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From head to tail: the myoseptal system in basal actinopterygians.

Experimental studies indicated that myomeres play several functional roles during swimming. Some of the functions in question are thought to change rostrocaudally, e.g., anterior myomeres are thought to generate forces, whereas posterior myomeres are thought to transmit forces. In order to determine whether these putative functions are reflected in myoseptal morphology we carried out an analysis of the myoseptal system that includes epaxial and hypaxial myosepta of all body regions for the first time. We combined clearing and staining, microdissections, polarized light microscopy, SEM technique, and length measurements of myoseptal parts to reveal the spatial arrangement, collagen fiber architecture, and rostrocaudal gradients of myosepta. We included representatives of the four basal actinopterygian clades to evaluate our findings in an evolutionary and in a functional context. Our comparison revealed a set of actinopterygian groundplan features. This includes a set of specifically arranged myoseptal tendons (epineural, epipleural, lateral, and myorhabdoid tendons) in all epaxial and postanal hypaxial myosepta. Only preanal hypaxial myosepta lack tendons and exclusively consist of mediolateral fibers. Laterally, myosepta generally align with the helically wound fibers of the dermis in order not to limit the body's maximum curvature. Medially, the relationship of myosepta to vertebrae clearly differs from a 1:1 relationship: a myoseptum attaches to the anterior margin of a vertebra, turns caudally, and traverses at least three vertebrae in an almost horizontal orientation in all body regions. By this arrangement, horizontal multiple layers of myosepta are formed along the trunk dorsal and ventral to the horizontal septum. Due to their reinforcement by epineural or epipleural tendons, these multiple layers are hypothesized to resist the radial expansion of underlying muscle fibers and thus contribute to modulation of body stiffness. Rostrocaudally, a dorsoventral symmetry of epaxial and hypaxial myosepta in terms of spatial arrangement and collagen fiber architecture is gradually developed towards the postanal region. Furthermore, the rostrocaudal extension of myosepta measured between anterior and posterior cones gradually increases. This myoseptal region is reinforced by longitudinal fibers of lateral tendons. Furthermore, the percentage of connective tissue in a cross section increases. These morphological data indicate that posterior myosepta are equipped for multisegmental force transmission towards the caudal fin. Anteriormost myosepta have reinforced and elongated dorsal posterior cones. They are adequately designed to transmit epaxial muscular forces to the neurocranium in order to cause its elevation during suction feeding.

Anatomy, Artistic↗

Imaging the physical and morphological properties of a multi-zone young articular cartilage at microscopic resolution.

PURPOSE: To characterize a number of physical and morphologic properties of young articular cartilage. These properties include the anisotropy of T(2) relaxation, optical retardation, orientation of the collagen fibrils, total thickness of the tissue, number of histologic sub-zones in the tissue, width of individual sub-zones, and correlation between the depths of the local T(2) maxima and the local retardation minima. MATERIALS AND METHODS: Microscopic magnetic resonance imaging (mciro MRI) and polarized light microscopy (PLM) were used to examine three side-by-side specimens from a humeral head of a three-month-old beagle that exhibited a unique topographic heterogeneity from three-zones centrally to multi-zone peripherally. RESULTS: The centrally located specimen showed that the collagen fibrils across the tissue depth have a pattern of the classic three histologic sub-zones (tangential, transitional, and radial). A much more complicated multi-zone structure was found in the specimen located peripherally, with a second transitional zone and a second tangential zone located at the deep part of the tissue. We also showed that the orientation of the collagen fibrils that form the cocoon-shaped territorial matrix surrounding the clusters of chondrocytes can be imaged by our PLM technique. CONCLUSION: The results from the young animal in this report, together with our observations from older animals, demonstrate that MRI and PLM can be used to study the epiphyseal expansion of cartilage in young animals during its growth and subsequent loss in older animals. An illustrative model for the structure of collagen fibrils in a humeral head is suggested as an extension to the classic three-zone model for young articular cartilage.

Animals↗

Structure and physical stability of hydrates and thermotropic mesophase of calcium benzoate.

The aim of this study is to investigate the hydration and the dehydration processes of calcium benzoate hydrates (trihydrate and monohydrate), thermotropic mesophases (dehydrated mesophase and lyophilized mesophase) and amorphous state, and the influence of their molecular order on those processes. X-ray analysis revealed that trihydrate has a planar structure composed of two types of planes-one from benzoic acid, water, and calcium ion and another from benzoic acid and water-and that both planes are linked by three water molecules. It was found that calcium benzoate was able to exist as thermotropic mesophases by dehydration of trihydrate and lyophilization. These mesophases were characterized by polarizing-light microscopy (PLM), X-ray powder diffraction (XRPD), differential thermal analysis (DTA), thermogravimetric analysis (TGA), and scanning electron microscopy (SEM). Both mesophases prepared by two procedures showed some similar physical properties, but lyophilized mesophase seemed to have molecular structure with higher order than dehydrated mesophase. The mesophases exhibited different hydration behavior. The dehydrated mesophase showed a stepwise rehydration process where it became monohydrate first and then trihydrate. The lyophilized mesophase became trihydrate without appearance of monohydrate. An amorphous form could also be prepared and it rehydrated first to the monohydrate and then trihydrate. The results suggest that the more disordered dehydrated mesophase and amorphous state change to monohydrate whereas the more ordered lyophilized mesophase cannot change to monohydrate but only to trihydrate.

Absorption↗

Phase equilibria of nafcillin sodium-water.

The phase diagram for the binary system nafcillin sodium-water, was determined using differential scanning calorimetry (DSC) and polarized light microscopy. In the temperature range of -20-30 degrees, three crystalline forms and an amphiphilic liquid crystalline phase were detected. The stable crystalline form of nafcillin sodium (form alpha) and water exhibit a eutectic mixture containing 28% drug (w/w) at -1 degrees. In more dilute solutions a lower temperature eutectic (-5 degrees) occurs. The composition and form of nafcillin sodium in this eutectic are not known. The form -5 degrees eutectic was found to be metastable above 28% concentration and converted to form alpha. Two other crystalline forms were observed at 9 degrees (form beta) and 22 degrees (form gamma) at concentrations above 40%. The crystalline forms could not be further characterized due to their transient nature and existence in highly concentrated mixtures. A lamellar mesophase is present near ambient temperature in mixtures containing more than 55% nafcillin sodium. The phase equilibria were highly susceptible to supercooling. Temperature cycling methods were devised which gave reproducible DSC data and allowed construction of the phase diagram.

Calorimetry, Differential Scanning↗

Phase behavior of tyloxapol/Triton X100/water mixtures.

The influence of admixture of Triton X100 to the polymer tyloxapol and the phase behavior of the mixtures in contact with water was investigated by viscometry, polarization microscopy, and X-ray scattering. The viscosity of tyloxapol/Triton X100 mixtures is lower than that of pure tyloxapol. With increasing Triton X100 content, the phase behavior of the surfactant mixtures in contact with water evolves from the complex behavior of tyloxapol to the simpler one of Triton X100. In contact with water, mixtures rich in tyloxapol form hexagonal, cubic, and lamellar lyotropic liquid crystalline phases over a wide range of concentration and temperature, whereas Triton X100/water mixtures form only a hexagonal phase in a limited concentration and temperature range. The polymeric nature of tyloxapol stabilizes the liquid crystalline structures, and the size of the supramolecular structures in the water-containing surfactant systems is independent of the tyloxapol/Triton X100 mixing ratio but depends highly on water content. The ability of tyloxapol to form stable lyotropic liquid crystalline phases at body temperature, which has been proposed as a basis for the development of novel peroral sustained-release systems, is not significantly impaired by the addition of appropriate amounts of Triton X100. Admixture of Triton X100 to tyloxapol may thus provide a way to circumvent processing problems during the preparation of pharmaceutical formulations based on the highly viscous tyloxapol.

Chemistry, Pharmaceutical↗

Peripheral thermal and mechanical damage to dentin with microsecond and sub-microsecond 9.6 microm, 2.79 microm, and 0.355 microm laser pulses.

BACKGROUND AND OBJECTIVES: It is desirable to minimize peripheral thermal damage during laser irradiation, since thermal damage to collagen and mineral compromises the bond strength to restorative materials in dentin and inhibits healing and osteointegration in bone. There were two primary objectives of this study. The first objective was to measure the degree of thermal damage peripheral to incisions in dentin produced with lasers resonant to the specific absorption bands of water, collagen, and hydroxyapatite with varying pulse duration using polarized-light microscopy (PLM). The second objective was to use synchrotron radiation infrared spectromicroscopy (SR-FTIR) to identify the specific chemical nature of the optical changes observed under PLM in the respective zones of thermal damage peripheral to the laser incisions. STUDY DESIGN/MATERIALS AND METHODS: Precise incisions were produced in 3 x 3 mm2 blocks of human dentin using CO2 (9.6 microm), Er:YSGG (2.79 microm), and Nd:YAG (355 nm) lasers with and without a computer controlled water-spray. Optical coherence tomography (OCT) was used to obtain optical cross-sections of each incision to determine the rate of ablation. The peripheral thermal damage zone around each incision was analyzed using PLM and SR-FTIR. RESULTS: Thermally induced chemical changes to both mineral and the collagen matrix were observed with SR-FTIR with a 10 microm spatial resolution and those changes were correlated with optical changes observed with PLM. Minimal (<10 microm) thermal damage was observed for pulse durations less than the thermal relaxation time (Tr) of the deposited laser energy, with and without applied water at 9.6 microm and with only applied water at 2.79 microm. For pulse durations greater than Tr, greater peripheral thermal damage was observed for both IR laser wavelengths with and without the water-spray. There was minimal thermal damage for 355 nm laser pulses less than Tr with and without applied water; however, extensive mechanical damage (cracks) was observed. CONCLUSIONS: High resolution SR-FTIR is well suited for characterization of the chemical changes that occur due to thermal damage peripheral to laser incisions in proteinaceous hard tissues. Sub-microsecond pulsed IR lasers resonant with water and mineral absorption bands ablate dentin efficiently with minimal thermal damage. Similar laser parameters are expected to apply to the ablation of alveolar bone.

Collagen↗

Lithium ion induced stabilization of the liquid crystalline DNA.

A comparative study of the effects of alkali metal ions Li(+), Na(+), K(+), Rb(+), and Cs(+) on the liquid crystalline organization of high-molecular-weight calf thymus DNA using polarized light microscopy was performed. Major differences in the behavior of Li(+) as compared to the other ions were found. Critical DNA concentration expected to exhibit anisotropic behavior was found to be the same for all the monovalent ions, except for Li(+). DNA initially showed cholesteric textures, which later changed to higher ordered columnar phase for all ions, with the cholesteric-columnar transition facilitated upon increasing the size of the counterion. For Li(+) ion, a nematic schlieren-like texture was formed initially, which after a few days changed to a highly stable (for more than 2 months) biphasic cholesteric-columnar arrangement. The observed differences between Li(+) and other alkali metal ions could be rationalized on the basis of the higher number of hydration water molecules of Li(+) and its complexation behavior. Highly stable DNA mesophases may find applications in the field of nanoelectronics, in designing biosensing units, and in DNA chips.

Animals↗

Poly(epsilon-caprolactone)/chitin and poly(epsilon-caprolactone)/chitosan blend films with compositional gradients: fabrication and their biodegradability.

Poly(epsilon-caprolactone) (PCL)/chitin and PCL/chitosan blend films with compositional gradients were successfully fabricated by a dissolution/diffusion method; that is, repeatedly pouring the PCL/chitin (or PCL/chitosan) blend solutions, with variable composition, onto polysaccharide layers. The compositional gradient structure in the resulting films was characterized by polarized optic microscopy, ATR-FT-IR and trans-FT-IR microscopic spectroscopy. Enzymatic degradability of the PCL/chitin and PCL/chitosan blend films with compositional gradients in the presence of lysozyme was compared with those of homogeneous films and two-layer films. It was found that the degradation rate of PCL/chitin blend films with a compositional gradient was far lower than that of the neat chitin film, whereas the degradation rate of PCL/chitosan blend films with a compositional gradient was close to that of the neat chitosan film. The suppression of the chitosan crystallization, which accelerates the enzymatic degradation, at the surface of PCL/chitosan films with a compositional gradient was much more severe than that for PCL/chitin films with a compositional gradient.

Biodegradation, Environmental↗

T2 and T1rho MRI in articular cartilage systems.

T2 and T1rho have potential to nondestructively detect cartilage degeneration. However, reports in the literature regarding their diagnostic interpretation are conflicting. In this study, T2 and T1rho were measured at 8.5 T in several systems: 1) Molecular suspensions of collagen and GAG (pure concentration effects): T2 and T1rho demonstrated an exponential decrease with increasing [collagen] and [GAG], with [collagen] dominating. T2 varied from 90 to 35 ms and T1rho from 125 to 55 ms in the range of 15-20% [collagen], indicating that hydration may be a more important contributor to these parameters than previously appreciated. 2) Macromolecules in an unoriented matrix (young bovine cartilage): In collagen matrices (trypsinized cartilage) T2 and T1rho values were consistent with the expected [collagen], suggesting that the matrix per se does not dominate relaxation effects. Collagen/GAG matrices (native cartilage) had 13% lower T2 and 17% lower T1rho than collagen matrices, consistent with their higher macromolecular concentration. Complex matrix degradation (interleukin-1 treatment) showed lower T2 and unchanged T1rho relative to native tissue, consistent with competing effects of concentration and molecular-level changes. In addition, the heterogeneous GAG profile in these samples was not reflected in T2 or T1rho. 3) Macromolecules in an oriented matrix (mature human tissue): An oriented collagen matrix (GAG-depleted human cartilage) showed T2 and T(1rho) variation with depth consistent with 16-21% [collagen] and/or fibril orientation (magic angle effects) seen on polarized light microscopy, suggesting that both hydration and structure comprise important factors. In other human cartilage regions, T2 and T1rho abnormalities were observed unrelated to GAG or collagen orientation differences, demonstrating that hydration and/or molecular-level changes are important. Overall, these studies illustrate that T2 and T1rho are sensitive to biologically meaningful changes in cartilage. However, contrary to some previous reports, they are not specific to any one inherent tissue parameter.

Animals↗

Gelsolin-related familial amyloidosis, Finnish type, in a Portuguese family: clinical and neurophysiological studies.

We report a Portuguese family with familial amyloid polyneuropathy related to gelsolin. There were no known Finnish ancestors, but the same mutation as described in Finnish patients (G654A) was carried. Clinical and neurophysiological investigations were performed in four patients. Corneal lattice dystrophy affected all four patients; an axonal lesion of the facial nerve occurred in three patients; visual tract involvement was documented in one case; and corticospinal and posterior column dysfunction was present in one patient. Polarizing microscopy of skin and muscle samples demonstrated amyloid deposits in two patients; anti-gelsolin immunohistochemistry was positive for amyloidogenic gelsolin. The Finnish mutation of gelsolin protein (G654A) was detected in five family members. The utility of neurophysiological testing in the evaluation and follow-up of this type of amyloidosis is discussed.

Adult↗

Changes in renal hemodynamics and urodynamics in rats with chronic hyperoxaluria and after acute oxalate infusion: role of free radicals.

AIMS: The aim of this study was to evaluate possible changes in renal hemodynamic and urodynamic parameters in rats with chronic hyperoxaluria and after acute oxalate challenge. We also evaluated the possible association between free radical (FR) production, hyperoxaluria, and calcium oxalate (CaOx) calculi formation. METHODS: Chronic hyperoxaluria was induced by adding 0.75% ethylene glycol (EG) to the drinking water of male Wistar rats. After 7, 21, and 42 days of treatment, urinary biochemistry, oxalate levels, and lipid peroxides were measured. Kidney calculi were examined by polarizing microscopy. In the second part of the experiments, 1, 10, 20, and 30 mg kg(-1) hr(-1) oxalate was infused, by means of an intrarenal arterial catheter (IRA), into normal rats sequentially. Superoxide dismutase (SOD) infusion by means of IRA, in addition to oxalate, was also performed to check its influence on the altered renal function after oxalate infusion. In both the acute and chronic groups, renal blood flow (RBF), cortical microvascular blood flow (CMVBF), glomerular filtration rate (GFR), urine flow (UV), and urinary sodium excretion (U(Na)V) were measured, and chemiluminescence (CL) was examined in the renal venous blood. RESULTS: Levels of urinary lipid peroxides and enzymuria had increased since day 7, and increased the size of numbers of CaOx crystals in the kidney were noted beginning on day 21, but elevated CL was detectable only on day 7 after 0.75% EG treatment. Decreased UV and U(Na)V were noted in the 42-day EG group, although the 24-hr creatinine clearance values were normal in all experimental groups. On the other hand, RBF, GFR, and CMVBF were attenuated with elevated FR when the oxalate concentration was higher than 10 mg kg(-1) hr(-1) in the acute oxalate infusion group. With SOD pretreatment, the decreased RBF, GFR, and CMVBF could be reversed at 10 mg kg(-1) hr(-1) of oxalate, and be partially reversed at 20. FR also could be reduced significantly at 10 and 20 mg kg(-1) hr(-1) of oxalate. CONCLUSIONS: Decreased urine flow and sodium excretion were the main renal functions affected by chronic hyperoxaluria. However, that only the 42-day EG group had a decreased tubular function cannot be fully explained by the persistent tubular enzymuria and increased lipid peroxides that began on day 7 after EG treatment. With acute oxalate infusion, the major insult to renal function was renal hemodynamics. Pretreated SOD could reverse the attenuated hemodynamics and reduce the elevated FR partly, which suggested that FR is responsible for oxalate toxicity.

Acute Disease↗

Interactions in the Thallium(I) Heptanoate and Heptanoic Acid System: Association, Aggregation, and Phase Behavior

The phase, association, and aggregation behavior of the binary system [TlO2C(CH2)5CH3 + HO2C(CH2)5CH3] have been investigated through temperature and enthalpy vs composition diagrams by determination of the thermal behavior of the pure components and 30 mixtures over an interval of nearly 300 K, and by differential scanning calorimetry and polarized microscopy. Total miscibility was observed in liquid and liquid crystalline phases but essentially total immiscibility among crystalline phases. The phase behavior exhibits: (1) Formation of two intermediate compounds which dissociate in the solid state at low temperature; (2) A eutectic reaction at salt molar fraction x = 0.08 about 3 K below the acid melting point; (3) Unimolecular salt:acid association by very strong hydrogen bonding, which undergoes a crystal/crystal transition prior to incongruent melting at 295.5 K, occurring by a highly energetic peritectic reaction in which the compound evolves more than the sum of the total transition enthalpies of both components; (4) Lyotropic mesophase formation at 390.0 K over the x >/= 0.66 composition range by aggregation of a solid and a liquid phase to yield, on heating, a continuous series of liquid crystalline solutions with mixed-lamellar structure and interlamellar spacings around 20 A as determined by powder X-ray diffractometry, their texture being focal conic, consistent with the neat thermotropic mesophase of the pure salt. A tridimensional phase diagram is presented. All these main reactions have been fully characterized by their nature, stoichiometry, and relevant thermodynamic data.

Journal Article↗

Ternary Phase Equilibria of the 1-Dodecylpyridinium Bromide-Dodecanol-Water System.

The isothermal ternary phase diagram for the 1-dodecylpyridinium bromide (1-DPB)-water-dodecanol system was determined at 40 degreesC, using 2H NMR, polarizing microscopy, and SAXS methods. All of the phases were characterized, and their ranges of existence were determined. The surfactant is easy to dissolve in water, yielding a normal micellar solution phase. After the normal micellar phase, on the binary surfactant-water axis, a normal hexagonal liquid crystalline phase is found at higher surfactant concentrations. On addition of dodecanol, four more phases are formed, i.e. a cubic, a lamellar, and a reverse hexagonal phase, followed by a reverse micellar solution phase. The lamellar liquid crystalline phase dominates the ternary phase diagram. The structures of the liquid crystalline phases were further examined using SAXS measurements, and the results are discussed in terms of the critical packing parameter, cpp, and electrostatic forces. The SAXS experiments show a pronounced swelling of the rods in the hexagonal phase, from 28.5 to 33 Å on addition of dodecanol, whereas the cylindrical aqueous core of the reverse hexagonal phase has a diameter of 18-21 Å, depending on sample composition. The average bilayer thickness of the lamellar phase is about 24 Å. Copyright 1997 Academic Press.

Journal Article↗

Phase Equilibria and Structure of the 1-Dodecylpyridinium Bromide-Dodecane-Water System.

The isothermal ternary phase diagram for the 1-dodecylpyridinium bromide/dodecane/water system was determined at 40 degreesC by 2H NMR and polarizing microscopy methods. Two liquid crystalline phases, a large cubic area and a normal hexagonal phase, and one isotropic normal micellar solution phase were characterized, and their ranges of existence were determined. The micelles were found to be probably small and spherical at lower concentrations of surfactant, and were found to grow at higher concentrations and on addition of oil. The two-phase areas, L1 + HI and HI + I, are both very narrow. The comparatively large cubic area, containing 43-63 wt% surfactant and 3-10 wt% dodecane, is probably consistent of more than one structure. SAXS experiments indicate two different structures built of discrete micellar aggregates. Copyright 1998 Academic Press.

Journal Article↗

Formation and Properties of Lamellar Phases in Systems of Cationic Surfactants and Hydroxy-Naphthoate.

We investigated the phase behavior and the phase transitions in aqueous solutions of 100 mM cetyltrimethylammonium hydroxide (CTAOH) with 3-hydroxy-2-naphthoic acid (HNC) and of 100 mM cetyltrimethylammonium bromide (CTAB) with sodium-3-hydroxy-2-naphthoate (SHNC). The naphthoate/surfactant ratio has been varied. As previously observed by the groups of C. Manohar and J. Candau we observed for the second system two viscoelastic gel-like regions, two liquid crystalline regions, and a precipitate region. For the CTAOH/HNC system one finds with increasing concentration of HNC a low viscous solution, a viscoelastic gel, and a viscoelastic liquid crystalline Lalpha-phase. In both surfactant systems the lamellar phase is formed around an equimolar ratio of cationic surfactant and naphthoate. The lamellar phases have been examined by polarization microscopy and freeze-fracture electron microscopy. The Lalpha-phase in the system CTAOH/HNC consists of densely packed multilamellar vesicles while the lamellar phase in the system CTAB/SHNC contains vesicles, as well as stacked bilayers and tubuli. Corresponding to their different microstructures the lamellar phases in the system, CTAOH/HNC and CTAB/SHNC have different rheological properties. The vesicular phase is highly viscoelastic and has a yield stress value while the bilayer phase has a much lower viscosity and no yield stress value. The transition from the micellar to the vesicle phase occurs for CTAOH/HNC over a two-phase region, where micelles and vesicles coexist. In the case of CTAB/SHNC the transition from the micellar to the lamellar phase occurs over a three-phase region, where a surfactant-poor phase coexists with a lamellar and a coacervate phase. In mixtures of CTAB and SHNC a thick precipitate is formed at an equimolar ratio of CTAB and SHNC. This precipitate consists of condensed multilamellar vesicles that contain little water and stick together, as the vesicles collapse due to the shielding of the repulsive forces by NaBr from an unbinding to a binding state. The precipitate can be retransformed to a swollen lamellar phase by charging the vesicles with an excess of ionic surfactant, by adding electrolyte in high concentrations, or by increasing the temperature. As predicted by C. Manohar et al. the vesicle phases show a phase transition at a critical temperature Tc of 46 degreesC. This transition was detected by us for the first time by DSC and by conductivity measurements. It occurs within a narrow temperature range of 2-3 degrees with an enthalpy change of 0.5 kJ/mol. The transition is observed both in the swollen and in the precipitated vesicle phase. It is well separated from the vesicle/rod transition at higher temperatures (>70 degreesC) and the liquid crystalline/crystalline transition at lower temperatures (25-30 degreesC) that has a melting enthalpy of 55 kJ/mol. It is conceivable that the observed transition at 46 degreesC is due to the melting of a two-dimensional solid-like lattice of the HNC-counterions on the vesicle interface. Copyright 1998 Academic Press.

Journal Article↗

Spermidine-induced aggregation of nucleosome core particles: evidence for multiple liquid crystalline phases.

We investigate the effect of the addition of a trivalent cation, spermidine, to dilute solutions of nucleosome core particles (NCP). In the presence of spermidine, part of the NCP segregates from the initial homogeneous solution, forming dense aggregates. We follow this precipitation process over a wide range of spermidine and NaCl concentrations and determine the conditions of aggregation of the particles. The structure of the dense phases is analyzed by means of polarizing light microscopy and cryo-electron microscopy. We report the existence of multiple supramolecular organizations. According to the relative concentrations of spermidine, monovalent salt and NCP, the particles may aggregate into amorphous phases, stack into randomly oriented columns, or form liquid crystalline phases. Two discotic liquid crystalline phases are identified and analyzed: a columnar nematic corresponding to columns of NCP simply aligned in parallel, and a columnar hexagonal phase in which the columns order into a transversal 2D hexagonal array. We discuss the nature and origin of the interactions possibly involved in the formation and maintenance of these different types of order.

Animals↗

Liquid crystalline ordering of procollagen as a determinant of three-dimensional extracellular matrix architecture.

The precise molecular mechanisms that determine the three-dimensional architectures of tissues remain largely unknown. Within tissues rich in extracellular matrix, collagen fibrils are frequently arranged in a tissue-specific manner, as in certain liquid crystals. For example, the continuous twist between fibrils in compact bone osteons resembles a cholesteric mesophase, while in tendon, the regular, planar undulation, or "crimp", is akin to a precholesteric mesophase. Such analogies suggest that liquid crystalline organisation plays a role in the determination of tissue form, but it is hard to see how insoluble fibrils could spontaneously and specifically rearrange in this way. Collagen molecules, in dilute acid solution, are known to form nematic, precholesteric and cholesteric phases, but the relevance to physiological assembly mechanisms is unclear. In vivo, fibrillar collagens are synthesised in soluble precursor form, procollagens, with terminal propeptide extensions. Here, we show, by polarized light microscopy of highly concentrated (5-30 mg/ml) viscous drops, that procollagen molecules in physiological buffer conditions can also develop long-range nematic and precholesteric liquid crystalline ordering extending over 100 microm(2) domains, while remaining in true solution. These observations suggest the novel concept that supra-fibrillar tissue architecture is determined by the ability of soluble precursor molecules to form liquid crystalline arrays, prior to fibril assembly.

Animals↗

The role of microtubule assembly dynamics in mitotic force generation and functional organization of living cells.

This article summarizes the author's presentation at the Baylor Medical School Symposium on the Biophysics of Microtubules, held April 12 to 14, 1996, in Houston, Texas. It presents a brief historical sketch and discusses the role that assembly/disassembly of microtubules is likely to be playing in force generation for chromosome movement and related organellar positioning in living cells. The article starts out with how polarized light microscopy of living cells had laid the foundation for this concept in the 1950s and 1960s, but was then eclipsed for some 2 decades following the discovery of force generation by microtubule sliding powered by an ATP-hydrolyzing motor protein, dynein. The intriguing recent discoveries: that microtubules undergo dynamic instability; that they both assemble and disassemble right at the kinetochore where they are attached to the chromosome; and that assembling and disassembling microtubules can of themselves push and pull reasonable loads in model experiments, even in the absence of hydrolyzable nucleotides, have refocused serious attention on the probable role played by assembly/disassembly of microtubules. This mode of force generation may well be intricately coupled, and interact, with force-generating and/or dynamic attachment roles played by "motor" proteins, especially at the kinetochore.

Animals↗