Jan Vincent

Computerized Wound Image Analysis

The concept is based on assessing macromorphological changes in wounds during the healing process. These changes are expressed in terms of colour characteristics and the area of the necrotic tissue and total wound. The colour shifts for the phases between black/yellow (necroses/fibrin) and the red tissue (granulation) are quantified. The wound is reproduced on Polaroid colour film by a special camera (Acmel) equipped with a ring strobe and a focusing aid, that renders sharp and well centered close-up photographs without need for a view finder or touching the wound. Reflections are minimized by tilting the camera at an angle of 30° to the normal wound surface plane. To make it possible to later correct for colour and optical/geometrical distortion in the photograph, a reference scale is incorporated alongside the wound. The photographs taken daily throughout the treatment period are collected and digitally processed via a video camera. The information obtained is then fed into an image processing system (Epsilon, Imtec AB, Uppsala). The colour of every single pixel in the wound image is automatically determined according to the standardized classification table and shown in tabular form. The method allows rapid and objective analysis of large series of wound images which considerably facilitates the evaluation of wound healing and markedly increases the reproducibility of result assessment.

Infrared spectroscopy for tracing of topically applied ointment vehicles and active substances on healthy skin

SummaryInfrared spectroscopy was used to trace active substances and ointment vehicles applied on the skin. Vaseline and lanoline could be traced after 8 hrs but not olive oil. From the active substances, ethyl-4-amino-benzoate (5%), clioquinol (5%), parabenes (15%), 5,7-dichlor-8-hydroxy-2-methyl-chinolin (5%), balsam of Peru (25%) and pyroleum pini (12%) could be traced 1 hr after application but had disappeared after 8 hrs. Ethylenediamine (1%), chlorcresol (1%), pyroleum lithantracis (5%), were not traceable after 1 hr, and curiously neither neomycine sulphate in spite of its high concentration (20%). The reaction of the skin surface lipids, after application of different substances, was deduced from the spectra. Clioquinol and pyroleum lithantracis seem to give rise to hydrolysis of the triglycerides, the free fatty acids being clearly identifiable. It is felt that infrared spectroscopy can be used as an effective method to trace different substances such as potent allergens on healthy or diseased skin.ZusammenfassungMit Hilfe von Infrarotspektroskopie wurde versucht aktive Substanzen wie auch Salbengrundlagen nach lokaler Applikation auf der Haut nachzuweisen. Im Gegensatz zum Olivenölkonnten Vaseline und Lanolin 8 Std nach Auftragung gefunden werden. Von den aktiven Substanzen wurden Äthyl-4-amino-benzoat (5%), Clioquinol (5%), Parabene (15%), 5,7-Dichlor-8-hydroxy-2-methyl-chinolin (5%), Perubalsam (25%) und Pyroleum pini (12%) 1 Std nach Auftragung nachgewiesen. Nach 8 Std waren diese Substanzen nicht mehr nachweisbar. Äthylendiamin (1%), Chlorkresol (1%) und Pyroleum lithantracis (5%) wurden nach 1 Std nicht gefunden, interessanterweise gelang auch der Nachweis von Neomycinsulfat trotz hoher Konzentration (20%) nicht. Durch Clioquinol und Pyroleum lithantracis werden wahrscheinlich Triglyceride hydrolisiert, die freien Fettsäuren lassen sich sicher nachweisen.Mit der Infrarotspektroskopie bietet sich eine wertvolle Methode an, mit der verschiedene Substanzen, beispielsweise starke Allergene, in kranker wie auch in gesunder Haut nachgewiesen werden können.

Topical Erythromycin for Acne vulgaris

The clinical response of topical erythromycin (1%) in a new penetrative vehicle was evaluated in a double-blind investigation on 28 patients with severe and moderate acne vulgaris. The verum preparation was superior to the placebo in 21 of the patients, while in 4 the reverse was true and in 3 no differences were seen. It is thus obvious that topical erythromycin in the penetrative vehicle is a potent agent for acne treatment. The risk for development of resistant Propionibacterium acnes strains must, however, be considered in topical as well as in systemic antibiotic therapy of acne vulgaris.

Antibodies against extractable components from Propionibacterium acnes in humans with and without acne vulgaris.

SummaryAcidic polysaccharides (AP) are released from Propionibacterium acnes I and II during growth. Similar antigens are released from two strains of P. avidum, but in a smaller amount. Furthermore, prostaglandin-like substances (PLS) have been isolated from the lipid fraction of P. acnes.The AP antigens give rise to a precipitation reaction against a large proportion of normal sera. However, significantly higher antibody titre levels towards the AP fraction were found in patients with severe acne vulgaris. No rise of antibodies was found towards PLS. These findings suggest that APs may contribute to the inflammatory response in acne vulgaris as antigens, whereas PLS, as one of the potential mediators trigger development of the initial lesions.ZusammenfassungSaure Polysaccharide (AP) werden von dem Propionibacterium acnes I und II während des Wachstums freigesetzt. Ähnliche Antigene werden von zwei Stämmen des P. avidum liberiert, jedoch in einem geringeren Ausmaß. Weiterhin wurden Prostaglandin-ähnliche Substanzen (PLS) von der Lipidfraktion von P. acnes isoliert. Die AP-Antigene führen zu einer präzipitierenden Reaktion gegen Normalserum. Es wurden jedoch signifikant höhere Antikörpertiter gegen diese AP-Fraktion bei Patienten mit schwerer Akne vulgaris gefunden. Eine Erhöhung von Antikörpern gegen PLS wurde nicht festgestellt. Diese Resultate lassen die Annahme zu, daß saure Polysaccharide an dem Zustandekommen der entzündlichen Reaktion bei der Akne vulgaris in Form von Antigenen beteiligt sind, während prostaglandin-ähnliche Substanzen als eine der potentiellen Mediatoren, die Entwicklung einer Frühreaktion stimulieren.

Effect of Systemic Erythromycin Stearate on the Inflammatory Lesions and Skin Surface Fatty Acids in Acne vulgaris

In a 6-week and double-blind study the efficacy of systemic administered erythromycin stearate and doxycycline was compared in 56 patients with facial acne vulgaris. With both antibiotics a statistically significant improvement was demonstrated without any difference between the two groups. No patient had to withdraw therapy due to side-effects. Another part of this investigation correlates the degree of inflammation in acne vulgaris with the fatty acid level in human skin surface lipids before and after systemic erythromycin stearate therapy. This antibiotic reduced the amount of fatty acids 25--50% after a month of medication. The reduction was correlated with a definite clinical improvement in 14 of the 17 patients.

Studies on the biological effects of deuteriated organic compounds

The antifugal activity of some perdeuteriated fatty acids, with normal chain 11 to 18 carbon atoms, was investigated on common dermatophytes Epidermophyton floccosum, Microsporum canis, Trichophyton mentagrophytes and T. rubrum under in vitro conditions. These studies were performed by dilution technique and with respiratory measurements. Perdeuteriation of some fatty acids increases their inhibitory effect on the dermatophyte growth. Perdeuteriated n-hendecanoic acid proved to be the most active of the substances tested. Possible mechanisms behind the enhanced antifungal activity due to the perdeuteriation of fatty acids are discussed.The present study investigates the antifungal properties of some perdeuteriated fatty acids on dermatophytes in vitro.Heavy water (deuterium oxide, D2O), the simplest deuterium containing compound differs from ordinary water (protium oxide, H2O) in many physical properties. In the same way molecular configuration, ionic equilibrium and vapour pressure of deuteriated substances are different from those possessing protium (hydrogen). Furthermore reaction rates for the rupture of bonds C-D, N-D and O-D are not the same as for those involving C-H, N-H and O-H. Therefore, very soon after the discovery of deuterium, in 1932 (42), Urey (41) and Lewis (32) suggested that these differences in properties might give rise to differences in the chemical and biological activities of deuteriated compounds. Since then, a large number of miscellaneous investigations covering the physical, chemical and biological properties of deuteriated compounds have appeared in the literature (26,27, 29, 31.).Early investigations and many subsequent studies have concluded that substantial replacement of protium by deuterium is incompatible with life (33, 38, 44, 45). While this is true for higher plants and animals which tolerate a maximum of about 35% D2O, it is not valid for e.g. algae, bacteria or fungi. Chroney et al. (4) were the first to report successful growth of the green algae Chorella vulgaris and Scenedesmus obliques in media containing 99.7% D2O with CO2 as the only carbon source. However, adaptation of algae to growth in so high concentration of deuterium oxide is a complex phenomenon illustrated by the occurrence of giant cells (3). Among a large number of microorganisms, even fungi have been studied in order to establish how their growth and morphology are affected by replacement of H2O by D2O in culture media (26).The effect of unlabelled fatty acids on fungi is well known. As long ago as 1899 Clark (5) reported how fatty acids affected mould germination. Also early systematic studies (28) have revealed the importance of chain length in determining the activity of fatty acids. Fatty acids were introduced as therapeutic agents for mycotic infections by Peck et al. (35, 36). These authors showed that human perspiration was protective against infections in general and dermatophytes in particular. They were able to demonstrate, that sweat was fungistatic due to its content of middle chain fatty acids (C10-C12). Since then many investigations have confirmed the therapeutic value of some fatty acids in dermatophytosis (40). However, the therapeutical value of the middle-chain acids, especially n-hendecen-10-oic (undecylenic) acid, is limited.Specific, partially and fully deuteriated organic compounds have been synthesised (7–9, 13–20, 23, 24). Methods of synthesis and biosynthesis for these substances are continuously developed in this laboratory. Most of these compounds have already been utilized for studies with mass spectrometry (1, 6, 9, 10, 18–22), gas-liquid chromatography (14, 16, 17) and infrared spectroscopy (11, 12).

New group of prostaglandin-like compounds in P. acnes

Abstract 1. 1. The lipid moiety of P. acnes contains vasoactive substances, which are prostaglandin-like. A survey of their pharmacological properties on smooth muscle preparations from different tissues, vessels, digestive tract or reproductive organs as well as their effects on cAMP, leucocyte migration and thrombocyte aggregation is presented. 2. 2. Recent structural data from a HPLC purified sample reveal the presence of diagnostic peaks characteristic for the aliphatic chain of prostaglandin family. The fact, that none of the known members of this group matched with the total spectra suggests, that the isolated compounds represent a new type of bacterial prostanoids.

Skin Surface Retention of an Applied Ointment

A routine method using quantitative thin-layer chromatography for the determination of the retention time of an applied ointment on healthy and psoriatic skin is described. The retention time of the o

Erythromycin stearate in acne vulgaris: its effect on the skin surface lipids and on the activity of purified pancreatic lipase.

To study the mechanism of action of erythromycin stearate in acne vulgaris, the composition of the skin surface lipids was analyzed before, during and after treatment (500 mg daily for 1 week, 250 mg daily for 2 months) in 17 patients. Quantitative thin-layer chromatography showed that the fatty acid fraction in the skin surface lipids decreased significantly during the treatment period. The clinical effect was good in 14 of 17 patients. Further investigations were performed on the effect of erythromycin stearate on purified pancreatic lipase. The enzyme activity was not inhibited by exposure to the drug.

Studies on the biological effects of deuteriated organic compounds. III. Antifungal activity of perdeuteriated fatty acids on dermatophytes in vivo experimental microsporie in guinea pigs.

Investigations on the antimycotic properties of perdeuteriated fatty acids were carried out on Microsporum cains infections in vivo. The study was performed on experimental microsporie in guinea pigs using four different methods, all based on the ability of M. canis to cause alopecia. Perdeuteriated n-hendecanoic acid showed in vivo a statistically significant enhanced antimycotic effect compared to its unlabelled analogue. This is in accordance with our previous observations in vitro conditions. The remaining perdeuteriated fatty acids (C12--C18) showed no statistically significant growth retarding effect on M. canis infections in guinea pigs when compared with their unlabelled analogues. The present study attempts to ascertain if some perdeuteriated fatty acids have any antifungal activity in vivo conditions. Our previous papers (3,4) concerning the evaluation of the antifungal activity of some perdeuteriated fatty acids on dermatophytes in vitro, demonstrated that the perdeuteriation of n-hendecanoic acid, lead to a pronounced antimycotic effect on common dermatophytes as e.g. E. floccosum, T. rubrum, M. canis and T. mentagrophytes. As our previous results indicate a decreasing of fungistasis with an increasing carbon-chain length of the perdeuteriated fatty acids, we have attempted to verify this observation in vivo conditions. A special interest was, of course, focused on the most promising compound, the perdeuteriated n-hendecanoic acid.