New proposal for the treatment of nodular basal cell carcinoma with intralesional 5-aminolevulinic acid.

Photodynamic therapy (PDT) is a treatment modality using a photosensitizer, light and oxygen to cause photochemically-induced selective cell death. Topical PDT is most suitable for thin lesions such as superficial basal cell carcinoma and actinic keratoses in dermatology. Results with PDT as treatment of thicker lesions such as nodular basal cell carcinoma appear to have a limited role because the photosensitizer or the light cannot penetrate deeply enough into the thicker tumor volume. In this preliminary study we use intralesional administration of 5-aminolevulinic acid to enhance the efficacy of the photosensitivity of nodular basal cell carcinomas, thus improving clinical cure.

Squamous cell carcinoma of the eyelid treated with photodynamic therapy.

The ocular tissues can be the site of a number of malignant tumors in adults. Approximately 5% to 10% of all skin tumors occur in the eyelid. Incidence studies indicate that basal cell carcinoma is the most frequent malignant eyelid tumor (90%) followed by squamous cell carcinoma (9%). A 55-year-old man presented a squamous cell carcinoma (SCC) of 8 mm diameter, localized in the middle third of the lower eyelid, 3 mm under the eyelid margin on the eyelids. The histopathologic examination of a biopsy specimen showed the typical features of squamous cell carcinoma. Photodynamic therapy (PDT) with topical 5-aminolevulic acid (ALA) after Frost suture was employed. Very good results were obtained with rapid healing, without invasiveness, and without anesthesia. There was no evidence of scar formation and no signs of recurrence at 6 months follow-up. Many therapeutic methods have been suggested for squamous cell carcinoma of the eyelid. We consider photodynamic treatment of eyelid skin malignancies to be of great interest and it may represent an interesting future perspective for their management especially when surgical intervention cannot be tolerated by the patient.

BCA-1, A B-cell chemoattractant signal, is constantly expressed in cutaneous lymphoproliferative B-cell disorders.

We analysed the immunophenotypic and molecular expression of BCA-1 (B-cell-specific chemokine) and CXCR5 (BCA-1 receptor) in normal skin and different cutaneous lymphoproliferative disorders (cutaneous T-cell lymphoma (CTCL); cutaneous B-cell lymphoma (CBCL); cutaneous B-cell pseudolymphoma (PCBCL)), with the aim of investigating their possible involvement in the pathogenesis of cutaneous B-cell disorders. BCA-1 and CXCR5 were constantly expressed in CBCL and PCBCL, but not in normal skin and CTCL. BCA-1 and CXCR5 were constantly coexpressed by CD22+ B-cells, while CD35+ follicular dendritic cells coexpressed BCA-1 in PCBCL cells only. In low grade CBCL, as compared with high grade CBCL, the intensity of CXCR5 expression on neoplastic CD22+ cells was lower than that of BCA-1. The image analysis of reverse transcriptase-polymerase chain reaction (RT-PCR) products showed a significant quantitative difference between PCBCL/low grade CBCL and high grade CBCL. The above findings, although only observed in a small series of patients, are in keeping with findings in MALT gastric and gastric MALT lymphomas, adding further evidence of the close similarities between CBCL and MALT lymphomas.

Monochromatic excimer light (308 nm): an immunohistochemical study of cutaneous T cells and apoptosis-related molecules in psoriasis.

BACKGROUND: Various types of UVB radiation source (290-320 nm) are used in treating psoriasis and their therapeutic mechanism has been attributed to immunosuppressive properties. Recently, a new UVB source generated by a 308-nm excimer laser has been introduced for the treatment of psoriasis. OBJECTIVE: In this study we investigated the immunohistochemical evaluation of T cells and the expression of various apoptosis-related molecules in the psoriatic hyperproliferative skin before and after treatment with 308-nm monochromatic excimer light (MEL). METHODS: Ten patients (three women and seven men), ranging in age from 29 to 79 years, affected by plaque-type psoriasis vulgaris, were treated with MEL. Biopsies from psoriatic lesions of MEL-treated sites were taken before, 24 h and/or 48 h after the first irradiation and analysed by the immunophosphatase alkaline technique (APAAP). RESULTS: MEL treatment was found to cause a significant decrease in the rate of proliferation of keratinocytes and a relevant depletion of T cells in all psoriatic lesions, 48 h after the first irradiation: 308 nm light eliminated T cells from the psoriatic epidermis and also from the dermis, highlighting the ability of this UVB source to penetrate the skin compared with normal UVB and establish direct cytotoxic action on T cells infiltrating skin lesions. Rapid clearing of psoriatic lesions involves potential molecular targets of UVB in T cells including p53, which is upregulated after direct irradiation with 308-nm UVB. Moreover, Bcl-2 expression in healing psoriasis epidermis after MEL treatment is significantly decreased compared with untreated skin and the TUNEL (TdT-mediated dUTP-biotin nick end labelling) technique revealed the presence of relevant apoptotic keratinocytes in the irradiated epidermis. CONCLUSIONS: These results indicate that psoriatic skin after monochromatic excimer light therapy is associated with significant T-cell depletion and alterations of apoptosis-related molecules accompanied by a decreased proliferation index and clinical remission.

Giant exophytic basal cell carcinoma treated with radiotherapy.

Basal cell carcinomas may attain giant proportions due primarily to recurrence or because the tumour is neglected. We report the case of a 66-year-old man who presented with a bleeding, polypoid, cutaneous tumour located on the left shoulder region of 13 years duration. The man had not received any previous treatment. The lesion was biopsied and histopathologically diagnosed as a solid type basal cell carcinoma with focal areas of squamous differentiation and keratinization. The man refused complete surgical removal and therefore was treated with roentgentherapy, with satisfactory results and no complications in the irradiated area. No recurrences had manifested after 1 year follow-up.

Immunophenotypical markers, ultrastructure and chemosensitivity profile of metastatic melanoma cells.

The survival of patients affected by cutaneous melanoma has improved dramatically in the last 10 years, because of earlier diagnosis. Despite this, the therapeutic results obtained in metastatic melanoma (MM) are very disappointing due to its poor responsiveness to cytotoxic agents. In this type of solid tumor, tumor chemosensitivity assays have been suggested to be an important tool to predict clinical responsiveness to therapy. Metastatic melanoma cells (MMCs) were obtained from subcutaneous melanoma metastases of five patients and cultured for several consecutive passages. An immunofluorescence and an electron microscopic study were performed in order to establish the ultrastructural and physiopathological features of MMCs. A sulphorodamine-B test was used to measure in vitro sensitivity of MMCs to temozolomide, cisplatin, vindesine, taxol and interpheron alpha-2a. Following a 72 h exposure, maximum activity was obtained with vindesine (median inhibitory concentration, IC(50), 0.23 nM) and taxol (median IC(50) 0.31 nM). Cisplatin median IC(50) values were higher (4.6 microM) than taxol and vindesine, but still in the range of clinically achievable plasma concentrations. Temozolomide inhibited cell proliferation only at very high concentrations (median IC(50) 228 microM). No significant cell growth inhibitory effects (

IGFBPs modulate IGF-I- and high glucose-controlled growth of human retinal endothelial cells.

Insulin-like growth factor binding proteins (IGFBPs) are important local factors in the development of proliferative diabetic retinopathy. We investigated the effects of IGF-I and increased glucose concentrations on the release of IGFBPs and the growth of human retinal endothelial cells (HRECs). HRECs secrete IGFBPs-2 to -5. IGF-I stimulated thymidine incorporation and modified the pattern of IGFBPs, decreasing the inhibitory IGFBP-4 through down-regulation of its mRNA, and increasing IGFBP-5 which, per se, was able to modulate HREC growth, exerting post-transcriptional control. Studies using an antibody (alpha IR3) against the IGF-I receptor, and compounds with low affinity for IGFBPs, such as insulin and des(1-3)IGF-I, showed that an interaction between IGF-I and IGFBP-5 was necessary to detach this IGFBP from its binding sites. The dose of IGF-I that significantly decreased the IGFBP-4/IGFBP-5 ratio was the same that stimulated HREC growth. Chronic exposure to high concentrations of glucose was able to reduce HREC mitogenesis, interacting with the IGF system through a decrease in the stimulatory IGFBPs-2, -3 and -5, leaving the concentration of the inhibitory IGFBP-4 constant. These results extend our previous observations in endothelial cells and suggest that the IGFBP-4/IGFBP-5 ratio regulates IGF-I-induced growth of HRECs, whereas a general decrease in IGFBPs (except for IGFBP-4) was the anti-proliferative effect of chronic exposure to high glucose concentrations.

Non-invasive analysis of melanoma thickness by means of dermoscopy: a retrospective study.

Epiluminescence microscopy (ELM), or dermatoscopy, is a non-invasive technique for the diagnosis of cutaneous melanoma that may play a role in the non-invasive, preoperative assessment of melanoma thickness. This study investigated the correlation between the frequency of appearance of some standard ELM criteria and the histological thickness of melanomas. In addition, the possible role of the total dermoscopic score (TDS) according to ABCD rule of dermoscopy as a predictor of melanoma thickness was analysed. The dermoscopic images of 84 cutaneous melanomas were retrospectively investigated to evaluate the presence of 10 standard ELM criteria, and for each lesion the TDS was established (with observers blinded as to the tumour thickness). A statistically significant association was found between the presence of an irregular pigment network and melanomas with a Breslow index equal to or lower than 0.75 mm (positive predictive value of 68%); in contrast, radial streaming, atypical vascular pattern and grey-blue areas were associated with melanomas > 0.75 mm (positive predictive values of 77%, 65% and 70%, respectively). Of the melanomas thinner than 0.76 mm, 82% showed a TDS lower than 6.80 (optimized cut-off point), while 79% of melanomas thicker than 0.75 mm had a TDS higher than 6.80 (chi2 = 30.815, P < 0.001); the positive predictive value of a TDS > 6.80 in the detection of lesions thicker than 0.75 mm was 79%. In conclusion, a statistically significant correlation does exist between some dermoscopic features and melanoma thickness. Both the mostly used dermoscopic methods (standard ELM pattern analysis and the ABCD rule of dermatoscopy) may provide useful information in the non-invasive assessment of melanoma thickness. However, their diagnostic performance is far from 100%; further studies are needed to investigate whether the combination of dermoscopy with other non-invasive approaches (e.g. sonometry) may result in an overall improvement in the diagnostic performance.

Raynaud's phenomenon (primary or secondary to systemic sclerosis). The usefulness of laser-Doppler flowmetry in the diagnosis.

BACKGROUND: Raynaud's phenomenon (RP) consists of painful vascular spasms of the extremities stressed by cold exposure. It is often a symptom, in many cases the onset, of systemic sclerosis (SSc). The aim of this study was to evaluate the role that laser-Doppler flowmetry (LDF) could have in the differential diagnosis of these disorders and to characterize the functional behaviour of the microvessels in RP. METHODS: In this study we examined 52 patients (five males and 47 females, aged 17-57) affected by primary RP, 21 patients (three males and 18 females, aged 31-63) affected by RP secondary to SSc as defined by Wouda in 1987 and 20 healthy subjects (three males and 17 females, aged 26-58) by means of LDF with an appropriate cold-test (11 degrees C for 2 min). RESULTS: We show that the cold test applied at hand level can distinguish RP patients from healthy subjects. Data show that basal blood flow of patients affected by SSc is diminished in its medium value and presents a typical tracing without vasomotion in comparison to healthy subjects and also to patients affected by primary RP. On the basis of our data among the primary RP patients we can also distinguish a group that show basal blood flow tracing very similar to that of sclerodermic patients. These subjects should be carefully followed-up. CONCLUSIONS: In conclusion, it could be stimulating for the future to introduce LDF as a complementary method for the diagnosis of RP and possibly for early discrimination between the primary disease and that secondary to SSc.

Topical 5-aminolevulinic acid and photodynamic therapy in dermatology: a minireview.

Photodynamic therapy (PDT) is a treatment modality using a photosensitizer, light and oxygen to cause photochemically induced selective cell death. When exposed to light with the proper wavelength, the topically applied photosensitizer or photosensitizer precursor can activate a biomolecule through electron transfer to yield free radicals or produce singlet oxygen from energy transferred from the excited sensitizer to molecular oxygen. The tissue damage is the result of the activation of reactive singlet oxygen or free radical production. Photodynamic therapy with topical application of 5-aminolevulinic acid (ALA) is a new technique and although it remains largely experimental, it has potential application for treatment of malignant skin tumors, various precancerous and selected benign skin diseases. This technique yields not only a high percentage of good therapeutic results but also excellent cosmetic outcome. This paper reviews the recently published data on clinical ALA-based PDT in dermatology.

CD30-CD30 ligand interaction in primary cutaneous CD30(+) T-cell lymphomas: A clue to the pathophysiology of clinical regression.

Primary CD30(+) cutaneous T-cell lymphomas (CTCLs) represent a spectrum of non-Hodgkin's lymphomas (NHLs) that have been well defined at the clinical, histologic, and immunologic level. This group, which includes 2 main entities (large cell lymphoma and lymphomatoid papulosis [LyP]) and borderline cases, is characterized by the expression of CD30 antigen by neoplastic large cells at presentation, possible spontaneous regression of the skin lesions, and generally favorable clinical course. Although the functional relevance of CD30 and its natural ligand (CD30L) expression in most cases of NHL is presently undefined, previous studies indicate that CD30L is likely to mediate reduction of proliferation in CD30(+) anaplastic large-cell NHL. No information is currently available concerning the expression of CD30L in primary CD30(+) CTCLs. In this study, we investigated the immunophenotypic and genotypic expression of CD30 and CD30L in different developmental phases of skin lesions (growing v spontaneously regressing). By immunohistochemistry, CD30L expression was detected in regressing lesions only; by molecular analysis, the expression of CD30L was clearly higher in regressing lesions than in growing ones. CD30L, while expressed by some small lymphocytes, was most often coexpressed by CD30(+) neoplastic large cells, as demonstrated by 2-color immunofluorescence and by immunohistochemistry on paraffin sections. Taken together, these data suggest that CD30-CD30L interaction may play a role in the pathobiology of primary cutaneous CD30(+) lymphoproliferative disorders. In particular, CD30L (over)expression might have a major role in the mechanism of self-regression of skin lesions, the most distinctive clinical feature of this cutaneous lymphoma subtype.

Human glomerular endothelial cells IGFBPs are regulated by IGF-I and TGF-beta1.

The release of insulin-like growth factor binding proteins (IGFBPs) and their regulation in human glomerular endothelial cells (GENC) was characterised. GENC produce IGFBP-4, IGFBP-2 and IGFBP-3 and express mRNA for IGFBP-2 to IGFBP-5. Due to the fact that IGF-I and TGF-beta1 modulate glomerular hypertrophy, their action on IGFBP release and GENC growth was studied. IGF-I increased IGFBP-3, IGFBP-2 and decreased IGFBP-4, while TGF-beta1 decreased IGFBP-3 and apparently increased IGFBP-4. All of the IGFBPs, except the TGF-beta1-regulated IGFBP-4, were modulated at mRNA level. IGF-I stimulated GENC proliferation, while TGF-beta1 inhibited their growth. It was demonstrated that an IGFBP-3 antibody reduced GENC proliferation. However, rhIGFBP-3 alone had no effect on GENC, but after 48 h pre-incubation the IGF-I stimulated GENC growth was increased, suggesting that IGFBP-3 could modulate the IGF-I induced GENC proliferation. It was concluded that the stimulatory IGFBP-3 and the inhibitory IGFBP-4 could regulate GENC growth, although the IGFBP-3 seems to have a predominant effect in this control.

The oligosaccharidic component of the glycoconjugates in lichen planus, granuloma annulare, seborrheic keratosis and plamoplantar keratoderma: lectin histochemical study.

It is well known that cell surface glycoconjugates play an important role in cell proliferation, adhesion and differentiation. The aim of this investigation was to define the changes of the glycoconjugate saccharidic moieties in the epidermis and derma of patients affected by several skin pathologies such as seborrheic keratosis, lichen planus, granuloma annulare and palmoplantaris keratoderma. Bioptical specimens from skin lesions as well as from normal skin were fixed in Carnoy's fluid and routinely processed. The sections were treated with HRP-lectins (PNA, DBA, SBA, WGA, ConA, LTA and UEAI). Cytochemical controls were performed for specificity of lectin-sugar reaction. Some sections were pre-treated with neuraminidase prior to staining with HRP lectins. In comparison with normal human skin, epidermal lectin binding pattern in the considered diseases showed considerable qualitative and quantitative variations. In general, in all the considered pathologies, a lack and/or a decrease in lectin binding at the epidermal layers was observed; among the various diseases, differences in cellular localisation of the sugar residues were also noted. In such respect, an exception was represented by seborrheic keratosis, where the cells of the basal layer showed PNA reactivity, which was absent in the basal layer of the normal skin. Although seborrheic keratosis and lichen planus have been studied by others authors, our findings are not in total accordance concerning lectin binding; this is probably due to the different fixatives employed. Our findings seem to reveal significant changes in keratinocyte glycoconjugate oligosaccharides in the previously mentioned diseases, providing clues to their pathogenesis.

Primary tuberculosis of the penis.

We present a case of primary tuberculosis of the inner lining of the prepuce in a 63-year-old man. The condition resembled carcinoma. The diagnosis was based on histopathology and on M. tuberculosis culture. Successful treatment was by a combined medical and surgical approach. The rarity of the case is emphasized.

Characterization of [3H]substance P binding sites in human skin.

Radioligand binding experiments were performed with crude homogenates from normal human skin in order to investigate substance P receptor density. Binding of [3H]substance P ([3H]SP) reached equilibrium after 20 min and was saturable analysis of saturation curves gave a significantly better fit using two-site binding compared to the single-site model. Competition studies employing some selective agonists for NK1, NK2 and NK3 receptors have demonstrated that only the NK1 selective agonist, [Sar9, Met(O2)11]-SP, was a competitor for [3H]SP binding. In addition, the non-hydrolyzable guanosine 5'-0-(3-thiotriphosphate) altered the dissociation of SP from NK1 receptors by increasing the number of low-affinity sites. These data show that in the skin [3H]SP binds to a single population of substance P high-affinity sites, which represent NK1-type receptors.

The role of histamine in platelet aggregation by physiological and immunological stimuli.

BACKGROUND: Platelets participate in allergic and inflammatory processes beside their role in haemostasis and thrombosis. This paper reports the level, the uptake, the metabolism and the release of histamine in human platelets. The effects of exogenous histamine, as well as the receptor and signal transduction of these effects, are also described. METHODS: Purified suspensions of platelets, prepared from healthy volunteers and from atopic patients, were exposed in vitro to physiological and immunological stimuli. Platelet aggregation was measured by the increase in light transmission; histamine content and release, as well as cytosolic free Ca2+ concentration, were measured fluorimetrically. Platelet histamine forming capacity, and the uptake of exogenous histamine, were measured with a radioisotopic method. RESULTS: Human platelets contain 72.5 +/- 9.6pmoles of histamine x 10(9) platelets, and their capacity to form histamine is 18.7 +/- 3.5pmoles h(-1)g(-1) protein, which is reduced by alpha-fluoromethylhistidine (10(-5) M) a selective inhibitor of the specific histidine decarboxylase. Human platelets take up the preformed amine by a calcium and energy-dependent process, and the uptake of histamine is reduced by mepyramine, an H1-receptor antagonist, and N,N-diethyl-2-[4-(phenylmethyl) phenoxyl] ethanamine (10(-6) M), a blocker of intracellular histamine receptors. Histamine is also metabolized by human platelets. The exposure of platelets to thrombin (10-60 mUml(-1)) produced a progressive aggregation, associated with histamine release. The same is observed in platelets isolated from atopic patients exposed to anti-IgE antibodies. Exogenous histamine dose-dependently potentiates the aggregation induced by physiological and immunological stimuli. In resting platelets cytosolic calcium level (207 +/- 4.2 nM/10(8) platelets) is increased by thrombin as well as by anti-IgE; this effect is potentiated by 10(-5) M histamine. CONCLUSIONS: The synergistic effect between histamine and other monoamines on platelet aggregation may explain some aspects of allergic vasculitis in which platelet aggregation is present.