bluuebunny/crossref_metadata_2025 huggingface dataset:
Applications of Spectral Imaging: Detection and Analysis of Human Melanoma and Its Precursors
Authors: FARKAS DANIEL L. , BECKER DOROTHEA
Published: March 2003

Light‐based imaging has extensive applications for medicine and biology, and recent advances in optical imaging modalities, such as confocal and multi‐photon scanning fluorescence microscopy, bioluminescence, optical coherence tomography, and spectral imaging, have opened new avenues for visualizing and recording over time dynamic changes in genetic, developmental, and disease mechanisms that cannot be captured by conventional light microscopy. In the present article, we focus on spectral imaging, and using human melanoma and its precursor lesions as an example, we describe the ability of spectral imaging to detect early‐stage disease, capture gene expression profiles in tissue specimens, and visualize gene functions in tumors growing in living animals.
UVB‐induced Decrease of p16/CDKN2A Expression in Skin Cancer Patients
Authors: Krähn G. , Leiter U. , Udart M. , Kaskel P. , Peter R. U.
Published: March 2003

The lack of p16 expression has been shown in cultured melanoma cells, however contradictory evidence for p16 expression in melanoma tissues exist. Ultraviolet (UV) C and UVB have been shown to affect p16 expression, which impairs cell cycle regulation in vitro and in vivo. In this study, p16/CDKN2A gene expression was determined by reverse transcription polymerase chain reaction in seven skin cancer patients, in one dysplastic nevus patient and in seven healthy individuals, prior to UVB exposure and at various times after application of one minimal erythema dose (MED). Five of the seven skin cancer patients showed a down‐regulation of p16/CDKN2A expression after UVB exposure, while controls remained unaltered. The UVB‐induced decline of p16/CDKN2A in skin cancer patients might offer new insights into photocarcinogenesis. The putative sequence of events could start with a down‐regulation of p16/CDKN2A expression, which would lead to impaired cell cycle regulation. Altered expression patterns of p16/CDKN2A following UVB exposure could be of value for identifying people with an increased risk of UV‐induced skin cancer.
Fibroblasts Play a Regulatory Role in the Control of Pigmentation in Reconstructed Human Skin from Skin Types I and II
Authors: Hedley Susan J. , Layton Christopher , Heaton Martin , Chakrabarty Kaushik H. , Dawson Rebecca A. , Gawkrodger David J. , Neil Sheila Mac
Published: March 2003

Human melanocytes in monolayer culture are extremely dependent on a wide range of soluble signals for their proliferation and melanogenesis. The advent of three‐dimensional models of reconstructed skin allows one to ask questions of how these cells are regulated within a setting which more closely approximates normal skin. The purpose of this study was to investigate to what extent melanocytes within a reconstructed skin model are sensitive to regulation by dermal fibroblasts, basement membrane (BM) proteins and the addition of α‐melanocyte‐stimulating hormone (α‐MSH). Sterilized acellular de‐epidermized dermis (prepared to retain BM proteins or deliberately denuded of BM by enzymatic treatment) from skin type I or II was reconstituted with fibroblasts, melanocytes and keratinocytes. In all but one case (9/10), cell donors were skin type I or II. The presence of BM antigens was found to be necessary for positional orientation of the melanocytes; in the absence of BM, melanocytes moved into the upper keratinocyte layer pigmenting spontaneously. Addition of fibroblasts suppressed the extent of spontaneous pigmentation of melanocytes within this model. Neither α‐MSH nor cholera toxin induced pigmentation in this model despite the fact that melanocytes clearly had the ability to synthesize pigment.
Morphological Studies on Microfilaments and their Organizing Center in Killifish (Fundulus heteroclitus L.) Melanophores
Authors: Kimler Victoria A. , Palazzolo Katherine L. , Anne Premchand , Haddad Maher M. , Lee James B. , Harkins Carmel , Vallarapu Bharat , Taylor John D.
Published: March 2003

Fish chromatophores serve as excellent study models for cytoskeleton‐dependent organelle translocations because the distribution of pigmentary organelles can be observed against a time frame by microscopy. In this study the distribution of microfilaments along with microtubules in cultured melanophores of the killifish (Fundulus heteroclitus Linneaus) are examined using whole‐cell transmission electron microscopy (WCTEM), fluorescence, and laser scanning confocal microscopy. Dispersing, dispersed, aggregating and aggregated states of pigment are induced by adding either caffeine (for dispersion) or epinephrine (for aggregation) to the cells in a standard culture medium. The cells that exhibited a random melanosome distribution in the standard culture media without these two reagents, served as the control. The results indicate that: (i) a structure considered to be the actin‐filament organizing center (AFOC) is in close proximity to the microtubule‐organizing center (MTOC); (ii) the radial layout of microfilaments remains similar over four physiological states of pigmentary response with the exception of epinephrine‐aggregated pigment, in which the aggregate blocks the viewing of the AFOC and central microfilament rays, yet radial microfilaments, whether central and/or peripheral, are apparent in all physiological states of distribution; and (iii) microfilaments serve, together with microtubules, as scaffolding for melanosomes which migrate in bi‐directional rows on cross‐bridges, thus shedding light on the mechanisms for orderly melanosome translocations in a structural continuum.
Approaches to Repigmentation of Vitiligo Skin: New Treatment with Ultrasonic Abrasion, Seed‐Grafting and Psoralen Plus Ultraviolet A Therapy
Authors: Tsukamoto Katsuhiko , Osada Atsushi , Kitamura Reiko , Ohkouchi Masayuki , Shimada Shinji , Takayama Osami
Published: March 2003

Vitiligo vulgaris is a common disease throughout the world although its pathogenesis is not yet known. The most frequent treatment used for vitiligo is PUVA (psoralen plus ultraviolet A) and topical steroids but against stable refractory vitiligo, various other surgical techniques have been developed such as autografting, epidermal grafting with suction blisters, epithelial sheet grafting, and transplantation of cultured melanocytes. We have discovered a new method using ultrasonic abrasion, seed‐grafting and PUVA therapy. The ultrasonic surgical aspirator abrades only the epidermis of recipient sites. This easily and safely removes only the epidermis, even on spotty lesions or intricate regions which are difficult to remove using a conventional motor‐driven grinder or liquid nitrogen. Epidermal seed‐grafting can cover more area than sheet‐grafting, and subsequent PUVA treatment can enlarge the area of pigmentation with coalescence of adjacent grafts. In this article, we provide a general overview of the current surgical therapies including our method for treating stable refractory vitiligo.
In Vitro Migration of Melanoblasts Requires Matrix Metalloproteinase‐2: Implications to Vitiligo Therapy by Photochemotherapy
Authors: Lei Tie Chi , Vieira Wilfred D. , Hearing Vincent J.
Published: March 2003

It is known that the migration of melanocyte precursors (melanoblasts) from the outer root sheath of hair follicles into clinically depigmented epidermis is crucial to the repigmentation of vitiliginous skin treated with photochemotherapy (PUVA), but such migratory cells must penetrate extracellular matrix tissue barriers in vivo. To test the hypothesis that matrix metalloproteinases (MMPs) are required for this process, we determined whether cultured melb‐a cells, an immortal line of melanoblasts isolated from neonatal mouse epidermis, express and secrete MMPs and whether a synthetic metalloproteinase inhibitor, GM6001 (Galardin), inhibits their migratory behavior in vitro. Reverse transcriptase‐polymerase chain reaction and Western blotting were used to determine the patterns of MMP expression by melanoblasts at the mRNA and protein levels, respectively. The proteolytic activities of MMPs secreted into the culture medium were assessed by gelatin zymography. The capacity of melanoblasts to migrate on fibronectin, laminin or laminin‐5 substrates was estimated using Transwell migration assays. The results show that MMP2, MMP9 and MT1‐MMP transcripts are expressed by these melanoblasts, but only MMP2 is secreted and activated in the extracellular environment. Although the therapeutic efficacy of PUVA in stimulating repigmentation of vitiliginous skin might derive from direct effects of UVA and/or 8‐methoxypsoralen (8MOP), recent studies have shown that keratinocyte‐derived factors induced by ultraviolet radiation, especially α‐melanocyte stimulating hormone (αMSH), play a major role in regulating melanocyte function. Therefore, we also examined whether 8MOP and/or αMSH are involved in the up‐regulation of MMP2 expression in melanoblasts. Western blotting and zymographic analyses revealed that MMP2 synthesis and secretion were induced by 8MOP and/or by αMSH. This induction of MMP2 resulted in significant increases of migration by melanoblasts on laminin or on laminin‐5 substrates, while concomitant treatment with GM6001 blocked that induced migration. Taken together, these results suggest the importance of MMP2 in melanoblast migration and in the response to PUVA therapy.
Maxadilan Activates PAC1 Receptors Expressed in Xenopus laevis Melanophores
Authors: Pereira Phyllis , Reddy Vermuri B. , Kounga Kounga , Bello Ysabel , Lerner Ethan
Published: March 2003

Functional interactions between ligands and their cognate receptors can be investigated using the ability of melanophores from Xenopus laevis to disperse or aggregate their pigment granules in response to alterations in the intracellular levels of second messengers. We have examined the response of long‐term lines of cultured melanophores from X. laevis to pituitary adenylate cyclase activating peptide (PACAP), a neuropeptide with vasodilatory activity, and maxadilan, a vasodilatory peptide present in the salivary gland extracts of the blood feeding sand fly. Pituitary adenylate cyclase activating peptide increased the intracellular levels of cyclic adenosine monophosphate (cAMP) and induced pigment dispersion in the pigment cells, confirming that melanophores express an endogenous PACAP receptor. Maxadilan did not induce a response in non‐transfected melanophores. When the melanophores were transfected with complementary DNA (cDNA) from the three different members of the PACAP receptor family, maxadilan induced pigment dispersion specifically and cAMP accumulation in melanophores transfected with the cDNA for PAC1 receptors but not VPAC1 or VPAC2 receptors. A melanophore line was generated that stably expresses the PAC1 receptor.
Co‐Culture of Mouse Epidermal Cells for Studies of Pigmentation
Authors: Yoon Tae‐Jin , Hearing Vincent J.
Published: March 2003

Interactions between melanocytes and keratinocytes in the skin suggest bi‐directional interchanges between these two cell types. Thus, melanocytes cultured alone may not accurately reflect the physiology of the skin and the effects of physiological regulators in vivo, because they do not consider possible interactions with keratinocytes. As more and more pigment genes are identified and cloned, the characterization of their functions becomes more of a challenge, particularly with respect to their roles in the processing and transport of melanosomes and their transfer to keratinocytes. Immortalized melanocytes mutant at these loci are now being routinely generated from mice, but interestingly, successful co‐culture of murine melanocytes and keratinocytes is very difficult compared with their human counterparts. Thus, we have now optimized co‐culture conditions for murine melanocytes and keratinocytes so that pigmentation and the effects of specific mutations can be studied in a more physiologically relevant context.
Mixed infection of Porphyromonas gingivalis and Bacteroides forsythus in a murine abscess model: involvement of gingipains in a synergistic effect
Authors: Yoneda Masahiro , Hirofuji Takao , Anan Hisashi , Matsumoto Akiko , Hamachi Takafumi , Nakayama Koji , Maeda Katsumasa
Published: March 2003

Several microorganisms including Porphyromonas gingivalis and Bacteroides forsythus have been implicated to be etiologically important agents of periodontal disease. In this study, we determined the ability of combinations of periodontopathogenic microorganisms to cause tissue destruction in a murine abscess model. Although all bacterial combinations used in this study produced larger abscesses than did monoinfection of each bacterium, the combination of P. gingivalis and B. forsythus showed a synergistic effect on abscess formation. Since these two bacteria have been frequently found together in lesions of periodontitis, these results suggest the significance of their co‐infection in the progression of periodontitis. P. gingivalis produces extracellular and cell‐associated cysteine proteinases (gingipains) which appear to be involved in its virulence. The rgpA rgpB double and kgp mutants induced significantly smaller abscesses than the wild type. Moreover, the rgpA rgpB kgp triple (gingipain‐null) mutant hardly showed lesion formation at all with the experimental conditions used in this study, indicating that these genes encoding gingipains are important for virulence of P. gingivalis. Mixed infection of these P. gingivalis mutants with B. forsythus showed an additive effect on abscess formation, indicating that the gingipains of P. gingivalis may play an important role in the pathological synergism between P. gingivalis and B. forsythus.
Chewing sticks: timeless natural toothbrushes for oral cleansing
Authors: Wu C. D. , Darout I. A. , Skaug N.
Published: March 2003

It is generally accepted that oral hygiene maintenance through regular removal of dental plaque and food deposits is an essential factor in the prevention of dental caries and periodontal disease. Methods for oral hygiene vary from country to country and from culture to culture. Despite the widespread use of toothbrushes and toothpastes, natural methods of tooth cleaning using chewing sticks selected and prepared from the twigs, stems or roots from a variety of plant species have been practised for thousands of years in Asia, Africa, the Middle East and the Americas. Selected clinical studies have shown that chewing sticks, when properly used, can be as efficient as toothbrushes in removing dental plaque due to the combined effect of mechanical cleaning and enhanced salivation. It has also been suggested that antimicrobial substances that naturally protect plants against various invading microorganisms or other parasites may leach out into the oral cavity, and that these compounds may benefit the users by protection against cariogenic and periodontopathic bacteria. Some clinical epidemiological studies are in support of this, and many laboratory investigations have suggested the presence of heterogeneous antimicrobial components extractable using different chemical procedures. A few recent studies have identified some of the active antimicrobial compounds. Today, chewing sticks are still used in many developing countries because of religion and/or tradition, and because of their availability, low cost and simplicity. The World Health Organization also encourages their use. The Year 2000 Consensus Report on Oral Hygiene states that chewing sticks may have a role to play in the promotion of oral hygiene, and that evaluation of their effectiveness warrants further research.
T‐cell‐receptor gene usage of Actinobacillus actinomycetemcomitans‐reactive periodontal CD4+ T cells from localized juvenile periodontitis patients and human peripheral blood leukocyte‐reconstituted NOD/SCID mice
Authors: Gao Xuijuan , Teng Yen‐Tung A.
Published: March 2003

We investigated the variable Vα and Vβ gene usage of Actinobacillus actinomycetemcomitans‐reactive periodontal CD4+ T cell receptors (TCR) from: (i) four A. actinomycetemcomitans‐infected localized juvenile periodontitis (LJP) patients, (ii) four groups of A. actinomycetemcomitans‐inoculated NOD/SCID mice engrafted with individual LJP‐derived HuPBL and (iii) HuPBL samples of four LJP patients and two healthy control subjects, by quantitative PCR analyses. The results show that: (i) the majority of the TCR genes (82.5% of Vα and 91.1% of Vβ) used by periodontal CD4+ T cells in A. actinomycetemcomitans‐inoculated HuPBL‐engrafted NOD/SCID mice overlap with those used by local periodontal T cells in LJP patients, (ii) although A. actinomycetemcomitans‐reactive periodontal CD4+ TCR repertoire is relatively widespread, there are a few dominant genes shared by the LJP patients, suggesting a limited number of antigens or epitopes commonly recognized and (iii) A. actinomycetemcomitans likely lacks superantigenic characteristics. These results suggest A. actinomycetemcomitans‐associated human CD4+ T cell repertoire established in HuPBL‐NOD/SCID mice provides a useful approach to study specific aspects of immune–parasite interactions in the periodontium.
Alterations of neutrophil f‐actin kinetics by tobacco smoke: implications for periodontal diseases
Authors: Ryder Mark I. , Wu Timothy C. , Kallaos Steven S. , Hyun William
Published: March 2003

Tobacco smoking is a major risk factor in the incidence and severity of periodontal diseases. Alterations of neutrophil function by short‐term high levels of smoke during the act of smoking (acute smoke exposure) as well as long‐term exposure to lower levels of tobacco substances in the bloodstream (chronic smoke exposure) may play a role in the pathogenesis of periodontal diseases in smokers. The polymerization and depolymerization of f‐actin in response to infectious agents or inflammatory mediators is a critical process in a variety of neutrophil functions. In this study, we examined the effects of in vitro smoke exposure on neutrophils from smokers and non‐smokers (which may be comparable to in vivo acute smoke exposure) and neutrophils from smokers not exposed to further in vitro smoke (which may be comparable to chronic smoke exposure) on f‐actin kinetics. Peripheral neutrophils were isolated from seven healthy smoking subjects and seven healthy age‐matched non‐smoking subjects and exposed to 1–5 min of acute smoke in a smoke box system or not exposed to further smoke (baseline controls). Selected aliquots of neutrophils from control and 5‐min exposures of acute smoke were then stimulated with the chemotactic peptide F‐met‐leu‐phe at 10−7m for an additional 30–360 s. Cells were fixed and permeabilized, stained for f‐ actin with NBD phallacidin, and analyzed by flow cytometry. From baseline to 5 min of invitro smoke exposure, there was a 38% decline in f‐actin stain in non‐smokers and a 30% decline in f‐actin stain in smokers ( p > 0.05) with f‐actin values slightly higher in smokers than‐non‐smokers ( p > 0.05). With F‐met‐leu‐phe stimulation, both smokers and‐non‐smokers demonstrated a characteristic rise in f‐actin stain from 0 to 120 s with a subsequent decline to baseline at 360 s and no significant differences in f‐actin levels at any time of stimulation between groups. After preincubation with 5 min of in vitro smoke, the magnitude of rise in f‐actin was less in both smokers and non‐smokers when compared to cells not incubated with 5 min of smoke ( p < 0.05 at 120 s for both smokers and non‐smokers). F‐actin values in smokers were higher than‐non‐smokers from 30 to 360 s of F‐met‐leu‐phe exposure ( p > 0.05). These results demonstrate that in vitro smoke exposure may impair normal f‐actin kinetics. These alterations in f‐actin kinetics may in turn affect other neutrophil functions which may impact on the pathogenesis of periodontal diseases in smokers.
Simple and rapid detection of serum antibody to periodontopathic bacteria by dot blotting
Authors: Kawashima Yoko , Shikawa Isao I
Published: March 2003

The aim of this study was to detect the specific immunoglobulin G antibodies against periodontopathic bacteria by dot blotting. In the procedure used, bacterial preparations were blotted on a nitrocellulose membrane. After blocking the non‐specific binding sites, the diluted serum was blotted onto the preparations. The membrane was immersed in secondary antibodies and then in substrate buffer. The colored blots were then evaluated. To test the reliability of this procedure, 20 serum samples were examined for antibody: ten for anti‐Porphyromonas gingivalis antibody, and the other ten for anti‐Actinobacillus actinomycetemcomitans antibody. Five samples out of each set of ten had previously been confirmed as having high enzyme‐linked immunosorbant assay (ELISA) titers to the antigen, while the other five had been confirmed as having average titer levels. Both whole‐cell sonic extracts and fimbriae of P. gingivalis were used as antigens in the dot blotting, in order to compare their use as antigens in assays of the patients' sera. ELISA was also used to measure anti‐P. gingivalis antibody titers. For the measurement of IgG antibodies against A. actinomycetemcomitans, formalin‐killed whole cells were used. Fifty serum samples were examined for IgG antibodies against A. actinomycetemcomitans by dot blotting and ELISA. With both antigens, after 4 h, coloration of blots was more clearly visible for the high‐titer sera than for the average‐titer sera. The intensity of coloration of the blots for P. gingivalis and A. actinomycetemcomitans showed correlation with the ELISA titers. A particularly significant correlation was shown when P. gingivalis fimbriae were used as antigen. These results suggest that this dot blot method is a simple and rapid means of detection of serum antibodies, and that it shows promise as a chair‐side assay method.
Determination of eight selected periodontal pathogens in the subgingival plaque of maxillary first molars in Japanese school children aged 8–11 years
Authors: Suda Reiko , Kurihara Chikako , Kurihara Masaki , Sato Taisuke , Lai Chern‐Hsiung , Hasegawa Kohji
Published: March 2003

To prevent the onset or progression of periodontitis, we must understand when periodontal pathogens are first harbored and how they develop the biofilm that causes periodontal disease. The purpose of this study was to determine the relationship between clinical status and selected periodontal pathogens in subgingival plaque in school children. This study was conducted with 95 school children, 8–11 years old. The presence and distribution of eight selected periodontal pathogens sampled from the maxillary right first molar were determined by an indirect immunofluorescent technique and compared with clinical parameters. Of the 95 sites sampled, only one site had all eight pathogens and five sites did not have any of the eight pathogens. The mean number of positive pathogens per site was 3.5 ± 1.8 and mean percentage of positive pathogens was 3.82 ± 4.22%. The number and total percentage of positive pathogens were strongly correlated with the Plaque Index (PI). In addition, the number of positive pathogens was correlated to the presence of subgingival calculus. The most frequently found pathogens were Campylobacter rectus (84.2%) and Eikenella corrodens (83.2%), and the least, Actinobacillus actinomycetemcomitans serotype c (7.4%). Of the eight pathogens, the frequency and distribution of Porphyromonas gingivalis were significantly correlated with PI and the presence of calculus. In addition, seven sites with both P. gingivalis and Bacteroides forsythus showed a correlation with gingival inflammation. In conclusion, the presence of P. gingivalis or P. gingivalis and B. forsythus may be a risk marker to be sought in screening for the onset of periodontal disease.
The Role of Myocardial KATP‐Channel Blockade in the Protective Effects of Glibenclamide against Ischaemia in the Rat Heart
Authors: Legtenberg Roger J. , Rongen Gerard A. , Houston Ralph J. F. , Oeseburg Berend , Smits Paul
Published: March 2003

Abstract: Glibenclamide preserves postischaemic myocardial function in the isolated, erythrocyte perfused, working rat heart model. This study addresses the possible involvement of KATP channels in this beneficial action of glibenclamide. We hypothesized that if glibenclamide improved postischaemic cardiac function by blocking of KATP channels, opening of these KATP channels should result in the opposite, namely detrimental effects on postischaemic heart function. Postischaemic functional loss and coronary blood flow were recorded during treatment with glibenclamide (4 μmol.l−1; n=5), the KATP channel openers pinacidil (1 μmol.l−1; n=5) and diazoxide (30 μmol.l−1; n=5), the combination of glibenclamide with pinacidil (n=5) and glibenclamide with diazoxide (n=5), and vehicle (n=8). Both pinacidil and diazoxide significantly increased coronary blood flow 2–3 times, which was abolished by glibenclamide pre‐ and postischaemically. This confirms that under both flow conditions glibenclamide significantly blocks KATP channels in the coronary vasculature. The 12 min. global ischaemic incident resulted in a cardiac functional loss of 22.2±2.9% during vehicle. Glibenclamide reduced the cardiac functional loss to 4.3±1.2% (P<0.01). Interestingly, both pinacidil and diazoxide reduced the cardiac functional loss to 4.0±1.5% (P<0.01) and 2.9±1.4% (P<0.001), respectively. The combination pinacidil+glibenclamide resulted in additional protection compared with the individual components (0.6±0.1 versus 4.0±1.5%, P<0.05). Thus, in contrast to its effect on coronary vascular tone, the glibenclamide‐induced improvement of postischaemic cardiac function may not be mediated through blockade of the KATP channel. Alternative mechanisms may be operative, such as uncoupling of the mitochondrial respiratory chain, thereby preconditioning the hearts against stunning.
Cellular Localization of Cholecystokinin Receptors as the Molecular Basis of the Periperal Regulation of Acid Secretion
Authors: Schmidt Wolfgang E. , Schmitz Frank
Published: March 2003

Abstract: Gastrin stimulates gastric acid secretion through direct activation of CCK‐B/gastrin receptors on parietal cells and indirectly through release of histamine from ECL cells. Cholecystokinin (CCK) is structurally closely related to gastrin and shares high affinity for CCK‐B/gastrin receptors. In contrast to gastrin, CCK also recognizes CCK‐A receptors. While CCK appears to be a negative regulator of gastric acid secretion and postprandial release of gastrin in the normal human gastrointestinal tract, its impact on the pathogenesis of acid hypersecretion in Helicobacter pylori‐infected individuals remains uncertain. This article will review the endocrine and paracrine regulatory pathways which are activated by CCK/gastrin peptides and which appear relevant in the pathogenesis of peptic ulcer disease in man.
The kinetics of the tanning response to tanning bed exposures
Authors: Caswell M.
Published: March 2003

No clinical studies have been published documenting the development of pigmentation following the use of the Food and Drug Administration (FDA) recommended exposures from a tanning bed. Panelists were exposed three times weekly for eight weeks (24 exposures) using the FDA recommended exposure schedule. The initial tan was noted after only six exposures and quantitatively increased through the remainder of the study.
Five cases of photocontact dermatitis due to topical ketoprofen:photopatch testing and cross‐reaction study
Authors: Matsushita Tetsuya , Kamide Ryoichi
Published: March 2003

Background: In parallel with the popular usage of topical ketoprofen, the number of reported cases of ketoprofen‐induced photoallergic contact dermatitis has been increasing. It is clinically important to know the cross‐reactivity of ketoprofen in order to avoid cross‐sensitization caused by several structurally similar non‐steroidal anti‐inflammatory drugs (NSAID) on the market.

Methods: To evaluate the spectrum of cross sensitization, photopatch testing was performed on five patients with ketoprofen‐induced photoallergic contact dermatitis using ketoprofen and other structurally similar chemicals, such as oxybenzone, tiaprofenic acid and suprofen.

Results: All five patients reacted positively to ketoprofen or ketoprofen plaster on photopatch testing. All four patients photopatch tested with related chemicals showed cross‐photosensitization with tiaprofenic acid and suprofen. However, none of the patients reacted positively to oxybenzone.

Conclusion: Either the diphenylketone moiety or a structurally similar tiophene‐phenylketone moiety is important as the antigenic determinants of ketoprofen photoallergy. The arylpropionic acid side chain would not be involved.
Comparison between seasons of the ultraviolet environment in the shade of trees in Australia
Authors: Parisi A. V. , Wong J. C. F. , Kimlin M. G. , Turnbull D. , Lester R.
Published: March 2003

Background/Purpose: This paper has considered the erythemal UV (UVery), UVA and visible irradiances in the shade of Australian trees for each season at a sub‐tropical southern hemisphere site.

Methods: The irradiances in tree shade have been measured with radiometers as a percentage of the irradiances in the sun for each season of the year.

Results: Although the solar irradiances are lower in winter, the percentages of the UV in tree shade compared to the UV in full sun are marginally higher (by up to 7%) in the winter compared to summer. The range of percentages for UVery was up to double that of the percentages of the visible waveband. The percentages for UVery were also higher than for the UVA waveband. The percentages of the irradiances in the tree shade compared to full sun are 8–14% lower at noon compared to the morning and afternoon for the UVery waveband. The ratio of UVA to UVery is lower in the tree shade compared to the full sun.

Conclusions: The UVA to UVery ratio is expected to be even lower in the tree shade as a result of ozone depletion. This, combined with the visible irradiances in the tree shade not being a reliable indication of the biologically damaging UV irradiances, has consequences for public health and skin cancer prevention.
001 UVA‐1 for the treatment of chronic hand dermatitis
Authors: Bergamo B. , Elmets C.
Published: March 2003

UVA‐1 has distinct immunological effects which has allowed it to be useful for a number of cutaneous diseases that have been relatively resistant to other treatment modalities. These include urticaria pigmentosa, morphea/scleroderma and T‐cell mediated diseases such as atopic dermatitis and cutaneous T‐cell lymphoma. Since chronic hand dermatitis is also mediated by T‐cells, we hypothesized that UVA‐ 1 would be an effective treatment for that disease as well. Five subjects with recalcitrant hand dermatitis caused by dyshidrosis and/or chronic irritancy were treated with a medium dose regimen of UVA‐ 1. This consisted of 15 treatments of 60 J/cm2 UVA 1 over 3 weeks. Subjects received a total of 900 J/cm2. A clinical scoring parameter, Dyshidrotic Area and Seventy Index Score (DASI), was administered to subjects prior to and following the UVA‐ 1 treatment course. Clinical photographs were also taken. All subjects showed improvement in hand dermatitis as demonstrated by clinical photographs and the DASI score (13 ± 5.35 before treatment; 6.3 ± 4.50 after treatment). UVA‐ 1 is an effective and rapid acting treatment for chronic hand dermatitis.
012 Photodynamic therapy of non‐melanoma skin cancers with verteporfin and red light – tumor response and cosmetic outcome
Authors: Lui H. , Hobbs L , Tope W. , Elmets C. , Provost N. , Lee P. , Herne K. , Mclean D. , Hamzavi I. , Jain H. , Bissonnette R.
Published: March 2003

Introduction and objectives: Photodynamic therapy (PDT) with verteporfin may be particularly well suited for patients with multiple low risk non‐melanoma skin cancers (NMSC) while providing good cosmetic outcomes as compared to standard treatments. The objectives of this study were to prospectively assess the tumor responses and cosmetic outcomes of verteporfin‐treated NMSC.

Patients and methods: This was a phase 11, open‐label, light‐dose ranging, multicenter study of 54 patients with multiple NMSC. A total of 421 biopsy‐proven tumors that were either basal cell carcinomas or in situ squamous cell carcinomas underwent PDT with intravenous verteporfin 14 mg/m2 followed by exposure to red light from LED diode arrays (658‐718 nm FWHM) at fluences of 60, 120, or 180 J/cm2. Each patient was randomly assigned to receive one of the three light doses to all their tumors. Treated tumors underwent follow up biopsies at 6 months after the initial PDT session to determine the pathologic complete response rate. In addition, the treated tumors were assessed clinically for up to 24 months. The cosmetic outcome of each treated tumor site was assessed by the investigators based on color, profile, and surface texture.

Results: The pathologic complete response rates at 6 months were 69, 79, and 93% for the 60, 120 and 180 J/cm2 light fluences, respectively. The clinical complete response rates by tumor at 6 months were 78, 89 and 98% at 60, 120 and 180 J/cm2, respectively, while at 24 months, the corresponding rates were 51, 79, and 95%. The cosmetic outcome by tumor at 24 months judged by the investigator to have achieved at least a satisfactory or higher outcome was 92, 76, and 86% at 60, 120 and 180 J/cm2, respectively.

Conclusions: PDT of NMSC with verteporfin provides effective tumor clearing that is dose‐dependent with satisfactory to excellent outcomes in the majority of patients.
005 Topical photodynamic therapy in dermatology – 3 years experience at Beaumont Hospital
Authors: Collins S. , Ahmadi S. , Murphy G.M.
Published: March 2003

Topical photodynamic therapy (PDT) involves selective photosensitization of a target tissue by means of a topical agent, which is then activated by light to effect an oxygen dependent cytotoxic reaction. Premalignant and malignant skin conditions including actinic keratosis, squamous cell carcinoma in situ (Bowen's disease), and superficial basal cell carcinoma (BCC) have been effectively treated with PDT. A number of studies support the use of topical PDT in the treatment of acne, warts, psoriasis and T‐cell lymphoma. There are many case reports and small series, with varying results, in a wide variety of other non‐malignant applications.

We report our experience of topical PDT over a 3 year period. We selected patients to participate in an open study of ALA‐PDT on the basis of failure of, or unsuitability for conventional therapy. One hundred and ten patients participated in the study, with a mean age of 73 years. One hundred and twenty‐nine lesions were treated, which consisted of 95 lesions of Bowen's disease, 21 superficial BCC's, nine patients with multiple actinic keratosis and four with porokeratosis. Patients were assessed at 3, 6 and 12 month intervals.

Clearance and recurrence rules, adverse events and patient preference (where lesions had been treated previously) were recorded, Clearance rates for actinic keratosis, Bowen's disease and BCC were 87.5%, 84%, and 57%, respectively. Seventy‐four percent of patients had a single treatment. Fourteen percent had two, and the remainder had up to four treatements in total.

Though topical PDT has not proven to be superior to conventional methods, it is well tolerated with excellent cosmetic results. It is of particular value in the treatment of large lesions, in those with poorly vascularised skin, and in areas of diffuse actinic damage. Careful patient selection, prior debulking of lesions, and newer more selective agents with enhanced penetration, promise to further improve outcome.
009 Skin aging and photoaging effects can be quantified in vivo by fluoresence excitation spectroscopy
Authors: Kolias N. , Stamatas G.
Published: March 2003

Fluoresence excitation spectroscopy of skin is a noninvasive method for objective evaluations of skin aging and photoaging. Fluoresence bands in the UV have been assigned to tryptophan moieties, pepsin digestible collagen cross‐links (PDCCL), collagenase digestible collagen cross‐links (CDCCL) and elastin cross‐links. We have studied the dependence of these bands to aging and photoaging in the hairless mouse model and in humans. In the mouse, gluorescence of the tryptophan moieties decreases linearly with age, while the fluorescence band due to PDCCL increases linearly with age. In contrast, chronically UVB exposed mice showed an increase in the tryptophan band and a dramatic reduction of the PDCCL band compared to age‐matched controls. The same phenomena were observed on mouse skin immediately after UVA exposure. The dependence of fluorescence of human facial skin with aging was studied in a large population sample (> 500 people in total) ages 15–70 at five geographic locations. Similar to the mouse model we observed a decrease in the tryptophan fluorescence signal, which is probably related to the reduction of the cell turnover rate with aging. Furthermore, observed increases in the fluorescence signals corresponding to collagen and elastin cross‐links with aging may be attributed to chronic accumulation of cross‐links in these long‐lived molecules. The fluorescence ratio of the elastin cross‐links signal to that of tryptophan moieties correlates strongly with age and is independent of geographical region and seasonal effects. The same fluorescence ratio has been used to monitor the antiaging effects of retinol treatment.
Nocturnal 6‐hydroxymelatonin sulfate excretion in female workers exposed to magnetic fields
Authors: Juutilainen Jukka , Stevens Richard G. , Anderson Larry E. , Hansen Norman H. , Kilpeläinen Marko , Kumlin Timo , Laitinen Jarmo T. , Sobel Eugene , Wilson Bary W.
Published: March 2003

The objective of this study was to determine whether daytime occupational exposure to extremely low frequency magnetic fields (MFs) suppresses nocturnal melatonin production. Sixty female volunteers were recruited. Thirty‐nine worked in a garment factory, and 21 office workers served as a reference group. Exposure assessment was based on the type of sewing machine used and MF measurements around each type of machine. Eye‐level MF flux density was used to classify the operators to higher (>1 μT) and lower (0.3–1 μT) exposure categories. A third group of factory workers had diverse MF exposures from other sources. The reference group had average exposure of about 0.15 μT. Urine samples were collected on Friday and Monday for three consecutive weeks. Melatonin production was assessed as urinary 6‐hydroxymelatonin sulfate (6‐OHMS) excretion. The ratio of Friday morning/Monday morning 6‐OHMS was used to test the hypothesis that melatonin production is suppressed after 4 days of occupational MF exposure with significant recovery during the weekend. Possible chronic suppression of melatonin production was evaluated by studying exposure‐related differences in the Friday values by multivariate regression analysis. The Monday/Friday ratios were close to 1.0, suggesting that there is no increase in melatonin production over the weekend. The average 6‐OHMS excretion on Friday was lower among the factory workers than in the reference group, but no monotonous dose–response was observed. Multivariate regression analysis identified MF exposure, smoking, and age as significant explanatory variables associated with decreased 6‐OHMS excretion.
Prolonged swim‐test immobility of serotonin N‐acetyltransferase (AANAT)‐mutant mice
Authors: Uz Tolga , Manev Hari
Published: March 2003

Serotonin N‐acetyltransferase (AANAT; EC 2.3.1.87) metabolizes serotonin into N‐acetylserotonin (NAS). AANAT mRNA is expressed in the pineal gland and retina, and also in the rat brain. It was proposed that NAS could be a mediator of the antidepressant action of drugs, and it was shown that chronic but not acute treatment of rats with the antidepressant fluoxetine increases the content of AANAT mRNA in the rat hippocampus. Consequently, AANAT deficiency might be involved in the pathobiology of depression. C57BL/6J mice have a mutant AANAT gene and are considered AANAT‐deficient, i.e., “knocked down” (compared with their normal counterparts, C3H/HeJ mice). In this study, we investigated whether AANAT mRNA is expressed in the brain of C57BL/6J and C3H/HeJ mice and whether those mice differ behaviorally, i.e., in a forced swimming test which is used to evaluate antidepressant drugs (such drugs shorten the time of immobility). We found that C3H/HeJ mice express in the brain normal AANAT mRNA, whereas C57BL/6J mice express mutated AANAT mRNA. The mutant, AANAT knocked down C57BL/6J mice displayed significantly longer times of immobility (“depression”). This difference was evident regardless of the circadian rhythm, i.e., both during the day and in the dark at night. Further studies are needed to fully characterize the behavioral significance of AANAT mutation and its possible link to depression.
Pineal control of the dopamine D2‐receptor gene and dopamine release in the retina of the chicken and their possible relation to growth rhythms of the eye
Authors: Ohngemach S. , Feldkaemper M. , Schaeffel F.
Published: March 2003

Retinal dopamine (DA) and the DA D2‐receptor have been implicated in the development of “deprivation myopia”, induced by frosted eye occluders. We have studied the changes in D2‐mediated dopaminergic transmission in the retina, their possible relations to eye growth rhythms and myopia, and their control by the pineal gland. (1) We found that the sensitivity of eye growth to retinal image degradation varied over the day. Intermittent periods of normal vision inhibited deprivation myopia more if they occurred in the evening than in the morning. (2) Diurnal growth rhythms in both eyes interacted even though it was previously shown that both deprivation myopia and the accompanying changes in retinal DA release can be monocularly induced. (3) The D2‐receptor mRNA concentration in the retina showed no systemic diurnal changes and was not affected by deprivation myopia, but was increased after 2 days in darkness. Since DA release varies over the day, the gain of dopaminergic transmission may also vary, which could explain the observation described in (1) above. (4) Depletion of retinal DA by intravitreal application of reserpine, which lowers DA content severely, had little effect on D2‐receptor mRNA concentration. (5) Selective illumination of the pineal gland reduced the D2‐receptor mRNA content in the retina to a similar level to full illumination, indicating that the pineal gland controls the D2‐receptor mRNA content in the retina. The pineal also controlled DA release in the retina. These results show that the pineal has a surprisingly large influence on both the retinal DA receptor gene transcription and DA release. It can probably control the gain of dopaminergic transmission in the retina and deprivation myopia and mediate the interactions of the growth rhythms in both eyes.
Daytime 50 Hz magnetic field exposure and plasma melatonin and urinary 6‐sulfatoxymelatonin concentration profiles in humans
Authors: Crasson M. , Beckers V. , Pequeux Ch. , Claustrat B. , Legros J.J.
Published: March 2003

Concern about the health effects of extremely low frequency (ELF) magnetic fields (MF) has been raised by epidemiological studies indicating an association between certain cancers and living near power lines or working in high electric field environments. Alterations in pineal function have been proposed as a mechanism through which power‐frequency MFs may interact with living organisms. A double blind laboratory study was performed to evaluate daytime exposure effects of 100 μT root mean square (rms) 50 Hz MF. Three head exposure sessions of 30 min each were performed: sham, continuous, and intermittent (15 s on/off cycles) MFs were presented to each subject in early or late afternoon (13:30 or 16:30 hr). Twenty‐one healthy male volunteers (20–27 yr old) participated in these 3‐weekly experimental conditions. Blood samples were drawn for serum melatonin measurement, hourly at night (from 20:00 to 07:00 hr) under controlled environmental conditions. Urinary excretion of 6‐sulfatoxymelatonin (aMT6s), the main melatonin metabolite, was measured for a 17 hr period, by means of urine samples taken at 19:00 hr (14:00–19:00 hr “afternoon period”), 23:00 hr (19:00–23:00 hr “evening period”), and 07:00 hr, day 2 (23:00–07:00 hr day 2 “night‐time period”). There were no significant differences in either plasma melatonin or in aMT6s excretion profiles in the three experimental conditions. However, a tendency for a smaller increase of night‐time urinary aMT6s after continuous MF exposure was found (P=0.08) particularly in men with the lower excretion rate of aMT6s (“Low Group”) (P=0.07). We conclude that this study does not indicate that daytime acute MF exposure influences either melatonin secretion or aMT6s excretion. Inter‐individual differences in pineal production of melatonin, however, have to be taken into account in further studies.
How important is stimulation of α‐adrenoceptors for melatonin production in rat pineal glands?
Authors: Tobin V. A. , McCance I. , Coleman H. A. , Parkington H. C.
Published: March 2003

The objective of this study was to determine the role of α‐adrenoceptors in melatonin production by rat pineal gland. Pineal glands were isolated from adult male rats and maintained in organ baths. The perfusate was sampled every 5 min, stored, and later assayed for melatonin. Exposure to norepinephrine (10 μM) or the β‐adrenoceptor agonist orciprenaline (2–10 μM) increased the glands' production of melatonin. The time courses of melatonin production in response to these agonists were unaffected by the rats' pretreatment in vivo with the α‐adrenoceptor antagonist prazosin (2 mg/kg i.p., three times). Rats that had had their superior cervical ganglia removed were primed with either orciprenaline (2 mg/kg i.p) or both orciprenaline and phenylephrine (1 mg/kg i.p) 1 hr before decapitation. Exposure of the pineal glands from these rats to orciprenaline evoked melatonin release that was similar in each group. These results lend weight to the suggestion that the marked potentiation by α‐adrenoceptor agonists of the stimulation of cAMP and N‐acetyltransferase (NAT) by β‐adrenoceptor agonists, demonstrated most readily in cultured glands or dispersed rat pinealocytes, does not carry over into significant augmentation of melatonin production in intact pineal glands.
Altered glial fibrillary acidic protein content and its degradation in the hippocampus, cortex and cerebellum of rats exposed to constant light: reversal by melatonin
Authors: Baydas Giyasettin , Reiter Russel J. , Nedzvetskii Viktor S. , Nerush Peter A. , Kirichenko Svetlana V.
Published: March 2003

Abstract: Reactive astrocytosis is a well‐known phenomenon that occurs rapidly after physical or metabolic injury to the brain. One of the important events during astrocyte differentiation is the increased expression of glial fibrillary acidic protein (GFAP), a member of the family of intermediate filament structural proteins. Free radicals are neurotoxic and free radical scavengers have been shown to protect the brain against neurotoxic damage. In the present study, we examined the effect of melatonin on astrocytic reactivity by determining the expression of the glial marker, GFAP, in different brain regions. Rats were exposed to constant light to reduce endogenous melatonin production; half of the animals were injected with melatonin during the exposure to constant light for 7 days. Western blots showed increases in total and degraded GFAP content in the brain of rats exposed to constant light. Melatonin admisnistration caused a reduction of degraded GFAP content. In addition, melatonin significantly reduced neural tissue lipid peroxidation while constant light significantly enhanced the breakdown of lipids in the brain. Brain glutathione levels decreased significantly as a result of constant light exposure; this reduction was reversed by melatonin administration. These results suggest that melatonin potentially protects both neurons and glial cells from free radicals; melatonin's protective actions are probably related to the antioxidant properties of the indole.