Reconstruction of stratum corneum in organotypically cultured canine keratinocyte-derived CPEK cells

The stratum corneum of epidermis is an essential barrier against the external environment and water loss. This study aimed to develop an organotypic culture model that targets the reconstruction of the stratum corneum using canine keratinocyte-derived CPEK cells. The CPEK cells cultured at the air-liquid interface became stratified and formed a stratum corneum-like layer on stratum spinosum- and stratum granulosum-like layers. The CPEK cells in the stratum granulosum-like layer expressed the cornified cell envelope (CCE)-related proteins loricrin and keratinocyte differentiation-associated protein. Organotypically cultured CPEK cells were considered to form a CCE at the stratum granulosum-like layer, allowing the formation of a stratum corneum-like layer. The organotypic culture of CPEK cells could be useful for studying the barrier function of canine stratum corneum.     

Micromorphology of the skin (epidermis, dermis, subcutis) of the dog

Grossly, the skin of the dog is characterized by varying thicknesses, density of the hair coat and the implantation angle of the hairs. Multiple folds varying in shape are formed by the epidermis and the papillary layer of the corium. The epidermis consists of a stratum profundum with a continuous layer of basal cells, a stratum spinosum consisting of 3-6 cell layers, a stratum superficiale with single granulosa and "lucidum" cells and a well-developed stratum corneum. The corium shows a narrow stratum papillare corli containing delicate fibres and a broad stratum reticulare corii with mainly coarse fibre bundles. The subcutis is divided into adipose and fibrous layers. The broad adipose layer displays breed and individual differences which vary with the region of the body. The fibrous layer corresponds to the connective tissue layer, formerly described as fascia trunci superficialis. The morphological and functional unity of the corium and subcutis is discussed.     

Dermatophilus-like infection in beluga whales, Delphinapterus leucas, from the St. Lawrence estuary

Six beluga whales ( Delphinapterus leucas ) found dead on the shores of the St. Lawrence estuary had multiple slightly depressed greyish round areas randomly distributed over the whole body. Histologically, the surface of these areas was covered with a thick layer of Dermatophilus -like organisms which invaded the stratum corneum. The underlying stratum spinosum had marked spongiosis and vacuolar degeneration. Minimal neutrophilic infiltration was present within the underlying dermal papillae. To the authors' knowledge, dermatophilosis in cetaceans has not been reported previously.     

Parapoxvirus infection in harbor seals (Phoca vitulina) from the German North Sea

In the summer of 2000, proliferative lesions of the skin and oral mucosa were observed in 26 young harbor seals (Phoca vitulina) from a rehabilitation center in Schleswig-Holstein, Germany. Verrucose, roundish nodules, approximately 1-2 cm in diameter, were presented in the oral cavity, especially on the tongue. Some animals developed similarly sized spherical dermal elevations with ulceration on flippers, chest, neck, and perineum. Necropsy of one animal showed multifocal, verrucose nodules in the oral cavity and a mild tonsillitis. Histologically, the nodules were characterized by ballooning degeneration of the outer parts of the spiny layer and stratum granulosum, with large eosinophilic cytoplasmic inclusions and a perivascular to interstitial lymphohistiocytic infiltration accompanied by fibroblastic proliferation and neovascularization. Negative staining of mucosal tissue homogenates demonstrated parapoxvirus-like particles. The presence of parapoxvirus was confirmed by polymerase chain reaction, using primers specific for parapoxvirus of ungulates. By in situ hybridization, using a parapox-specific, digoxigenin-labeled DNA probe, abundant parapoxvirus DNA-positive epithelial cells were detected in the stratum granulosum and the outer parts of the spiny layer. There was no parapoxvirus-positive signal in the adjacent submucosa. Although DNA analysis revealed that the causative agent can clearly be distinct from terrestrial parapoxviruses, lesions resembled parapoxvirus infections in other terrestrial species, and the pattern of virus DNA distribution indicated a direct effect of the virus on keratinocytes. In contrast, changes in the corium may be considered an indirect response mediated by the virus or the immune system.     

Effect of 7-nitroindazole, a selective neuronal nitric oxide synthase inhibitor, on parvalbumin immunoreactivity after cerebral ischaemia in the hippocampus of the Mongolian gerbil

Previous studies have demonstrated that a loss of parvalbumin-immunoreactive (PV-ir) neurones is observed in the hippocampus after transient cerebral ischaemia. However, whether the loss of parvalbumin (PV) immunoreactivity is related to the over-production of nitric oxide (NO) during cerebral ischaemia has not been evaluated. This study was designed to test the effect of 7-nitroindazole pre-treatment (7-NI, 50 mg/kg), a selective neuronal NO synthase inhibitor, on PV immunoreactivity and its cellular activity following forebrain ischaemia. PV-ir neurones in the hippocampus of the control group were widely distributed in the pyramidal cell layer and stratum oriens of CA1 and CA3, and the granular cell layer of dentate gyrus. 7-NI pre-treatment completely suppressed the reduction of PV immunoreactivity in CA1 that was observed in the ischaemia-induced group. Subsequently, 7-NI pre-treatment also protected against the structural loss of microtubule-associated protein 2 (MAP2) immunoreactivity in CA1 after ischaemic insult. In addition, the Fos-defined neuronal activity of PV-ir neurones was slightly increased by the 7-NI pre-treatment 3 h after ischaemia. Based on these data, we conclude that the neuronal toxicity of NO may be involved in the loss of PV-ir neurones after cerebral ischaemia.     

Histochemical localization of adenosine triphosphatase activity in bovine ruminal epithelium.

A fine-structural histochemical technique was used to localize magnesium-dependent adenosine triphosphatase (Mg-ATPase) activity in ruminal mucosa. Precipitate appeared on the cytoplasmic surface of the plasmalemma in cells of the upper stratum spinosum, the stratum granulosum, and the deepest layer of the stratum corneum. This ATPase activity was sensitive to glutaraldehyde fixation and possibly to ouabain, but was unaffected by sodium and potassium. The preponderance of Mg-ATPase activity in bovine ruminal epithelium may make it impossible to detect sodium-potassium-activated adenosine triphosphatase ((Na + K)-ATPase) activity histochemically. A Mg-ATPase activity also occurred in mitochondria of the stratum spinosum and stratum granulosum. None of the ruminal sections hydrolyzed adenosine diphosphate, inosine triphosphate, or beta-glycerophosphate when these compounds were used as substitute substrates for adenosine triphosphate. When adenosine-5'-monophosphate was the available substrate, a reaction product appeared in the same layers as Mg-ATPase activity, but the reaction product was confined to the intercellular space.     

Changes in the microscopic anatomy of the bovine teat canal during mammary involution

Changes in the microscopic anatomy of the bovine teat canal were examined during mammary involution. Morphometric analyses revealed a significant (P less than 0.05), temporary dilatation of the teat canal lumen on day 7 of the nonlactating period. Additionally, the teat canal epithelium physiologically atrophied as evidenced by decreased cross-sectional area and thickness during the first 30 days of the nonlactating period, significantly so (P less than 0.05) between days 0 and 7. This physiologic atrophy was due mainly to a reduction in area and thickness of the stratum granulosum and may have resulted from continuing keratinization, a process that led to increased thickness of the keratin layer and formation of a functional plug during later stages of involution. Changes in cells of the stratum granulosum indicated a decrease in the rate of epithelial cell maturation during involution. The mitotic index (percentage of basal cells in mitosis) of the teat canal epithelium decreased significantly (P less than 0.05) between days 0 and 7 of the nonlactating period. Bacteria, observed in histologic sections, appeared to colonize only certain regions of the keratin layer. Seemingly, changes in the teat canal during mammary involution may be important factors in changing susceptibility to new intramammary infection during the early and mid-nonlactating periods.     

Microscopic studies of the skin of the Turkish Angora goat. I. Skin layers

The study describes basic structural aspects of the integumental layers (epidermis, dermis, hypodermis) of the Turkish Angora goat. The results have been obtained by light- and electronmicroscopical methods. Central topics are measurements of layer thickness as related to annual season and life period, as well as typical criteria of integumental layer construction as connected with the hair cycle. Remarkable is a trimerous organization of the dermis and cyclic changes in the thickness of the stratum adiposum of the hypodermis. The findings are discussed with regard to comparable observations from other domestic mammals.     

Alteration in the apoptosis process of rat esophageal epithelium with hyperproliferation of indigenous bacteria under a physiological condition

The apoptosis process in rat esophageal epithelium was investigated using enzyme-immunohistochemistry and transmission electron microscopy. As a result, Fas and Fas-L were expressed in the epithelial cell membrane and cytoplasm from the stratum spinosum (SS) to the stratum granulosum (SG). No TNF-R1 show immunopositivity in the cell membranes. TNF-α and caspase-8 were not observed in any layer. Caspase-10, cleaved caspase-3, XIAP and DNase-1 were found in the epithelial cytoplasm from the SS to the SG, whereas Bid, Apaf-1 and cleaved caspase-9 were detected only in the SG. Cytochrome c was observed as cytoplasmic granular positivity from the stratum basale (SB) and altered into homogeneous immunopositivity in the SG. Bcl-2 and Bcl-X immunopositivity was detected in cytoplasm from the SB to the SG. Immunoreactions of Bak in the cytoplasm and Bax beneath the cell membrane were observed from the upper portion of the SS with increasing intensity toward the SG. In the sites with the hyperproliferation of indigenous bacteria, TNF-R1, TNF-α and caspase-8 were detected in the SG and the immunopositive intensities of Bid, Apaf-1 and cleaved caspase-9 were altered to be strong. Prominently swollen cells and decreased mitochondria were ultrastructurally confirmed in the uppermost layers of stratum corneum. These findings suggest that the Fas-Fas-L-interaction initially induces apoptosis through a mitochondria-independent pathway and secondarily through a mitochondria-dependent pathway, leading to eventual epithelial cell death in the rat esophageal epithelium. The bacterial stimuli probably enhance the mitochondria-dependent pathway through the TNF-R1-TNF-α interaction.     

Structure of the epidermis in Ayrshire bullocks.

The living epidermis and stratum corneum of cattle, when measured on cryostat sections, were both found to be approximately 30 micrometer thick. However, the corneum contained about 30 cell layers, whereas the living epidermis was composed of only four. Histological processing involving fixation, dehydration and paraffin embedding, resulted in the loss of half of the stratum corneum and proved to be unsuitable for study of this layer. Lipid derived mainly from the sebaceous gland was present between the squames in the outer two thirds of the corneum probably in the form of an emulsion with sweat.     

Morpho- und Histogenese der Colliculi rostrales beim Rind

Based upon macroscopic and light microscopic examinations the development of the superior colliculus of bovine embryos and fetuses with crown-rump lengths (CRL) ranging from 0.8 to 90.0 cm was studied. Macroscopically, the mesencephalon can be recognized for the first time at 0.8 cm CRL, whereas the superior colliculus can be clearly differentiated in embryos of 4.5 cm CRL. The macroscopic features of this brain area have reached adult conditions at 80.0 cm CRL. The light microscopic examination reflected the beginning of layer formation at 0.8 cm CRL, which is induced by the proliferating activity of the ventricular zone. Up to 3.4 cm CRL the primordium of the tectum opticum exhibits still a trilaminate pattern (ventricular-, intermediate- and marginal zone), but at 4.5 cm CRL, the formation of the specific tectal layers is marked by the origin of the stratum profundum and intermedium. With the appearance of the tenth layer named stratum opticum at 8.0 cm CRL the laminar pattern corresponds to the characteristics of adult animals.     

In vitro development and characterization of canine epidermis on a porcine acellular dermal matrix

The aim of this study was to develop and to characterize a canine skin epidermal model able to form a proper epidermis on a porcine acellular dermal matrix (PADM). In addition, the role of fibroblasts in skin barrier formation was studied by incorporating or omitting canine dermal fibroblasts in the PADM. Canine epidermal composites were developed by seeding keratinocytes onto the surface of PADM that were previously seeded or non-seeded with dermal fibroblasts. After 14days of culture under air-exposed conditions and in a special growth medium, skin composites were histologically processed and immunohistochemically characterized to determine the expression of cytokeratins and of vimentin and the presence of basement membrane. In all composites, keratinocytes underwent differentiation to a multilayer epidermis with 5-7 viable cell layers. The stratum basalis, stratum spinosum, stratum granulosum and stratum corneum were identified. The expression of cytokeratins was similar to that described in healthy canine epidermis. Laminin and collagen IV immunostaining revealed a homogeneous layer in the epidermal-dermal junction only when the matrix had been seeded by canine dermal fibroblasts. The model may become a simple, useful and cost-effective tool to investigate the biology and pathology of canine epidermis and could partially replace animal testing in several areas of dermatological research.     

Afferent and efferent connections of the nucleus geniculatus lateralis ventralis demonstrated by WGA-HRP in the chick

Fibre connections of the chick nucleus geniculatus lateralis ventralis (GLv) were investigated using the axonal tracing method with wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP). After an injection of WGA-HRP into the GLv, many labelled neurons were observed in layer i of the stratum griseum et fibrosum superficiale (SGFS) in the ipsilateral tectum opticum (TO) and in the nucleus lentiformis mesencephali (LM). In the TO-GLv projection, cells of origin were located in the deeper part of layer i of the TO and were topographically distributed along the direction from the rostrodorsal part to the caudoventral part of the TO relating to a rostrocaudal axis of the GLv. In the LM-GLv connection, the dorsal and ventral parts of the LM connected reciprocally with the rostral and caudal halves of the GLv, respectively. In contrast, in the GLv efferent connection, labelled axon terminals spread widely in the ipsilateral area pretectalis without any clear topographical arrangement.     

Sarcoptic mange in free-ranging raccoon dogs (Nyctereutes procyonoides) in Japan

Sarcoptes scabiei infestation was diagnosed in three freshly dead free-ranging raccoon dogs (Nyctereutes procyonoides) in Kanagawa Prefecture, Japan. The dogs presented with an alopecic pruritic skin disease, with signs of alopecia on the ears, muzzle, around the eyes, elbow, thigh and the neck, and hyperpigmented and crusted skin lesions, which had a severe malodour. Skin scrapings revealed the presence of the mite Sarcoptes scabiei. Histopathology of lesions demonstrated marked acanthosis, hyperkeratosis, parakeratosis and fungal elements, which were subsequently identified as Acremonium sp., Alternaria sp. and an unknown fungus. Mite segments were located mainly in the stratum corneum and also in the stratum granulosum. Tunnels could be observed in the hyperkeratotic stratum corneum. Scanning electron microscopy (SEM) revealed the tortoise-like Sarcoptes scabiei with four long bristles, suckers and blade-like claws on legs 1 and 2, cuticular spines, prominent body striations and a terminal anus. SEM also revealed an adult female mite digging a tunnel with the head wedged into the very end of the closed burrow. Tunnels filled with eggshells, corneocyte debris and faecal pellets were also observed.     

盐胁迫下2种樱桃叶片解剖结构的变化

为了筛选耐盐樱桃砧木,用于天津市周边轻度盐碱地区栽培,本试验通过徒手切片的方法,研究了两种樱桃砧木在盐胁迫下的叶片解剖结构变化。结果表明：在盐胁迫下,四川樱桃的叶片增厚了3.99%,大青叶的减少了2.68%;大青叶的表皮层细胞增厚了23.6%,四川樱桃的减少了3.84%;大青叶的角质层增厚3.57%,四川樱桃的减少了18.31%;四川樱桃的栅栏组织层变化不大,大青叶的减少了16.42%;四川樱桃的海绵组织层增厚了31.96%,大青叶的增厚了21.27%;四川樱桃的栅栏组织和海绵组织层厚度的比值变小、减少了29.17%,大青叶的减少了20.80%。盐胁迫使大青叶叶片表皮细胞层变厚,角质层加厚,栅栏组织层变薄,海绵组织层加厚,整个叶片变薄。盐胁迫使四川樱桃表皮细胞层变薄,角质层变薄,海绵组织层加厚,整个叶片变厚。从盐胁迫下的2种樱桃叶片结构的变化综合分析认为大青叶的耐盐性明显高于四川樱桃。     

Zur Wasseranpassung der Walhaut

In the whale the superficial layer of the epidermis (stratum paracorneum) is incompletely cornified and is characterized by high contents of phospholipids, neutral fats, and glycoproteins. This finding, and especially the structure of the dermis (hypodermis) in relation to its function, are discussed as to the role they play in the adaptation of whales to the aquatic environment.     

Development of monitoring procedures for the herbaceous layer on the Northeastern Transvaal escarpment

Abstract

A step‐wise approach was followed towards the design of a monitoring system for the herbaceous layer on the escarpment of the Drakensberg mountains in the Transvaal, South Africa. The parameter of concern was proportional species composition. In a first phase, the plot size and number of point observations required to achieve a repeatable assessment of a single plot at a predetermined precision level were investigated. The Chapman‐Richards three‐parameter model was fitted to similarity values obtained from comparisons between successively larger sets of points. This resulted in 140 points being required within a 30 × 30 m plot in order to recover 95% of the internal association of the composition data. In a second phase, the number of plots required for a repeatable assessment of a large stratum was determined. Successively larger nested areas were sampled. Monitoring of a large stratum at 95% of asymptotic replicate similarity would require sets of 21 randomly selected plots. The third and final phase was concerned with the development of a procedure for detecting change. A similarity test made it possible to detect change between successive surveys with a known degree of precision.     

Experimental bovine Trichophyton verrucosum infection. Comparison of the rate of epidermal cell proliferation and keratinisation in non-infected and reinoculated cattle.

The rate of epidermal cell renewal in normal bovine skin and that reinoculated with Trichophyton verrucosum was measured using a radioautographic technique. The transit times of both nucleated and fully keratinised cells were measured in sequential biopsy samples removed at predetermined periods after intradermal inoculation with radio-labelled isotopes. The total time taken for cells of the basal layer to travel to the point of desquamation in the stratum corneum was 18 days in normal cattle. In similar areas on cattle that had been reinoculated with T verrucosum the total epidermal cell renewal time was reduced to 12 days. Increased protein synthesis, as measured by incorporation of radio-labelled nucleoside was evident in basal cells within 24 h of reinoculation with the fungus. The nucleated epidermal cell thickness had almost doubled in areas of reinoculated skin within 72 h and increased cell proliferation was maintained for at least 10 days. Desquamation of the thickened stratum corneum had occurred within seven days of reinoculation with the fungus.