 8.0 seconds were sufficient both in vitro and in vivo to prevent a significant rise in baseline temperatures at the weld site over time.

Animals↗

PubMed1999

Mechanisms underlying the anti-inflammatory actions of central corticotropin-releasing factor.

Immune activation of hypothalamic corticotropin-releasing factor (CRF) provides a negative feedback mechanism to modulate peripheral inflammatory responses. We investigated whether central CRF attenuates endothelial expression of intercellular adhesion molecule 1 (ICAM-1) and leukocyte recruitment during endotoxemia in rats and determined its mechanisms of action. As measured by intravital microscopy, lipopolysaccharide (LPS) induced a dose-dependent increase in leukocyte rolling, adhesion, and emigration in mesenteric venules, which was associated with upregulation of endothelial ICAM-1 expression. Intracisternal injection of CRF abrogated both the increased expression of ICAM-1 and leukocyte recruitment. Intravenous injection of the specific CRF receptor antagonist astressin did not modify leukocyte-endothelial cell interactions induced by a high dose of LPS but enhanced leukocyte adhesion induced by a low dose. Blockade of endogenous glucocorticoids but not alpha-melanocyte-stimulating hormone (alpha-MSH) receptors reversed the inhibitory action of CRF on leukocyte-endothelial cell interactions during endotoxemia. In conclusion, cerebral CRF blunts endothelial upregulation of ICAM-1 and attenuates the recruitment of leukocytes during endotoxemia. The anti-inflammatory effects of CRF are mediated by adrenocortical activation and additional mechanisms independent of alpha-MSH.

Adrenalectomy↗

PubMed1995

Quantal puffs of intracellular Ca2+ evoked by inositol trisphosphate in Xenopus oocytes.

1. Ca2+ liberation induced in Xenopus oocytes by a poorly metabolized derivative of inositol 1,4,5-trisphosphate (3-deoxy-3-fluoro-D-myo-inositol 1,4,5-trisphosphate; 3-F-InsP3) was visualized using a video-rate confocal microscope to image fluorescence signals reported by the indicator dye calcium green-1. 2. Low (10-30 nM) intracellular concentrations of 3-F-InsP3 evoked Ca2+ release as localized transient 'puffs'. Progressively higher concentrations (30-60 nM) gave rise to abortive Ca2+ waves triggered by puffs, and then (> 60 nM) to a sustained elevation of Ca2+ followed by the appearance of propagating Ca2+ waves. At concentrations up to that giving waves, the frequency of puffs increased as about the third power of [InsP3], whereas their amplitudes increased only slightly. 3. The rise of cytosolic Ca2+ during a puff began abruptly, and peaked within about 50 ms. The peak free Ca2+ level was about 180 nM, and the total amount of Ca2+ liberated was several attomoles (10(-18) mol), too much to be accounted for by opening of a single InsP3-gated channel. The subsequent decline of Ca2+ occurred over a few hundred milliseconds, determined largely by diffusion of Ca2+ away from the release site, rather than by resequestration. Lateral spread of Ca2+ was restricted to a few micrometres, consistent with an effective diffusion coefficient for Ca2+ ions of about 27 microns2 s-1. 4. The peak amplitudes of puffs recorded at a given site were distributed in a roughly Gaussian manner, and a small proportion of sites consistently gave puffs much larger than the main population. Intervals between successive puffs at a single site were exponentially distributed, except for a progressive fall-off in puffs seen at intervals shorter than about 10 s. Thus, triggering of puffs appeared to be stochastically determined after recovery from a refractory period. 5. There was little correlation between the occurrence of puffs at sites more than a few micrometres apart, indicating that puff sites can function autonomously, but closely (ca 2 microns) adjacent sites showed highly correlated behaviour. 6. Puffs arose from sites-present at a density of about 1 per 30 microns2 in the animal hemisphere, located within a narrow band about 5-7 microns below the plasma membrane. 7. We conclude that Ca2+ puffs represent a 'quantal' unit of InsP3-evoked Ca2+ liberation, which may arise because local regenerative feedback by cytosolic Ca2+ ions causes the concerted opening of several closely clustered InsP3 receptor channels.

Animals↗

PubMed1999

Recovery of the vestibulocolic reflex after aminoglycoside ototoxicity in domestic chickens.

Avian auditory and vestibular hair cells regenerate after damage by ototoxic drugs, but until recently there was little evidence that regenerated vestibular hair cells function normally. In an earlier study we showed that the vestibuloocular reflex (VOR) is eliminated with aminoglycoside antibiotic treatment and recovers as hair cells regenerate. The VOR, which stabilizes the eye in the head, is an open-loop system that is thought to depend largely on regularly firing afferents. Recovery of the VOR is highly correlated with the regeneration of type I hair cells. In contrast, the vestibulocolic reflex (VCR), which stabilizes the head in space, is a closed-loop, negative-feedback system that seems to depend more on irregularly firing afferent input and is thought to be subserved by different circuitry than the VOR. We examined whether this different reflex also of vestibular origin would show similar recovery after hair cell regeneration. Lesions of the vestibular hair cells of 10-day-old chicks were created by a 5-day course of streptomycin sulfate. One day after completion of streptomycin treatment there was no measurable VCR gain, and total hair cell density was approximately 35% of that in untreated, age-matched controls. At 2 wk postlesion there was significant recovery of the VCR; at this time two subjects showed VCR gains within the range of control chicks. At 3 wk postlesion all subjects showed VCR gains and phase shifts within the normal range. These data show that the VCR recovers before the VOR. Unlike VOR gain, recovering VCR gain correlates equally well with the density of regenerating type I and type II vestibular hair cells, except at high frequencies. Several factors other than hair cell regeneration, such as length of stereocilia, reafferentation of hair cells, and compensation involving central neural pathways, may be involved in behavioral recovery. Our data suggest that one or more of these factors differentially affect the recovery of these two vestibular reflexes.

Aminoglycosides↗

PubMed2001

Objective laparoscopic skills assessments of surgical residents using Hidden Markov Models based on haptic information and tool/tissue interactions.

UNLABELLED: Laparoscopic surgical skills evaluation of surgery residents is usually a subjective process, carried out in the operating room by senior surgeons. By its nature, this process is performed using fuzzy criteria. The objective of the current study was to develop and assess an objective laparoscopic surgical skill scale using Hidden Markov Models (HMM) based on haptic information, tool/tissue interactions and visual task decomposition. METHODS: Eight subjects (six surgical trainees: first year surgical residents 2 x R1, third year surgical residents 2 x R3 fifth year surgical residents 2 x R5; and two expert laparoscopic surgeons: 2 x ES) performed laparoscopic cholecystectomy following a specific 7 steps protocol on a pig. An instrumented laparoscopic grasper equipped with a three-axis force/torque sensor located at the proximal end with an additional force sensor located on the handle, was used to measure the forces and torques. The hand/tool interface force/torque data was synchronized with a video of the tool operative maneuvers. A synthesis of frame-by-frame video analysis was used to define 14 different types of tool/tissue interactions, each one associated with unique force/torque (F/T) signatures. HMMs were developed for each subject representing the surgical skills by defining the various tool/tissue interactions as states and the associated F/T signatures as observations. The statistical distance between the HMMs representing residents at different levels of their training and the HMMs of expert surgeons were calculated in order to generate a learning curve of selected steps during laparoscopic cholecystectomy. RESULTS: Comparison of HMM's between groups showed significant differences between all skill levels, supporting the objective definition of a learning curve. The major differences between skill levels were: (i) magnitudes of F/T applied (ii) types of tool/tissue interactions used and the transition between them and (iii) time intervals spent in each tool/tissue interaction and the overall completion time. The objective HMM analysis showed that the greatest difference in performance was between R1 and R3 groups and then decreased as the level of expertise increased, suggesting that significant laparoscopic surgical capability develops between the first and the third years of their residency training. The power of the methodology using HMM for objective surgical skill assessment arises from the fact that it compiles enormous amount of data regarding different aspects of surgical skill into a very compact model that can be translated into a single number representing the distance from expert performance. Moreover, the methodology is not limited to in-vivo condition as demonstrated in the current study. It can be extended to other modalities such as measuring performance in surgical simulators and robotic systems.

Animals↗