Simulating the wrinkling and aging of skin with a multi-layer finite element model

One of the outward signs of the aging process of human skin is the increased appearance of wrinkles on its surface. Clinical studies show that the increased frequency of wrinkles with age may be attributed to changes in the composition of the various layers of skin, leading to a change in mechanical properties. A parameter study was performed on a previously proposed multi-layer finite element model of skin. A region of skin was subject to an in-plane compression, resulting in wrinkling. A number of physical properties of the skin model were changed and the effects these changes had on the size of the subsequent wrinkles were measured. Reducing the moisture content of the stratum corneum by 11% produces wrinkles 25–85% larger. Increasing the dermal collagen fibre density by 67%, results in wrinkles, which are 25–50% larger. A reduction and change in the pre-stress distribution in the skin model, which represents the natural tension and relaxed skin tension lines in real skin, also influences the wrinkle height in a similar manner to real aging skin. Typically, there can be up to a 100% increase in the height of wrinkles as skin ages. This model would be of benefit in the development of cosmetic moisturisers and plastic-surgery techniques to reduce the appearance of aging.     

The surface structure of spores and aerial hyphae in Streptomyces viridochromogenes

Summary The fine structure of the surface membrane which covers the cell walls of spores and aerial hyphae of S. viridochromogenes was investigated by means of electron microscopy of air-dried whole mounts, thin sections, negative stainings and freeze-etchings. The delicate sheath of the spores, which is responsible for the surface features, exhibits conspicuous cone shaped protrusions called spines. They are composed of 5–12 rodlike units, each apparently consisting of a bundle of fine fibrils. Non-sporulating hyphae have no spines; their surface membrane shows an irregular pattern consisting of long tubules.     

Internal ultrastructure of spores of microsporidans isolated from the Silkworm,Bombyx mori

Nosema bombycis, two Nosema spp., and a Pleistophora sp. were propagated in the silkworm and the fine structures of their spores were studied. The morphology of the polaroplast, the appearance of the nucleus, and the number of coils in the polar filament differed among the spores of the four species. The spores of the three Nosema species, however, had several identical components; e.g., the polaroplast was made up of two parts, they had two nuclei, and the ribosome arrangement was similar. On the other hand, the spore of Pleistophora sp. had a polaroplast composed of three parts, a single nucleus, and ribosomes arranged around the polar filament. Thus the fine structures of the spore differentiate microsporidan species.     

Scanning Electron and Phase-Contrast Microscopy of Bacterial Spores

The three-dimensional immages of free and intrasporangial spores produced by scanning electron microscopy show surface structures not visible by phase-contrast microscopy. Although fine surface detail is not elucidated by scanning electron microscopy, this technique does afford a definitive picture of the general shape of spores. Spores of Bacillus popilliae, B. lentimorbus, B. thuringiensis, B. alvei, B. cereus, and Sarcina ureae have varying patterns of surface ridge formation, whereas spores of B. larvae, B. subtilis, and B. licheniformis have relatively smooth surfaces.     

Examination of the cyst of Malamoeba locustae and of the spore of Mattesia dispora with the scanning microscope

The fine structure of the surface of cysts of the amoeba Malamoeba locustae and that of spores of the neogregarine Mattesia dispora have been examined with the scanning microscope. Freeze-drying was found to be the most convenient of the three methods tested for the preparation of the specimens. The surface of both mature and immature cysts of M. locustae is smooth, without a microrelief. By contrast, spores of M. dispora are equipped with a small, circular structure at the poles indicating the site of the future pore through which the sporozoites will emerge in the intestine of the new host. In addition, a flat relief forming small depressions is discernable on the surface of these spores at a high magnification.     

Why Some Women Look Young for Their Age

The desire of many to look young for their age has led to the establishment of a large cosmetics industry. However, the features of appearance that primarily determine how old women look for their age and whether genetic or environmental factors predominately influence such features are largely unknown. We studied the facial appearance of 102 pairs of female Danish twins aged 59 to 81 as well as 162 British females aged 45 to 75. Skin wrinkling, hair graying and lip height were significantly and independently associated with how old the women looked for their age. The appearance of facial sun-damage was also found to be significantly correlated to how old women look for their age and was primarily due to its commonality with the appearance of skin wrinkles. There was also considerable variation in the perceived age data that was unaccounted for. Composite facial images created from women who looked young or old for their age indicated that the structure of subcutaneous tissue was partly responsible. Heritability analyses of the appearance features revealed that perceived age, pigmented age spots, skin wrinkles and the appearance of sun-damage were influenced more or less equally by genetic and environmental factors. Hair graying, recession of hair from the forehead and lip height were influenced mainly by genetic factors whereas environmental factors influenced hair thinning. These findings indicate that women who look young for their age have large lips, avoid sun-exposure and possess genetic factors that protect against the development of gray hair and skin wrinkles. The findings also demonstrate that perceived age is a better biomarker of skin, hair and facial aging than chronological age.     

Cytology of Spore Formation in Clostridium perfringens

The sequential morphological events in spore formation by Clostridium perfringens type D were observed in Ellner's medium where 80 to 100% of the cells formed spores. Gross structural changes were studied with the light microscope under phase-contrast, and in fixed cells by the use of both nigrosin and Giemsa preparations. Fine structure was examined with the electron microscope in both thin sections and frozen-etched preparations. During the first 3 hr of incubation, the original rod-shaped cells became ellipsoid to ovoid in shape; by 5 to 6 hr, subterminal spores had developed within these enlarged cells. The fine structural sequence was in most respects identical to that in other Bacillaceae, although some stages were illustrated with particular clarity. A unique feature was the development of a convoluted, membranous exosporium which adhered to the outer surface of the two coats and had an unusual fine structure resembling a rectangular array of subunits.     

毛子蕨孢子形态的研究

利用光镜和扫描电镜对毛子蕨的孢子进行了详细研究,对该种孢子的生长重要形态特征,有了一些新的认识和发现。     

Scanning electron microscopy of sporulating cultures of the myxomycete Physarum polycephalum

Scanning electron microscopy of sporulating cultures of Physarum polycephalum shows in detail the fine structure of the stalk, peridium, capillitia and spores. The peridium has a wrinkled structure and funnel-like cavities (5-10 μ in diameter) which are the capillitia openings. The capillitia are a network of tubes with differing diameters. The capillitia are covered with fine granules, and contain various amounts of bead-like granules of up to 2 μ in diameter. The spherical spores (10 μ in diameter) are covered with fine granules similar to those covering the capillitia.     

Theory of deformable substrates for cell motility studies.

Linear theory is used to relate the tractions F applied by a cell to the resulting deformation of fluid, viscoelastic, or solid substrates. The theory is used to fit data in which the motion of a fluid surface in the neighborhood of a motile keratocyte is visualized with the aid of embedded beads. The data are best fit by modeling the surface layer as a two-dimensional, nearly incompressible fluid. The data favor this model over another plausible model, the planar free boundary of a three-dimensional fluid. In the resulting diagrams for the distribution of F, it is found that both curl F and div F are concentrated in the lateral extrema of the lamellipodium. In a second investigation, a nonlinear theory of weak wrinkles in a solid substrate is proposed. The in-plane stress tensor plays the role of a metric. Compression wrinkles are found in regions where this metric is negative definite. Tension wrinkles arise, in linear approximation, at points on the boundary between positive definite and indefinite regions, and are conjectured to be stabilized by nonlinear effects. Data for the wrinkles that would be produced by keratocyte traction are computed, and these agree qualitatively with observed keratocyte wrinkles.     

Adhesion and development of the root rot fungus (Heterobasidion annosum) on conifer tissues: effects of spore and host surface constituents

The objective of this study was to correlate the occurrence of particular root and woody stump surface components with the ability of spores of the root rot fungus (Heterobasidion annosum) to adhere, germinate and establish on conifer tissues. With the aid of high performance liquid chromatography, several sugars (pinitol, xylitol, dulcitol, mannitol, D-glucose, mannose, fructose) were detected on both stump and fine root surfaces of Scots pine and Norway spruce. Of all the sugars observed, xylose and arabinose were poorly utilized for initiation of germ tube growth whereas spore germination was enhanced in the presence of D-glucose, mannose or fructose. Oxidation of these sugars by pretreatment of wood discs or roots with periodic acid abolished the ability of the spores to germinate. Non-sugar components such as long chain fatty acids on spores and root surfaces as detected with nuclear magnetic resonance were found to have a significant influence on adhesion and initiation of germ tube development. Removal of these aliphatic compounds from the root surface increased spore germination by 2-fold, whereas similar treatment on spores led to a 5-fold decrease in adhesiveness to root material. In vitro studies revealed that the di-ethyl ether extract from the roots had no long term adverse effect on spore germination which suggests that the fungus may possess the capability to detoxify this substance. Similarly, adhesion of spores was affected by low and freezing temperatures. The role of significant levels of mannitol and trehalose accumulated in spores and hyphae of the fungi on viability, survival and tolerance to adverse conditions such as oxidative stress, freezing and desiccation are discussed.     

Lower Devonian coalified sporangia from Shropshire: Salopella Edwards & Richardson and Tortilicaulis Edwards

A diverse assemblage of coalified sporangia from Lochkovian/Gedinnian, fluvial rocks imicrornatus-newportensis Spore Biozone) contains fusiform forms assignable to Salopella Edwards & Richardson and Tortilicaulis Edwards. Both show bifurcation within some sporangia. In specimens assigned to Salopella cf. marcensis , longitudinal dehiscence produces two equal valves revealing spores resembling Aneurospora , although trilete marks may be difficult to distinguish. The spores are in two forms with distal ornament of either coarse or fine coni, each occurring in separate sporangia. A further sporangium of Salopella shape contains distally apiculate, hilate cryptospores. Elongate sporangia exhibiting spiralling of superficial cells and sometimes gross twisting, are placed in a new species of Tortilicaulii, T. offaeus. Anatomical data include details of irregularly thickened epidermal cells, sporangial wall and in situ spores. The latter are trilete, equatorially thickened and highly distinctive, because the entire exospore surface is covered with grana, a feature not common in dispersed spores of this age. The nomenclatural and curatorial problems associated with these small, coalified fossils, whose anatomical examination by scanning electron microscope necessitates elimination of gross morphology, are discussed.     

Microcycle conidiation in Penicillium urticae: an ultrastructural investigation of spherical spore growth.

A cultivation system has been developed for Penicillium urticae (NRRL 2159A) which yields 'microcycle' conidiation in submerged culture. Spherical growth of spores was initiated by incubation at 37 degrees C in a growth-favoring medium. Transfer of these enlarged spores to a nitrogen-poor medium at 35 degrees C resulted in synchronous germination and limited outgrowth followed by roughly synchronous conidiation. An ultrastructural study of the spherical growth stage indicates a significant loss of cell envelope fine structure and a general increase in the number of cellular organelles. Loss of the complex pattern of rodlets on the spore surface, and the trench-like invaginations of the plasma membrane were most prominent. From an ultrastructural point of view these large spores (about 6 mum in diameter) appeared to be perfectly normal.     

Comparison of coats and surface-dependent properties of Bacillus cereus T prepared in two sporulation environments

The surface or coat-associated properties of Bacillus cereus T spores produced from modified G medium (MGM) and fortified nutrient agar (FNA) were compared. The two populations appeared structurally similar by transmission electron microscopy. Spores prepared on FNA were more susceptible to ozone inactivation than MGM-prepared spores. When activated by heating for 15 min at 70–85°C, FNA-prepared spores were optimally activated at 85°C and did not become hydrophilic on heat activation while MGM spores were optimally activated at 70°C and became hydrophilic on activation. Susceptibility to removal of coat and outer membrane by chemical and enzymatic extraction treatments was measured by monitoring reduced ability to germinate in nutrients and acquired ability to germinate in the presence of lysozyme. Bacillus cereus T MGM-prepared spores germinated in lysozyme upon<1 h exposure to sodium dodecyl sulphate-dithiothreitol. FNA-prepared spores were lysozyme sensitive after > 2 h treatment. Thus, B. cereus T FNA spore coats and outer membranes were more resistant to these denaturing agents. Transmission electron micrographs revealed no change in appearance of extracted spores. Sporulation environment must be considered when laboratory-prepared spores are used to assess or predict the effect of control procedures on spores present in nature.     

Synthesis of a spore-specific surface antigen during sporulation of Saccharomyces cerevisiae

Antisera raised against purified yeast ascospores caused agglutination of both ascospores and vegetative cells. A spore-specific activity was obtained by absorbing out anti-vegetative activity with vegetative cells. The anti-vegetative cell activity was directed against mannan, and was probably due to exposure of some spore coat mannan at the spore surface since concanavalin A and lentil lectin also caused agglutination of ascospores. The spore-specific activity was probably determined by a protein or proteins, since extraction of spores with a mixture of sodium dodecyl sulphate and dithiothreitol markedly affected their agglutination by the spore-specific serum. The spore-specific antigen was synthesized in a soluble form during sporulation several hours before the appearance of the spore surface and the pool of soluble antigen declined as the spore was assembled. Synthesis of the soluble antigen was inhibited by adding cycloheximide at all times up to its first appearance in the sporulating cell.     

Cytoskeletal alterations in Dictyostelium induced by expression of human cdc42

The rho family of small G proteins has been shown to be involved in controlling actin filament dynamics in cells. To evaluate the functional overlap between human and Dictyostelium G proteins, we conditionally expressed constitutively active human cdc42 (V12-cdc42) in Dictyostelium cells. Upon induction, cells adopted a unique morphology: a flattened shape with wrinkles running from the cell edge toward the center. The appearance of these wrinkles is highly dynamic so that the cells cycle between the wrinkled and relatively normal morphologies. Phalloidin staining indicates that the stellate wrinkles contain dense actin structures and also that numerous filopods project vertically from the center of these cells. Consistent with the hypothesis that cdc42 induces actin polymerization in vivo, cells expressing V12-cdc42 show an increase in the amount of F-actin associated with the cytoskeleton. This is accompanied by an increase in the association of the actin-binding proteins 34-kDa bundler, ABP-120 and alpha-actinin with the cytoskeleton. In conclusion, human cdc42 has various effects on the Dictyostelium actin cytoskeleton consistent with a conserved role of small GTPases in control of the cytoskeleton.     

The measurement of Bacillus mycoides spore adhesion using atomic force microscopy,simple counting methods,and a spinning disk technique

An atomic force microscope has been used to study the adhesion of Bacillus mycoides spores to a hydrophilic glass surface and a hydrophobic-coated glass surface. AFM images of spores attached to the hydrophobic-coated mica surface allowed the measurement of spore dimensions in an aqueous environment without desiccation. The spore exosporium was observed to be flexible and to promote the adhesion of the spore by increasing the area of spore contact with the surface. Results from counting procedures using light microscopy matched the density of spores observed on the hydrophobic-coated glass surface with AFM. However, no spores were observed on the hydrophilic glass surface with AFM, a consequence of the weaker adhesion of the spores at this surface. AFM was also used to quantify directly the interactions of B. mycoides spores at the two surfaces in an aqueous environment. The measurements used "spore probes" constructed by immobilizing a single spore at the apex of a tipless AFM cantilever. The data showed that stretching and sequential bond breaking occurred as the spores were retracted from the hydrophilic glass surface. The greatest spore adhesion was measured at the hydrophobic-coated glass surface. An attractive force on the spores was measured as the spores approached the hydrophobic-coated surface. At the hydrophilic glass surface, only repulsive forces were measured during the approach of the spores. The AFM force measurements were in qualitative agreement with the results of a hydrodynamic shear adhesion assay that used a spinning disk technique. Quantitatively, AFM measurements of adhesive force were up to 4 x 10(3) times larger than the estimates made using the spinning disk data. This is a consequence of the different types of forces applied to the spore in the different adhesion assays. AFM has provided some unique insights into the interactions of spores with surfaces. No other instrument can make such direct measurements for single microbiological cells.     

Synthesis of teichoic acids. VII. Synthesis of teichoic acids during spore germination

The synthesis of teichoic acids has been examined during germination in Bacillus licheniformis ATCC 9945 and in B. subtilis W-23. Teichoic acids are absent from the spores of both organisms. B. licheniformis spores lack the enzymes responsible for teichoic acid synthesis. The appearance of these enzymes during germination is correlated with the appearance of teichoic acids in the cell. The appearance of teichoic acid-synthesizing enzymes and of teichoic acids in the cell are inhibited by the addition of chloramphenicol to the germination medium. In B. subtilis W-23 the situation is similar for the synthesis of polyribitolphosphate. The synthesis of glucosyl polyribitolphosphate is only partially inhibited by chloramphenicol, puromycin, and penicillin, and uridine diphosphate-d-glucose polyribitol-phosphate glucosyl transferase can be demonstrated in spores. The possible implications of some of these observations are discussed.     

SPORE WALL ULTRASTRUCTURE IN FOUR SPECIES OF THE LIVERWORT RICCIA

Mature spores of Riccia californica, R. campbelliana, R. sorocarpa, and R. trichocarpa (Marchantiales, Ricciaceae) were examined with both scanning and transmission electron microscopes. The thick-walled spores have species-specific surface sculpturing and similar internal structure. Major surface features include ridges and depressions in areolate or irregular configurations. Finer surface features such as tubercles or papillae may ornament the ridges or depressions. Internally, the sporoderm consists of a thin intine and a three-layered, lamellate exine. The innermost exine lamellae are thin and closely spaced, giving this layer a fibrillar appearance. Lamellae of the middle region are thick in section and have a relatively thick, electron-dense coating. The outermost lamellae are of intermediate thickness and also have a relatively thick electron-dense coating. Lamellae of all regions frequently split, branch and anastomose. The outermost lamella forms the surface features of the spore. Interior lamellae follow the contour of the outermost lamella. Pores usually occur in these four species at the junctures of the triradiate ridge arms and equatorial rim. Pores do not penetrate the entire exine, and they represent localized areas where the outermost group of lamellae have not been formed.