Lamellar Units Research Articles

Epidermal Steroid Sulfatase and Cholesterol Sulfotransferase Are Regulated During Late Gestation in the Fetal Rat

Lipids in the stratum corneum (SC) are organized into lamellar membrane unit structures that provide the permeability barrier. Cholesterol sulfate, a SC membrane lipid, is synthesized by cholesterol sulfotransferase (CSTase) in the lower epidermis and hydrolyzed to cholesterol by steroid sulfatase (SSase) in the SC. To determine whether these enzymes are induced during barrier ontogenesis, we examined their activity in epidermis of fetal rats before (gestational day 17), during (day 19), and after (day 21) barrier formation. CSTase activity increased approximately 10-fold between day 17 and day 19, then declined between day 19 and day 21. In contrast, SSase activity reached its peak activity on day 21, increasing >5-fold. Fetal rat skin explants develop a SC and barrier over the same time course in vitro as in utero. Likewise, CSTase and SSase activities during in vitro ontogenesis precisely mirrored those obtained in utero. Moreover, hormones that accelerate barrier ontogenesis (e.g. glucocorticoids, thyroid hormone, and estrogen) accelerated the increase in CSTase and SSase activities during in vitro ontogenesis. mRNA levels of SSase increased in parallel with enzymatic activity, suggesting that these developmental changes are regulated at the genomic level. Finally, addition of exogenous cholesterol sulfate to explants in vitro did not accelerate either SC development or barrier formation. These studies suggest that induction of the cholesterol sulfate cycle enzymes during SC ontogenesis is a component of the fetal epidermal differentiation program and that the synthetic and degradative enzymes of this pathway are differentially regulated.

Rotated plywood structure of primary lamellar bone in the rat: Orientations of the collagen fibril arrays

A basic structural motif of lamellar bone is the arrays of parallel collagen fibrils, with successive arrays having different orientations to form a plywood-like structure. Measurements of the angles between adjacent arrays from cryomicrotomed and vitrified thin sections of demineralized rat bone, cut approximately parallel to the lamellar boundary plane, show that most angles are around 30°, although a subset are around 70°. A structural model for collagen organization based on these measurements is proposed in which an individual lamellar unit (thick and thin lamellae together with transition zones) is composed of five arrays of parallel collagen fibrils, each offset by 30°.

The mechanical properties of fin whale arteries are explained by novel connective tissue designs.

The aortic arch and the descending aorta in the fin whale (Balaenoptera physalus) are structurally and mechanically very different from comparable vessels in other mammals. Although the external diameter of the whale's descending thoracic aorta (approximately 12 cm) is similar to that predicted by scaling relationships for terrestrial mammals, the wall thickness:diameter ratio in the whale (0.015) is much smaller than the characteristic value for other mammals (0.05). In addition, the elastic modulus of the thoracic aorta (12 MPa at 13 kPa blood pressure) is about 30 times higher than in other mammals. In contrast, the whale's aortic arch has a wall thickness/diameter ratio (0.055) and an elastic modulus (0.4 MPa) that are essentially identical to those for other mammals. However, the aortic arch is unusual in that it can be deformed biaxially to very large strains without entering a region of high stiffness caused by the recruitment of fully extended collagen fibres. Chemical composition studies indicate that the elastin:collagen ratio is high in the aortic arch (approximately 2:1) and that this ratio falls in the thoracic (approximately 1:2) and abdominal (approximately 1:3) aortas, but the magnitude of the change in composition does not account for the dramatic difference in mechanical properties. This suggests that there are differences in the elastin and collagen fibre architecture of these vessels. The descending aorta contains dense bands of tendon-like, wavy collagen fibres that run in the plane of the arterial wall, forming a fibre-lattice that runs in parallel to the elastin lamellae and reinforces the wall, making it very stiff. The aortic arch contains a very different collagen fibre-lattice in which fibres appear to have a component of orientation that runs through the thickness of the artery wall. This suggests that the collagen fibres may be arranged in series with elastin-containing elements, a difference in tissue architecture that could account for both the lower stiffness and the extreme extensibility of the whale's aortic arch. Thus, both the structure and the mechanical behaviour of the lamellar units in the aortic arch and aorta of the whale have presumably been modified to produce the unusual mechanical and haemodynamic properties of the whale circulation.

Congenital stenotic arteriopathy with medial dysplasia of aorta, pulmonary artery, and their major branches

This report describes the morphological findings in a young child with congenital stenotic arteriopathy who died suddenly following arteriography. Hyperplasia of all of the medial components had produced severe thickening of the wall of the aorta (mean number of lamellar units = 133 in the thoracic aorta and 125 in the abdominal aorta), the pulmonary artery, and their major proximal branches, resulting in significant luminal narrowing. Bilateral renal artery stenosis, attributable mainly to intimal longitudinal smooth muscle hyperplasia associated with fibroelastosis, was the cause of her systemic hypertension. The left ventricle showed healed subendocardial infarction.

Epidermal Barrier Ontogenesis: Maturation in Serum-Free Media and Acceleration by Glucocorticoids and Thyroid Hormone but Not Selected Growth Factors

Because the cutaneous permeability barrier develops late in gestation, prematurity may result in increased morbidity and mortality due to barrier incompetence. The purpose of the present study was to develop an in vitro model of barrier ontogenesis in order to identify those factors critical for fetal barrier formation. Skin explants from gestational day 17 fetal rats (term is 22 days) were incubated in hormone- and serum-free media. After 4 d in culture, a multi-layered stratum corneum (SC) developed that demonstrated a membrane pattern of fluorescence using the hydrophobic probe, nile red, and the deposition of mature lamellar unit structures throughout the SC interstices, ultrastructurally. Transepidermal water loss rates declined during explant culture such that after 4 d a competent barrier was present. Similarly, lanthanum permeation studies showed tracer penetration into all cell layers in 2-d explants, whereas it did not penetrate above the stratum granulosum in 4-d explants. Thus, the chronology of epidermal development in the explants precisely mirrored that observed in utero. Treatment with either 10 nM dexamethasone or 10 nM triiodothyronine accelerated SC development and barrier formation by 2 d. These results indicate that (i) the late events of fetal epidermal development progress in vitro under serum- and growth factor-free conditions, culminating in the formation of a functional barrier, and (ii) both dexamethasone and triiodothyronine accelerate barrier development.

Stress and strain distribution in hypertensive and normotensive rat aorta considering residual strain.

The effects of hypertension on the stress and strain distributions through the wall thickness were studied in the rat thoracic aorta. Goldblatt hypertension was induced by constricting the left renal artery for 8 weeks. Static pressure-diameter-axial force relations were determined on excised tubular segments. The segments were then sliced into thin ring specimens. Circumferential strain distributions were determined from the cross-sectional shape of the ring specimens observed before and after releasing residual stresses by radial cutting. Stress distributions were calculated using a logarithmic type of strain energy density function. The wall thickness at the systolic blood pressure, P(sys) significantly correlated with P(sys). The mean stress and strain developed by P(sys) in the circumferential direction were not significantly different between the hypertensive and control aortas, while those in the axial direction were significantly smaller in the hypertensive aorta than in the control. The opening angles of the stress free ring specimens correlated well with P(sys). The stress concentration factor in the circumferential direction was almost constant and independent of P(sys) although the stress distributions were not uniform through the wall thickness. Histological observation showed that the wall thickening caused by hypertension is mainly due to the hypertrophy of the lamellar units of the media, especially in the subintimal layer where the stress increase developed by hypertension is larger than in the other layers. These results indicate that: (a) the aortic wall adapts itself to the mechanical field by changing not only the wall dimensions but also the residual stresses, (b) this adaptation is primarily related to the circumferential stress but not to the axial stress, and (c) the aortic smooth muscle cells seem to change their morphology in response to the mechanical stress.

Microstructure and Phase Separation of Pekk, Pekk and their Blends

ABSTRACTMicrostructures of PEEK|PEKK blends at both structural and lamellar levels were studied using simultaneously time-resolved small-angle and wide-angle x-ray scattering (SAXS and WAXS) with synchrotron radiations. Lamellar dimensions and unit cell parameters were determined, and phase separation within the blends were observed. A suitable two-phase growth model is proposed. It is concluded that when crystallized from the melt, PEEK and PEKK lamellae formed in separate lamellar bundles within a common spherulite.

Lyotropic Phases of Dodecylbenzenesulfonates with Different Counterions in Water

The lyotropic phase behavior of (technical grade) dodecylbenzenesulfonates (DoBS) is strongly influenced by the type of counterion and the relative amount of water. Phase diagrams are composed for the following systems: HDoBS/water, NaDoBS/water, (HDoBS + NaDoBS 1:1)/water, LiDoBS/water, KDoBS/water, CsDoBS/water, Ca(DoBS)2/water, NaDoBS/water/NaCl, and NaDoBS/water/CsCl. The phases were characterized by light microscopy, freeze-fracture electron microscopy, X-ray diffraction, and macroscopic appearance. The phase diagrams all contain large areas of lamellar phases. The appearance of the lamellar phases differs along this series, in particular regarding swelling behavior, either with or without a micellar phase next to the lamellar phase, and formation of large, rather irregular lamellar units versus smaller, perfectly spherical lamellar topologies (so-called lamellar droplets). LiDoBS shows, in addition, a hexagonal phase between 25 and 50 wt %. Explanations for the occurrence of the different phases are given in molecular terms and in terms of interactions between bilayers and between aggregates.

Interlamellar Transition Mechanism in Model Membranes

Time-resolved X-ray diffraction was used to monitor structure changes during a partial lamellar liquid crystal (smectic A)/lamellar gel (smectic B) transition in lipid multilayer vesicles in response to small pressure oscillations covering 4 decades in frequency. The data suggest that hydrocarbon chain ordering/disordering is tightly coupled to changes in lamellar unit cell length that occur during the phase change and are consistent with a bidirectional layer-by-layer transition mechanism best described by the Kolmogorov−Avrami kinetic model having an effective dimensionality of 1.

Permeability barrier requirements regulate epidermal beta-glucocerebrosidase.

The intercellular spaces of the outermost layers of the epidermis (stratum corneum, SC) of terrestrial mammals contain a mixture of lipids, enriched in ceramides that are critical for the epidermal permeability barrier. Whereas glucosylceramides (GlcCer) are synthesized in abundance in the epidermis, they disappear coincident with an increase of ceramides (Cer) in the SC. Hence, hydrolysis of GlcCer to Cer by beta-glucocerebrosidase (GlcCer'ase), may be required for permeability barrier homeostasis. We determined first whether modulations in epidermal GlcCer'ase activity and mRNA levels occur in response to barrier disruption; and second, how GlcCer'ase inhibitors influence barrier function and SC membrane ultrastructure. Barrier disruption significantly increased epidermal GlcCer'ase mRNA levels, with a 2.8-fold increase over untreated control levels at 8 h (P < 0.01). GlcCer'ase activity was increased in whole epidermis (34%; P < 0.02) 24 h after barrier disruption. Localization of GlcCer'ase activity showed an increase (33%; P < 0.05) in the outer epidermis (SC and stratum granulosum), without a change in lower epidermal activity (stratum spinosum and stratum basale). Furthermore, a single topical application of the GlcCer'ase inhibitor, bromoconduritol-B-epoxide (BrCBE), inhibited enzyme activity (98%) and significantly delayed permeability barrier recovery after acetone treatment. In addition, BrCBE treatment disrupted SC intercellular lamellar bilayers, without evidence of cellular toxicity. These results indicate that epidermal processing of GlcCer to Cer by GlcCer'ase is required for barrier homeostasis, and that this important enzymatic step is regulated by barrier requirements.

Reversed circulation in acardiac fetuses is associated with anatomic inversions in the aortic wall.

Blood vessel anatomy is probably related to hemodynamic factors which change during development. This principle has been postulated as the basis for differences in the numbers of elastic lamellae in the tunica media of the proximal and distal human aorta. Recent studies of human fetuses at varying stages confirmed that the number of elastin lamellae in the aortic wall varied along its length (spatially) and with age (temporally). These findings suggest that hemodynamic influences during prenatal development induce structural changes in the aortic wall. Acardiac fetuses provide a model for studying the effects of hemodynamic changes on the structure of elastic arteries because blood flow through the aorta in such fetuses is reversed and greatly reduced. Also, analysis of the vascular structure of acardiac fetuses would further define the characteristic features of this congenital disorder. In the present study, we have examined the gross and histological anatomy of the aorta from seven acardiac fetuses. In each case, the microscopic architecture of the aorta was deranged and the normal proximal-distal differences in arterial caliber and number of elastin lamellae were reversed. In the proximal aorta, medial lamellar units were thin, fragmented, and irregular. In some segments, only traces of medial lamellar units existed. We believe that these anatomic abnormalities represent degenerative or dysplastic responses to reversed flow and pressure gradients in the aorta. These findings are consistent with the hypothesis that aortic wall structure is influenced by hemodynamic factors during development.

Glucocorticoids accelerate fetal maturation of the epidermal permeability barrier in the rat.

The cutaneous permeability barrier to systemic water loss is mediated by hydrophobic lipids forming membrane bilayers within the intercellular domains of the stratum corneum (SC). The barrier emerges during day 20 of gestation in the fetal rat and is correlated with increasing SC thickness and increasing SC lipid content, the appearance of well-formed lamellar bodies in the epidermis, and the presence of lamellar unit structures throughout the SC. Because glucocorticoids accelerate lung lamellar body and surfactant maturation in man and experimental animals, these studies were undertaken to determine whether maternal glucocorticoid treatment accelerates maturation of the epidermal lamellar body secretory system. Maternal rats were injected with betamethasone or saline (control) on days 16-18, and pups were delivered prematurely on day 19. Whereas control pups exhibited immature barriers to transepidermal water loss (8.16 +/- 0.52 mg/cm2 per h), glucocorticoid-treated pups exhibited competent barriers (0.74 +/- 0.14 mg/cm2 per h; P < 0.001). Glucocorticoid treatment also: (a) accelerated maturation of lamellar body and SC membrane ultrastructure; (b) increased SC total lipid content twofold; and (c) increased cholesterol and polar ceramide content three- to sixfold. Thus, glucocorticoids accelerate the functional, morphological, and lipid biochemical maturation of the permeability barrier in the fetal rat.

Preservation of Permeability Barrier Ontogenesis in the Intrauterine Growth-Retarded Fetal Rat

The epidermal permeability barrier is provided by intercellular lipids forming multiple membrane bilayers in the stratum corneum. In the fetal rat, the barrier to transepidermal water loss forms during the 20th d of gestation and is accompanied by 1) increasing stratum corneum thickness; 2) increasing stratum corneum lipid content, particularly nonpolar ceramide and cholesterol content; and 3) the formation of lamellar unit structures throughout the stratum corneum interstices. In this report, we demonstrate that among pups of 20 d gestational age increasing barrier competence is correlated with increasing fetal weight. It has been previously demonstrated that fetal rats subjected to intrauterine growth retardation (IUGR) exhibit a thinner stratum corneum and decreased content of differentiation-specific epidermal structural proteins. To determine whether IUGR fetal rats also exhibit immaturity of barrier function and the barrier membrane system, maternal rats underwent unilateral uterine vessel ligation on d 17 or 18 of gestation and IUGR and control littermates were harvested on d 20, 21, or 22 of gestation for determination of transepidermal water loss. Despite significant somatic growth retardation and a thinner stratum corneum, barrier function in IUGR fetal rats did not significantly differ from that in control littermates at any gestational age. In both IUGR and control fetal rat epidermis at 21 d gestational age, lipids were deposited in a membrane pattern as visualized by nile red fluorescence microscopy and formed lamellar unit membrane structures throughout the stratum corneum intercellular domains as observed by electron microscopy.(ABSTRACT TRUNCATED AT 250 WORDS)

Observations on fluorinite and fluorescent vitrinite with the transmission electron microscope

Transmission electron microscopy (TEM) has been used to study fluorinite occurrences in four coals, ranging in rank from soft brown coal to high volatile bituminous. The fluorinite has a substructure of lipoid-rich lamellar and nodular units about 0.1–0.5 μm across. These components, together with enclosing humic matter and (in some cases) thin cuticle, appear to derive from leaves or leaf-like material from vascular plants. Fluorescent vitrinite in one of the coal samples (of bituminous rank) was also studied with TEM. This fluorescence is attributable to exsudatinite-like inclusions of petroleum origin within the humic matter of the vitrinite.

The relationship between wall tension, lamellar thickness, and intercellular junctions in the fetal and adult aorta: Its relevance to the pathology of dissecting aneurysm

It is known that the distribution of stress and strain in the vessel wall is not uniform. We believe that this explains the location of the plane of dissection in dissecting aneurysms of large elastic arteries. We have investigated the effects of non-uniformity of stress and strain on the thickness of each elastic lamella and on the distribution of intercellular junctions in the media of developing and adult rats, to seek evidence to support this hypothesis. Intercellular junctions were identified by transmission electron microscopy of whole wall sections. A morphometric study of elastic tissue distribution was made on an image analysis computer. Differences were analysed using one-way analysis of variance. There are between six and eight elastic lamellae in the aorta of rats. In the fetus, only the internal elastic lamella is complete; the others were not fully formed by term. In the adult, the inner five elastic lamellae were thicker than the remaining two or three, and smooth muscle cells in the thicker lamellar units had more cell-cell contacts of all types examined. These data support the concept of a difference in stress-resisting properties of the aortic wall on the junctions between the inner two-thirds and the outer third of the media. The findings indicate that, as proposed in theoretical models the innermost lamellae support the high tension. In the adult aorta, the structure is modified to enhance the capacity to resist stress in the internal two-thirds of the media.(ABSTRACT TRUNCATED AT 250 WORDS)

Can we predict the long-term function of the subclavian flap angioplasty?

Neonatal operations have improved the prognosis for newborn children with aortic coarctation. The 30-day mortality of 123 neonates with isolated coarctation of the aorta collected from nine series was found to be 0.8%. The subclavian flap angioplasty was the most frequently used surgical procedure in this collected series. This technique is relatively new, however, and many questions have yet to be answered. In this study we have done subclavian flap repair in newborn pigs and followed them up to adult ages. The pigs were killed 28 or 44 weeks postoperatively, and the aortas were reexamined. All flaps had grown symmetrically in width and length and parallel to the growth of the descending thoracic aorta. The flaps were macroscopically intact. Signs of degenerative processes were not found. The wall thickness of the subclavian flap increased by growth of the individual fibroelastic lamellar units in the tunica media. This adaptation to the increased wall stress occurred early in life. The wall strength of the flap also increased by thickening of the intimal layer. We conclude that the subclavian flap is well suited to function as a part of the aorta in adult life.

Biogenesis of myeloid bodies in regenerating newt (Notophthalmus viridescens) retinal pigment epithelium.

Myeloid bodies are believed to be differentiated areas of smooth endoplasmic reticulum membranes, and they are found within the retinal pigment epithelium in a number of lower vertebrates. Previous studies demonstrated a correlation between phagocytosis of outer segment disc membranes and myeloid body numbers in the retinal pigment epithelium of the newt. To test the hypothesis that myeloid bodies are directly involved in outer segment lipid metabolism and to further characterize the origin and functional significance of these organelles, we examined the effects on myeloid bodies of eliminating the source of outer segment membrane lipids (neural retina removal) and of the subsequent return of outer segments (retinal regeneration) in the newt Notophthalmus viridescens. Light- and electron-microscopic analysis demonstrated that myeloid bodies disappeared from the pigment epithelium within six days of neural retina removal. By week 6 of regeneration, rudimentary photoreceptor outer segments were present but myeloid bodies were still absent. However, at this time, the smooth endoplasmic reticulum in some areas of the retinal pigment epithelial cells had become flattened, giving rise to small (0.5 micron long), two-to-four layer-thick lamellar units, which are myeloid body precursors. Small myeloid bodies were first observed one week later at week 7 of retinal regeneration. This study revealed that newt myeloid bodies are specialized areas of smooth endoplasmic reticulum. It also showed that a contact between functional photoreceptors and the retinal pigment epithelium is essential to the presence of myeloid bodies in the epithelial cells.

Ontogeny of the epidermal barrier to water loss in the rat: correlation of function with stratum corneum structure and lipid content.

The mammalian epidermal permeability barrier is provided by highly hydrophobic lipids forming multiple membrane bilayers within the extracellular domains of the outer, cornified cell layers. To characterize the critical events associated with barrier maturation, we correlated the emergence of a competent barrier to transepidermal water loss with development of the lamellar body secretory system, the organization of stratum corneum membrane bilayers, and the lipid composition of these membranes in the perinatal rat. Whereas pups of 19 d estimated gestational age had no measurable barrier (transepidermal water loss greater than 10 mg/cm2/h), by 21 d the barrier was well established (mean transepidermal water loss 0.41 mg/cm2/h). Development of a functional barrier correlated with increasing thickness of the stratum corneum, as well as with development of a membrane pattern of lipid deposition, visualized with the hydrophobic fluorescent probe nile red. At 19 d estimated gestational age, the stratum corneum intercellular domains exhibited an abundance of secreted lamellar body contents, but they were not organized into basic bilayer unit structures. Lamellar unit structures became evident by 20 d and extended throughout the stratum corneum interstices by 22 d (term). The quantity of lipid in isolated stratum corneum increased significantly between 19 and 20 d (34.08 versus 50.08 mean micrograms lipid/cm2, respectively; p less than 0.02) and still further between 20 and 21 d estimated gestational age (74.49 micrograms lipid/cm2; p less than 0.001). This increase was due to progressive accumulation of neutral lipids, particularly cholesterol, as well as nonpolar ceramides, as shown by thin-layer chromatography/scanning densitometry. These studies imply that in the development of cutaneous barrier function in the fetal rat both the generation of sufficient quantities of hydrophobic lipids and the organization of these lipids into bilayer unit structures are required.

Lamellar Body Secretory Response to Barrier Disruption

Abundant evidence points to an important role for epidermal lamellar body secretion in permeability-barrier maintenance. However, the response of the lamellar body secretory system to barrier disruption has not been examined. Hence, we examined the lamellar body secretory response at various points after acetone-induced barrier abrogation in hairless mice in air-exposed animals and those occluded with impermeable versus vapor-permeable membranes. Tape-stripped animals served as a control for chemical toxicity. Barrier perturbation with either acetone or tape stripping was followed by rapid secretion of lamellar body contents from the uppermost granular cell layer, leaving the cytosol largely devoid of lamellar bodies. The newly secreted lamellar body contents comprised pleated sheets (not "discs," as previously thought), which unfurled in the intercellular spaces at the granular-cornified cell interface. At this time (15-30 min), the basic unit structure of the lamellar bilayers in the mid-to-upper stratum corneum appeared disorganized and interspersed with large lacunae, reflecting solvent extraction. Nascent lamellar bodies began to reappear in the granular cell cytosol by 30 min and by 360 min the cells displayed a full complement of normal-appearing lamellar bodies. Between 60 and 360 min, the density of lamellar body sheets at the granular-cornified cell interface increased, whereas the membrane bilayers of the outer stratum corneum remained disorganized. New lamellar bilayer units first appeared in the lower stratum corneum between 60 and 180 min, as a result of the transformation of secreted lamellar body sheets and over time these lamellae appeared at more apical locations. Occlusion with a water vapor-impermeable but not a vapor-permeable membrane resulted in a) decreased quantities of lamellar bodies and lamellar body-derived intercellular products; b) formation of lamellar bodies with abnormal internal contents; c) inhibition of lamellar body secretion; and d) inhibition of transformation of lamellar body-derived sheets into lamellar bilayer units. These results demonstrate the central role of the lamellar body-secretory system in barrier repair and homeostasis.

Watanabe Hyperlipidemic Rabbit as a Model of Aortic Degeneration of the Medial Lamellar Elastin Unit

At age 3 years, WHHL rabbits are near the end of their lifespan, frequently dying from the progression of their hyperlipidemic disease from events such as myocardial infarction. Out of a colony of 20 three-year-old WHHL rabbits raised as part of a NIH breeding project, 2 rabbits actually died of a ruptured thoracic aortic aneurysm. The need for a model to study abdominal aortic aneurysm formation led us to explore further the abdominal aortic pathology in aged WHHL rabbits. Six rabbit abdominal aortas from 3-year-old WHHL rabbits were preserved in formalin, sectioned, and stained for elastin. These were compared to the same sections of six normolipidemic age matched New Zealand white (NZW) rabbits. There was significant (P less than or equal to .001) destruction of the medial lamellar elastin unit in the aorta of the WHHL rabbits compared with the control NZW rabbits. Severe cholesterol deposits appeared to destroy the medial lamellae from the inside out. No definite aneurysm formation was seen in the abdominal aorta despite the significant changes in the medial lamellar elastin units. Thus, this model could be used to study the elastin degeneration of the media, but not necessarily abdominal aortic aneurysm formation.