Detection of urinary TGF-alpha by HPLC and western blot in patients with melanoma.

Transforming growth factor-alpha (TGF-alpha) secretion in the urine may reflect autonomous production by neoplasms. Previous measurements of TGF-alpha in the urine were limited by the large volumes of urine required for detection. Sensitive assays are required to detect TGF-alpha in small urine samples from individual patients with melanoma. An assay which detects ng quantities of immunoreactive TGF-alpha in 10 ml of urine using dialysis, reverse-phase high-performance liquid chromatography (HPLC), and Western blot of the HPLC fractions was used to study the urine from 6 melanoma patients and 6 normal human volunteers. No TGF-alpha was detectable in the first voided urine specimens from the normal volunteers. In contrast, TGF-alpha was detected in the urine specimens of two of three patients with primary melanomas and two of three patients with metastatic melanoma. The sensitivity and reliability of the assay were most affected by methods of urine collection, sample preparation, and storage. TGF-alpha was less often found in randomly voided urine samples from melanoma patients (1 of 3 detectable) as compared to first voided morning urine samples (3 of 3 detectable). This data indicates urinary TGF-alpha assay may be useful in the diagnosis and management of patients with melanoma.

Expression and distribution of transforming growth factor-alpha within melanocytic lesions.

Stimulation of epidermal growth factor (EGF) receptor by ligands such as transforming growth factor (TGF) alpha may be associated with cell proliferation or transformation in both nevocytes and keratinocytes. Previously, EGF receptors have been identified within a variety of pigmented lesions, suggesting a possible responsiveness to ligands such as TGF alpha. In the present study, we characterize the intralesional expression and distribution of immunoreactive TGF alpha protein by avidin-biotin immunoperoxidase localization in benign nevi, congenital nevi, dysplastic nevi, and malignant melanomas. In situ hybridization techniques with TGF alpha riboprobes confirmed the constitutive production of TGF alpha in all types of pigmented lesions. The localization of TGF alpha expression to nevocytes when coupled with the previous reports of expression in basal keratinocytes suggests the possibility of either an autocrine mechanism of action for TGF alpha or a paracrine interplay of TGF alpha between keratinocytes and nevocytes within melanocytic lesions. An increase in immunoreactive TGF alpha in nevocytes was noted in both benign and dysplastic nevi from dysplastic nevus patients, as compared to benign nevi from normal patients. Congenital nevi and malignant melanomas showed an intermediate and variable level of TGF alpha immunoreactivity. When coupled with previous studies the data suggest linkage of the TGF alpha/EGF receptor pathway in the evolution of melanocytic lesions.

Recent advances in tissue synthesis in vivo by use of collagen-glycosaminoglycan copolymers.

Biologically active analogues of the extracellular matrix (ECM) are synthesized by grafting glycosaminoglycan (GAG) chains onto type I collagen, and by controlling the physicochemical properties of the resulting graft copolymer. Collagen-GAG ECM analogues have previously been shown to induce regeneration of the dermis in humans and the guinea pig, and of the rat sciatic nerve. Current studies have emphasized elucidation of the molecular mechanism through which tissue-specific ECM analogues induce regeneration. The contribution of the GAGs to the biological activity of the skin regeneration template was confirmed by studying the contribution of several GAGs to the inhibition of wound contraction in guinea pigs. The interaction between cells and the porous structure of an ECM analogue was studied with emphasis on the deformation of pores which occurs during wound contraction. The synthesis of scar, as well as of partly regenerated tissue which has a morphology between that appropriate for scar and for normal dermis, was quantitatively assayed for the first time using a laser light scattering technique. An ECM analogue which has been shown to be capable of inducing regeneration of functional sciatic nerve in the rat over a gap larger than 10 mm was incorporated in the design of a biodegradable implant for peripheral nerve regeneration.

Estrogen and progesterone receptors in melanocytic lesions. Occurrence in patients with dysplastic nevus syndrome.

Estrogen and progesterone binding studies in a series of 22 melanocytic lesions from 14 patients with the dysplastic nevus syndrome were done using a fluorescent estrogen and progesterone binding technique. Melanocytic lesions from these patients, including primary cutaneous melanomas, dysplastic nevi, and benign nevi, contained large numbers of estrogen and progesterone binding cells. Comparison is made to a series of control intradermal nevi that had little or no detectable estrogen or progesterone binding. Increased hormonal binding, and possibly responsiveness, is seen in melanomas, melanoma precursor lesions such as dysplastic nevi and congenital nevi, as well as benign nevi from patients with the dysplastic nevus syndrome.

Porphyrialike bullous dermatosis after chronic intense tanning bed and/or sunlight exposure.

Five fair-skinned patients had porphyrialike blisters and mechanical fragility of skin chronically exposed to intense radiation from tanning devices and/or the sun, but they had normal red blood cell, plasma, urine, and stool porphyrin levels. Use of several weak photosensitizing drugs during the year before or while developing lesions was identified or suspected in four patients.

Optimizing fluence and debridement effects on cutaneous resurfacing carbon dioxide laser surgery.

OBJECTIVE: To develop methods to compare carbon dioxide (CO2) resurfacing lasers, fluence, and debridement effects on tissue shrinkage and histological thermal denaturation. DESIGN: In vitro human or in vivo porcine skin samples received up to 5 passes with scanner or short-pulsed CO2 resurfacing lasers. Fluences ranging from 2.19 to 17.58 J/cm2 (scanner) and 1.11 to 5.56 J/cm2 (short pulsed) were used to determine each laser's threshold energy for clinical effect. Variable amounts of debridement were also studied. MAIN OUTCOME MEASURES: Tissue shrinkage was evaluated by using digital photography to measure linear distance change of the treated tissue. Tissue histological studies were evaluated using quantitative computer image analysis. RESULTS: Fluence-independent in vitro tissue shrinkage was seen with the scanned and short-pulsed lasers above threshold fluence levels of 5.9 and 2.5 J/cm2, respectively. Histologically, fluence-independent thermal depths of damage of 77 microns (scanner) and 25 microns (pulsed) were observed. Aggressive debridement of the tissue increased the shrinkage per pass of the laser, and decreased the fluence required for the threshold effect. In vivo experiments confirmed the in vitro results, although the in vivo threshold fluence level was slightly higher and the shrinkage obtained was slightly lower per pass. CONCLUSIONS: Our methods allow comparison of different resurfacing lasers' acute effects. We found equivalent laser tissue effects using lower fluences than those currently accepted clinically. This suggests that the morbidity associated with CO2 laser resurfacing may be minimized by lowering levels of tissue input energy and controlling for tissue debridement.

Mesenteric venous thrombosis in two women taking oral contraceptives.

PIP: 2 cases of mesenteric venous thrombosis in women taking oral contraceptives (48-year old gravida 5 taking norethindrone with mestranol for 10 years, a 33-year old gravida 2 taking Enovid and C-Quens since 1962 followed by norethindrone with mestranol 1 month prior to admission) were treated at the Jewish Hospital of St. Louis. Both patients had acute abdominal pain, vomiting, and bloody and diarrheal stools. In order to remove nonviable portions of the bowel which are viable at initail operation, 2 operations are necessary in the treatment of mesenteric venous thrombosis. Both of these patients underwent 2 operations and both had extensive segments of bowel removed. The post-operative courses of both patients were long (85 and 40 days respectively) and difficult. No predisposing or etiologic factor could be determined in either patient. A relationship of mesenteric venous thrombosis to oral contraceptives is suggested, but no definite causal relationship can be established.^ieng

A comparison of scar revision with the free electron and carbon dioxide resurfacing lasers.

Laser scar revision was studied to measure the effects of targeting extracellular matrix protein versus tissue water on scar revision. We compared the free electron laser used at 7.7 microm (the amide III protein absorption band) to the carbon dioxide (CO2) laser and dermabrasion.Nude mice (n = 40) that had rejected skin grafts on their dorsal surface and developed mature scars were used as a model for scar revision. One-half of each scar was revised with either the free electron laser at 7.7 microm (32 to 38 mJ, nonoverlapping pulses delivered with a computerized adjustable pattern generator at 30 Hz, and two to three passes), a 100-microsec CO2 resurfacing laser (500 mJ, 5.0 Hz, and two to five passes), or dermabrasion. The untreated portion of each scar served as an internal control. Evaluation was by measurement of the clinical size of the scar using photography with quantitative computer image analysis to compare the data and histology to evaluate the quality and depth of the scars. The free electron laser at 7.7 microm was significantly better than the CO2 laser and dermabrasion for scar size reduction (p < 0.046 and p < 0.018). The CO2 laser and a highly skilled dermabrader were not statistically significantly different (p < 0.44). The result seen with less skilled dermabraders was significantly worse than all other methods (p < 0.009). The free electron laser at 7.7 microm, which is preferentially absorbed by the proteins of the extracellular matrix, provided better scar reduction than the CO2 resurfacing laser and dermabrasion. Dermabrasion by a skilled operator resulted in improvement similar to the results obtained with the CO2 resurfacing laser, but less skilled operators had significantly poorer results.

Pregnancy and sex steroid hormone effects on nevi of patients with the dysplastic nevus syndrome.

Female patients with the dysplastic nevus syndrome who were pregnant or taking sex steroid hormones were prospectively studied to evaluate the effectiveness of photography and close clinical follow-up in detecting nevus change or melanoma development. Seventeen patients, who served as their own controls, were studied during 22 pregnancies. Twenty-four patients who used oral contraceptives and seven who took hormone supplements were similarly studied. This clinical management method provided timely biopsy of changing nevi with a small number of biopsies required per patient. One melanoma occurred during pregnancy, but neither patients who were taking sex steroid hormones nor those in the control groups had melanomas. The rate of nevus change observed clinically was 3.9 times higher when patients with dysplastic nevus syndrome were pregnant than when they were not, whereas no differences were observed whether or not women took oral contraceptives or hormone supplements. The rate of histologically proven dysplastic nevi that changed was 2.0-fold higher when women were pregnant; 1.4-fold higher with the use of hormone supplements and 1.1-fold higher with the use of oral contraceptives. These preliminary data suggest pregnancy and hormone supplements may be temporally associated with an increased rate of dysplastic nevus change.

Treatment of telangiectasia macularis eruptiva perstans with the 585-nm flashlamp-pumped dye laser.

BACKGROUND: Telangiectasia macularis eruptiva perstans (TMEP) is a form of cutaneous mastocytosis characterized by truncal telangiectases that are refractory to treatment. OBJECTIVE: The safety and efficacy of treating TMEP with laser surgery was tested in a patient with extensive truncal lesions. METHODS: The patient was treated with the 585-nm flashlamp-pumped dye laser. Diphenhydramine and ranitidine or doxepin were used pre- and postoperatively to block the effects of mast cell mediator release during surgery. RESULTS: All treated cutaneous lesions resolved completely, without scarring, after one treatment. Approximately 70% of the truncal lesions recurred 14 months postoperatively. Doxepin provided the best mast cell mediator blockade. Postoperative biopsy of a treated lesion demonstrated focal dermal vascular fibrosis with minimal telangiectasia and upper level of normal numbers of mast cells on the biopsy. CONCLUSIONS: An excellent therapeutic result was obtained by treating TMEP with the 585-nm flashlamp-pumped dye laser, although the response was temporary. The therapeutic effect of the laser appears to be secondary to reduction of the vasculature with no apparent effect on the mast cells. Physicians treating TMEP with laser therapy must use proper H1 and H2 receptor blockade to avoid potential complications from laser-induced mediator release.

Identification of an androgen receptor in the adult chicken oviduct.

The chicken oviduct androgen receptor was characterized by sucrose density gradient centrifugation, Scatchard analysis, competition studies, and affinity labeled with dihydrotestosterone 17 beta-bromoacetate. A specific 8.5 S peak was seen on 0.01 M KCl sucrose density gradients when the receptor was labeled with [3H]5 alpha-dihydrotestosterone. Specific 4.6 S peaks were seen when receptor labeled with [3H]5 alpha-dihydrotestosterone or [3H]dihydrotestosterone 17 beta-bromoacetate was analyzed on 0.3 M KCl sucrose density gradients. Scatchard analysis of [3H]5 alpha-dihydrotestosterone binding by oviduct cytosol was consistent with two binding sites. A Kd of 0.13 nM was found for the high affinity androgen receptor. Competition studies showed the following order of ligand affinity: 5 alpha-dihydrotestosterone greater than dihydrotestosterone 17 beta-bromoacetate greater than progesterone greater than estradiol. A 61.2 kDa protein was specifically covalently labeled with [3H]dihydrotestosterone 17 beta-bromoacetate. The chicken oviduct androgen receptor possesses characteristics similar to other androgen receptors, and provides a good source of androgen receptor for physicochemical studies of the native receptor protein.

Increased nevus estrogen and progesterone ligand binding related to oral contraceptives or pregnancy.

Melanocytic nevi excised from women who were pregnant or had given birth less than 1 month previously or from women who were taking oral contraceptives were studied with the use of a fluorescent estrogen and progesterone ligand binding assay. All nevi from these patients had markedly increased numbers of estrogen- and progesterone-binding cells, similar to those observed in melanoma and melanoma precursor lesions. In contrast, nevus cells in acquired intradermal nevi from women who were not pregnant nor taking oral contraceptives had few or no detectable estrogen- or progesterone-binding cells. The induction of increased hormonal receptors by pregnancy or oral contraceptives may be related to an increased risk of melanoma and melanoma precursor lesions in specific subsets of patients, as well as to the pigmentary changes observed in female patients in general.

Increased epidermal growth factor receptors in seborrheic keratoses and acrochordons of patients with the dysplastic nevus syndrome.

Seborrheic keratoses and acrochordons may change in biologic behavior with pregnancy or during treatment with sex steroids. Because epidermal growth factor receptor localization may change in hyperproliferative skin diseases, we compared epidermal growth factor receptor immunolocalization in seborrheic keratoses and acrochordons from women who were or were not pregnant or taking oral sex steroid hormones. Epidermal growth factor receptor was further investigated in growing and quiescent seborrheic keratoses and acrochordons from women with and without the dysplastic nevus syndrome. The epidermal growth factor receptor concentration was strikingly elevated in suprabasilar keratinocytes of growing seborrheic keratoses and acrochordons from patients with the dysplastic nevus syndrome who were pregnant or taking sex steroid hormones and less elevated in growing seborrheic keratoses from other patients. In contrast, the epidermal growth factor receptor distribution pattern in acrochordons did not correlate as well with the history of growth of these lesions in normal patients. These data suggest sex steroids may affect epidermal growth factor receptor metabolism in benign epidermal hyperproliferative lesions, particularly in patients with the dysplastic nevus syndrome.

Estrogen and progesterone receptors in primary cutaneous melanoma.

Using a variety of techniques, estrogen and progesterone receptors have previously been identified in variable percentages of malignant melanomas. We examined 10 primary superficial spreading melanomas (SSM) with a fluorescent hormone-binding technique for estrogen and progesterone cytoplasmic receptors. Of these 6 SSM were markedly positive for estrogen and progesterone binding. Patients with dysplastic nevus syndrome (DNS) or a family history of DNS were markedly positive for estrogen and progesterone binding. A single patient with lentigo maligna and another patient with lentigo maligna melanoma were negative for estrogen and progesterone binding. None of the 21 control intradermal nevi examined for estrogen and progesterone binding exhibited marked positivity.

Optimizing free electron laser incisions at 6.45 microm with computer-assisted surgical techniques and the utilization of a heat-conducting template.

BACKGROUND AND OBJECTIVE: We sought to optimize laser incisions by evaluating the effects of varying the slit width of a heat-conducting template and the step size with the Computer-Assisted Surgical Techniques (CAST) system on free electron laser (FEL) incisions at 6.45 microm. STUDY DESIGN/MATERIALS AND METHODS: Stainless steel calipers were used as a heat-conducting template on human skin in vitro. The CAST system made the incisions as a series of spot ablations with set step sizes of 50 microm or 1,250 microm. At each step size, incisions were made with or without the calipers, by using varying slit widths. Histologic specimens were analyzed for lateral thermal damage over the entire depth of the incision and over the superficial 150 microm of dermis. RESULTS: Lateral thermal damage over the superficial 150 microm of dermis was most significantly reduced with the calipers at a slit width approximating the FEL's beam diameter (636 microm +/- 100 microm). Also, incisions made with the larger step size (1,250 microm) had significantly less lateral thermal damage over the entire depth of the incision. CONCLUSIONS: The use of a heat-conducting template with an aperture approximating the FEL's beam diameter and larger step size improved FEL incisions at 6.45 microm.

Childhood bullous lichen planus.

A child with previous lichenoid lesions had bullae refractory to antistaphylococcal antibiotics. Biopsy of a bulla showed histologic changes of bullous lichen planus. Indirect immunofluorescence using the patient's serum on a perilesional bulla biopsy showed positive staining for IgA and IgG in the stratum granulosum and stratum corneum, confirming the diagnosis. The lesions resolved with dapsone treatment.

Increased epidermal growth factor receptors in melanocytic lesions.

BACKGROUND: Epidermal growth factor receptors (EGF/R) have been reported to be absent in melanomas or, in contrast, to be markers for potential malignancy in melanocytic lesions. OBJECTIVE: Our purpose was to evaluate the literature discrepancies regarding the presence of EGF/R in melanocytic lesions and to determine whether EGF/R presence correlates with the potential for malignancy of melanocytic lesions. METHODS: An EGF/R-specific polyclonal antibody was used to study melanomas, dysplastic nevi, congenital nevi, and nevocellular nevi. RESULTS: All melanocytic cell types (nevus and melanoma cells) in the lesions studied had immunoreactive EGF/R. EGF/R immunoreactivity was also observed throughout the epidermal basal to granular cell layers overlying the melanocytic lesions, although dermal fibroblasts were negative. CONCLUSION: The pattern of increased immunoreactive EGF/R in both benign and malignant nevocellular lesions suggests that although EGF/R are not a specific marker of potential malignancy in melanocytic lesions, they may mediate or coordinate growth of keratinocytes and nevus cells.