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Plectin is one of the largest and most versatile cytolinker proteins known. In basal keratinocytes it links the intermediate filament network to cell membrane-associated hemidesmosomes. Several mutations in its gene have been identified that lead to the recessive disease epidermolysis bullosa with muscular dystrophy. We report here a mutation that leads to a dominant form of the disease, epidermolysis bullosa simplex Ogna. We found that the epidermolysis bullosa simplex Ogna phenotype is due to a site-specific missense mutation within plectin's rod domain. Further, we show that epidermolysis bullosa simplex Ogna is not restricted to a single Norwegian kindred as previously believed. A German family with the phenotypic hallmarks of epidermolysis bullosa simplex Ogna was found to carry an identical de novo mutation. These two mutations arose about 200 y apart in time. Consistent with the absence of muscular symptoms in these patients, muscle biopsies from several epidermolysis bullosa simplex Ogna members of the Norwegian kindred showed normal staining patterns using antibodies to plectin. Skin changes in epidermolysis bullosa simplex Ogna patients are documented on the ultrastructural level.

We describe a patient with severe generalized dystrophic epidermolysis bullosa (EBD) and a novel combination of compound heterozygous mutations in the COL7A1 gene. The maternal mutation was an A-to-G transition (425-A --> G) at position -2 of the donor splice site within exon 3 that causes aberrant splicing of two abnormal transcripts. One includes intron 3, and one excludes both exon 3 and intron 3. Both splice variants contained a premature termination of the translation. The paternal mutation is a 25-bp deletion in exon 20 (2638de125) that leads to a frameshift and a premature termination codon 133 bp downstream from the site of deletion. This combination of mutations allowed expression of collagen VII mRNA. Immunofluorescence staining of the patient's skin and cultured keratinocytes with domain-specific collagen VII antibodies, however, demonstrated markedly reduced levels of alpha1(VII) polypeptides, and no stable collagen VII protein could be extracted from the patient's cells. Electron microscopy showed severely hypoplastic fibrils below the lamina densa, without evidence of normal anchoring fibrils. The clinically unaffected parents were heterozygous for the mutations, suggesting that both COL7A1 gene defects were recessively inherited disease-causing mutations that are "silent" in heterozygous carriers but in combination can severely interfere with the dermal-epidermal adhesion and lead to severe EBD.

The present article discusses specific, directly gene-dependent ultrastructural markers of dominantly inherited epidermal disorders that serve as clues to their underlying molecular genetic abnormalities. These are epidermolysis bullosa simplex Koebner and Weber-Cockayne with rupture or non-assembly of basal cell keratins and point mutations in keratins 5 and 14. Clumping of basal cell keratins is pathognomonic of EB Dowling-Meara and caused by mutations in hot spots of the rod domain of K 5 and K 14. Clumps and aggregates of basal keratins occur side by side in the same cell and thus do not indicate specific different types of mutations. Similar clumping of suprabasal keratins in bullous CIE Brocq and in palmoplantar keratoderma Voerner have been assigned to identical types of mutations in the same critical position of the rod domain in K 1, K 10, and K 9, respectively. Highly unusual tubular keratins are pathognomonic of another dominant palmoplantar keratoderma type the genetic basis of which still awaits elucidation. Shell formation of (low molecular weight?) keratins in ichthyosis hystrix Curth-Macklin is not linked to the keratin gene clusters on chromosomes 12 and 17 and might be related to regulatory genes of keratin expression. Suprabasal shells in congenital reticular ichthyosiform erythroderma do not consist of keratins but resemble glycoprotein networks. Finally, the keratohyalin abnormality in ichthyosis vulgaris was the clue for the identification of a filaggrin deficiency, at the same time giving evidence to the heterogeneity of keratohyalin proteins.

In random pattern skin flaps of mice, tissue fluorescence measured by means of videodensitometry at 24 h after flap elevation significantly correlates with intravital microscopically measured functional vessel density and viability of skin tissue as assessed by transmission light and electron microscopy. The correlation was found reproducible in non-ischaemic flaps (r = 0.86) and flaps being rendered ischaemic for 6 h after elevation (r = 0.98), indicating that increased microvascular permeability as a result of ischaemia/reperfusion does neither affect tissue fluorescence nor the accuracy of the fluorescein dye test. In addition, tissue fluorescence at 24 h after flap elevation accurately predicts ultimate flap survival on the 7th postoperative day with a sensitivity of 0.89 and a specificity of 0.85. These results suggest that in non-ischaemic as well as in ischaemic random pattern skin flaps tissue fluorescence can be used for assessment of nutritive blood flow, viability of skin tissue and ultimate tissue survival in these flaps.

Epidermolysis bullosa simplex (EBS) is caused by defective assembly of keratin intermediate filaments in basal keratinocytes and recent studies indicated causal mutations in the keratin KRT5 and KRT14 genes. In this study, we describe a novel KRT5 mutation in a German sporadic case of EBS Dowling–Meara. Transition of G to T (nucleotide position 2334) leads to a premature stop codon (E477stop, residue 93 of the 2B helix) in the last residue of the highly conserved helix-termination peptide K/LLEGE of the 2B rod domain of keratin K5. This represents the first premature stop codon mutation identified within the K/LLEGE motif of any disorder reported so far that is caused by keratin mutations.

The ultrastructure of epidermolysis bullosa simplex type Weber-Cockayne has as yet not been studied systematically. Therefore biopsies of blisters freshly produced by friction after a hot bath were investigated by electron microscopy in 8 subjects of 6 families. Blister formation was found always to start in the basal layer of the epidermis. At first the cytoplasm with the cell organelles appears to become diluted, then holes develop in the cytoplasm which merge and finally form cytolytic blisters between the dermo-epidermal junction and the nucleus. The cell organelles remain remarkably intact. The cause of blister formation is still unknown, however, no structural abnormality was observed.

Comparative ultrastructural investigations on the pathomorphogenesis of inherited ichthyoses and epidermolyses have shown that such heterogeneous skin disorders may serve as model systems for genetic interactions with developmental processes, such as keratinization, or functional systems, such as dermal-epidermal junctional integrity. Most interesting from the morphologic point of view are dominantly inherited skin disorders in the ichthyosis and epidermolysis bullosa groups in which primary structural defects of structural proteins have been demonstrated that seem to be under the direct control of the mutant gene. Such structural abnormalities concern keratohyalin in autosomal-dominant ichthyosis vulgaris, the tonofilament system in hystrix-like ichthyoses, and the anchoring fibrils in dominant dystrophic epidermolyses. Taking bullous congenital ichthyosiform erythroderma (epidermolytic hyperkeratosis) as a central example, we discuss the stability of such structural defects, the heterogeneity in the ultrastructural abnormalities of clinically closely similar entities (ichthyosis hystrix Curth-Macklin, congenital reticulate ichthyosiform erythroderma), and, in the latter keratinization disorder, the presence of an unusual filament system of unknown biochemical composition in the abnormal keratinocytes. Expression of mutant genes during fetal life and fetal manifestation of such abnormalities are a precondition for the prenatal diagnosis of genetic skin disorders (bullous ichthyosiform erythroderma, epidermolysis bullosa dystrophica Hallopeau-Siemens, Herlitz syndrome). Finally, problems related to the differentiation of mutant keratinocytes and of amniotic fluid cells of fetuses at risk of genetic skin disorders under the in vitro conditions of primary cell cultures are briefly discussed.

Total replacement of the esophagus by colonic interposition has been recommended as the treatment of esophageal obstruction in recessive dystrophic epidermolysis bullosa. We report our experience in the conservative management of esophageal blisters, strictures, and complete occlusion in 5 patients (aged 2-61 yr). Our therapy consists of a combination of the following principles: (a) inhibition of collagenase formation by oral phenytoin to reduce epithelial detachment; (b) pureed or semiliquid food because minor trauma by hard food particles may induce blistering and result in scarring of the upper esophagus, and larger food particles may obstruct an esophageal stricture; (c) avoidance of tangential shearing forces induced by bougienage and endoscopy and instead use of inflatable dilatator balloons which produce vertical pressure that seems to be less harmful; and (d) long-term nasogastric tube feeding, which may relieve even tight strictures. Our observations suggest that successful long-term conservative management of esophageal stenoses in dystrophic epidermolysis bullosa is possible.

In the cytoplasm of several Epilobium-hybrid plasmotypes belonging to the unstable cytoplasmic alteration “irregulare,” the occurrence of unusual spherical structures could be demonstrated by means of electron microscopy. These structures are absent in parents and reciprocal hybrids. Moreover, such structures have, so far as is known, not yet been described for plant or animal cells as cytoplasmic constituents occurring either permanently or under special conditions. They are therefore considered to be peculiarities of the irregulare plasmotypes. Similarities and differences in their morphological features to spherical viruses and virus-like particles are briefly discussed.