Search

Terminal hair follicles of the human scalp of all ages showed apoptotic pocket-like structures in the outer root sheath of the bulge area at anagen, but not telogen phase. The occurence of these hole structures was roughly estimated in 15% of anagen terminal hair follicles of the human scalp. The size of these apoptotic pockets was variable, ranging from pin hole-like spaces to larger structures filled with homogeneous black materials. These unusual variations were often co-localized with apoptotic degenerations and exclusively present in the presumptive bulge of anagen terminal hair follicles where arrector pili muscles were seen in the vicinity. In fact, these vacuolated structures tended to be present on the side where the major part of the arrector pili muscles anchored.

Etiology of connective tissue disease is unknown. The association of infectious agents has been suspected serologically, ultrastructurally and recently by means of molecular biological techniques. We extracted RNA from lesions of 25 discoid lupus erythematosus, 9 systemic LE, and 3 systemic sclerosis biopsies, and reverse transcription-polymerase chain reaction was performed for all paramyxoviruses known to infect human beings. None of the samples yielded positive signal for any of the viruses. The existence of paramyxovirus in the skin of connective tissue disease is very unlikely.

Extracted human terminal and vellus hair follicles were prepared by EDTA and examined by light and electron microscopes. These samples were morphologically well preserved and enabled us to observe 3-dimensional (3-D) views of the outer surfaces of hair follicles. Interestingly, the bulge areas of terminal and vellus hair follicles showed several morphological variations, such as knob-like swellings and villous projections, independent of hair cycle. Moreover, skirt-like structures were furnished in the small vellus hair follicles independent of hair cycle, but not terminal and large vellus hair follicles. These morphological variations of human hair follicles were confirmed to genuinely represent 3-D views by means of the conventional transverse and vertical sections of the human skin.

Mouse monoclonal antibodies (MoAbs), IKH-1 and IKH-2, were produced against cloned human melanoma cells, KHm-6, which were cultured with 12-O-tetradecanoylphorbol-13-acetate (TPA) and processed by formalin fixation and alcohol dehydration (FFAD). According to the biochemical analysis, antigenic substances which reacted with IKH-1 were 34.0-60.0 kDa glycoproteins, and those which reacted with IKH-2 were 33.5, 34.5 and 36.0 kDa glycoproteins. Immunoelectron microscopy revealed that reaction products of IKH-1 were seen in some membranous vesicles, premelanosomes and cell membrane of TPA-treated KHm-6 cells, while IKH-2 recognized only premelanosomal structures. Immunohistochemical tests revealed that IKH-1 and IKH-2 have a high sensitivity (94.0% and 85.0%, respectively) to formalin-fixed, paraffin-embedded (FFPE) tissue sections of malignant melanomas. IKH-1 had a high specificity and IKH-2 and 100% specificity to FFPE tissue sections of melanocytic lesions.

Using Merkel cell specific antikeratin antibodies and neurofilament antibody the nerve-Merkel cell relationship was studied with a double staining method on frozen sections. Merkel cells were stained with monoclonal anti-cytokeratin CK-5 and CAM 5.2 which react against human cytokeratin polypeptide 45 kDa and 52.5 kDa, respectively. Peripheral nerves were stained with monoclonal murine antibody Neurofilament 70 and 200 kDa. Epidermal Merkel cells were recognized in 12-week fetuses in the plantar skin. In 15-week fetuses dermal Merkel cells were found. Most of the dermal Merkel cells initially lacked a close association with immunoreactive nerve fibers. In 16-week fetuses immunoreactive small nerves reached the epidermis and a few dermal Merkel cells became heavily entangled with the meshwork of nerve endings. Nerve-Merkel cell complex in the dermis was confirmed by electron microscopy. The appearance of epidermal Merkel cells preceded the attachment of immunoreactive nerve endings to the epidermis. It an analysis of 448 dermal Merkel cells the nerve-Merkel cell complex became more frequent as the age of fetus advanced. It was concluded that Merkel cells do not arrive at the epidermis with peripheral nerves. Rather, the peripheral nerves are attracted to the dermal Merkel cells which originated in the epidermis.

Crystalloid inclusion bodies (CIB) of the endothelial cells (EC) were investigated in blood vessels of human fetal skin and the umbilical cord by electron- and immunoelectron microscopy. They were found in up to 15% of the investigated EC in various types of vessels. Their sizes ranged from 0.2 μm to 0.6 μm in the largest diameter. Most frequently we observed a laminated pattern of the crystalloid structure with a regular periodicity of dark and light bands. Additionally a honeycomblike pattern was also seen. Measurement of the CIB laminated structure revealed similar dimensions to Weibel-Palade bodies (WPB). In EC of all vessel types we found numerous WPB differing in electron density, shape and size from those of WPB found in adult blood vessels. WPB were found much less frequently in EC with CIB, suggesting that CIB is a precursor of WPB. After incubation with monoclonal antibody against von Willebrand factor (vWf) both WPB and large organelles were labeled. Because of their shape and size the labeled large organelles seemed to represent inadequately preserved CIB. After incubation with anti-lysozyme only the large organelles were labeled. A possible relationship of CIB to WPB is thus suggested. The presence of lysosomal enzymes such as lysozyme suggest that CIB are lysosomal organelle and participate in the uptake of vWf. The crystalloid pattern of CIB may represent an accumulation of a highly condensed form of vWf.

Dermal reactions to primary intradermal or appendageal sensitization are compared to predominantly dermal reactions to standard patch tests and to intradermal antigen tests. In contrast to epicutaneous spongiotic contact dermatitis, HLA-DR was only seen on skin appendages and nearby basal keratinocytes in indurated tissue reactions with the exception of the reactions with focal basal cell layer disruption and an indurated patch test performed one week post angry back syndrome. Other intradermal skin tests showed only minimal epidermal HLA-DR expression despite spongiotic epidermal changes. Predominantly dermal hypersensitivity reactions can be induced by intradermal or epicutaneous routes. They can evoke hypersensitivity responses which do not cause most epidermal keratinocytes to express HLA-DR.

Desmoplakin I and II are important components of the attachment plaque of the desmosome which mediates cell to cell adhesion, in epithelial cells. In this study we used well-characterized antibody against desmoplakin I and II immunohistochemically and immunoelectron microscopically on two cases of pemphigus vulgaris and one case of pemphigus erythematosus and two cases each of Hailey-Hailey's disease and Darier's disease. In the normal human epidermis the desmosomes were demonstrated in a dotted pattern along cell periphery. In pemphigus vulgaris and pemphigus erythematosus acantholytic cells and the perilesional cells exhibited normal dotted pattern along the cell periphery. In Hailey-Hailey's disease and Darier's disease, the dotted pattern is lost in acantholysed and perilesional areas and anti-desmoplakin I + II positive proteins were observed diffusely in the cytoplasm. Immunoelectron microscopical findings correspond to these light microscopical observations. It is concluded that in autoimmune acantholytic disease such as pemphigus vulgaris and pemphigus erythematosus, desmoplakins are intact even in acantholytic cells, whereas in genodermatoses such as vulgaris and pemphigus erythematosus, desmoplakins are intact even in acantholytic cells, whereas in genodermatoses such as Hailey-Hailey's disease and Darier's disease primary or secondary abnormalities abnormalities of desmosomes may be involved in their pathogenesis.

The present study was performed to investigate the ultrastructural localization of the binding sites of the lectin Ulex europaeus agglutinin, Type I (UEA) and the antibody against factor VIII related antigen (F VIII RAG), both widely used as vascular markers in human skin capillaries at light microscopy level. In addition the ultrastructural localization of vimentin, the major protein of the endothelial intermediate filaments, was demonstrated in human vascular endothelial cells (EC) in situ. The low temperature postembedding technique with Lowicryl K4M with subsequent application of gold labelled antibodies provided both well preserved antigenicity and morphology. The antigenic site of UEA, alpha- l -fucose containing glycohydrate residues, was primarily found in or near the luminal plasma membrane of EC. Gold particles representing the sites of F VIII RAG were located in the Weibel-Palade bodies (WPB) of EC. The ultrastructural staining pattern of the anti-vimentin antibody showed an exclusive labelling of the intermediate filaments of EC.

A case of papillary eccrine adenoma was analysed with immunohistochemical and ultrastructural methods regarding their direction of differentiation. It was found that the majority of the structures show either eccrine ductal or glandular differentiation. There were some segments, particularly in those exhibiting papillary growth, where cells similar to eccrine secretory (clear) cells or cells with characteristics of both ductal basal cells and glandular myoepithelial cells were present.