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Biomedical subjects

S Arase

Publications and source records attributed to S Arase.

At least 19 recordsLinked to original sources

Immunohistological analysis of P53 expression in human skin tumors.

The p53 expression in various skin tumors was immunohistologically evaluated using two mouse monoclonal anti-p53 antibodies, PAb421 and PAb1801. The p53 expression was not detected in the normal epidermal cells. Nuclear staining suggested that the p53 expression was observed in 10 of 26 squamous cell carcinomas (SCCs) from 24 patients, in one undifferentiated carcinoma, one proliferating trichilemmal cyst, one malignant proliferating trichilemmal tumor and in one metastatic carcinoma of breast cancer. None off four cases of Bowen's disease (SCC in situ) showed nuclear staining. In the SCCs, five of 20 primary lesions, three of four recurrent lesions and both of two metastatic lesions had positive nuclei. There was one case of SCC in which a primary lesion was negative but a recurrent lesion was positive. Thus, p53 expression was more frequently observed in SCCs at more clinically advanced stages. This may suggest that p53 has some relevance to progression of SCC. Nuclear staining was not detected in any of the following cases: two cases of seborrheic keratosis, one eccrine poroma, one keratoacanthoma, 11 basal cell epitheliomas, two mammary Paget's disease, three genital Paget's disease, one sebaceous carcinoma, four malignant melanomas, six lymphomas, two leukemia cutis and two angiosarcomas.

Antibodies, Monoclonal

The effect of 1,25-dihydroxyvitamin D3 on the growth and differentiation of cultured human outer root sheath cells from normal subjects and patients with vitamin D-dependent rickets type II with alopecia.

We examined the effect of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) on the growth and differentiation of cultured human hair outer root sheath cells (ORSC) from normal subjects and patients with vitamin D-dependent rickets type II (DDR-II) with alopecia. 1,25(OH)2D3 dose-dependently suppressed the plating efficiency, clonal growth, and DNA synthesis of normal ORSC. It enhanced the cornified envelope formation and caused morphological changes in the cells. All results indicated the existence of specific receptors for 1,25(OH)2D3 in the ORSC, and suggest that 1,25(OH)2D3 is a potent inhibitor of proliferation of ORSC as well as a stimulator of terminal differentiation. However, the cells from DDR-II patients with alopecia did not respond to 1,25(OH)2D3, suggesting a lack of the specific receptors in the cells. The differences in the cellular response to the hormone between the normal ORSC and those from the patients were apparent and easily distinguishable, therefore this experiment may be a rapid and simple diagnostic test for DDR-II patients with alopecia. Large number of hairs were difficult to obtain from patients with alopecia, and we developed a new culture method to accomplish these studies from a few plucked hair follicles. Our system may be useful in the culture of ORSC from limited number of follicles, and could be utilized to analyse the cellular characteristics of ORSC in patients with hair diseases.

Alopecia

Culture of human outer root sheath cells from plucked hair follicles in serum-free conditions.

We succeeded in culturing human outer root sheath cells (ORSC) in serum-free conditions without a biological feeder layer. The combination of collagen type IV substrate and modified MCDB 153 medium supplemented with bovine pituitary gland extract (K-GM medium) could support the growth of ORSC. These cells can be serially cultivated for at least 4 passages and stored in liquid nitrogen with good recovery. Thus, a large series of experiments using ORSC may be run simultaneously.

Cell Division

Transcutaneous PO2 and PCO2 measurements in various skin lesions.

Low tcPO2 levels were observed in various skin lesions, indicating that most pathological changes in the skin induce such a reduction. In contrast, the tcPCO2 changed less, with the exception of marked elevations in bullous or prenecrotic lesions. This procedure may have practical applications in clinical dermatology, particularly in quantitating the response to therapy and in predicting a necrotic change.

Adolescent

The influence of cutaneous factors on the transcutaneous pO2 and pCO2 at various body sites.

The transcutaneous partial pressures of oxygen (tcpO2) and carbon dioxide (tcpCO2) were measured at eight different sites in 10 adult male subjects with an electrode at a temperature of 44 degrees C. The mean tcpO2 values (mmHg) were significantly lower on the face (forehead 26.6, cheek 29.6) and the palm (26.4) than at other sites (60.6-69.6). The tcpCO2 values (mmHg) were only slightly higher on the face. Removal of the stratum corneum produced an average increase of the tcpO2 on the palm of 37.6 mmHg and on the forehead of 19.6 mmHg. However, in 10 children with an age range of 3-9 years, the difference in the mean pO2 between the cheek and forearm was very small. There was no significant difference in the cutaneous blood flow at 44 degrees C between the cheek, palm and forearm.

Adult

Immunohistochemical demonstration of peptidylarginine deiminase in human sweat glands.

Human skin is known to contain protein-bound citrulline. This is the product of enzymatic deimination of arginine residues catalyzed by peptidylarginine deiminase. We probed frozen sections of human skin with a rabbit antiserum raised to rat skeletal muscle peptidylarginine deiminase using the avidin-biotin-peroxidase complex technique. This led us to interesting findings. No staining was observed in epidermis, inner root sheaths of hair follicles, sebaceous glands, and hair erector muscle. However, we noticed specific staining of the cytoplasm of secretory and myoepithelial cells of both eccrine and apocrine sweat glands. The procedure also stained neoplastic cells present in specimens dissected from extramammary Paget's disease. The data mean that peptidylarginine deiminase may be used as a new marker in the classification of skin neoplasms showing sweat gland differentiation. Possible localization of multiple types of peptidylarginine deiminases in human skin is discussed.

Antigens, Differentiation

A new method for culturing human hair follicle cells on floating mixed collagen membranes.

A new method for culturing human hair follicle cells without a biological feeder layer has been developed. Dispersed hair follicle cells obtained from plucked hairs have been successfully grown on floating mixed collagen membranes (collagen types I and IV) and have reached a confluent state. They could be subcultured up to twice. Differentiation of the cells was maintained throughout a month in culture. Stratification of the cells and continuous epithelial pavement formation were seen, as in cultured epidermal keratinocytes.

Collagen

Eosinophilic polymyositis induced by tranilast.

A 38-year-old man with asthma developed eosinophilic polymyositis following the administration of Tranilast, an antiasthmatic agent. Low grade fever, erythematous rashes on the entire body, dysphagia, blood eosinophilia, elevations of serum creatine phosphokinase and myoglobin levels, and inverted T waves in the electrocardiogram were noted. A muscle biopsy showed focal degeneration of muscle fibers with an infiltrate of eosinophils and lymphocytes. A rechallenge with Tranilast resulted in erythema formation, blood eosinophilia, and elevations of some serum muscle enzymes and myoglobin levels. Tranilast was considered to be the causative agent. This is the first reported case of Tranilast-induced eosinophilic polymyositis.

Asthma

Co-culture of human hair follicles and dermal papillae in a collagen matrix.

Human hair follicles, either alone or in combination with dermal papillae, were cultured in a collagen matrix. When plucked hair follicles were cultured alone, spike-like structures composed of outer root sheath cells started growing around the follicle and then radiated into the gel. When isolated dermal papillae were embedded close to the follicles, spikes started growing earlier and grew more rapidly than without the papillae. In cultures of excised follicles from which the dermal papilla had been removed, epithelial cells (possibly hair bulb cells) started growing out from the bulbous portion and then also formed spikes. In the presence of a papilla, the spikes elongated toward the papilla, finally reaching and surrounding it. These findings suggest that dermal papilla cells produce a factor(s) that enhances growth of follicular epithelial cells and also attracts those cells. In cultures of whole excised follicles, two major characteristic patterns of cellular growth were recognized. When the dermal papilla remained inside the bulb in contact with the hair bulb matrix, the hair matrix cells proliferated and differentiated in the normal manner, resulting in elongation of the hair shaft and follicle. But when the papilla was detached from the hair bulb matrix, epithelial cells proliferated from the bulbous portion and finally formed hair follicle-like structures. Thus, attachment of the dermal papilla to the hair bulb matrix in the bulbous portion appears to be necessary for growth of the hair and follicle in the normal manner. Our model may be useful for examining the interaction between follicular epithelial cells and dermal papillae and for studying the growth of hair and follicles in vitro.

Adult

[Three-dimensional culture of plucked human hair follicles inside the collagen gel matrix].

Plucked human hair follicles were cultured in collagen gel matrix. Epithelial cells, possibly outer root sheath keratinocytes, appeared from the outer root sheath 4 to 5 days after culturing and continually grew into the gel to form spike-like structures for next 3 weeks. The number and size of the spikes differed in each follicle. Autoradiographically, many DNA-synthesizing cells were seen at the outer cell layer in the enlarged outer root sheath and at the edges in the newly formed spike-like structures. The culture method described here appears to be suitable to study the three-dimensional growth, morphogenesis and differentiation of the outer root sheath cells in vitro.

Adult

[Difference between the hair follicle cell and epidermal keratinocyte in vitro].

We cultured human hair follicle cells (HFC) and epidermal keratinocytes (EK) using mixed collagen membranes as the substrate for the culture, and studied the degree of differentiation of these cells. Morphologically, polygonal epithelioid arrangements were seen in both cultures. However, in the culture of EK compared with HFC, large denucleated cells appeared within shorter periods of culture and stratification of cells were more prominent. In the culture of HFC, keratohyalin granules were not observed and cornified envelope formation was suppressed compared with that in EK. Then it is suggested that HFC and EK are different in the degree of keratinization and of other nature.

Adult

[The effect of 1, 25-dihydroxyvitamin D3 on cultured human hair follicle cells from a patients with vitamin D-dependent rickets type II with alopecia].

We examined the effects of 1, 25-dihydroxyvitamin D3 on the growth and differentiation of cultured human hair follicle cells from a patient with vitamin D-dependent rickets type II [DDR-II] with alopecia. In the presence of 100 nM 1, 25-dihydroxyvitamin D3, the clonal growth of normal hair follicle cells was inhibited and terminal differentiation was markedly stimulated. However, the cells from a patient with DDR-II with alopecia did not respond to the hormone, suggesting a lack of the specific receptors in the cells. The difference in the cellular response to the hormone between the normal hair follicle cells and those from a patient were clear and easily distinguished, and these studies may provide a basis for the rapid diagnosis of DDR-II. Large number of hairs were difficult to obtain from patients with alopecia, and we developed a new culture method to accomplish these studies. Our system may be useful in the culture of HFC from only a few hair follicles.

Alopecia

Two siblings with vitamin-D-dependent rickets type II: no recurrence of rickets for 14 years after cessation of therapy.

Rickets in a 3-year-old boy and his 1-year-old sister, both with alopecia, was cured by treatment with 50,000 IU of vitamin D2 daily for 2 years and did not recur within 14 years after cessation of therapy. A diagnosis of vitamin-D-dependent rickets type II was made in these patients at the ages of 20 and 18 years based on the findings that 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] did not inhibit DNA biosynthesis in phytohaemagglutinin-stimulated lymphocytes and that cultured skin fibroblasts showed impaired nuclear uptake and normal cytosol binding of [3H]1,25(OH)2D3. Surprisingly, the serum 1,25(OH)2D levels of these patients were high and their serum 24,25-dihydroxyvitamin D levels were low, although neither patient showed any symptoms except alopecia. The presence of vitamin D metabolite imbalances in the absence of rickets in these patients might be explained by differences in sensitivity to 1,25(OH)2D3 of bone formation and vitamin D metabolism. In addition, changes of sensitivity to treatment with vitamin D derivatives might be a consequence of differentiation of target cells. From the present findings, it is suggested that in this disease treatment with a sufficient dose of vitamin D derivatives should be initiated in the active phase of rickets.

Alopecia

Vimentin-positive squamous cell carcinoma arising in a burn scar. A highly malignant neoplasm composed of acantholytic round keratinocytes.

Two patients had unusual squamous cell carcinoma (SCC) arising in a burn scar. The SCCs rapidly recurred and metastasized after radical operation, and the patients died of disseminated metastases. Histopathologically, the SCC was poorly differentiated and consisted of acantholytic round cells that diffusely proliferated into the deep dermis. However, small, solid nests composed of squamoid cells were focally observed. Immunohistochemical studies revealed that the acantholytic round neoplastic cells expressed not only keratin but also vimentin, and the coexpression was substantiated with double immunostaining. Vimentin-positive SCC composed of acantholytic round neoplastic cells may be a highly malignant subset of cutaneous SCC.

Burns

Decreased high-affinity epidermal growth factor receptors in psoriatic epidermis.

Using 125I-EGF the distribution of EGF receptors in psoriatic epidermis was autoradiographically examined. Binding sites of 125I-EGF (20 ng/ml) were mainly located at basal and suprabasal cells in normal and uninvolved psoriatic epidermis, whereas they were located at spinous cells as well as at basal and suprabasal cells in affected psoriatic epidermis. Binding of 125I-EGF (1 ng/ml), on the other hand, was observed in normal and uninvolved psoriatic epidermis but not in affected psoriatic epidermis. The majority of EGF receptors in affected psoriatic epidermis were featured by low affinity for EGF.

Autoradiography

Culture of dispersed hair follicle cells from plucked out hairs without a feeder layer.

Dispersed hair follicle cells from plucked out hairs have been successfully grown on collagen type IV without a biological feeder layer. The optimal calcium concentration for the culture of these cells was also studied and decided to be 0.3-0.5 mM. In this calcium range and on collagen type IV, colony formation and colony growth were best and big colonies with a paving stone-like cell arrangement were formed. The availability of such cultured cells for the treatment of missing skin is proposed instead of cultured epidermal keratinocytes which have been commonly used for wound dressing.

Calcium