Effects of adrenergic stimulating and blocking agents on the eccrine sweat secretion in atopic dermatitis and psoriasis

Summary Quantitative measurements of eccrine sweat secretion following stimulation with adrenaline and terbutaline sulphate, a-stimulator, have been performed in patients with atopic dermatitis and psoriasis by means of the electrolytic water analyzer, Meeco. Seasonal variations were demonstrated, the values being lower in the late autumn. The response to adrenaline could be blocked by phentolamine, an -inhibitor, while propranolol, a-inhibitor, had no effect.—The response to terbutaline was blocked by atropine and partly by practolol, a-inhibitor. Terbutaline induced a larger sweat response than isoprenaline, another-stimulator. A-receptor mechanism, in some way related to cholinergic receptors, is suggested.

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Zusammenfassung Die ekkrine Schweißabgabe nach intradermaler Infektion von Adrenalin und Terbutalin sulfat, einem-Stimulator, wurde mittels des elektrolytischen Wasser-Analysators Meeco quantitativ gemessen. Saison-abhängige Variationen wurden festgestellt, wobei eine signifikante Verminderung im Herbst beobachtet wurde. Die adrenalininduzierte Schweißabgabe wurde mittels Phentolamin, einem -Inhibitor, aber nicht mittels Propranolol, einem-Inhibitor, blockiert. — Das Terbutalin-induzierte Schwitzen wurde durch Atropin eindeutig, aber durch Practolol, ein-Inhibitor, nur teilweise blockiert. Terbutalin induzierte eine größere Schweißaktivität als Isoprenalin, ein anderer-Stimulator. Die Resultate deuten auf einen-Receptoren-Mechanismus in den ekkrinen Schweißdrüsen hin, der vielleicht in engem Zusammenhang mit cholinergischen Receptoren steht.

     

Effects of interferon-γ treatment on the cutaneous DTH reaction in rats

Summary The capacity of interferon- (IFN-) to induce class II histocompatibility antigens on different cell types including keratinocytes, is well known, but the impact of IFN- on the immune response is still unclear. Lewis rats sensitized with dinitrofluorobenzene (DNFB) were injected with recombinant rat IFN- (10⁵ U) or phosphate-buffered saline (PBS) once daily on 3 successive days at the bases of the ears either before or after they were challenged on the ears. As expected, the PBS-treated animals showed about a 30% increase in ear thickness and there was an induced expression of class II antigens on the keratinocytes as judged by immunohistochemistry 72 h after challenge. Exogenously added IFN- prior to DNFB challenge resulted in a significantly reduced ear swelling at 24 (p<0.01) and 48 h (p<0.05) after challenge. In this case the keratinocytes expressed class II antigens already at the time of challenge. When IFN- injections were given during the contact allergic reaction there was no significant reduction of ear swelling until 72 h (p<0.01). At that time point there was a more pronounced expression of class II antigens on the keratinocytes compared with PBS-injected animals, due to the IFN- treatment. These in vivo data support our previous observations that IFN- may play a self-limiting role in certain immune responses.     

Human interferon-γ induces expression of HLA-DR on keratinocytes and melanocytes

Summary Primary human epidermal cell cultures composed of keratinocytes and melanocytes were exposed to supernatants of phytohaemagglutinin (PHA)-stimulated T cells, various lymphokines and interferon-, and checked for the emergence of HLA-DR antigen using immunofluorescence and immunoelectron microscopy. HLA-DR expression was induced by the supernatants and human recombinant interferon- (rIFN-), whereas recombinant ₂, interleukin-2 and non-recombinant human interferon- had no such effect. The threshold concentration of rIFN- required to induce this phenomenon was 10 IU/ml; no further increase of reaction intensity was observed using doses of more than 100 IU/ml. Maximum reaction intensity was achieved after 72 h of incubation; a minimum of 3 h of incubation with rIFN- followed by 72 h incubation in rIFN--free medium proved sufficient to induce HLA-DR expression. The inductive effect of the supernatants and rIFN- could be completely abrogated by pretreatment with excess doses of the monoclonal antibody GZ4 specific for human IFN-. Keratinocytes and melanocytes reacted in an identical fashion both qualitatively and quantitatively in all experiments. These data indicate that IFN- possesses specific signal functions in the induction of HLA-DR expression on epidermal cells.Abbreviations IFN- interferon- - rIFN- recombinant interferon- - r IFN-₂ recombinant interferon-₂ - nrIFN- nonrecombinant interferon- - IL-2 interleukin-2 - EC epidermal cells - K keratinocytes - M melanocytes     

Distribution of carbohydrate residues in normal skin

Summary Sections of biopsies of normal skin obtained from 11 individuals were incubated with 8 lectins using an avidin-biotin complex (ABC). All sections when incubated with the appropriate lectin showed the presence of the following carbohydrate residues: l-fucose, -(1–4)-d-GlcNAc)₂ (N-acetylglucosamine), acetylneuraminic acid, Gal--(1–3)-GalNAc (N-acetyl-galactosamine), -d-galactose, -d-glucose, and -d-mannose. In addition, sections of individuals with blood group A showed -d-GalNAc and sections of individuals with blood group B showed -d-galactose. In the stratum (str.) basale, carbohydrates were present in small quantities, but as the cells matured and moved upward, the incorporation of carbohydrates into the cell membranes increased considerably. In the str. granulosum, lectin reactivity was absent in many sections, probably due to masking by phospholipids. The dark cells in the eccrine glands showed reactivity with all lectins except in the one nonsecretor with blood group A₁, whose dark cells showed no l-fucose and -d-GalNAc. The endothelial cells of the blood vessels showed lectin reactivity except when incubated with concanavalin A. The sebaceous glands showed both cytoplasmic and membrane staining when incubated with various lectins.     

A plea to expunge the word “eczema” from the lexicon of dermatology and dermatopathology

Summary Despite its common use for centuries, the term eczema has never been defined in a repeatable way. Although there have been innumerable attempts to do that, the word eczema still lacks specific meaning. Dermatology and dermatopathology will come of age when the word eczema is no longer used.Eczema is any non-contagious eruption of small vesicles. Willan (1817) Eczema is that which looks like eczema. Hebra, Kaposi (1874) The definition of eczema depends to a large extent on a particular school of dermatology and the concept of individual dermatologist. Sulzberger, Wolf (1952) The word eczema has never been successfully defined... Pillsbury (1952) Eczema is a dermatitis of an indetermined cause, whereas dermatitis always has a cause. Sutton (1956) Eczema is a self-perpetuating process in which itching necessitates scratching. Bobroff (1962) Eczema is not a disease, but a characteristic inflammatory response of the skin to multiple stimuli. Soter and Fitzpatrick (1971) The term eczema and dermatitis are used by many dermatologists as synonyms. Lever (1977)     

Activity of 3β-hydroxysteroid dehydrogenase Δ4–5 isomerase in the human skin

Summary The activity of 3-hydroxysteroid dehydrogenase ^4–5 isomerase (3-HSD) was assayed in various tissues microdissected from the freeze-dried human skin of 13 subjects. The sebaceous gland possessed the highest activity of 3-HSD in the skin, while the apocrine sweat gland showed only one fourth of that activity. The vellus hair follicle showed nearly one half of the activity of the sebaceous gland, whereas the terminal hair follicle exhibited much lower activity. The activity of the epidermis and of the dermis were negligible. Incubation of fresh whole skin of the forehead with 7-³H-DHA and subsequent isolation of various tissues revealed that the metabolites identified in the sebaceous gland were androstanedione and androstenedione, whereas testosterone and/or dihydrotestosterone were not detected.Supported by a grant from Shiseido Company for basic dermatological research     

The “strip” patch test: results of a multicentre study towards a standardization

Background The strip patch test (SPT) is a variant of patch testing which is used for substances with a poor percutaneous penetration. Penetration of the substances is enhanced by repeated applications of adhesive tape prior to their application to the skin. However, no guidelines exist for standardized performance of the SPT.Objectives The aim of this multicentre study was to obtain a first practical approach towards a standardized SPT procedure.Methods Intact noninflamed skin of the upper back of 83 healthy volunteers was tape-stripped. For sequential strips, a 25-mm diameter 3M Blenderm surgical tape was vertically applied and gently pressed downward using the fingertips for about 2 s. The tape was removed in one quick movement at an angle of 45° in the direction of adherence. Each strip was performed with a new piece of tape on exactly the same skin area.Results In each subject, we first determined the number of strips (A) until the skin surface started to glisten and calculated the median number of strips () in the sample (=26 strips). We then ascertained the median number of strips () in the sample that was necessary to achieve a statistically significant and twofold increase in TEWL (=11 strips), revealing a critical stratum corneum strip depth. The unknown number of strips (a) for each subject was finally calculated from the formula a/A=/, i.e. the individual number of strips (A) until the skin surface started to glisten was multiplied by a derived tape-specific correction factor (cf=/=11/26=0.4). The increase in percutaneous penetration in strip patch testing by performing a strips versus conventional patch testing was shown by scoring of clinical and subjective SLS irritant reactions.Conclusions The present multicentre study outlines an experimentally derived approach for a uniform SPT procedure, which does not require the use of complex technical equipment. This first approach now requires validation by a study involving the application of allergens to obtain evidence of enhancement in the sensitivity of patch testing.This work is presented on behalf of the German Contact Dermatitis Research Group (DKG).Parts of the work have been presented before in oral presentations at the 42nd Meeting of the German Dermatology Society (DDG) in Berlin, Germany, in May 2003, at the 7th Meeting of the Working Group on Occupational and Environmental Dermatology (ABD) of the DDG in Heidelberg, Germany, in September 2003, and at the 7th Congress of the European Society of Contact Dermatitis (ESCD) in Copenhagen, Denmark, in June 2004.We regret to report the recent death of Hans Joachim Schwanitz.     

Haar und Haarcuticular im Raster-Elektronenmikroskop

Zusammenfassung Die Oberfläche eines gesunden Körperhaares wird mit dem Raster-Elektronenmikroskop bis zu einer Vergrößerung von 1:3000 untersucht. Die Haare treten zwischen den Hautfeldern hervor. Im Gegensatz zu den epidermalen Hornzellen, die während der Präparation schrumpfen, bleiben die Cuticulahornzellen glatt, flach und fest aufeinander geheftet. Die dachziegelartige Anordnung führt zur Stufenbildung, dabei entspricht die Stufenhöhe von 0,4 m etwa der Dicke einer Cuticulazelle. Der Abstand zwischen den Stufen ist weitgehend regelmäßig und beträgt ca. 5,5 m. Dies entspricht der Länge des freibleibenden Anteils jeder Cuticulahornzelle, der ca. 1/7 der Gesamtlänge der Zelle ausmacht.

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Observations on hair and hair-cuticle employing a Stereoscan electronmicroscope

Summary The surface of a normal body hair is enlarged up to 1:3000 and examined with the aid of a Stereoscan electron microscope. The hairs project from deep craters between epidermal fields. Unlike the superficial epidermal horny cells, which shrink during treatment, the cuticular horny cells remain flat, smooth, and firmly fixed one on another. This imbricate formation consists of several steps, each one about 0.4 m high, corresponding to the thickness of a cuticular horny cell. Generally the distance between these steps is constant, measuring about 5.5 m. This is the length of the uncovered part of each cuticular horny cell, which amounts to about one seventh of the total length of the cell.

     

Variations in percutaneous absorption of testosterone in the rhesus monkey due to anatomic site of application and frequency of application

Summary This study determines if the anatomic region affected percutaneous absorption in the rhesus monkey, an animal model with some relevance to man. Percutaneous absorption of testosterone (13.3 g/cm²) from the ventral forearm was 8.8±2.5%. Absorption from the chest was slightly less (5.3 ±0.6%) while that from the cheek was about the same (9.6±0.2%). Absorption from the scalp was greatly increased (20.4±2.7%), that from the vagina was the greatest (63.1±2.6%). As previously noted in man, anatomic variation in skin absorption exists in the rhesus. The ratio of scalp absorption to ventral forearm absorption in the rhesus was similar to that in man.The next objective was to determine the percutaneous absorption of testosterone when applied as a single dose or on a repetitive basis. There was no substantial difference in total absorption when 13.3 g/cm² was applied as a single dose or when the 13.3 g/cm² was applied three times, totaling 40 g/cm². However, when 40 g/cm² was applied as a single dose, absorption was substantially increased over 13.3 g/cm² applied either once or three times. These results confirm previously reported results done with single versus repetitive doses of hydrocortisone.     

Monoclonal antibodies to the β-andrenergic receptor: Modulation of catecholamine-sensitive adenylate cyclase by the antibody

Summary We have developed two types of hybridomas producing monoclonal antibodies to the turkey erythrocyte ₁-adrenergic receptor in order to study the -adrenergic-cAMP system of epidermis. Splenic cells from BALB/c mice immunized with partially purified turkey erythrocyte ₁-adrenergic were fused with mouse myeloma cell line SP2/0-Ag14. Five hybridomas of 17 positive cells producing antibodies which could precipitate soluble turkey erythrocyte ₁-receptors were cloned by the limiting dilution method. The antibodies cross-reacted with - and ₂-adrenergic receptors and stained epidermal basal cells with immunocytochemical techniques. Neither type of antibody interfered with the antagonist binding, i.e., all antibodies bound to sites other than the ligand binding site on the surface. One type of antibody inhibited epinephrine-stimulated adenylate cyclase activity in our leaky epidermal cell system. The data suggest that the antibody interferes with the coupling of the receptor to the regulatory protein.     

Nachweis und Herkunft der unspezifischen Carboxylesterase mit ψ-Mobilität (ψ-Organ-Esterase) an der Oberfläche der menschlichen Haut

Zusammenfassung Schweiß erwachsener Männer und Frauen, welcher von der Hautoberfläche entnommen wird, enthält in der Regel eine unspezifische Carboxylesterase mit -Mobilität, die sowohl -Naphthylacetat als auch Indoxylacetat hydrolysiert. Mit einem Antiserum, das spezifisch gegen Spermaplasmaproteine gerichtet ist, konnte nachgewiesen werden, daß die -Esterase aus dem Schweiß mit einer Organesterase identisch ist, die auch in den oberflächlichen Schichten der Haut und in Komedonen, sowie in zahlreichen inneren Organen und Körperflüssigkeiten vorkommt. Da der Esterasegehalt des Schweißes anscheinend von der regionalen Verteilung der Talgdrüsen abhängig ist, wird angenommen, daß die -Organesterase nicht mit dem Schweiß, sondern mit dem Talg an die Hautoberfläche gelandt.Im Körperschweiß eines Patienten wurde ein Isoenzym mit ₂-Mobilität gefunden, das mit der -Organesterase immunologisch identisch ist. Vereinzelt konnten auch Spuren einer ₁-Esterase, sowie der ₂-Esterase des Serums nachgewiesen werden.

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Demonstration and origin of the nonspecific carboxylesterase with -mobility (-organesterase) on the human skin surface

summary The sweat of adult men and women, which has been collected from the skin surface usually contains a non-specific carboxylesterase with -mobility, which hydrolyzes -napthyl acetate as well as indoxyl acetate. Using an antiserum specifically directed against seminal plasma proteins, we found that the -esterase in the sweat is identical with an esterase occurring in the superficial layers of the skin and in comedones as well as in numerous internal organs and body fluids. The esterase content of the sweat seems to depend on the regional distribution of sebaceous glands. We therefore assume, that the -organ esterase reaches the skin surface not with sweat but with sebum.In sweat from the trunk of one patient we found an isoenzyme with ₂-mobility which is immunologically identical with the -organ esterase. Traces of an ₁-esterase, as well as traces of the ₂-esterase from the serum were detected in a small number of samples.

     

Blood group substance a prepared from human seminal plasma

Summary The blood group A substance occurring in the human seminal plasma has been isolated and purified to a certain degree by phenol/saline extraction. Immunological and biochemical investigations have shown that it is a glycoprotein with the electrophoretic mobility of a-globulin containing 7.38% N-acetylneuraminic acid.Immunization of a rabbit with pooled human seminal plasma proteins yielded an antiserum of high quality containing incomplete agglutinating antibodies against human red blood cells but no precipitating antibodies against the blood group A substance.

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Zusammenfassung Die Blutgruppensubstanz A des menschlichen Spermaplasma wurde durch Phenol-Kochsalz-Extraktion isoliert und weitgehend gereinigt. Immunologische und biochemische Analysen ergaben, daß es sich um ein Glykoprotein mit der elektrophoretischen Beweglichkeit eines-Globulins handelt, das 7,38% N-acetyl-neuraminsäure enthält.Die Immunisierung eines Kaninchens mit gepooltem menschlichem Spermaplasma ergab ein sehr gutes Antiserum, das inkomplette agglutinierende Antikörper gegen menschliche Erythrocyten enthielt, aber keine präcipitierenden Antikörper gegen die Blutgruppensubstanz A.

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This investigation was in part supported by the Deutsche Forschungsgemeinschaft.

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With technical assistance of I. Weyland.     

Immunoelektrophoretische Untersuchungen bei Sklerodermie

Zusammenfassung Immunoelektrophoretische Untersuchungen wurden in 16 Fällen von diffuser Sklerodermie bei Anwendung von normalem, homologem und heterologem Immunserum LE durchgeführt. In einem Teil der Fälle führten wir auch die Adsorption des homologen Immunserums mittels normalem Menschenserum durch, wobei das Fehlen von pathologischen Antigenen aufgezeigt wurde. Außerdem führten wir in einigen Fällen, in denen die Spaltung der -Globulinlinie festgestellt wurde, die Adsorption mittels dem Handelspräparat von menschlichem -Globulin durch, was den Beweis erbrachte, daß diese Spaltung nicht die Folge des Vorhandenseins von pathologischem Protein ist.Das untersuchte Material wurde aufgrund des klinischen Zustands und des Verlaufs in 3 Gruppen eingeteilt. Alle Fälle wiesen jedoch viscerale Veränderungen auf, die für Sklerodermie charakteristisch sind. In der Gruppe mit verhältnismäßig gutartigem Verlauf waren die Abweichungen von der Norm nicht groß, dagegen trat in den schweren Fällen eine Zunahme des ₁-Seromucoids und der Immunglobuline auf. In allen Gruppen war eine der ₁-Globulinfraktionen vermindert, was ein gemeinsames Merkmal der sogenannten Kollagenosen-Gruppe ist.Die Anwendung von homologem Immunserum ist für die Feststellung von pathologischen Antigenen zweckmäßig; es ermittelt auch besser Immunglobuline.Die Anwendung des Gruppenimmunserums LE ist nicht zweckmäßig, da die Ergebnisse denen ähnlich sind, die mit normalem Immunserum erzielt werden.Herrn Prof. Dr. A. Memmesheimer zum 70. Geburtstag gewidmet.     

Allergic contact dermatitis to tulips: An example of enantiospecificity

Summary Allergic contact dermatitis (ACD), an immunological reaction of the skin resulting from contact with reactive compounds occurring in plants was shown to the enantiospecific (animals sensitized to a compound do not react to its nonsuperimposable mirror image). Thus, when guinea pigs were experimentally sensitized to (+)-tulipalin B (a compound present in tulip bulbs) they did not react to its enantiomer, (-)-tulipalin B. This was also true for (+)- and (-)--hydroxy--methyl--methylene--butyrolactones.     

Tierexperimentelle Untersuchungen zur Pathophysiologie der “gemischten Alopecie”

Zusammenfassung Die pathophysiologischen Vorgänge beim nicht-narbigen Haarausfall sind abhängig von der Intensität der einwirkenden Schädigung. Zwei Intensitätsschwellen: Telogenschwelle und Dystrophieschwelle bestimmen die Reaktion des einzelnen Haarfollikels. Die Qualität der auslösenden Noxe spielt nur eine untergeordnete Rolle. Daß die gleiche Noxe an der Haarwurzel verschiedene Reaktionen hervorrufen kann, wird durch die vorgelegten experimentellen Ergebnisse bewiesen.Da bei Einwirkung eines alopecieauslösenden Faktors nicht alle Haarwurzeln in gleicher Stärke betroffen werden, kann die Stärke der Schädigung teilweise oberhalb der Telogenschwelle, teilweise oberhalb der Dystrophieschwelle liegen. Entsprechend reagieren die Follikel bei diesen gemischten Alopecien (Braun-Falco u. Zaun) teils mit vorzeitiger Kolbenhaarbildung, teils mit Matrixdystrophie und Entstehung dystrophischer Haare. In ein pathophysiologisches System, das die Schadensintensität als reaktionsbestimmender Faktor nicht hinreichend berücksichtigt (anagenes Effluvium — telogenes Effluvium Kligman, sind solche Alopecieformen vom pathomechanischen Standpunkt aus nicht einzuordnen. Das erweiterte System: Kolbenhaaralopecie — gemischte Alopecie — dystrophische Alopecie erlaubt demgegenüber bei den bisher bekannten Reaktionsmöglichkeiten der Haarwurzel eine Zuordnung aller nicht-narbigen Haarverluste. In den beschriebenen Versuchen konnte die klinisch überaus häufige gemischte Alopecie erstmals experimentell hervorgerufen werden.     

Interleukin 2, soluble interleukin 2 receptor,and interferon-γ in the suction blister fluids from psoriatic skin

Summary Psoriasis represents inflammatory skin disorders characterized by significant changes in cellular immunity, particularly exhibiting alterations in T lymphocyte-related functions. Early psoriatic lesions have been reported to show an infiltration of activated helper T cells. Elevated levels of interleukin 2 (IL-2), IL-2 receptor (IL-2R), and interferon- (INF-) are associated with an early activation of T cells. To examine local activation of T cells in psoriatic skin, the amounts of activated T cell products, IL-2, secretory form of IL-2R (sIL-2R) and INF- were measured in the fluids of suction blisters raised on psoriatic skin. sIL-2R levels were significantly elevated in the suction blister fluids raised on psoriatic involved skin compared with those on normal and psoriatic uninvolved skin. On the other hand, neither IL-2 or IFN- was detected in the suction blister fluids either from normal, psoriatic uninvolved, or involved skin. However, we could detect IFN- and IL-2 in the psoriatic scale extracts. Although we failed to detect IL-2 and IFN- in the suction blister fluids, the increased levels of sIL-2R in the suction blister fluids from the psoriatic lesional skin indicate local activation of T cells in psoriatic lesional skin.This work was supported by grants-in-aid for scientific research nos. 01570558 and 63480243 from the Ministry of Education, Science and Culture, Japan and a grant from the Lydia O'Leary Memorial Foundation     

γδ T-cell receptor-positive cells of human skin. II. Appearance in delayed-type hypersensitivity reaction

In order to investigate the distribution and involvement of human T-cell receptor-positive (TCR⁺) cells in delayed-type hypersensitivity reactions of the skin, we examined the occurrence and kinetics of TCR⁺ cells during skin reactions of allergic contact dermatitis. In normal human skin sections, TCR⁺ cells were scarce. In allergic contact dermatitis from DNCB, increased TCR⁺ cells were observed both in the epidermis and in the dermis from 48 h after the challenge. Most of the TCR⁺ cells were TCR1⁺ TCS1^– BB3⁺ TiA⁺ (V1^– V2⁺ V9⁺). The percentage of TCR⁺ cells in the peripheral blood remained unchanged and a few TCR⁺ cells in the skin lesions proliferated in situ. It is suggested that the TCR⁺ cells in skin lesions of allergic contact dermatitis may not be involved in initiation of delayed-type hypersensitivity but may have some other roles responding to factors induced in the reaction.     

Immunoelektrophoretische Studien bei Dermatomyositis

Zusammenfassung Immunoelektrophoretische Untersuchungen wurden bei 8 Fällen von Dermatomyositis durchgeführt; 2 davon hatten gleichzeitig Tumoren. Kontrolluntersuchungen fanden in 4 Fällen mit Tumoren und anderen Hautveränderungen statt. Außer normalem Immunserum wurden auch die Gruppenimmunseren Erythematodes, Sklerodermie und Dermatomyositis verwendet.Es wurden immunoelektrophoretische Unterschiede zwischen der idiopathischen und der die Tumoren begleitenden Form festgestellt, die hauptsächlich auf der Vermehrung der ₁-Lipoproteinfraktion bei Tumoren sowie auf dem höheren Immunglobulinniveau bei der idiopathischen Form beruhen.Die Gruppenimmunseren erwiesen sich als nicht geeignet, da sie die Unterschiede zwischen den Dermatomyositis-Formen verwischten, die mittels normalem Immnserum festgestellt wurden.Herrn Prof. Dr. Dr. Dr. h. c. O. Gans zum 75. Geburtstag gewidmet.     

Immunologische Charakterisierung der Carboxylesterasen des menschlichen Spermaplasma und der Organe des Urogenitaltraktes

Zusammenfassung Das Spermaplasma des Mannes enthält zwei unspezifische Carboxylesterasen, welche nicht aus dem Serum, sondern aus dem Genitaltrakt stammen. Die eine hat eine relative elektrophoretische Wanderungsgeschwindigkeit von 0,25 (Bereich der -Globuline), die andere eine relative Mobilität von 0,7 (₂-Bereich). Das Molekulargewicht der -Esterase liegt in der Größenordnung von ca. 150 000, das der ₂-Esterase um 900 000. Beide Enzyme vermögen sowohl -Naphthylacetat als auch Indoxylacetat zu spalten. Die enzymatische Aktivität gegenüber -Naphthylacetat wird durch Kupferionen (bis 0,005 m) nicht gehemmt. Die Hydrolyse des Indoxylacetats durch die -Esterase wird durch Eserin (bis 0,01 m) nicht gehemmt; es handelt sich daher nicht um eine Cholinesterase. Organextrakte aus Samenbläschen, Hoden, Nebenhoden und Niere zeigen nach elektrophoretischer Trennung je eine, die Prostata 2 und die Leber 3 esterspaltende Eiweißbanden mit -Mobilität. Diejenigen Esterasen der Prostata, welche mit verschiedener Mobilität im -Bereich wandern, bilden mit dem spezifischen Antikörper nur eine Präcipitationslinie; sie sind also immunologisch identische Isoenzyme. Im -Bereich zeigen Prostata, Samenbläschen und Nebenhoden je eine, Hoden 2, Niere und Leber 2–3 esterase-reaktive Zonen.Mit Antiseren, welche spezifisch gegen Spermaplasmaproteinen, bzw. gegen Eiweißkörper der Prostata gerichtet sind, konnte nachgewiesen werden, daß die -Esterase des menschlichen Spermaplasma mit einer Organesterase identisch ist, welche in zahlreichen Organen und Körperflüssigkeiten vorkommt.Bei doppelseitigem Verschluß der ductus ejaculatorii enthält das Ejaculat nur die -Esterase, während die ₂-Esterase fehlt. Die -Esterase stammt daher mit größter Wahrscheinlichkeit aus der Prostata.

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Immunological characterization of the carboxylesterases of human seminal plasma and the urogenitary organs

Summary Human seminal plasma contains two non-specific carboxylesterases, originating in the genital tract. They are not, however, serum proteins. One of them has a relative electrophoretic mobility of 0.25 (mobility of -globulin), the other one migrates with the relative electrophoretic mobility of 0.7 (mobility of ₂). The molecular weight of the -esterase is approximately 150,000, that of the ₂-esterases about 900,000.Both enzymes can hydrolyse -naphthyl acetate as well as indoxylacetate. Their enzymatic effect on -naphthyl acetate is not inhibited by copper-ions (conc. up to 0.005 m). The hydrolysis of indoxylacetate by the -esterase is not inhibited by eserine (conc. up to 0.01 m); it is not, therefore, a specific cholinesterase.The electrophoretic pattern of organ extracts from seminal vesicles, testis, epididymis and kidney reveals one esterse-reactive component with -mobility; the prostate gland has two, the liver two to three. The two -esterases of the prostate gland which migrate with different electrophoretic mobilities, form one single precipitation line when they react with a specific antibody.In the region of -globulins we found one ester-splitting component in extracts from epididymis, seminal vesicles, and prostate gland; two in the extract from testis, and two to three in extracts from the kidney and liver.Using antisera specifically directed against seminal plasma proteins or proteins of the prostate gland, respectively, we found that the -esterase of the seminal plasma is identical with an esterase occurring in numerous organs and body fluids.The ejaculates of patients with bilateral occlusion of the ductus ejaculatorii contain only the -esterase whereas the ₁-esterase is absent. This indicates that the -esterase is most probably derived from the prostate gland.

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Mit Unterstützung des Landesamtes für Forschung des Landes Nordrhein-Westfalen.     

Assessment of TGF-β1-mediated growth inhibition of HPV-16- and HPV-18-transfected foreskin keratinocytes during and following immortalization

The responsiveness to transforming growth factor-1 (TGF-1) of two human keratinocyte cell lineages (FK16A and FK18B) generated after transfection with HPV-16 and HPV-18, respectively, was investigated. Both cell lineages revealed loss of heterozygosity (LOH) at 18q and/or 3p associated with the acquisition of the immortal phenotype. These loci harbour genes (TGF- receptor II gene at 3p, and Smad2 and Smad4 genes at 18q) encoding products involved in the TGF-1 signalling pathway. Mortal and early immortal stages of both cell lineages displayed growth reduction upon exposure to TGF-1 concentrations in the range 100 pg/ml to 1 ng/ml. However, the late immortal stages were resistant to TGF-1 at concentrations up to 10 ng/ml. TGF-1 receptors type I and II were expressed at all stages in both cell lineages. Moreover, mRNA levels of Smad2 and Smad4 genes were nearly constant throughout. TGF-1 expression and secretion, which were demonstrated in all analysed stages, may provide selective conditions underlying unresponsiveness to TGF-1 upon prolonged monolayer culturing. Thus, LOH at 3p and/or 18q seen during HPV-mediated immortalization of human keratinocytes was not associated with resistance to TGF-1-mediated growth inhibition or a marked reduction in TGF- 1 receptors and mRNA levels of Smad2 or Smad4. Therefore, alternative events are likely to underlie unresponsiveness to TGF- 1 in late-passage FK16A and FK18B cells.This work is dedicated to the memory of Jan Walboomers, who passed away on 2 February 2000.This work has not been published elsewhere either whole or in part. There exists no financial or other relationship that might lead to a conflict of interest.