Donor Specific Response of Estrogen and Progesterone on Cultured Human Melanocytes

The mechanisms of estrogen and progesterone in human cutaneous pigmentation are largely unknown. The molecular identification of estrogen receptor (ER) and progesterone receptor (PR) in the human melanocytes is of great importance to understand the mechanisms. We performed immunocytochemistry analysis and demonstrated that ER and PR were expressed in the cytoplasms and nuclei of human melanocytes. Reverse transcriptase-polymerase chain reaction (RT-PCR) and sequence analysis confirmed the expression of ER and PR at the transcriptional level. Despite of the presence of ER and PR, the physiological and pregnant levels of estrogen and progesterone showed inconsistent effects on the proliferation and tyrosinase activity of cultured human melanocytes. These results suggest that human melanocytes express ER and PR, which have a donor-specific action in human pigmentation. Further studies are needed to elucidate the induction mechanism and functions of these receptors, and the role of estrogen and progesterone in melanocytes.

Expression of progesterone receptor in human keratinocytes

Despite the various responses of human skin to female sex hormones, cellular and subcellular targets and the mechanisms of action of estrogen and progesterone in human skin are not well understood. The detection of estrogen receptor (ER) and progesterone receptor (PR) in the skin is of great importance to understand the effect of estrogen and progesterone. In primary cultures of human keratinocytes, expression of ER and PR was monitored by immunocytochemistry and reverse transcriptase polymerase chain reaction (RT-PCR). Paraffin embedded skin tissues were stained with monoclonal antibodies to human ER and PR by immunohistochemistry. Cultured human keratinocytes expressed cytoplasmic PR protein and PR mRNA transcripts. By contrast, ER was detected only at the mRNA level. Suprabasal keratinocytes from samples of pruritic urticarial papules, plaques of pregnancy (PUPPP) and psoriasis were stained positively only for PR, while those from samples of erythema nodosum were negative for both ER and PR. Lesional epidermis of PUPPP showed positive PR immunoreactivity, while nonlesional epidermis did not. No other cells in the normal human skin were stained with ER and PR. The present study suggests that by expressing PR human keratinocytes act as targets for progesterone action.

Assignments of the tyrosinase related protein-1 and -2 genes to human chromosome bands 9p23 and 13q32.1 by in situ hybridization

To determine the precise chromosomal localization of tyrosine related protein-1 and -2 (TRP-1 and TRP-2) genes by fluorescence in situ hybridization, we used DNAs isolated from human bacterial artificial chromosome clones. They contain genomic sequences with approximately 120 kb inserts for TRP-1 and TRP-2. The TRP-1 and TRP-2 genes were assigned to human chromosome bands 9p23 and 13q32.1, respectively. These results confirmed the previously mapped location for the TRP-1 gene and more precisely located the TRP-2 gene, which had previously been mapped to chromosome 13q31-q32.

A Case of Reticular Erythematous Mucinosis (REM) Syndrome

A 43-year-old male patient visited our clinic complaining of persistent erythematous skin eruptions on the anterior chest, abdomen, and back for 10 years. Reticular erythematous mucinosis (REM) syndrome was diagnosed by the clinical and histopathologic features. Mucin deposition is much more conspicuous in papular lesions than in plaque lesions. Therefore, we suggest that the papular lesions may show the characteristic changes of REM and that a biopsy specimen should be obtained from a papular lesion for proper diagnosis of REM syndrome.

The Effects of Cytoskeletons on the Cultured Human Melanocytes

BACKGROUND: Cytoskeletons, the complex set of protein fibers found in the cytoplasm, have important roles in the movement of cells and subcellular structures and the generation of shapes. Melanocytes have numerous dendritic processes which are in direct contact with many keratinocytes and transfer the melanosomes into the neighboring keratinocytes. Little information is available on the structure and function of cytoskeletons, and the effects of ultraviolet light on the cytoskeletons of the melanocytes. OBJECTIVE: The purpose of this study was to investigate the general cytoskeletal system of cultured melanocytes and to find out the effects of the cytoskeletal antagonists and UVB on the cytoskeletal system of the cultured melanocytes. METHODS: Melanocytes were cultured from adult foreskin and then exposed to various cytoskeletal antagonists and UVB radiation. The changes of the cultured melanocytes were evaluated by using phase contrast microscopy, immunofluorescence staining methods and electron microscopic examinations. RESULTS: Colchicine produced shortening of dendrites, stellate cellular contour and granular fluorescence of the tubulin. Cytochalasin D produced round cellular contour and granular fluorescence of the actin. Acrylamide produced disorganization of cytoplasmic constituents, but no specific fluorescent change was observed. Colchicine also had inhibitory effects on the vimentin. Cellular responses induced by these agents were reversible. UVB caused morphological changes of the melanocytes, but their effects on the organization of the cytoskeletal system could not be detected in this method. CONCLUSION: Microtubules are related to the dendritic movement of the melanocytes. Vimentin may be involved in the transfer of cellular organelles, probably including the melanosomes. Cytoskeletal antagonists produce their characteristic morphological changes to cultured melanocytes.

Clinical Study of Vitiligo

BACKGROUND: The clinical behavior of vitiligo has not been clearly understood and hypothesis concerning the pathogenesis of the disease has been confusing and contradictory though autoimmune mechanisms have been considered important by many authors. OBJECTIVE: The purpose of this study was to develop a better understanding of the clinical features and pathogenesis of vitiligo. METHODS: We investigated clinical features of vitiligo in 1315 patients, and also compared the clinical course and features of non-segmental type(type A) and segmental type(type B) vitiligo patients to see whether the two types of vitiligo have a different pathogenic mechanism. RESULTS: Previously reported clinical patterns of the disease were reviewed and compared with our data, and the different clinical findings between the two types which supported the hypothesis of Koga et al. that type A and type B vitiligo had a different pathogenesis and autoimmune mechanisms played a role only in type A were shown. CONCLUSION: We investigated the clinical characteristics of vitiligo in Korea and showed that the type A vitiligo might have a different pathogenic mechanism with type B.

Unilateral Keratosis Lichenoides Chronica

Keratosis lichenoides chronica is rare chronic dermatosis characterized by progressive development of licheniod papulonodules especially on the extremities and trunk. A 15-year-old male patient had erythematous to violaceous scaly patches and plaques on the left side of trunk and lower extremity along Blaschko's lines. Clinical and histologic findings were compatible with keratosis lichenoides chronica showing unilateral distribution.

Erythropoietic Protoporphyria in a Family

Erythropoietic protoporphyria (EPP) is an autosomal dominant condition due to decreased activity of ferrochelatase. The disease is characterized by a wide range of photocutaneous changes and occasionally by liver disease. The level of protoporphyin is raised in erythkocytes and it may also be increased in the feces. We report herein a case of EPP present in a family which was diagnosed by a high free erythrocyte protoporphyrin (FEP) count.

Erythropoietic Protoporphyria in a Family

Erythropoietic protoporphyria (EPP) is an autosomal dominant condition due to decreased activity of ferrochelatase. The disease is characterized by a wide range of photocutaneous changes and occasionally by liver disease. The level of protoporphyin is raised in erythkocytes and it may also be increased in the feces. We report herein a case of EPP present in a family which was diagnosed by a high free erythrocyte protoporphyrin (FEP) count.

Artificial Reproduction of Lupus Erythematosus by Provocative Phototesting

Sunlight is one of the well-established factors which play key roles in the induction and exacerbation of lupus erythematosus. In two patients of discoid lupus erythematosus, we have experimentally reproduced skin lesions by provocative phototesting. Both UVA (100 joules/cm²) and UVB (80 millijoules/cm²) radiation induced the skin lesions. The reproduced skin lesions were clinically and histopathologically consistent with lupus erythematosus.

Culture of melanocytes obtained from normal and vitiligo subjects

The development of human melanocyte culture in vitro from normal adult skin and uninvolved skin of vitiligo patients is essential to investigate the mechanism of depigmentation in vitiligo and other pigmentary dermatoses. By using selective growth and long-term maintenance conditions, we selectively cultured melanocytes derived from normal foreskins and arm skins, and uninvolved foreskins and arm skins of vitiligo patients. The melanocytes of the arm skins were successfully cultured from the roofs of suction blisters. Melanocyte Growth Media (MGM) consisting of MCDB-153 formulation with basic fibroblast growth factor (bFGF), bovine pituitary extract (BPE), insulin, hydrocortisone, phorbol 12-myristate 13-acetate (PMA) and 10% human AB serum was sufficient to grow the melanocytes from normal and vitiligo donors. Melanocytes from uninvolved skin of vitiligo donors showed no different morphologic features, initial seeding capacity and population doubling time compared with those from normal skin. Melanocytes from both cell types grew without any lag period for more than 6 months (6-11 passages). Melanocytes obtained from foreskins had higher initial seeding capacity and shorter population doubling time than those obtained from arm skins using suction-blistered roofs. Our results suggest that the culture method using suction blisters may be a simple and easy way to obtain melanocytes. In addition, vitiligo melanocytes can be successfully cultured with appropriate growth conditions and may show no defective growth patterns. This culture system will be applied to investigate the basic pathophysiology of vitiligo and other various pigmentary dermatoses.

Allergic contact dermatitis due to 8-methoxypsoralen(8-MOP) / 대한피부과학회지

Allergic contact dermatitis due to 8-MOP is a rarely known si(ie effect of this widely used drug. Other known adverse reactions due to 8-MOP such as the oallergic dermatitis as well as some isolated cases of exanthema, papular eruptions, and astloma like symptoms are also sporadically reported. A 52-year-old man with vitiligo developed erythema to the UVA exposed 0.3% Oxoralen cream applied area. Prior to this episode, the patient had history of generalized burns after systernic PUVA therapy in 1983. Even after this experience, the patient had few more episodes of erythema at the site of 0.3%. Oxoralen cream application. We performed patch test and photopatch tests with Scandinavian series, 0.3% Oxoraler or am (as is), and diluted 8-MOP, 5-MOP, TMP solution. The result showed positive reactivity to 6-methylcoumarin, 8-MOP, as well as to 0.3% Oxoralen cream. The size of erythema was same in both irradiated areas which indicates an allergic contact dermatitis rather than photoallergic dermatitis or phototoxic dermatitis.

Persistent light reaction / 대한피부과학회지

Persistent light reaction is a condition of chronic photodermatitis in which photosensitive reaction persists even after the rernoval of all photosensilizers. A 56-year-old man had experienced a recurrent dermatitis involving primarily the face, neek, forearms and hands for 9 years, this condition was aggravated by sunexposure. Photopatch testing disclosed a strongly positive reaction to chloropromazine, promethazine, acid trichlorocarbanilide, Phototesting also revealed lowered MED with UVA and UVB thar norrmal mean value.

The skin concentration and minimal phototoxic dose following administration of phototoxic drugs as a function of time / 대한피부과학회지

No abstract available.

High Doses of UVA Suppress Contact Hypersensitivity

Contact hypersensitivity (CH) responsiveness to 24-dinitro-l-fluorobenzene(DNFB)is depressed in mice sensitized through unexposed skin sites after exposure to high dose of ultraviolet B radiation(UVB). Exposure of mice to ultraviolet A(UVA) radiation in combination with 8-methoxypsoralen(8-MOP) also results in a systemic suppression of CH. Our study was designed to determine whether a high dose of UVA radiation alone can induce a systemic suppression of CH, and if so, which phase of CH response is influenced by UVA radiation. Relatively large doses of UVA(400, 600, 800J/cm²) induced significant systemic suppression of CH when DNFB was applied to UVA-unirradiated abdominal skin. The duration of the rest period after UVA exposure did not cause any significant change in systemic suppresion of CH. Functional analyses showed that lymph node cells(LNCs) obtained from donors that were sensitized on the unirradiated skin site with DNFB 5 days after UVA treatment transferred normal ear-swelling responsiveness to non-primed recipients, thus implying that high doses of UVA can induce systemic suppression which is not affected in the induction phase of CH but affected in the elicitation phase of CH. UVA irradiation de-creased Langerhans cell(LC) numbers significantly with a dose of 100J/cm² or greater. LNCs obtained from donors that were sensitized on the irradiated skin site with DNFB 5 days after UVA treatment did not transfer normal ear-swelling responsiveness to non-primed recipients. This phenomenon may be related to the decreased number of LC after UV treatment. To look for possible mediators impairing the elicitation phase of the CH reaction, we checked prostaglandin E(PGE) levels in serum after 800J/cm² irradiation. A high dose of UVA did not increase the serum PGE level in mice as much as UVB irradiation, in which a significant increase of PGE may affect CH response.

High Doses of UVA Suppress Contact Hypersensitivity

Contact hypersensitivity (CH) responsiveness to 24-dinitro-l-fluorobenzene(DNFB)is depressed in mice sensitized through unexposed skin sites after exposure to high dose of ultraviolet B radiation(UVB). Exposure of mice to ultraviolet A(UVA) radiation in combination with 8-methoxypsoralen(8-MOP) also results in a systemic suppression of CH. Our study was designed to determine whether a high dose of UVA radiation alone can induce a systemic suppression of CH, and if so, which phase of CH response is influenced by UVA radiation. Relatively large doses of UVA(400, 600, 800J/cm²) induced significant systemic suppression of CH when DNFB was applied to UVA-unirradiated abdominal skin. The duration of the rest period after UVA exposure did not cause any significant change in systemic suppresion of CH. Functional analyses showed that lymph node cells(LNCs) obtained from donors that were sensitized on the unirradiated skin site with DNFB 5 days after UVA treatment transferred normal ear-swelling responsiveness to non-primed recipients, thus implying that high doses of UVA can induce systemic suppression which is not affected in the induction phase of CH but affected in the elicitation phase of CH. UVA irradiation de-creased Langerhans cell(LC) numbers significantly with a dose of 100J/cm² or greater. LNCs obtained from donors that were sensitized on the irradiated skin site with DNFB 5 days after UVA treatment did not transfer normal ear-swelling responsiveness to non-primed recipients. This phenomenon may be related to the decreased number of LC after UV treatment. To look for possible mediators impairing the elicitation phase of the CH reaction, we checked prostaglandin E(PGE) levels in serum after 800J/cm² irradiation. A high dose of UVA did not increase the serum PGE level in mice as much as UVB irradiation, in which a significant increase of PGE may affect CH response.

The ultraviolet B protection effects of topically applied melanosomes onto human skin

Melanosome is a cellular organelle that is composed of a melanosomal matrix and a brown biochrome, melanin which is formed by tyrosine-tyrosinase reactions. The melanosome is formed within the melanocyte and transferred to the surrounding keratinocytes through dendritic processes. Human skin color is related to the number, size, type and distribution of melanosomes, and the major role of melanosomes is to prevent skin from injurious nonionizing ultraviolet radiation. Controlled NaOH hydrolysis and centrifugation of human hair make it possible to isolate large amounts of melanosomes which are synthesized within the follicular melanocytes and transferred to hair matrix cells. In this study, the sun protection factors of topically applied melanosomes isolated from human hair were evaluated using ultraviolet B phototesting. Topically applied melanosomes increased the minimal erythemal doses. And the sun protection factors of each 50% and 25% melanosomal preparation were 12.3 +/- 5.5 and 3.1 +/- 1.3 respectively, and these ultraviolet B protection effects showed statistically significant differences from 10%, 5% and 1% melanosomal preparations and vehicle. Form these results, the dose-related photoprotective role of melanosomes was confirmed.