Carcinoma-associated 38-kD membrane glycoprotein MH 99/KS 1/4 is related to proliferation and age of transformed epithelial cell lines.

A 38-kD cell-surface glycoprotein (MH 99, KS 1/4) appears strongly elevated in epithelial malignancies, although it also occurs in a few immature, germ-cell phenotypic tissues in human epidermis. In this study, the expression and synthesis characteristics of the MH 99 antigen were examined in several transformed epithelial cell lines. Marked increase of MH 99 (by 100-200%) was found in highly proliferative cell populations, as demonstrated by radioimmunoprecipitation, fluorescence-activated cell sorter analysis, and proliferation experiments in cultures of spontaneously immortalized keratinocytes (HaCaT) squamous cell carcinoma lines (SCL 1 and SCL 2), and SV-40 transformed keratinocytes (130, 425, and HaSV). The relation of the MH 99 antigen to cell proliferation was underscored by a 70-75% decrease of synthesis in nonproliferating HaCaT cells treated with mitomycin C. Blocking of two distinct epitopes of the molecule with two different monoclonal antibodies (MH 99 and MM 104) decreased cell proliferation by approximately 25% (F = 519.6; df = 1,145; p < 0.0001), as compared to untreated cells and to cells treated with unrelated antibodies (BT 15 and J 143). Analysis of MH 99 synthesis in aging transformed keratinocytes cultured without passage revealed downregulation by approximately 50-60% after 60 d, indicating inverse correlation of the MH 99 antigen with increasing cell age. It seems, therefore, that the MH 99 antigen is directly correlated with cell proliferation and inversely correlated with increasing age of transformed epithelial cells, possibly playing an active role in the process of cell proliferation.

Vellus hair follicle-derived keratinocyte culture: a new experimental model in human hair research.

Biological and biochemical mechanisms of hair growth are difficult to study in vivo; therefore the development of in vitro models is of great interest. Today, we have of our disposal reliable techniques to cultivate cell populations and entire components of terminal hair follicles; however, in vitro culture models for cells derived from vellus hair follicles have not yet been established. In this study, we present a technique for cultivating vellus hair follicle-derived keratinocytes (VHK) and we present first findings on their characterization. Primary cultures of VHK were obtained as outgrowths of cultured intact vellus hair follicles prepared by microsurgical means after incubation of full-thickness human skin with dispase. (1) VHK cultures reached confluency after 16-20 days and 3-4 subcultures were possible. (2) VHK were characterized as epithelial cells by light and electron microscopy. (3) A multi-layered stratified epithelium with 8-10 cell layers was observed by electron microscopy presenting abundant keratinosomes in individual cells in contrast to outer root sheath keratinocytes. (4) Synthesis studies of two glycoproteins characteristic for undifferentiated (gp 38) and for differentiated (gp 80) keratinocytes revealed higher synthesis levels for gp 80 and lower levels for gp 38 in VHK as compared to normal epidermal keratinocytes in vitro. These findings suggest a distinct morphologic and differentiation pattern of VHK in culture. This experimental model provides a new tool to study mechanisms of hair growth regulation in vellus hair follicles and to compare them to those of terminal hair follicles.

Tumors of the pilosebaceous unit.

Appendage tumors of the pilosebaceous apparatus are relatively rare, and their clinical aspect is usually nonspecific. In most cases pilosebaceous tumors are benign; however, malignant neoplasias (e.g. trichilemmal carcinoma, malignant pilomatricoma) have been described. In the present review we propose the classification of pilosebaceous tumors by degree and by direction of differentiation and by histological configuration. Three different histological patterns have to be distinguished: tumors with central dilated hair follicle, tumors consisting of epithelial islands and keratotic cysts in the corium and neoplasias with predominantly mesenchymal components surrounding epithelial formations. Careful histopathological examination of pilar appendage tumors will help to classify the wide range of clinically and histologically different tumors and to establish the exact diagnosis.

Outcomes of cryosurgery in keloids and hypertrophic scars. A prospective consecutive trial of case series.

BACKGROUND AND DESIGN: A variety of therapeutic regimens has been used in keloids and hypertrophic scars with unsatisfactory final results. Application of cryosurgery could be beneficial since it was reported to produce less scarring after treatment of skin tumors compared with other surgical procedures. We performed a prospective consecutive trial of randomly enrolled case series including 32 months of average follow-up to assess the outcomes of cryosurgical treatment in keloids and hypertrophic scars and to determine factors influencing the therapeutic result. Ninety-three white patients, aged 10 to 79 years (median age, 26 years) with keloids (n = 55; median duration, 36 months; median size, 4 cm2) and with hypertrophic scars (n = 38; median duration, 12 months; median size, 2 cm2) were treated using the contact method. One freeze-thaw cycle of 30 seconds per lesion and session was employed, and, if needed, treatment was repeated every 20 to 30 days. RESULTS: Excellent responses (ER) were recorded in 30 subjects (32.3%), good responses (GR) in 27 (29.0%), poor responses (PR) in 27 (29.0%), while nine subjects (9.7%) did not respond at all. Hypertrophic scars responded better (n = 38; GR to ER, 76.3%) than keloids (n = 55; GR to ER, 50.9%; P < .005; odds ratio, 6.92). No progressions or recurrences occurred. Histologically, neovascularization, loss of the anarchic arrangement of collagen bundles, increased fibroblasts in a stroma running parallel to the skin surface, and mononuclear cells mostly arranged at the perivascular area were found in clinically responding lesions. Improved responses were detected in subjects treated with three or more sessions (n = 57; GR to ER, 78.9%) compared with subjects treated once or twice (n = 36; GR to ER, 33.3%; P < .001; odds ratio, 13.68). The cryosurgical treatment was generally well tolerated, with local pain during freezing and/or shortly after treatment (n = 32, 34.4%) and hypopigmentation or hyperpigmentation (n = 11, 11.8%) being the most frequent side effects. CONCLUSIONS: Cryosurgery was found effective and safe in keloids and, especially, in hypertrophic scars, in an open, uncontrolled study. Satisfactory results could be obtained after at least three cryosurgical sessions of 30 seconds each, applied once monthly using the contact method of treatment.

Culture of hair matrix and follicular keratinocytes.

In recent years, a variety of in vitro models for the cultivation of hair follicles and their constituents have been developed. Outer root sheath (ORS) keratinocytes (KC) have been mainly studied in explant cultures, planted on bovine eye lens capsules, collagen substrata, 3T3 cell feeder layers, or dermal equivalents, yielding outgrowth of a multilayered stratified epithelium with some biochemical and ultrastructural characteristics of keratinocytic differentiation. More recently, ORS KC cultures have also been initiated from single cell suspensions, and organotypic cultures have been obtained by recombination with dermal cells, inducing a higher degree of epidermal differentiation. Presumptive human hair matrix cells have been isolated from plucked anagen hair follicles and have been successfully propagated on 3T3 cell or normal human fibroblast feeder layers, giving rise to multilayered stratified KC cultures. In contrast, only preliminary data exist concerning the cultivation of bulge cells that have been suggested to represent follicular stem cells. In conclusion we dispose of several in vitro models today to cultivate ORS KC and hair matrix cells that have increased our knowledge on the regulation of the human hair cycle by soluble factors and dermal-epidermal interactions. Further comparative studies on ORS KC, bulge cells and matrix cells have to be carried out to confirm the distinct character of these hair KC subsets.

The vellus hair follicle in acne: hair growth and sebum excretion.

In this study we investigated the activity of the vellus hair follicle in acne. Hair growth and sebum excretion in vellus hair follicles were measured on the forehead and back of men, and on the forehead, cheek, and back of women with acne. Hair growth was assessed by computerized image analysis (phototrichogram), and sebum excretion by computer analysis using Sebutape. In patients with acne, marked differences were revealed when results were compared with recent data from healthy persons. In particular, the mean growth rate of vellus hairs was higher, whereas the percentage of anagen hairs was lower, and the duration of the anagen phase shorter in patients with acne than in healthy individuals. Hair growth and sebum excretion depended significantly (P < 0.01) on the anatomical site (forehead 414 hairs/cm2, 0.053 mm/day, 34%; back 93 hairs/cm2, 0.16 mm/day, 21%). In addition, analysis of hair growth revealed significantly higher values in females than in males for (i) percentage of anagen hairs (P > 0.01), (ii) for vellus hair length (P < 0.05), and (iii) for the duration of the anagen phase (P < 0.01). The present study demonstrates that the activity of the vellus hair follicle is influenced by acne, and vice versa, and therefore its role in the aetiopathogenesis of acne should be reconsidered.

Comparative activity of the topical quinolone OPC-7251 against bacteria associated with acne vulgaris.

The antibacterial activity of the topical quinolone OPC-7251 against bacteria commonly found in acne vulgaris was tested in vitro by an agar dilution method. The MIC50 was 0.25 mg/l for Propionibacterium acnes, 0.125 mg/l for Propionibacterium granulosum, 0.03 mg/l for Staphylococcus aureus and 0.06 mg/l for coagulase-negative staphylococci. Compared with seven other antibiotics tested (ciprofloxacin, penicillin, erythromycin, tetracycline, clindamycin, fusidic acid and gentamicin), OPC-7251 had potent activity against both propionibacteria and staphylococci and the lowest incidence of resistant strains.

Physiology of the vellus hair follicle: hair growth and sebum excretion.

The growth of vellus hair and the secretion of sebum from vellus hair follicles were measured on the forehead, cheek, chest, shoulder and back of healthy men and women aged 15-30 years. Hair growth was assessed by computerized image-analysis of photographs and sebum excretion by the use of Sebutape followed by image analysis. The density of vellus hairs and the percentage of growing hairs were higher on the face than on the thorax (439 hairs/cm2 with 49% growing hairs on the forehead compared with 85 hairs/cm2 with 31.5% growing hairs on the back). The rate of growth ranged from 0.03 mm/day on the forehead to 0.13 mm/day on the back. The maximum length of vellus hair significantly decreased with age; otherwise hair growth was not affected by age or sex. Some variations in hair growth and sebum secretion were observed over a period of 3 months, but no consistent rhythms were detected. There was no obvious link between vellus hair growth and sebum excretion.

[Adverse drug reactions on hair]. / Unerwünschte Arzneimittelwirkungen am Haar.

Adverse drug reactions on the hair are common side effects. The spectrum of loss of hair reaches from diffuse telogen effluvium up to circumscribed or diffuse alopecia on the scalp, sometimes also including other body areas. Other side effects observed are the stimulation of hair growth with hypertrichosis and induction or worsening of hirsutism, as well as changes of the structure or color of the hair. In most cases--other than those treated with cytostatics, androgens, or hormonal contraceptives with some androgenic effect--it is rather difficult to determine the time of onset and primary cause, since the diffuse types often start subclinically, and other factors may play an additional part. As a rule, adverse drug reactions are reversible provided the causative drug is avoided.

[In vitro evaluation of adsorbents used in haemoperfusion (author's transl)]. / Vergleichende in vitro Untersuchungen zur Hämoperfusion mit verschiedenen Adsorbentien.

The properties of six adsorbents for Haemoperfusion were investigated in vitro: Amberlite XAD-2; Amberlite XAD-4; Haemocol; collodion coated charcoal; albumine-collodion encapsulated charcoal and activated carbon fibers. Carbromal and diazepam are most quickly removed from aqueous solutions by Amberlite XAD-2 and XAD-4. Of the charcoal containing materials, Haemocol eliminates most rapidly carbromal and diazepam; while activated carbon fibers remove phenobarbital faster. When heparinized blood is perfused, a remarkable reduction in platelet count is observed. The drop is most pronounced with Amberlite XAD-2 and XAD-4, but even when using charcoal preparations the platelet depletion can reach 10% of the original number over 5 hours. Haemolysis is most pronounced with Amberlite XAD-2, XAD-4 and Haemocol. As the efficiency of the adsorbents towards the various drugs differ and as their adverse effects also vary, the decision for a particular material will have to be made according to the individual case.

[Hemoperfusion by means of encapsuled charcoal for the treatment of exogenous and endogenous intoxications]. / Hämoperfusion durch verkapselte Aktivkohle zur Therapie exogener und endogener Intoxikationen

The method developed by Chang for using in collodion and albumin encapsuled charcoal as artificial cells for hemoperfusion has been studied in vitro. The capacity for the absorption of more or less dialysable exogenous substances (barbiturate, Diazepam, Parathion) and an endogenous toxic metabolite (p-hydroxyphenyl acetic acid) has been examined. With the absorption unit one can eliminate 80 to 90 per cent of the initial concentration of barbiturate, Diazepam and p-hydroxyphenyl acetic acid in about 90 min, of Parathion about 50 per cent in the same time, whereas bromide and ammonia are not absorbed. Attempts to avoid regional heparinisation by the incorporation of heparin into the capsule of charcoal granules were not successful. Assuming correct preparation of the hemoperfusion system, observance of steril precoutions and regional heparinisation, the use of the "detoxication shunt" is a promising means in the treatment of severe exogenous and endogenous intoxications which are not accessible by conventional therapeutic means.