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[The spatial structure and interrelation of plerocercoids of Digramma interrupta (Cestoda, Ligulidae) and the bream (Abramis brama) in the Kuibyshev reservoir].

The character of the distribution of the plerocercoid D. interrupta in the bream Abramis brama from the Kuibyshev water reservoir was studied. The spread of the parasites in A. brama population is of complex character and changes with the increase of fish body length. The factors affecting the maintenance and regulation of the relationships in the host-parasite system (size composition of fishes, peculiarities of the host's genotype, values of occurrence of different numbers of the parasites, variability of plerocercoids) are discussed.

Animals↗

Responses of fibroblasts to anchorage of dorsal extracellular matrix receptors.

Fibroblasts in 2D cultures differ dramatically in behavior from those in the 3D environment of a multicellular organism. However, the basis of this disparity is unknown. A key difference is the spatial arrangement of anchored extracellular matrix (ECM) receptors to the ventral surface in 2D cultures and throughout the entire surface in 3D cultures. Therefore, we asked whether changing the topography of ECM receptor anchorage alone could invoke a morphological response. By using polyacrylamide-based substrates to present anchored fibronectin or collagen on dorsal cell surfaces, we found that well spread fibroblasts in 2D cultures quickly changed into a bipolar or stellate morphology similar to fibroblasts in vivo. Cells in this environment lacked lamellipodia and large actin bundles and formed small focal adhesions only near focused sites of protrusion. These responses depend on substrate rigidity, calcium ion, and, likely, the calcium-dependent protease calpain. We suggest that fibroblasts respond to both spatial distribution and mechanical input of anchored ECM receptors. Changes in cell shape may in turn affect diverse cellular activities, including gene expression, growth, and differentiation, as shown in numerous previous studies.

Acrylic Resins↗

Morphological and elemental integrity of freeze-fractured, freeze-dried cultured cells during ion microscopic analysis.

The effects of progressive ion beam bombardment on freeze-fractured, freeze-dried cultured cells during ion microscopic (SIMS) analysis were studied with scanning electron microscopy (SEM) and ion microscopy. The freeze-fracture, freeze-dry sample preparation method was generally found to preserve cell morphology to a level far exceeding the spatial resolution of the ion microscope, with splitting at the nuclear envelope being the most commonly observed artefact. SEM monitoring of surface topography of an NRK-49F fibroblast after various ion bombardment doses showed relatively uniform erosion of cellular material, with some apparent selective retention of small cytoplasmic granules. Prolonged bombardment produced no detectable lateral elemental translocation. 41K+/24Mg+ signal ratios from Swiss 3T3 fibroblasts and RBL rat basophilic leukaemia cells were shown to vary generally by less than 10% during the course of extended ion bombardment. GM0415 human skin fibroblasts containing engorged lysosomes characteristic of Hurler's Syndrome were used to evaluate the effects of ion bombardment during a typical analysis session, where ion images of 39K+, 23Na+, 40Ca+ and 24Mg+ are sequentially recorded. This cell line was chosen as a worst-case system, because these cells are often thinly spread and possess extreme surface topography. Thin cell edges were shown sometimes to sputter away during analysis, giving misleadingly low ion signals from these regions in some 24Mg+ micrographs. Various non-uniform sputtering phenomena occurring in the submicrometre spatial domain had little or no measurable impact on local intensities in ion micrographs, indicating that freeze-dried, freeze-fractured cells are sampled in a sufficiently uniform fashion that quantitative ion microscopic evaluations of intracellular elemental levels in the general cytoplasmic or nuclear regions are feasible.

Animals↗

Spatial correlation between primary redox components in reaction centers of Rhodopseudomonas sphaeroides measured by two electrical methods in the nanosecond range.

Relative distances between the the primary donor P, the intermediary pheophytin acceptor H, and the iron-quinone acceptor Q of bacterial reaction centers were determined by recording laser flash-induced photovoltages in two experimental systems with nanosecond time resolution. In one system a suspension of chromatophores was subjected to a light gradient and in the other system chromatophores were spread at a heptane/water interface. The 10-ns back reaction occurring in reaction centers with reduced Q could be time resolved. The initial photovoltage amplitude under conditions in which the charge separation proceeded up to the state [P(+)H(-)] was about (2/3) of that when it proceeded up to the state [P(+)HQ(-)]. If the amplitude of the photovoltage is considered to be proportional to the spatial displacement of charges, this result means that pheophytin lies closer to Q than to P.

Journal Article↗

Spatiotemporal structure of genetic variation of a spreading plant metapopulation on dynamic riverbanks along the Meuse River.

Long-distance seed dispersal is a crucial determinant of within-population genetic variability and among-population genetic differentiation in plant metapopulations undergoing recurrent local extinctions and (re-)colonization. We investigated the spatial and temporal structure of genetic variation in a metapopulation of Sisymbrium austriacum located along a dynamic river system using dominant AFLP markers. Data on riverbank dynamics and colonization history allowed separating populations based on their age (< or =5 vs >5 years old). Bayesian analysis of population genetic structure indicated that populations were significantly differentiated from each other, but Mantel tests revealed that there was no relationship between pairwise geographic and genetic distances, suggesting that long-distance seed dispersal partly determines spatial genetic structure. Recent populations were less differentiated from each other than old populations. Analysis of molecular variance (AMOVA) indicated that both spatial factors and population age significantly determined genetic diversity, the effects of age being more important than spatial location. Clustering analysis revealed five large clusters, which were related primarily to population age and to a minor extent to geographical location. Our results indicate that the recurrent formation and destruction of riverbank habitats following peak flow events have a large impact on genetic diversity of riparian plant species.

Belgium↗

Polarized infrared spectroscopy of oriented purple membrane.

Polarized Fourier transform infrared spectroscopy has been used to study the structure of purple membrane from Halobacterium halobium. Membranes were oriented by drying a suspension of membrane fragments onto Irtran-4 slides. Dichroism measurements of the amide I, II and A peaks were used to find the average spatial orientation of the bacteriorhodopsin alpha-helices. By deriving a function that relates the observed dichroism to the orientational order parameters for the peptide groups, helical axis distribution, and mosaic spread of the membranes, the average orientation of the alpha-helices was found to lie in a range of less than 26 degrees away from the membrane normal, agreeing with electron microscopic measurements. The frequency of the amide I and A peaks is at least 10 cm-1 higher than values found for most alpha-helical polypeptides and proteins. This may indicate that bacteriorhodopsin contains distorted alpha-helical conformations.

Bacteriorhodopsins↗

Functional MRI of the motor cortex using a conventional gradient system: comparison of FLASH and EPI techniques.

Gradient echo (GE) and echo planar imaging (EPI) techniques are two different approaches to functional MRI (fMRI). In contrast to GE sequences, the ultra short EPI technique facilitates fMRI experiments with high spatial and temporal resolution or mapping of the whole brain. Although it has become the method of choice for fMRI, EPI is generally restricted to modern scanners with a strong gradient system. The aim of our study was to evaluate the applicability of EPI for fMRI of the motor cortex using a 1.5 T scanner with a conventional gradient system of 10 mT/m (rise time: 1 ms). Therefore, EPI was compared with a well-established high-resolution fast low-angle shot (FLASH) technique (matrix size 1282). The FLASH technique was applied additionally with a 64(2) matrix size to exclude influences caused by different spatial resolution, because the EPI sequence was restricted to a 64(2) matrix size. A total of 35 healthy volunteers were included in this study. The task consisted of clenching and spreading of the right hand. FLASH and EPI techniques were compared regarding geometric distortions as well as qualitative and quantitative fMRI criteria: Mean signal increase between activation and rest and the area of activation were measured within the contralateral, ipsilateral, and supplementary motor cortex. The quality of subtraction images between activation and rest, as well as the quality of z-maps and time course within activated regions of interest, was evaluated visually. EPI revealed significant distortions of the anterior and posterior brain margins; lateral distortions (relevant for the motor cortex) could be neglected in most cases. The mean signal increase was significantly higher using FLASH 1282 compared to FLASH 64(2) and EPI 64(2), whereas the activated areas proved to be smaller in FLASH 1282 functional images. Both results can be explained by well-documented partial volume effects, caused by different voxel size. Similar quality of the subtraction images and of the time courses in different regions of interest were found for all techniques under investigation, but slightly reduced quality of z-map in FLASH 1282. Within the limits of reproducibility and measurement accuracy, the location of contralateral activation was similar using FLASH and EPI sequences. In conclusion, EPI proved to be a reliable technique for fMRI of the motor cortex, even on an MR scanner with a conventional gradient system.

Adult↗

Epidemiological tracing of Salmonella enterica serotype Abortusovis from Spanish ovine flocks by PFGE fingerprinting.

Salmonella enterica serotype Abortusovis, an ovine host-specific serotype, rare in most countries, is responsible for epidemic abortion episodes in Spain. With the aim of surveillance and detection of the spread of specific clones, 55 Abortusovis isolates collected during 1996-2001 from flocks in 11 provinces, were studied using XbaI-PFGE. Despite the fact that the strains were geographically and spatially related, PFGE demonstrated an epidemiologically acceptable discriminating power, identifying 20 clones (similarity, 52-96%). Clones Sabv6, 1, 5 were disseminated in seven, five and two areas respectively, while another 17 clones appeared in single places. Clones from nearby geographic regions showed a high relatedness (one band of difference in the PFGE profile) Sabv1-2-3, Sabv5-6, Sabv7-8, and Sabv13-14, suggesting a common ancestor. Co-isolation in the same flock (Sabv5-6, Sabv1-3, Sabv1-6) was detected. PFGE surveillance detected the predominance and widespread distribution of clone Sabv6 in 21 out of the 55 Abortusovis serotype episodes studied in Spain.

Abortion, Veterinary↗

Cytomimetic engineering of hepatocyte morphogenesis and function by substrate-based presentation of acellular E-cadherin.

Although cadherin-mediated intercellular contacts can be integral to the maintenance of functionally competent hepatocytes in vitro, the ability to engineer hepatocellular differentiated function via acellular E-cadherin has yet to be thoroughly explored. To investigate the potential of substrate-presented, acellular E-cadherin to modulate hepatocellular self-assembly and functional fate, rat hepatocytes were cultured at sparse densities on surfaces designed to display recombinant E-cadherin/Fc chimeras. On these substrates, hepatocytes were observed to recognize microdisplayed E-cadherin/Fc and responded by modulating the spatial distribution of the intracellular cadherin-complexing protein beta-catenin. Substrate-presented E-cadherin/Fc was also found to markedly alter patterns of hepatocyte morphogenesis, as cellular spreading and two-dimensional reorganization were significantly inhibited under these conditions, leading to multicellular aggregates that were considerably more three-dimensional in nature. Increasing cadherin exposure was also associated with elevated levels of albumin and urea secretion, two markers of hepatocyte differentiation, over control cultures. This suggested that cell-substrate cadherin engagement established more functionally competent hepatocellular phenotypes, coinciding with the notion that E-cadherin is a differentiation-inducing ligand for these cells. The morphogenetic and function-promoting effects of substrate-bound E-cadherin/Fc were further enhanced under conditions in which protein A was utilized as an anchoring molecule to present cadherin molecules, suggesting that ligand mobility may play an important role in the effective establishment of cell-to-substrate cadherin interactions. Interestingly, the percent increase in function detected for conditions of high cadherin exposure versus control cultures was found to be substantially higher at extremely low cell densities. This observation indicated that hepatocytes respond to substrate-presented E-cadherin even in the absence of native intercellular interactions and associated juxtacrine signaling. The incorporation of acellular E-cadherin on biomaterial substrates may thus potentially present a means to prevent hepatocellular dedifferentiation by maintaining liver-specific function in otherwise severely functionally repressive culture conditions.

Animals↗

An effective method to verify line and point spread functions measured in computed tomography.

This study describes an effective method for verifying line spread function (LSF) and point spread function (PSF) measured in computed tomography (CT). The CT image of an assumed object function is known to be calculable using LSF or PSF based on a model for the spatial resolution in a linear imaging system. Therefore, the validities of LSF and PSF would be confirmed by comparing the computed images with the images obtained by scanning phantoms corresponding to the object function. Differences between computed and measured images will depend on the accuracy of the LSF and PSF used in the calculations. First, we measured LSF in our scanner, and derived the two-dimensional PSF in the scan plane from the LSE Second, we scanned the phantom including uniform cylindrical objects parallel to the long axis of a patient's body (z direction). Measured images of such a phantom were characterized according to the spatial resolution in the scan plane, and did not depend on the spatial resolution in the z direction. Third, images were calculated by two-dimensionally convolving the true object as a function of space with the PSF. As a result of comparing computed images with measured ones, good agreement was found and was demonstrated by image subtraction. As a criterion for evaluating quantitatively the overall differences of images, we defined the normalized standard deviation (SD) in the differences between computed and measured images. These normalized SDs were less than 5.0% (ranging from 1.3% to 4.8%) for three types of image reconstruction kernels and for various diameters of cylindrical objects, indicating the high accuracy of PSF and LSF that resulted in successful measurements. Further, we also obtained another LSF utilizing an inappropriate manner, and calculated the images as above. This time, the computed images did not agree with the measured ones. The normalized SDs were 6.0% or more (ranging from 6.0% to 13.8%), indicating the inaccuracy of the PSF and LSE We could verify LSFs and PSFs for three types of reconstruction kernels, and demonstrated differences between modulation transfer functions (MTFs) derived from validated LSFs and inaccurate LSFs. Our technique requires a simple phantom that is suitable for clinical scanning, and does not require a particular phantom containing some metals or specific fine structures, as required in methods previously used for measurements of spatial resolution. Therefore, the scanned image of the phantom will be reliable and of good quality, and this is used directly as a confident reference image for the verification. When one obtains LSF, PSF or MTF values, verification using our method is recommended. Further, when another method for the measurement of LSF and PSF is developed, it could be validated using our technique, as illustrated in the method proposed by Boone [Med. Phys. 28, 356-360 (2001)] and used in this paper.

Algorithms↗

The line spread function and modulation transfer function of a computed tomographic scanner.

A simple method to measure the line spread function of computed tomographic scanners has been developed. The line spread function of the EMI-Scanner was measured and the MTF of the scanner was calculated. The reproducibility of the method indicated this technique could be utilized to compare the spatial resolution of various computed tomographic scanners.

Diagnosis, Computer-Assisted↗

[Structure of chromatin and chromosomes in preparations of interphase nucleus derivatives, prepared by removal of the nucleuar envelopes. II. Structure of chromatin and associations of chromosomes in stretched amembranous nuclei and mitotic figures].

Preparations of surface stretched amembranous nuclei and mitotic figures were used for revealing the high order nuclear and chromosomal structures. The preparations were obtained by dropping amembraneous nuclei and mitotic figures suspension in methanol-glacial acetic acid mixture (3:1) on wetted superclean slides. Amembraneous nuclei and mitotic figures were isolated from intact murine and human cells (lines L1210, SK-UT-1B, PHA-stimulated lymphocytes) by means of their 1-5 min prefixational capillary pipetting with freshly prepared 0.018-0.06% Triton X-100 solution in the conditional cultural medium. Stretched amembraneous nuclei and mitotic figures had no features of induced chromatin dispersion and compaction. Stretched interphase amembraneous nuclei showed spatially separated individual structures (thin chromatin fibres, nucleoli, intranuclear bodies), polymorphous pattern of perinucleolar chromatin aggregation and episodically expressed beaded thick chromatin fibres and a chromocenter. The chromomeric pattern of the spread chromosomes of mitotic figures was quite similar but hardly identical with that of G-banding. The stretched prometaphase mitotic figures in all tested cell types always contained loose "residual" nucleoli looking like typical prophase nucleoli as concerns their shape and number per cell (mitotic figure). The majority of chromosomes of stretched mitotic figures and of prophase amembraneous nuclei were attached to the nucleolar material. All tested cell lines showed almost the same variation in number of nucleolus-attached chromosomes, per both prophase amembraneous nucleus and prometaphase mitotic figure. Some chromosomes of stretched mitotic figures were colocated with "residual" nucleoli and looked shortened and strongly condensed. Other chromosomes, locally associated with "residual" nucleoli, were straight and oriented radially to these. Mutual chromosomal arrangements in mitotic cells on smears and in stretched mitotic figures were analogous. Equatorial plates from PBS-washed SK-UT-1B cells displayed a better stretching capacity than those from untreated cells. In the former case metaphase chromosomes were seen more uniformly stretched and well identified after GTG-banding procedure. The number of interchromosomal (mainly telomere-telomeric and telomere-centromeric) connections per stretched mitotic figure (or per stretched prophase amembraneous nucleus) was minimum in late prometaphase, maximum in prophase and early prometaphase, and intermediate in metaphase. The obtained data are discussed in terms of topology and longitudinal heterogeneity of mitotic chromosomes.

Animals↗

Optimal sensor distribution for measuring the tangential field components in MCG.

We discuss the optimal arrangement of detectors for a 52-channel magnetocardiogram (MCG) system measuring tangential components of the cardiac magnetic fields. Nowadays, most MCG instruments are designed to cover the whole heart area to maximize the information available from the myocardial magnetic field in a simultaneous measurement. In such a system, detectors should be spread over a sufficiently wide area. However, an increased diameter of the cooling dewar will result in more heat-loss and higher production and maintenance costs. Therefore, we reviewed the spatial sampling theory to determine the proper interval between detectors, and we decided on the number of channels to cover the whole heart area. In order to fit the detector array on the cylindrical dewar economically, we removed the detectors at the corners of the square array. Through simulations using the confidence region method, we verified that our design of the detector array is enough to obtain adequate information from the heart. Simulations also suggested that tangential-component MCG measurement can localize deep current dipoles better than normal-component measurement with the same confidence volume; therefore, we conclude that measurement of the tangential component is more suitable to an MCG system than the normal component.

Body Surface Potential Mapping↗

Laser-Doppler flowmetry.

Laser-Doppler flowmetry is a new method of the continuous and noninvasive measuring of the tissue blood flow, utilizing the Doppler shift of laser light as the information carrier. The method has already proved its potential usefulness in the clinical assessment of blood flow within disciplines such as dermatology, plastic surgery, and gastrointestinal surgery. In experimental medicine, laser-Doppler flowmetry has been used in the study of spontaneous rhythmical variations as well as in the study of spatial and temporal fluctuations in human skin blood flow. The method has facilitated further investigations of the nature of blood flow in a variety of tissue types. The method is rapidly spreading to many new applications in microvascular blood flow measurements. Several commercial laser Doppler instruments are available. This review article is a presentation of the state of the art of laser-Doppler flowmetry including theory, technology, and applications.

Animals↗

Cerebrovascular regulation and vasoneuronal coupling.

Maintenance of cerebral perfusion pressure is a prerequisite for the prevention of cerebral ischemia. Physiological fluctuations in systemic perfusion pressure are compensated by cerebrovascular autoregulation. Cerebral hypoperfusion could result from (1) systemic hemodynamic failure (eg, distal to severe arterial stenosis), overcharging the vasoregulatory capacity; (2) dysfunction and exhaustion of cerebrovascular autoregulation; or (3) both. Ultrasound offers an excellent temporal resolution, is noninvasive, and is easily applicable for follow-up investigations. Despite its poor spatial resolution, transcranial Doppler sonography has been used for determination of cerebral perfusion reserve studies measuring cerebral blood flow velocity (CBFV) during hypercapnia or application of vasoactive agents (eg, acetazolamide). This approach evaluates vasomotor regulation in patients with hemodynamic compromise distal to severe stenosis or occlusion of the brain supplying arteries. Monitoring CBFV during tilt table examinations directly measures cerebral autoregulation. In patients with systemic orthostatic hypotension, maintainance or failure of cerebrovascular compensation and, even more importantly, cerebrovascular dysautoregulation, despite normal systemic blood pressure regulation, may be demonstrated. Vasoneuronal coupling is reflected by CBFV variations during appropriate neuronal stimulation. Neuronal dysfunction is associated with CBFV abnormalities as exemplified by preconditions of focal cerebral dysfunction in the posterior cerebral artery (PCA) in migraineurs with aura, where massive alteration of vasoneuronal coupling and ischemia is threatening during spreading depression. A highly significant asymmetric gain of vasoneuronal coupling in the interictal state may act as a trigger mechanism in these patients. Testing for vasoneuronal coupling within the middle cerebral artery (MCA) territory is more difficult due to the poor spatial resolution with various neuronal stimuli (eg, motorsensory or cognitive paradigms), only eliciting local neuronal areas underrepresented in the MCA CBFV global changes. However, motor stimulation evoked CBFV may be used to indicate dysintegration of vasoneuronal coupling in the course of acute cerebral ischemia with sensorimotor hemiparesis and, moreover, seems to be of prognostic value regarding the motor deficit.

Blood Flow Velocity↗

A novel interpolation method for electric potential fields in the heart during excitation.

In mapping the electrical activity of the heart, interpolation of electric potentials plays two important roles. First, it permits the estimation of potentials in regions that could not be sampled or where signal quality was poor, and second, it supports the construction of isopotential lines and surfaces for visualization. The difficulty in developing robust interpolation techniques for cardiac applications lies in the abrupt change in potential in the vicinity of the activation wave front. Despite the resulting nonlinearities in spatial potential distributions, simple linear interpolation methods are the current standard and the resulting errors due to aliasing can be large if electrode spacing does not lie on the order of 0.5-2 mm--the thickness of the activation wave front. We have developed a novel interpolation method that is based on two observations specific to the spread of excitation in the heart: (1) that propagation velocity changes smoothly within a region large enough to contain several measurement electrodes and (2) that electrogram morphology varies very little in the neighborhood of each sample point except for a time shift in the potential wave forms. The resulting interpolation scheme breaks the interpolation of one highly nonlinear variable--extracellular potential--into two separate interpolations of variables with much less drastic spatial variation--activation time and electrogram morphology. We have applied this method to potentials originally recorded at 1.5 mm spacing and then subsampled at a range of densities for testing of the interpolation. The results based both on reconstruction of isopotential contour maps and statistical comparison showed significant improvement of this novel approach over standard linear techniques. The applications of the new method include improved determination of electrophysiological parameters such as spatial gradients of potential and the path of cardiac activation and recovery, estimation of electrograms at desired locations, and visualization of electric potential distributions.

Algorithms↗

The evolution of parasite virulence and transmission rate in a spatially structured population.

If the transmission occurs through local contact of the individuals in a spatially structured population, the evolutionarily stable (ESS) traits of parasite might be quite different from what the classical theory with complete mixing predicts. In this paper, we theoretically study the ESS virulence and transmission rate of a parasite in a lattice-structured host population, in which the host can send progeny only to its neighboring vacant site, and the transmission occurs only in between the infected and the susceptible in the nearest-neighbor sites. Infected host is assumed to be infertile. The analysis based on the pair approximation and the Monte Carlo simulation reveal that the ESS transmission rate and virulence in a lattice-structured population are greatly reduced from those in completely mixing population. Unlike completely mixing populations, the spread of parasite can drive the host to extinction, because the local density of the susceptible next to the infected can remain high even when the global density of host becomes very low. This demographic viscosity and group selection between self-organized spatial clusters of host individuals then leads to an intermediate ESS transmission rate even if there is no tradeoff between transmission rate and virulence. The ESS transmission rate is below the region of parasite-driven extinction by a finite amount for moderately large reproductive rate of host; whereas, the evolution of transmission rate leads to the fade out of parasite for small reproductive rate, and the extinction of host for very large reproductive rate.

Animals↗

Time frame of mitral cell development in the mice olfactory bulb.

Along with tufted cells, mitral cells are the principal projection neurons in the olfactory bulb (OB). During the development of the OB, mitral cells migrate from the ventricular zone to the intermediate zone, where they begin to send axons along the lateral olfactory tract (LOT) to the cortical olfactory zones. Subsequently, they lose their tangential orientation, enabling them to make contact with the axons of the olfactory sensory neurons (OSN) that innervate the whole OB. Here, we investigated the distinct morphological features displayed by developing mitral cells and analyzed the relationship between the changes undertaken by these neurons and the arrival of the OSN axons. Immunostaining for specific markers of developing axons and dendrites, coupled with the use of fluorescent tracers, revealed the morphological changes, the continuous reorientation, and the final refinement that these cells undergo. We found that some of these changes are dependent on the arrival of the OSN axons. Indeed, we identified three main chronological events: 1) newly generated neurons become established in the intermediate zone and project to the LOT; 2) the cells reorient and spread their dendrites at the same time as OSN axons penetrate the OB (this is a sensitive period between embryonic day (E)15-16, in which the arrival of afferents establishes a spatial and temporal gradient that facilitates protoglomerulus and glomerulus formation); and 3) final refinement of the radially orientated cells to adopt a mature morphology. These results suggest that both afferent inputs and intrinsic factors participate to produce the well-defined sensory system.

Aging↗