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B A Dale

Publications and source records attributed to B A Dale.

At least 73 records · Page 4Linked to original sources
PubMed1990

Filaggrin production by cultured human epidermal keratinocytes and its regulation by retinoic acid.

Filaggrin is a basic protein normally present in the stratum corneum of epidermis. It derives from a high-molecular-weight precursor synthesized in the stratum granulosum of epidermis. This precursor, called profilaggrin, is thought to be associated with the keratohyaline granules of granular cells. It is known that profilaggrin, but not filaggrin, is present in conventional cultures of human keratinocytes grown on plastic petri dishes. In this study, we show that cultured human keratinocytes can convert profilaggrin into filaggrin, when they are grown on a collagen matrix and raised at the liquid-air interface in order to induce terminal differentiation. Moreover, the presence of terminally differentiating keratinocytes above the granular layer is necessary, but not sufficient, for the accumulation of filaggrin. Finally, we show that the accumulation of filaggrin in the outermost layers of submerged cultured human keratinocytes can be triggered by extensive removal (double delipidization) of retinoids from the serum supplement and inhibited when small concentrations (10(-11)-10(-10) M) of retinoic acid are readded to the culture medium. Altogether, the data reported suggest that not only the synthesis of profilaggrin, but also the conversion of profilaggrin into filaggrin are negatively controlled by retinoic acid. Further, it seems that retinoic acid acts directly on the conversion of profilaggrin into filaggrin rather than on the production of terminally differentiating cells capable of accumulating this protein.

Blood Physiological Phenomena↗
PubMed1990

Epidermal growth factor receptors in genetically induced hyperproliferative skin disorders.

The presence and morphologic distribution of epidermal growth factor receptor (EGF-R) were investigated in a variety of genetic disorders that affect human epidermis. These diseases included various forms of ichthyoses as well as restrictive dermopathy and the CHILD syndrome (congenital hemidysplasia-ichthyosiform erythroderma-limb defects). The distribution of EGF-R was detected by immunohistochemical techniques. Increased staining of immunoreactive EGF-R was frequently, but not always, seen in lesions with experimental or clinical evidence of hyperproliferation, suggesting an increased potential to respond to endogenous levels of either transforming growth factor-alpha or EGF. The finding that EGF-R levels are not always increased in congenital epidermal disorders indicated that the presence of this receptor pathway is not simply a marker for aberrant epidermis.

Adult↗
PubMed1990

New approaches and concepts in the study of differentiation of oral epithelia.

Epithelial structural proteins, the keratins and keratin-associated proteins, are useful as markers of differentiation because their expression is both region-specific and differentiation-specific. In general, basal cells in all stratified oral epithelia express similar keratins, while the suprabasal cells express a specific set of markers indicating commitment to a distinct program of differentiation. Critical factors in the regulation of epithelial protein expression are now under investigation. The promoter regions of keratin genes are being characterized to determine what sequences within the genes are responsible for differential expression. One important extracellular factor that influences epithelial protein expression is retinol (vitamin A), which exerts its effects via a group of nuclear receptor proteins that may also be expressed in a region-specific manner. These molecular biological approaches enhance our understanding of the mechanisms regulating differentiation of oral epithelia and its regional complexity.

Animals↗
PubMed1990

Lyme disease.

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Family Practice↗
PubMed1989

Identification of proteolytic cleavage sites in the conversion of profilaggrin to filaggrin in mammalian epidermis.

Profilaggrin consists of multiple filaggrin domains joined by linker segments which are removed during proteolytic conversion to filaggrin. Analysis of tryptic peptides of filaggrin defined a 26-residue linker segment when aligned on the amino acid sequence of one repeat unit of mouse profilaggrin deduced from a cDNA sequence (Rothnagel, J. A., Mehrel, T., Idler, W. W., Roop, D. R., and Steinert, P. M. (1987) J. Biol. Chem. 262, 15643-15648). Two types of linker segments were distinguished by their different susceptibility to thermolysin and by the presence of a Phe-Tyr-Pro-Val sequence in only one type. These data led to a model of profilaggrin in which the two types of linker segments alternate along the length of profilaggrin. This model provides a structural basis for the two stages of proteolytic processing seen in vivo. In the first stage intermediates accumulate which have several filaggrin domains still joined by linker segments lacking Phe-Tyr-Pro-Val. In the second stage, the other linker segments are cleaved and mature filaggrin domains are released. Proteolytic activity with specificity consistent with first stage cleavage was partially purified from rat epidermis. Chymostatin inhibited both the in vitro enzymatic activity and the processing of profilaggrin in a cultured rat keratinocyte cell line. The products formed in vitro were 3-5 kDa larger than intermediates produced in vivo, suggesting that the linker segments are cleaved at one end only. This implies the existence of a third protease which completes the removal of the linker segments.

Amino Acid Sequence↗
PubMed1989

Immunohistochemical analysis of stratum corneum components in oral squamous epithelia.

The generation of a stratum corneum in squamous epithelia involves marked changes in morphology and in the expression of cell products. We have examined the expression of some of the components involved in this process in oral squamous epithelia with different terminal differentiation patterns by use of immunofluorescent techniques. Involucrin and transglutaminase are involved in formation of cornified envelopes consistently seen in the stratum corneum. Both components were present in keratinized oral epithelia (palatal epithelium and hyperkeratinized buccal epithelium). The nonkeratinized normal buccal epithelium stained positive as well. Filaggrin, a protein derived from a precursor present in keratohyalin granules, is proposed to aggregate keratin filaments in the cornified layer. Although the staining differed markedly in quantity, this component was likewise detected in both keratinized and nonkeratinized epithelia. The staining patterns for different keratin polypeptides, however, showed qualitative differences between the different epithelia. Thus, it seems that the keratin composition shows differentiation-specific characteristics, whereas the presence of other important components needed to generate a stratum corneum is not as closely related to the terminal differentiation pattern of oral epithelia.

Antibodies, Monoclonal↗
PubMed1989

Changes in cell phenotype during regeneration of junctional epithelium of human gingiva in vitro.

The relationship between cell attachment and the phenotype of the attached oral epithelial cells was studied by comparing junctional epithelium (JE) with a culture model for JE in which epithelial cells form an equivalent organization of tissues. Gingival explants were cultured on either a high or a low protein-binding membrane. The cut edge of epithelium and connective tissue was placed on the membrane; epithelial cells migrated to form a sheet of tissue between the explant and the membrane substratum. Cells which grew in contact with the high protein-binding membrane attached to the substratum and assumed a cuboidal shape. With time in culture these cells showed a decrease in reactivity with antibodies to psi-3 antigen (an antigen associated with epithelial migration) and an increase in reactivity with antibodies to cytokeratin 19 (a marker for JE). Cells grown on the low protein-binding membrane did not exhibit changes in shape or antigens. Because similar features were found in the JE in vivo, it was concluded that the junctional cells which are directly attached to the tooth (DAT cells) have a nonmigratory phenotype that develops as a response to the tooth surface. Because the cells are in contact with a metabolically inert material the changes appear to be largely self-induced. The culture method allows studies on putative inducer molecules and on mechanisms which may control the phenotype of epithelial cells at the dentogingival interface.

Adult↗
PubMed1988

The expression of congenital ichthyosiform erythroderma in second trimester fetuses of the same family: morphologic and biochemical studies.

The first born offspring of first-cousin parents was affected with a keratinization disorder thought to be nonbullous congenital ichthyosiform erythroderma (CIE). In each of three subsequent pregnancies, the parents elected to have prenatal diagnosis based on evaluation of fetal skin biopsies. The epidermis of fetus 1 was identical to normal 21-wk estimated gestation age (EGA) fetal epidermis, but because keratinization begins normally around 24 wk EGA, the procedure was repeated 4 wk later. A thin epidermis with a few layers of stratum corneum indicated a normal fetus and a healthy infant was born at term. Skin biopsy samples from fetus 2 gave conflicting results; the epidermis of one sample appeared normal but the second had 5-15 layers of incompletely keratinized cells superficial to basal and intermediate layers. The hair canals of both samples were hyperkeratotic. Pelleted amniotic fluid cells contained aggregates of incompletely keratinized epidermal cells and concentric rings of keratinized cells. The fetus was thought to be affected and the pregnancy terminated. Regional variation in epidermal thickness and keratinization was noted upon gross examination of the fetus and by histology of the skin. Marked hyperkeratinization of follicles was evident in all regions. No abnormal keratins were expressed in the affected epidermis but epidermal lipids analyzed from two body regions had a lower triglyceride content and a higher content of free sterols compared with age-matched, normal fetal epidermis. Immunolabeling for markers of differentiation revealed variable stages of epidermal differentiation according to region. Four structurally identical biopsy samples were obtained from a third fetus. The epidermis appeared normal for age and hair canals were keratinized to various extents. The pregnancy was continued and at 33 wk a male infant was born with a severe ichthyosis of the face and scalp and fine, white scaling on the body. The epidermis of both the severely and mildly affected regions of the newborn had a thick, compact stratum corneum and other features of CIE. Scars from all four fetal biopsies were identified on the trunk, in areas which appeared less affected clinically. This study reports, for the first time, the criteria for prenatal diagnosis of CIE and the variable expression of this disorder in the midtrimester fetus. More importantly, it demonstrates the risks and pitfalls of this in utero diagnosis based on epidermal morphology.

Adult↗
PubMed1988

Characterization of an epidermal phosphatase specific for filaggrin phosphorylated by casein kinase II.

During cellular remodeling that accompanies cornification of epidermal cells, the highly phosphorylated protein, profilaggrin, is dephosphorylated and proteolytically cleaved to filaggrin, the keratin matrix protein. Using rat filaggrin phosphorylated by bovine casein kinase II (CK II) as a substrate, we have partially purified a phosphatase from rat epidermis which dephosphorylates rat profilaggrin in vitro. Anion exchange, hydroxylapatite, and gel filtration chromatography yielded a 100-fold purification of phosphatase from a low-salt extract. Further purification led to loss of activity; therefore, only the partially purified phosphatase was characterized. Two forms of the phosphatase, with molecular weights of approximately 170 and 40 kDa, were resolved during gel filtration. The 170-kDa form could be converted to the 40-kDa form in the presence of dithiothreitol. Both forms had pH optima of 6.6, and were strongly inhibited by NaCl (50% inhibition at 35-40 mM). Neither form hydrolyzed para-nitrophenylphosphate or dephosphorylated casein or the synthetic peptide arg3-glu3-thr-glu3, which were phosphorylated by casein kinase II. The two forms were similarly inhibited by known inorganic phosphatase inhibitors, with 22%-36% inhibition by 0.1 mM Na+/K+ tartrate, 55%-60% inhibition by 0.1 mM NaF, and 75% inhibition by 0.1 mM Na pyrophosphate. Para-chloromercuribenzoate also inhibited the activity, suggesting that reduced thiols may be important in catalysis. One mM calcium chloride altered the activity in a complex manner depending on the pH, suggesting a possible role for calcium in regulating enzyme activity.

Acid Phosphatase↗
PubMed1988

Palmar-plantar keratoderma. A clinical, ultrastructural, and biochemical study.

The hereditary palmar-plantar keratodermas are a heterogeneous group of disorders characterized by hyperkeratosis of the palms and soles and distinguishable by other clinical characteristics, associated abnormalities, and mode of inheritance. We report a family with a new autosomal dominant condition with clinical similarities to mal de Meleda and Greither's disease. Biochemical characteristics before and during treatment with oral isotretinoin were typical of hyperproliferative epithelium with expression of the 48-kd keratin. Ultrastructural abnormalities were qualitatively reduced during treatment, but the abnormal keratohyaline aggregations were still seen. Clinical improvement with treatment was marked and directly correlated with dosage.

Adult↗
PubMed1987

Characterization of two monoclonal antibodies to human epidermal keratohyalin: reactivity with filaggrin and related proteins.

Two monoclonal antibodies (AKH1 and AKH2) were elicited with partially purified human filaggrin and characterized by immunohistochemistry on normal and abnormal skin biopsies, immunoblotting techniques, and antigen purification. Both antibodies react strongly with the granular cell layer consistent with the distribution of keratohyalin and show a more diffuse reaction with the stratum corneum in normal skin biopsies. Reaction in cultured human keratinocytes is limited to immunofluorescent granules in flattened, well-differentiated cells in confluent cultures, in which we have previously demonstrated keratohyalin. On immunoblots AKH1 reacts with filaggrin (37 kD) and profilaggrin (400 kD), while AKH2 primarily stains bands of 150 and 300 kD. The AKH2 antigens were identified in the cationic protein fraction used for immunization and were purified by gel permeation and high-performance liquid chromatography. Amino acid composition of these proteins differs only slightly from filaggrin. Immunohistochemical staining patterns of the two antibodies are very similar in the genetic disorders of keratinization tested, except for ichthyosis vulgaris, and reflect the presence and distribution of keratohyalin. In ichthyosis vulgaris, AKH1 staining is weak, consistent with the morphology and with biochemical absence of profilaggrin/filaggrin; however, AKH2 staining is positive, although weaker than normal, suggesting the presence of the AKH2 antigens even when keratohyalin is absent or abnormal. Antibodies AKH1 and AKH2 may be useful as differentiation markers for keratinization in tissues and for cells in culture. Antibody AKH1 can be used specifically for detection of profilaggrin/filaggrin in tissues, cultured keratinocytes, and extracts.

Antibodies, Monoclonal↗