Study of intestinal motility. Contribution of electromyography]

The longitudinal and circular deformations of the intestinal wall have been studied simultaneously with the intestinal electrical activity. Tracings were obtained for several weeks by means of a compact device which was implanted on the jejunum in 6 dogs and 4 rabbits. The results showed that the transverse movements of the intestine corresponded to the muscular activity of the circular layer. The longitudinal movements were observed permanently in the rabbit. In the dog they could correspond to a contraction or a relaxation movement. In the two cases, no electrical activity was related to the longitudinal mechanical activity of the intestine.     

The effect of hydration on mechanical anisotropy,topography and fibril organization of the osteonal lamellae

The effect of hydration on the mechanical properties of osteonal bone, in directions parallel and perpendicular to the bone axis, was studied on three length scales: (i) the mineralized fibril level (~100 nm), (ii) the lamellar level (~6 µm); and (iii) the osteon level (up to ~30 µm).We used a number of techniques, namely atomic force microscopy (AFM), nanoindentation and microindentation. The mechanical properties (stiffness, modulus and/or hardness) have been studied under dry and wet conditions. On all three length scales the mechanical properties under dry conditions were found to be higher by 30–50% compared to wet conditions. Also the mechanical anisotropy, represented by the ratio between the properties in directions parallel and perpendicular to the osteon axis (anisotropy ratio, designated here by AnR), surprisingly decreased somewhat upon hydration. AFM imaging of osteonal lamellae revealed a disappearance of the distinctive lamellar structure under wet conditions. Altogether, these results suggest that a change in mineralized fibril orientation takes place upon hydration.     

Probing the role of water in lamellar bone by dehydration in the environmental scanning electron microscope

Water, collagen and mineral are the three major components of bone. The structural organization of water and its functions within the bone were investigated using the environmental scanning electron microscope and by analyzing dimensional changes that occur when fresh equine osteonal bone is dehydrated and then rehydrated. These changes are attributed mainly to loss of bulk and weakly bound water. In longitudinal sections a contraction of 1.2% was observed perpendicular to the lamellae, whereas no contraction occurred parallel to the lamellae. In transverse sections a contraction of 1.4% was observed both parallel and perpendicular to the lamellae. SEM back scattered electron images showed that about half of an individual lamella is less mineralized, and thus has more water than the other half. We therefore propose that contractions perpendicular to lamellae are due to the presence of more water-filled rather than mineral-filled channels within the mineralized collagen fibril arrays. As these channels are also aligned with the crystal planes, the crystal arrays, oriented as depicted in the rotated plywood model for lamellar bone, facilitate or hinder contraction in different directions.     

THE FINE STRUCTURE OF THE CILIA FROM CTENOPHORE SWIMMING-PLATES

The ctenophore swimming-plate has been examined with the electron microscope. It has been recognized as an association of long cilia in tight hexagonal packing. One of the directions of the hexagonal packing is parallel to the long edge of the swimming-plate and is perpendicular to the direction of the ciliary beat. All the cilia in the swimming-plate are identically oriented. The effective beat in the movement of the swimming-plate is directed towards the aboral pole of the animal, and this is also the side of the unpaired peripheral filament in all the cilia. The direction of the ciliary beat is fixed in relation to the position of the filaments of the cilia. The swimming-plate cilium differs from other types of cilia and flagella in having a filament arrangement that can be described as 9 + 3 as opposed to the conventional 9 + 2 pattern. The central filaments appear in a group of two "tubular" filaments and an associated compact filament. The compact filament might have a supporting function. It has been called "midfilament." Two of the peripheral nine filaments (Fig. 1, Nos. 3 and 8) are joined to the ciliary membrane by means of slender lamellae, which divide the cilium into two unequal compartments. These lamellae have been called "compartmenting lamellae." Some observations of the arrangement of the compartmenting lamelae indicate that they function by cementing the cilia together in lateral rows. The cilia of the rows meet at a short distance from each other, leaving a gap of 30 A only. The meeting points are close to the termini of the compartmenting ridges. An electron-dense substance is sometimes seen bridging the gap. Some irregularities are noted with regard to the arrangement of the compartmenting lamellae particularly at the peripheral rows of cilia. In many cilia in these rows there are small vesicles beneath the ciliary membrane.     

Mechanical properties of the human gastrointestinal tract

The tensile properties of the human esophagus, stomach, small and large bowel were examined on an Instron 1221 tensiometer. The values of maximal stress and destructive strain were the following: for esophagus-1.2 MPa and 140%, respectively, for stomach axial specimens-0.7 MPa and 190%, for stomach transversal specimens-0.5 MPa and 190%, for small bowel transversal specimens-0.9 MPa and 140% and for large bowel transversal specimens-0.9 MPa and 180%. Tests conducted on small and large bowel axial specimens permitted examination of the intestinal wall as a multi-layered structure. The mechanical properties of tested bowels in axial and transversal directions were qualitatively different. The submucosa and muscular layers condition the mechanical strength of bowel wall, while the serosa and mucosa showed no significant strength. Reproducible results were generated for cadaveric and surgically removed stomach and small intestine, which showed their mechanical properties similar under certain storage conditions. The data received could be used for monitoring of the mechanical properties of bowel wall layers under different conditions and for checking of bowel distension sequences.     

Orientation of bone mineral and its role in the anisotropic mechanical properties of bone--transverse anisotropy

An orientation of hydroxyapatite (HAP) crystals in bovine femur mineral was investigated by means of X-ray pole figure analysis (XPFA). It was found that the c-axis of HAP generally orients parallel to the longitudinal axis of bone (bone axis) and a significant amount of c-axis was oriented in other directions, in particular, perpendicular to the bone axis. Comparing these results with those of the small angle X-ray scattering (SAXS) investigation by Matsushima et al. (Jap. J. appl. Phys. 21, 186-189, 1982) at least two types of morphology of bone mineral were found; rod like bone mineral having the c-axis of HAP crystal parallel to the longitudinal axis of the rod and that having the c-axis not parallel, in a particular case, perpendicular to its longitudinal axis. Transverse anisotropy in mechanical properties of bone was reproduced by the estimation of Young's moduli by using the structural results mainly from XPFA. It is concluded that the anisotropy in mechanical properties of bone is well explained by taking account of the non-longitudinal (off-bone) axial distribution of orientation of bone mineral.     

Structural differences between "dark" and "bright" isolated human osteonic lamellae

This investigation presents new insights into the structure of human secondary lamellae. Lamellar specimens that appear dark and bright on alternate osteon transverse sections under circularly polarizing light were isolated using a new technique, and examined by polarizing light microscopy, synchrotron X-ray diffraction, and confocal microscopy. A distribution of unidirectional collagen bundles and of two overlapping oblique bundles appears on circularly polarizing light microscopy images in relation to the angle between the specimen and the crossed Nicols' planes. The unidirectional collagen bundles observed at 45 degrees run parallel to the osteon axis in the dark lamellar specimens and perpendicular to it in the bright ones. Small and wide-angle micro-focus X-ray diffraction indicates that the dark lamellae are structurally quite homogeneous, with collagen fibers and apatite crystals preferentially oriented parallel to the osteon axis. Bright lamellar specimens exhibit different orientation patterns with the dominant ones bidirectional at +/-45 degrees with respect to the osteon axis. Accordingly, confocal microscopy evidences the presence of longitudinal bundles in dark lamellar specimens and oblique bundles in the bright ones. Radial bundles are evidenced in both lamellar types. The alternate osteon structure is described by a rather continuous multidirectional pattern, in which dark and bright lamellae display different mechanical and possibly biological functions.     

Size-Dependent Diffusion of Dextrans in Excised Porcine Corneal Stroma

Delivery of therapeutic agents to the eye requires efficient transport through cellular and extracellular barriers. We evaluated the rate of diffusive transport in excised porcine corneal stroma using fluorescently labeled dextran molecules with hydrodynamic radii ranging from 1.3 to 34 nm. Fluorescence correlation spectroscopy (FCS) was used to measure diffusion coefficients of dextran molecules in the excised porcine corneal stroma. The preferential sensitivity of FCS to diffusion along two dimensions was used to differentially probe diffusion along the directions parallel to and perpendicular to the collagen lamellae of the corneal stroma. In order to develop an understanding of how size affects diffusion in cornea, diffusion coefficients in cornea were compared to diffusion coefficients measured in a simple buffer solution. Dextran molecules diffuse more slowly in cornea as compared to buffer solution. The reduction in diffusion coefficient is modest however (67% smaller), and is uniform over the range of sizes that we measured. This indicates that, for dextrans in the 1.3 to 34 nm range, the diffusion landscape of corneal stroma can be represented as a simple liquid with a viscosity approximately 1.5 times that of water. Diffusion coefficients measured parallel vs. perpendicular to the collagen lamellae were indistinguishable. This indicates that diffusion in the corneal stroma is not highly anisotropic. Our results support the notion that the corneal stroma is highly permeable and isotropic to transport of hydrophilic molecules and particles with hydrodynamic radii up to at least 34 nm.     

Hornet cuticle-a composite structure comprised of a series of duplex lamellae attenuating toward the interior of the body

Our study deals with the ultramicroscopic structure of the yellow pigmented cuticular stripes on the abdomen of the oriental hornet Vespa orientalis (Hymenoptera, Vespinae). The abdominal cuticle is composed of numerous (more than 25) lamellae which progressively attenuate as one proceeds from the outside interiorly. On a logarithmic scale the pattern of lamellar attenuation is a linear drop to almost asymptotic thickness with the innermost lamellae at least one order of magnitude less than that of the outermost one and describable by a mathematic equation. Each lamella is a composite structure consisting of two primary elements, namely, a skeleton and filling material. The skeleton is composed of chitin and comprised of parallel ‘plates’ which visibly separate between the lamellae. These ‘plates’ are interconnected by: (1) perpendicular ‘pillars’ which are distributed intermittently at more or less uniform intervals; and (2) by a layer of chitin shaped like a horizontal helix or at times even a double helix whose diameter equals the distance between two successive lamellae. In the space within the skeletal element there is a filling material of variable nature, mostly proteinaceous, which dissolves out and vacates the cuticle during its processing for microscopic observation, leaving behind the skeletal element. Such structure of the cuticle, wherein two different materials combine serially into one duplex structure is typical of composite materials. This duplex structuring of the lamellae, combined with their gradual attenuation and compacting towards the interior, lends the cuticle in the abdominal region the shape of a thin-walled laminated beam endowed with a wide array of mechanical, thermal, electric, optic, acoustic and probably numerous other properties which render the cuticle efficient for multiple purposes.     

Relationship between transmural potential difference and smooth muscle slow waves and contractility in the rabbit small intestine in vitro

The relationship between transmural potential difference (PD) and smooth muscle electrical and mechanical activity was investigated in the rabbit ileum in vitro. Transmural PD was monitored using agar salt bridge electrodes connected via calomel half cells to an electrometer. Force displacement transducers recorded predominantly longitudinal smooth muscle activity. Concurrently, predominantly circular muscle activity was recorded at three sites using intraluminal pressure probes. At the same sites, suction electrodes monitored electrical activity of the smooth muscle. In all experiments, fluctuations in transmural PD were temporally linked to smooth muscle mechanical and electrical activity. The frequency of PD oscillations, electrical slow waves, and cyclic pressure changes were identical within each segment. Adrenaline abolished smooth muscle electrical spiking, all mechanical activity, and transmural fluctuations in PD. However, the slow waves were not abolished, though their frequency was increased. Phentolamine but not propranolol reversed the effects of adrenaline, thus slow wave frequency is influenced by alpha-adrenergic stimulation in the rabbit ileum. In conclusion, oscillations in transmural PD are unrelated to the ionic processes associated with the slow wave. However, they are in some way linked to smooth muscle contractile activity, possibly via an intrinsic neural mechanism as observed in the guinea pig.     

Kinetics of D-histidine diffusion across rat intestine in vitro.

A five-compartment linear model for diffusion in vitro across rat jejunum has been proposed for the study of the kinetic constants of D-histidine transport. Once preliminary experiments using 2,4-dinitrophenol and L-methionine have proved that D-histidine gives rise to passive transport only, the validity of the model was tested and its parameters estimated through a best-fitting procedure by using experimental data concerning D-histidine transport. D-Histidine diffusion was studied in everted and unreverted loops mounted in an oxygenated bath system. Both mucosa to serosa and seroa to mucosa movements of D-histidine (3-30 mM) were evaluated by measuring chemically the amount of D-histidine transported into intestinal lumen every 5 min for 60 min. Results obtained proved that D-histidine transport in each direction (mucosa to seroa or seroa to mucosa) was dependent-concentration process. Nevertheless different values of gain and time constants were estimated for the transport in the two directions.     

Development of the intestines. Human embryos]

In the present study an attempt was made to think over anew the development of the peritoneal situs, guided by the comparison of the "root-area" (the connective tissue area free of peritoneum between the left and right transition line of the peritoneum parietale to the peritoneum viscerale) of the so-called "primitive mesentery" of an embryo (10 mm CRL) and of an older fetus (250 mm CRL). The following results were obtained: The so-called primitive sagittal mesentery cannot be regarded as the anlage of the adult mesentery, in the sense that the latter develops out of the former dy displacement of rotation. The original "root area" widens with the broadening of the back and can still be demonstrated as an homogeneous "root area" of the "intestinal bulge", after the typical adult situs has developed. There is a connection between the development of the mesenteries and the formation of the folds of the embryonic serosa. Their position is determined through the recesses of the intestinal bulge. They exist because of the foldings and are therefore secondary characteristics. Under these aspects it is possible to consider the development of the situs as a uniform folding process of the embryonic serosa, without references to as yet unproved secondary adhesions of the peritoneal layers.     

Spatial and temporal coupling between slow waves and pendular contractions

In contrast to the mechanisms of segmental and peristaltic contractions in the small intestine, not much is known about the mechanism of pendular contractions. High-resolution electrical and mechanical recordings were performed from isolated segments of the rabbit ileum during pendular contractions. The electrical activities were recorded with 32 extracellular electrodes while motility was assessed simultaneously by video tracking the displacements of 20-40 serosal markers. The electrical activities consisted of slow waves, followed by spikes, that propagated in either the aboral or oral direction. The mechanical activity always followed the initial electrical activity, describing a contraction phase in one direction followed by a relaxation phase in the opposite direction. Pendular displacements were always in rhythm with the slow wave, whereas the direction of the displacements was dictated by the origin of the slow wave. If the slow wave propagated aborally, then the pendular displacement occurred in the oral direction, whereas if the slow wave propagated in the oral direction, then the displacement occurred in the aboral direction. In the case of more complex propagation patterns, such as in the area of pacemaking or collision, direction of displacements remained always opposite to the direction of the slow wave. In summary, the direction and pattern of propagation of the slow wave determine the rhythm and the direction of the pendular motility. The well-known variability in pendular movements is caused by the variability in the propagation of the underlying slow wave.     

Study of dielectric behavior of DNA in shear gradient

The dielectric behavior of the DNA molecule is investigated in the presence of mechanical force as well as the electrical field. In the present experiment, the direction of the electrical field is perpendicular to that of the mechanical force. The dipole moment of polar molecules manifest itself as dielectric increment at low frequencies or as the conductance increment at high frequencies. These two quantities are closely related to each other by Eq. (1) in the text. Because of the difficulty due to electrode polarization at low frequencies, no useful information was obtained by investigating the dielectric increment in the present system. Therefore, the effect of shear gradient was studied by measuring the conductance increment at high frequencies with various velocity gradients. The conductance increment decreased when the shear was applied perpendicular to the electrical field. The conductance change is converted into the unit of dielectric constant; it was found that the dielectric increment of DNA solution decreases by as much as 85 percent. From these observations, it is concluded that the direction of the dipole moment in DNA is longitudinal rather than transverse. The same experiment was repeated with the random coil DNA and no anisotropy in the dielectric increment was observed.     

Elastic anisotropy of bone lamellae as a function of fibril orientation pattern

In this study, the homogenized anisotropic elastic properties of single bone lamellae are computed using a finite element unit cell method. The resulting stiffness tensor is utilized to calculate indentation moduli for multiple indentation directions in the lamella plane which are then related to nanoindentation experiments. The model accounts for different fibril orientation patterns in the lamellae—the twisted and orthogonal plywood pattern, a 5-sublayer pattern and an X-ray diffraction-based pattern. Three-dimensional sectional views of each pattern facilitate the comparison to transmission electron (TEM) images of real lamella cuts. The model results indicate, that the 5-sublayer- and the X-ray diffraction-based patterns cause the lamellae to have a stiffness maximum between 0° and 45° to the osteon axis. Their in-plane stiffness characteristics are qualitatively matching the experimental findings that report a higher stiffness in the osteon axis than in the circumferential direction. In contrast, lamellae owning the orthogonal or twisted plywood fibril orientation patterns have no preferred stiffness alignment. This work shows that the variety of fibril orientation patterns leads to qualitative and quantitative differences in the lamella elastic mechanical behavior. The study is a step toward a deeper understanding of the structure—mechanical function relationship of bone lamellae.     

Vasoactive intestinal polypeptide (VIP) inhibits prostaglandin-F2α-induced activity of the rabbit myometrium

Seven female rabbits had myometrial autografts transplanted into ear-chambers. The electrical activity of the graft was recorded and mechanical activity observed. Vasoactive intestinal polypeptide (VIP) dose-dependently inhibited myometrial activity induced by prostaglandin-F_2α. This evidence and the occurrence of “VIPergic” nerve fibres, which seem to innervate the myometrial cells, suggest that VIP may play a physiological role in the local control of the myometrial activity.     

A microscale anisotropic biaxial cell stretching device for applications in mechanobiology

A multi-layered polydimethylsiloxane microfluidic device with an integrated suspended membrane has been fabricated that allows dynamic and multi-axial mechanical deformation and simultaneous live-cell microscopy imaging. The transparent membrane’s strain field can be controlled independently along two orthogonal directions. Human foreskin fibroblasts were immobilized on the membrane’s surface and stretched along two orthogonal directions sequentially while performing live-cell imaging. Cyclic deformation of the cells induced a reversible reorientation perpendicular to the direction of the applied strain. Cells remained viable in the microdevice for several days. As opposed to existing microfluidic or macroscale stretching devices, this device can impose changing, anisotropic and time-varying strain fields in order to more closely mimic the complexities of strains occurring in vivo.     

Assessment of composition and anisotropic elastic properties of secondary osteon lamellae

Measurement of the elastic properties of single osteon lamellae is still one of the most demanding tasks in bone mechanics to be solved. By means of site-matched Raman microspectroscopy, acoustic microscopy and nanoindentation the structure, chemical composition and anisotropic elasticity of individual lamellae in secondary osteons were investigated. Acoustic impedance images (911-MHz) and two-dimensional Raman spectra were acquired in sections of human femoral bone. The samples were prepared with orientations at various observation angles theta relative to the femoral long axis. Nanoindentations provided local estimations of the elastic modulus and landmarks necessary for spatial fusion of the acoustic and spectral Raman images. Phosphate nu(1) (961 cm(-1)) and amide I (1665 cm(-1)) band images representing spatial distributions of mineral and collagen were fused with the acoustic images. Acoustic impedance was correlated with the indentation elastic modulus E(IT) (R(2)=0.61). Both parameters are sensitive to elastic tissue anisotropy. The lowest values were obtained in the direction perpendicular to the femoral long axis. Acoustic images exhibit a characteristic bimodal lamellar pattern of alternating high and low impedance values. Since this undulation was not associated with a variation of the phosphate nu(1)-band intensity in the Raman images, it was attributed to variations of the lamellar orientation. After threshold segmentation and conversion to elastic modulus the orientation and transverse isotropic elastic constants were derived for individual ensembles of apparent thin and thick lamellae. Our results suggest that this model represents the effective anisotropic properties of an asymmetric twisted plywood structure made of transverse isotropic fibrils. This is the first report that proves experimentally the ability of acoustic microscopy to map tissue elasticity in two dimensions with micrometer resolution. The combination with Raman microspectroscopy provides a unique way to study bone and mineral metabolism and the relation with mechanical function at the ultrastructural tissue level.     

Propulsive action of a snake pushing against a single site: Its combined analysis

The simultaneous use of electromyography (EMG), strain gauges, and cinematography show that the capacity of continuous displacement from a single peg is based on the following: sequential activity of the tested muscles from front to rear; activity restricted to the short portion of the body in contact with the peg; alternate action of the muscle longissimus dorsi on the two sides, the transition between one side to the other occurring at the site of contact with the peg; unilateral activity of the muscle supracostalis ventralis responsible for a bulging against the peg; a great stability in the direction of the resultant force, which makes only a small angle with the directio of the motion.     

Effects of mechanical loading and hypercapnia on inspiratory muscle EMG.

The electromyograms of the diaphragm and an external intercostal muscle were analyzed to see if the effects of hypercapnia on inspiratory muscle electrical activity could be distinguished from those of mechanical loading and to determine whether changes in inspiratory muscle electrical activity were a sueful measure of CO2 response during mechanical loading. Anesthetized dogs were studied: 1) during progressive hypercapnia without mechanical loading, 2) during flow-resistive and elastic loading at constant PCO2, and 3) during progressive hypercapnia and mechanical loading. Both mechanical loading and hypercapnia increased total inspiratory diaphragmatic and intercostal muscle electrical activity. However, inspiratory duration was increased by mechanical loads but reduced by hypercapnia. Because of these changes in inspiratory duration, the average rate of diaphragmatic electrical activity remained unaffected by mechanical loading before and after vagotomy but was increased by hypercapnia. In contrast, both hypercapnia and mechanical loading increased the average rate of intercostal muscle electrical activity. There was a greater increase in both total and average rate of intercostal muscle electrical activity during hypercapnia in the presence of mechanical loading than during unloaded breathing. However, the change in total and average rate of diaphragmatic electrical activity with PCO2 was unaffected by added mechanical loads. These results suggest that diaphragmatic but not intercostal muscle electrical activity can be used as an index of CO2 response even during mechanical loading.