Air embolism during liver procurement: an underestimated phenomenon? A pilot experimental study.

BACKGROUND: Intrahepatic air embolism can occur during liver transplantation, jeopardizing the posttransplant outcome. Until now, the role of the procurement in the origin of intrahepatic air remains unclear; it might be underestimated. In this pilot study using magnetic resonance imaging (MRI), we observed a substantial amount of air trapped in porcine livers during multiorgan procurement. We quantified the amount of air, examining whether it could be reduced by avoiding direct contact of air with the lumen of the hepatic vasculature during procurement and back-table preparation. METHODS: Five livers (control group) were procured according to standard techniques for comparison with 6 livers (modified group) where air could not enter into the livers due to clamping of the vasculature. MRI was performed during continuous machine perfusion (MP) preservation there after. We counted the number of black signal voids on T(2)*-weighted images, which were indicative of air bubbles within the hepatic contour. Additionally, an MRI contrast agent (gadolinium-diethylene triamine pentaacetic acid [Gd-DTPA]) was injected into the hepatic artery and circulated by MP. Insufficiently perfused areas with less contrast enhancement were analyzed quantitatively in T(1)-weighted images and expressed as the percentage of total liver volume. RESULTS: The images of the control livers showed more air bubbles compared with the modified group (45 ± 27 vs 6 ± 3; P = .004). The percentage of insufficiently perfused areas was higher among the control compared with the modified group (28.0 ± 15.8% vs 2.6 ± 4.6%; P = .047) on first-pass postcontrast T(1)-weighted images. After recirculating the contrast agent, insufficiently perfused areas showed similar localizations and contours within every liver. CONCLUSION: These data suggested that a substantial amount of air enters into the hepatic microcirculation through direct contact of air with the hepatic vasculature during standard procurement and back-table preparation. Avoiding opening the hepatic vessels to air substantially reduced this phenomenon.

Can apparent diffusion coefficient discriminate ischemic from nonischemic livers? A pilot experimental study.

PURPOSE: Using magnetic resonance imaging, the apparent diffusion coefficient (ADC) is an indicator to assess cerebral ischemia. The aim of this porcine study was to evaluate whether ADC assessed hepatic ischemia during ex vivo hypothermic machine perfusion (HMP) as well as in vivo. METHODS: Ex vivo: ADC of normal versus warm ischemic (WI) livers was assessed during HMP and subsequent rewarming to mimic ischemia-reperfusion injury. As the preservation solution, we used either an acellular solution or diluted blood. WI was induced in the left lobe or in the whole liver and compared 2-hour WI and non-WI. In vivo: One liver was scanned with the left lobe vessels occluded for 2-hour WI and subsequently for 3 hour reperfusion to compare with the right lobe without WI. Aspartate aminotransferase (AST) in the perfusate and morphology were used as surrogates of WI. RESULTS: In all WI livers, AST reached high levels and histology showed severe injury. Ex vivo ADC during acellular perfusion showed negligible differences between the livers with versus without WI, namely, 0.75 x 10(-3) or 0.88 x 10(-3) mm(2)/s during HMP. Ex vivo ADC using sanguineous perfusion showed 1.11 x 10(-3) or 0.83 x 10(-3) mm(2)/s during HMP in regions with versus without WI, respectively, a difference that remained stable during the whole experiment. ADC in vivo decreased from the physiological level of 1.07 x 10(-3) mm(2)/s to 0.75 x 10(-3) mm(2)/s in the first 30 minutes of WI, whereas ADC in the non-WI liver remained constant. CONCLUSION: ADC in vivo decreased during hepatic ischemia, as previously seen in cerebral ischemia. However, the effect of WI on ADC was less clear during ex vivo HMP.

Evaluation of intramyocellular lipid breakdown during exercise by biochemical assay, NMR spectroscopy, and Oil Red O staining.

The study compared the net decline of intramyocellular lipids (IMCL) during exercise (n = 18) measured by biochemical assay (BIO) and Oil Red O (ORO) staining on biopsy samples from vastus lateralis muscle and by (1)H-MR spectroscopy (MRS) sampled in an 11 x 11 x 18-mm(3) voxel in the same muscle. IMCL was measured before and after a 2-h cycling bout ( approximately 75% V(.)(O(2) peak)). ORO and MRS measurements showed substantial IMCL use during exercise of 31 +/- 12 and 47 +/- 6% of preexercise IMCL content. In contrast, use of BIO for IMCL determination did not reveal an exercise-induced breakdown of IMCL (2 +/- 9%, P = 0.29) in young healthy males. Correlations between different measures of exercise-induced IMCL degradation were low. Coefficients were 0.48 for MRS vs. ORO (P = 0.07) and were even lower for BIO vs. MRS (r = 0.38, P = 0.13) or ORO (r = 0.08, P = 0.78). This study demonstrates that different methods to measure IMCL in human muscles can result in different conclusions with regard to exercise-induced IMCL changes. MRS has the advantage that it is noninvasive, however, not fiber type specific and hampered by an at least 30-min delay in measurements after exercise completion and may overestimate IMCL use. BIO is the only quantitative method but is subject to variation when biopsies have different fiber type composition. However, BIO yields lower IMCL breakdown compared with ORO and MRS. ORO has the major advantage that it is fiber type specific, and it therefore provides information that is not available with the other methods.

Efficacy of gene therapy-delivered cytosine deaminase is determined by enzymatic activity but not expression.

The potential utility of tumour-selective 5-fluorouracil treatment using attenuated Salmonella serovar typhimurium recombinant for cytosine deaminase (TAPET-CD) has been documented in experimental settings. The present data demonstrate that in vivo (19)F-magnetic resonance spectroscopy measurements allow the outcome prediction of this prokaryotic-based therapy, demonstrating the necessity of non-invasive real-time imaging techniques for treatment monitoring.

Model order selection for quantification of a multi-exponential magnetic resonance spectrum.

Magnetic resonance spectroscopic signals analyzed by time-domain models in order to retrieve estimates of the model parameters usually require prior knowledge about the model order. For multi-exponential signals where a superposition of peaks occurs at the same resonance frequency, but with different damping values, model order selection criteria from information theory can be used. In this study, several generalized versions of information criteria are compared using Monte-Carlo simulation signals. The best criterion is further applied for selecting the model order of experimental glycogen signals.

Attentional responses to unattended stimuli in human parietal cortex.

Right-sided parietal lesions lead to lateralized attentional deficits which are most prominent with bilateral stimulation. We determined how an irrelevant stimulus in the unattended hemifield alters attentional responses in parietal cortex during unilateral orienting. A trial consisted of a central spatial cue, a delay and a test phase during which a grating was presented at 9 degrees eccentricity. Subjects had to discriminate the orientation of the grating. The unattended hemifield was either empty or contained a second, irrelevant grating. We carried out a series of functional MRI (fMRI) studies in 35 healthy volunteers (13 men and 22 women, aged between 19 and 30 years) as well as a behavioural and structural lesion mapping study in 17 right-hemispheric lesion patients, 11 of whom had neglect. In the patients with but not in those without neglect, the addition of a distractor in the unattended hemifield significantly impaired performance if attention was directed contralesionally but not if it was directed ipsilesionally. In the healthy volunteers, we discerned two functionally distinct areas along the posterior-anterior axis of the intraparietal sulcus (IPS). The posterior, descending IPS segment in both hemispheres showed attentional enhancement of responses during contralateral attentional orienting and was unaffected by the presence of an irrelevant stimulus in the ignored hemifield. In contrast, the right-sided horizontal IPS segment showed a strong attentional response when subjects oriented to a stimulus in the relevant hemifield and an irrelevant stimulus was simultaneously present in the ignored hemifield, compared with unilateral stimulation. This effect was independent of the direction of attention. The symmetrical left-sided horizontal IPS segment showed the highest responses under the same circumstances, in combination with a contralateral bias during unilateral stimulation conditions. None of the six patients without neglect had a lesion of the horizontal IPS segment. In four of the 11 neglect patients, the lesion overlapped with the horizontal IPS activity cluster and lay in close proximity to it in another four. The remaining three patients had a lesion at a distance from the parietal cortex. Our findings reconcile the role of the IPS in endogenous attentional control with the clinically significant interaction between direction of attention and bilateral stimulation in right parietal lesion patients. Functional imaging in neglect patients will be necessary to assess IPS function in those cases where the structural lesion spares the middle IPS segment.

Subspace-based MRS data quantitation of multiplets using prior knowledge.

Accurate quantitation of Magnetic Resonance Spectroscopy (MRS) signals is an essential step before converting the estimated signal parameters, such as frequencies, damping factors, and amplitudes, into biochemical quantities (concentration, pH). Several subspace-based parameter estimators have been developed for this task, which are efficient and accurate time-domain algorithms. However, they suffer from a serious drawback: they allow only a limited inclusion of prior knowledge which is important for accuracy and resolution. In this paper, a new method is presented: KNOB-SVD and its improved variant KNOB-TLS. KNOB-SVD is a recently proposed method, based on the Singular Value Decomposition (SVD), which allows the use of more prior knowledge about the signal parameters than previously published subspace-based methods. We compare its performance in terms of robustness and accuracy with the performance of three commonly used methods for signal parameter estimation: HTLS, a subspace-based method which does not allow any inclusion of prior knowledge, except for the model order; HTLSPK(Delta fd(eq)), a subspace-based method obtained by incorporating in HTLS the prior information that the frequency differences between doublet components are known and the damping factors are equal; and AMARES, an interactive maximum likelihood method that allows the inclusion of a variety of prior knowledge. Extensive simulation and in vivo studies, using (31)P as well as proton MRS signals, show that the new method outperforms HTLS and HTLSPK(Delta fd(eq)) in robustness, accuracy, and resolution, and that it provides parameter estimates comparable to the AMARES ones.

Cerebellar and premotor function in bimanual coordination: parametric neural responses to spatiotemporal complexity and cycling frequency.

In the present functional magnetic resonance imaging (fMRI) study, we assessed the neural network governing bimanual coordination during manipulations of spatiotemporal complexity and cycling frequency. A parametric analysis was applied to determine the effects of each of both factors as well as their interaction. Subjects performed four different cyclical movement tasks of increasing spatiotemporal complexity (i.e., unimanual left-right hand movements, bimanual in-phase movements, bimanual anti-phase movements, and bimanual 90 degrees out-of-phase movements) across four frequency levels (0.9, 1.2, 1.5, and 1.8 Hz). Results showed that, within the network involved in bimanual coordination, functional subcircuits could be distinguished: Activation in the supplementary motor area, superior parietal cortex (SPS), and thalamic VPL Nc was mainly correlated with increasing spatiotemporal complexity of the limb movements, suggesting that these areas are involved in higher-order movement control. By contrast, activation within the primary motor cortex, cingulate motor cortex (CMC), globus pallidus, and thalamic VLo Nc correlated mainly with movement frequency, indicating that these areas play an important role during movement execution. Interestingly, the cerebellum and the dorsal premotor cortex were identified as the principal regions responding to manipulation of both parameters and exhibiting clear interaction effects. Therefore, it is concluded that both areas represent critical sites for the control of bimanual coordination.

Changes in brain activation during the acquisition of a new bimanual coodination task.

Motor skill acquisition is associated with the development of automaticity and induces neuroplastic changes in the brain. Using functional magnetic resonance imaging (fMRI), the present study traced learning-related activation changes during the acquisition of a new complex bimanual skill, requiring a difficult spatio-temporal relationship between the limbs, i.e., cyclical flexion-extension movements of both hands with a phase offset of 90 degrees. Subjects were scanned during initial learning and after the coordination pattern was established. Kinematics of the movements were accurately registered and showed that the new skill was acquired well. Learning-related decreases in activation were found in right dorsolateral prefrontal cortex (DLPFC), right premotor, bilateral superior parietal cortex, and left cerebellar lobule VI. Conversely, learning-related increases in activation were observed in bilateral primary motor cortex, bilateral superior temporal gyrus, bilateral cingulate motor cortex (CMC), left premotor cortex, cerebellar dentate nuclei/lobule III/IV/Crus I, putamen/globus pallidus and thalamus. Accordingly, bimanual skill learning was associated with a shift in activation among cortico-subcortical regions, providing further evidence for the existence of differential cortico-subcortical circuits preferentially involved during the early and advanced stages of learning. The observed activation changes account for the transition from highly attention-demanding task performance, involving processing of sensory information and corrective action planning, to automatic performance based on memory representations and forward control.

MRI and x-ray CT study of spatial distribution of core breakdown in 'Conference' pears.

Two non-destructive tomographic techniques, X-ray CT imaging and magnetic resonance imaging (MRI), were applied to study the development of core breakdown disorder in 'Conference' pears (Pyrus communis cv. Conference). This disorder, which is characterized by brown discoloration of the tissue and development of cavities, is induced by elevated CO(2) and decreased O(2) levels during controlled atmosphere storage. Tomographic images of pears stored for 10 months under disorder inducing conditions, were acquired with both techniques and compared to the actual slices. Both X-ray and MRI were able to differentiate between unaffected tissue, brown tissue and cavities. A simple image-processing program, based on threshold values, was developed to determine the area percentage of affected and unaffected tissue as well as the cavity and core area per slice. For all three imaging techniques the area percentage brown tissue per slice increased with the diameter of the pear, but was systematically underestimated by 12% and 6% for, respectively, X-ray and MRI, compared to the actual slices. The area percentage cavity corresponded very well for all techniques. It was also found that the contours of the brown tissue were parallel to the fruit boundaries, suggesting a relation between the disorder symptoms and gas diffusion properties of the fruit. It was concluded that MRI is the most appropriate technique to study the development of core breakdown disorder during postharvest storage in future experiments.

Non-invasive 19F MR spectroscopy of 5-fluorocytosine to 5-fluorouracil conversion by recombinant Salmonella in tumours.

The aim of this study was to evaluate the applicability of fluorine-19 magnetic resonance spectroscopy ((19)F MRS) for monitoring in vivo the conversion of 5-fluorocytosine (5-FC) to 5-fluorouracil (5-FU) after using an attenuated Salmonella Typhimurium strain recombinant to provide cytosine deaminase (TAPET-CD). The (19)F MRS measurements were done on mice bearing the human colon tumour xenograft (HCT116). The intratumoural conversion is greater when TAPET-CD/5-FC is delivered intratumourally (i.tu.) than when TAPET-CD is delivered intravenously (i.v.) and 5-FC intraperitoneally (i.p.). Repeat measurements of the same tumour also yielded important information on the tumour colonization by TAPET-CD through the correlated 5-FC to 5-FU conversion efficacy. The in vivo MRS spectra were confirmed by in vitro (19)F MRS of perchloric acid extracts of the tumour tissue. No 5-FU metabolites were detectable in vivo in the tumours. However, the in vitro measurements revealed, besides 5-FC and 5-FU, the presence of small amounts of catabolites. Finally, spectra obtained in vitro from liver extracts of tumour-bearing mice treated i.tu. with TAPET-CD/5-FC showed no 5-FU and only little amounts of catabolites. Our data illustrate most importantly the potential of (19)F MRS to monitor biologically-based treatments involving cytosine deaminase.

Quantification of the glycogen 13C-1 NMR signal during glycogen synthesis in perfused rat liver.

We studied glycogen synthesis from glucose in perfused livers of fed (n = 4) and 24 h starved (n = 7) rats. Glycogenolysis was inhibited by BAY R3401 (150 microM) and proglycosyn (100 microM). After 60 min, we replaced 99% (13)C-1 glucose by natural abundance glucose. This pulse-chase design allowed us to recognize residual ongoing futile glycogen turnover from the release of initially deposited (13)C-label, into the (13)C-free chase medium. Net residual turnover was less than 2 +/- 0.7% and 0.6 +/- 0.2% of 1-(13)C glycogen deposition rates of 0.31 +/- 0.04 and 0.99 +/- 0.04 micromol glucose g(-1) min(-1), in starved and fed livers, respectively. The 1-(13)C glycogen signal was monitored throughout the experiment with proton-decoupled (13)C NMR spectroscopy and analyzed in the time domain using AMARES. We noticed progressive line-broadening in any single experiment in the chase phase. One or a sum of two to three overlapping Lorentzians, with different exponential damping factors, were fitted to the signal. When the S/N was better than 40, the fit always delivered a small and a broad component. In the chase phase, the fit with a single Lorentzian resulted in a decline of glycogen signal by about 15 +/- 4 and 12 +/- 2% in starved and fed rats, respectively. This apparent decline in 1-(13)C glycogen signal could not be accounted for by the appearance of equivalent amounts of (13)C-labeled metabolites in the perfusate. The fit with a sum of two Lorentzians resulted in a decline of glycogen signal intensity of 7 +/- 5 and 5 +/- 3% in starved and fed rats, respectively, which reduced the apparent turnover to 8 +/- 9% and 6 +/- 4%, respectively. Quantification of the growing (13)C-1 glycogen signal requires a model function that accommodates changes in line shape throughout the period under study.

Improved Lanczos algorithms for blackbox MRS data quantitation.

Magnetic resonance spectroscopy (MRS) has been shown to be a potentially important medical diagnostic tool. The success of MRS depends on the quantitative data analysis, i.e., the interpretation of the signal in terms of relevant physical parameters, such as frequencies, decay constants, and amplitudes. A variety of time-domain algorithms to extract parameters have been developed. On the one hand, there are so-called blackbox methods. Minimal user interaction and limited incorporation of prior knowledge are inherent to this type of method. On the other hand, interactive methods exist that are iterative, require user involvement, and allow inclusion of prior knowledge. We focus on blackbox methods. The computationally most intensive part of these blackbox methods is the computation of the singular value decomposition (SVD) of a Hankel matrix. Our goal is to reduce the needed computational time without affecting the accuracy of the parameters of interest. To this end, algorithms based on the Lanczos method are suitable because the main computation at each step, a matrix-vector product, can be efficiently performed by means of the fast Fourier transform exploiting the structure of the involved matrix. We compare the performance in terms of accuracy and efficiency of four algorithms: the classical SVD algorithm based on the QR decomposition, the Lanczos algorithm, the Lanczos algorithm with partial reorthogonalization, and the implicitly restarted Lanczos algorithm. Extensive simulation studies show that the latter two algorithms perform best.

Brain areas involved in interlimb coordination: a distributed network.

Whereas behavioral studies have made significant contributions toward the identification of the principles governing the coordination of limb movements, little is known about the role of higher brain areas that are involved in interlimb coordination. Functional magnetic resonance imaging (fMRI) was used to reveal the brain areas activated during the cyclical coordination of ipsilateral wrist and foot movements. Six normal subjects performed five different tasks that were presented in a random order, i.e., isolated flexion-extension movements of the right wrist (WRIST) and right foot (FOOT), cyclical coordination of wrist and foot according to the isodirectional (ISODIR) and nonisodirectional (NON-ISODIR) mode, and rest (REST). All movements were auditory paced at 66 beats/min. During the coordination of both limb segments, a distributed network was identified showing activation levels in the supplementary motor area (SMA), cingulate motor cortex (CMC), premotor cortex (PMC), primary sensorimotor cortex (M1/S1), and cerebellum that exceeded the sum of the activations observed during the isolated limb movements. In addition, coordination of the limb movements in different directions was associated with extra activation of the SMA as compared to movements in the same direction. It is therefore concluded that the SMA is substantially involved in the coordination of the nonhomologous limbs as part of a distributed motor network. Accordingly, the long-standing exclusive association that has been made between this medial frontal area and bimanual (homologous) coordination needs to be abandoned and extended towards other forms of interlimb coordination (nonhomologous).

Visual motion processing investigated using contrast agent-enhanced fMRI in awake behaving monkeys.

To reduce the information gap between human neuroimaging and macaque physiology and anatomy, we mapped fMRI signals produced by moving and stationary stimuli (random dots or lines) in fixating monkeys. Functional sensitivity was increased by a factor of approximately 5 relative to the BOLD technique by injecting a contrast agent (monocrystalline iron oxide nanoparticle [MION]). Areas identified as motion sensitive included V2, V3, MT/V5, vMST, FST, VIP, and FEF (with moving dots), as well as V4, TE, LIP, and PIP (with random lines). These regions sensitive for moving dots are largely in agreement with monkey single unit data and (except for V3A) with human fMRI results. Moving lines activate some regions that have not been previously implicated in motion processing. Overall, the results clarify the relationship between the motion pathway and the dorsal stream in primates.

Oral creatine supplementation facilitates the rehabilitation of disuse atrophy and alters the expression of muscle myogenic factors in humans.

1. We investigated the effect of oral creatine supplementation during leg immobilization and rehabilitation on muscle volume and function, and on myogenic transcription factor expression in human subjects. 2. A double-blind trial was performed in young healthy volunteers (n = 22). A cast was used to immobilize the right leg for 2 weeks. Thereafter the subjects participated in a knee-extension rehabilitation programme (3 sessions x week(-1), 10 weeks). Half of the subjects received creatine monohydrate (CR; from 20 g down to 5 g daily), whilst the others ingested placebo (P; maltodextrin). 3. Before and after immobilization, and after 3 and 10 weeks of rehabilitation training, the cross-sectional area (CSA) of the quadriceps muscle was assessed by NMR imaging. In addition, an isokinetic dynamometer was used to measure maximal knee-extension power (Wmax), and needle biopsy samples taken from the vastus lateralis muscle were examined to asses expression of the myogenic transcription factors MyoD, myogenin, Myf5, and MRF4, and muscle fibre diameters. 4. Immobilization decreased quadriceps muscle CSA (approximately 10 %) and Wmax (approximately 25 %) by the same magnitude in both groups. During rehabilitation, CSA and Wmax recovered at a faster rate in CR than in P (P < 0.05 for both parameters). Immobilization changed myogenic factor protein expression in neither P nor CR. However, after rehabilitation myogenin protein expression was increased in P but not in CR (P < 0.05), whilst MRF4 protein expression was increased in CR but not in P (P < 0.05). In addition, the change in MRF4 expression was correlated with the change in mean muscle fibre diameter (r = 0.73, P < 0.05). 5. It is concluded that oral creatine supplementation stimulates muscle hypertrophy during rehabilitative strength training. This effect may be mediated by a creatine-induced change in MRF4 and myogenin expression.

The filtering approach to solvent peak suppression in MRS: a critical review.

Suppressing the solvent peak is important in many applications of biomedical NMR spectroscopy in order to quantify the metabolites with a great accuracy. Among the postprocessing methods proposed in the literature, many deal with the concept of filtering. However, several proposals lack a theoretical perspective and some have not been explicitly applied to quantification problems. The present article is intended to bridge this gap: five methods are analyzed from a theoretical perspective. Subsequently the different methods are applied to the same set of data, and then the latter are quantified using the model fitting method AMARES. With our set, the scheme proposed by T. Sundin et al. (J. Magn. Reson. 139(2), 189-204 (1999)) proved to be the most reliable method.

Land-cover change: quantification metrics for perforation using 2-D gap features.

Perforation or gap formation in a vegetation is a major process in landscape transformation. The occurrence of gaps profoundly alters the microclimatical conditions in a vegetation. A method is proposed to quantify perforation by using the three main 2-D characteristics of the gaps: area, number and boundary length. New measures are developed by normalizing the observed values to the reference status of minimum and maximum perforation. As minimum perforation status, the presence of one single gap with area equal to the map resolution is assumed. The new measures are combined using a 3-D Euclidean distance to visualize the process and to detect changes. The method is exemplified using a field case of gaps in a tropical terra firme rainforest at Tiputini, Ecuador.

Eosinophilic rhinitis accompanies the development of lower airway inflammation and hyper-reactivity in sensitized mice exposed to aerosolized allergen.

BACKGROUND: Allergic rhinitis is a risk factor for the development of asthma. About 80% of asthmatic patients also have rhinitis. However, the pattern of induction of allergic rhinitis and asthma remains unclear. OBJECTIVE: The purpose of this study was to investigate the development of upper airway inflammation in mice during the development of an asthma-like disease and after an acute allergen provocation. METHODS: BALB-c mice were sensitized intraperitoneally (i.p) to ovalbumin (OA, days 1-13) and were challenged with aerosols of either OA or saline on 8 consecutive days (days 33-40). In a second experiment, chronic exposure for 8 days was followed by 10 days of rest and then an acute nebulized allergen provocation was performed (day 50). Inflammatory parameters were investigated at different time-points. RESULTS: Upper and lower eosinophilic airway inflammation were simultaneously induced in the course of repeated inhalations of nebulized OA, as shown by analyses of nasal and broncho-alveolar lavage fluids and histological sections of the nose and bronchi. Mice that developed bronchial hyper-responsiveness also had increased thickness of the nasal mucosa on magnetic resonance image (MRI) scans. When chronic exposure was followed by acute allergen provocation, the latter caused a systemic increase in IL-5 levels, with a concomitant rise in blood and airway eosinophils, primarily in the nose. CONCLUSIONS: Simultaneous induction of eosinophilic inflammation in the nose and lungs was found in a mouse model of respiratory allergy. These findings support the viewpoint that upper and lower airway disease represent a continuum of inflammation involving one common airway and provide evidence for the concept of global airway inflammation after inhalation of allergen.

The interior-to-edge breakpoint distance as a guideline for nature conservation policy.

A method is proposed to quantify disturbance impact on isolated habitats. For every landscape patch, the breakpoint distance, defined as the penetration distance for which equality of interior and edge habitat is observed, can be calculated. Disturbance with equal impact at all patch sides is assumed. Effects of patch compactness, size, convolution, and perforation are discussed. The potential use of the measure for nature reserve design is discussed. The breakpoint distance follows the reserve design guidelines for individual patches, based on island biogeography and is consistent with the form and function principle. A large breakpoint distance is preferred for natural habitats. Small size, small compactness, intense convolution, and the occurrence of many gaps depress the breakpoint distance.