cysteinyldopa and Skin-Neoplasms

Melanoma is one of the most lethal and malignant cancers and its incidence is increasing worldwide, and Japan is not an exception. Although there are numerous therapeutic options for melanoma, the prognosis is still poor once it has metastasized. The main concern after removal of a primary melanoma is whether it has metastasized, and early detection of metastatic melanoma would be effective in improving the prognosis of patients. Thus, it is very important to identify reliable methods to detect metastases as early as possible. Although many prognostic biomarkers (mainly for metastases) of melanoma have been reported, there are very few effective for an early diagnosis. Serum and urinary biomarkers for melanoma diagnosis have especially received great interest because of the relative ease of sample collection and handling. Several serum and urinary biomarkers appear to have significant potential both as prognostic indicators and as targets for future therapeutic methods, but still there are no efficient serum and urinary biomarkers for early detection, accurate diagnosis and prognosis, efficient monitoring of the disease and reliable prediction of survival and recurrence. Levels of 5-

Topics: Biomarkers, Tumor; Cysteinyldopa; Drug Monitoring; Humans; Melanoma; Metabolome; Skin Neoplasms

Topics: Animals; Biomarkers, Tumor; Cysteinyldopa; Humans; Indoles; Melanoma; Skin Neoplasms

The biosynthesis of melanin from tyrosine is reviewed as the basis for assessment of laboratory tests that might potentially aid in the diagnosis and management of patients with malignant melanoma. These tests include qualitative and quantitative assays for the intermediates in metabolism of melanin and catecholamines, enzyme assays, metal ion analyses, and, most recently, immunoassays. Although currently no role exists for the clinical laboratory in the early diagnosis of malignant melanoma, serial quantitative analyses of total or individual melanogens or of catecholamine metabolites in urine or plasma specimens may be of value in the management of patients with this disorder. Immunologically based methods for the diagnosis and management of malignant melanoma hold some promise for the future.

Topics: Antigen-Antibody Complex; Antigens, Neoplasm; Catechols; Clinical Enzyme Tests; Cysteinyldopa; Humans; Immunologic Tests; Indoles; Melanins; Melanoma; Proteinuria; Skin Neoplasms; Trace Elements

5-S-Cysteinyldopa (5-S-CD) has been used as a biochemical marker of melanoma progression. Recently we have shown that the serum level of 5-S-CD is a sensitive and specific marker in predicting distant metastases. In melanocytes and melanoma cells, cysteinyldopa isomers are oxidized to phaeomelanin, the yellow to reddish melanin pigment. In this study we have developed a new method to measure levels of phaeomelanin in serum samples and have evaluated its clinical significance. The method is based on the production of 4-amino-3-hydroxyphenylalanine (4-AHP) and 3-amino-4-hydroxyphenylalanine (3-AHP) on reductive hydrolysis of phaeomelanin with hydriodic acid. 3-AHP is also derived from 3-nitrotyrosine-containing proteins. The isomeric 4-AHP and 3-AHP can be separated by high performance liquid chromatography. The mean +/- SD serum levels of 5-S-CD in control subjects (n = 36), in melanoma patients without recurrence (n = 92) and in melanoma patients with metastases (n = 24) were 2.7 +/- 1.2 nM (median 2.3 nM), 4.0 +/- 1.6 nM (median 3.8 nM) and 72 +/- 105 nM (median 35 nM), respectively. The serum levels of 4-AHP in these three groups were 45 +/- 21 nM (median 31 nM), 80 +/- 75 nM (median 53 nM) and 306 +/- 627 nM (median 133 nM), respectively. The serum levels of 4-AHP in patients with metastases (100 samples from 15 patients with progressive disease) correlated well (r = 0.887) with serum levels of 5-S-CD, while serum levels of 3-AHP did not (r = 0.240). The serum 5-S-CD and 4-AHP levels were serially analysed in the 15 patients with progressive disease. In two patients (13%), serum 4-AHP levels were elevated to abnormal levels before the serum 5-S-CD levels exceeded the cut-off value of 10 nM. In five patients (33%), the serum 4-AHP levels rose concurrently with the serum 5-S-CD levels. In the remaining eight patients (54%), serum 4-AHP levels were of less diagnostic value. Thus, the serum phaeomelanin level appears to be less sensitive than the serum 5-S-CD level in detecting distant metastases.

Topics: Adult; Aged; Biomarkers, Tumor; Chromatography, High Pressure Liquid; Cysteinyldopa; Female; Humans; Male; Melanins; Melanoma; Middle Aged; Serum; Skin Neoplasms; Tyrosine

The urinary excretion of 5-S-cysteinyldopa (5-S-CD) is known to be increased in certain patients with melanoma. To evaluate its diagnostic and prognostic utility, we measured the urinary excretion of 5-S-CD on at least three different occasions in 50 patients with melanoma. No significant increase was found in 26 patients without metastases, in 10 patients with regional lymph node metastasis and 2 patients with amelanotic melanoma. However, all the 12 patients with distant metastases demonstrated a significant increase. The patients with 5-S-CD > 1,000 micrograms/day survived for a mean of 8.1 +/- 5.6 months, while those with 5-S-CD > 10,000 micrograms/day survived for 3.5 +/- 3.7 months. All the 4 patients with a maximum excretion of 5-S-CD > 40,000 micrograms/day had multiple liver metastases. In conclusion, while data on the urinary excretion of 5-S-CD was not useful in the detection of early regional lymph node metastases, its increase indicated the presence of distant metastases and also provided prognostic information.

Topics: Adult; Aged; Biomarkers, Tumor; Cysteinyldopa; Female; Follow-Up Studies; Humans; Liver Neoplasms; Lymphatic Metastasis; Male; Melanoma; Middle Aged; Physical Examination; Prognosis; Radiography; Sensitivity and Specificity; Skin Neoplasms; Survival Rate

Topics: Clinical Trials as Topic; Cysteinyldopa; Dacarbazine; Dihydroxyphenylalanine; Drug Evaluation; Humans; Melanoma; Skin Neoplasms; Triazenes

Early diagnosis of malignant melanoma is critical for effective treatment and reduced patient mortality. However, current clinical and histological variables show limited accuracy in diagnosis. Serum or urine level of 5-S-cysteinyldopa (5-S-CD) is a commonly used melanoma biomarker in Japan owing to its increased sensitivity compared with other melanoma markers. However, its use as a diagnostic marker has shown some limitations. Therefore, here we examined the combination of 5-S-CD with melanoma inhibitory activity, which showed sensitivity in detecting melanoma comparable with that of 5-S-CD, and interleukin-8, a cytokine linked with melanoma progression, in a cohort of Japanese patients with melanoma. Our results revealed that the triple combination of 5-S-CD, melanoma inhibitory activity, and interleukin-8 showed high diagnostic accuracy in detecting melanoma compared with each of the individual factors. Importantly, the triple marker showed specificity and utility in detecting early-stage melanoma. Our results suggest the utility of the triple marker as a diagnostic biomarker for melanoma patients.

Topics: Biomarkers, Tumor; Cysteinyldopa; Cytokines; Humans; Interleukin-8; Melanoma; Melanoma, Cutaneous Malignant; Skin Neoplasms

Along with the expansion of therapeutic options for metastatic melanoma, the development of useful biomarkers is urgently required to predict and monitor treatment response. Serum 5-S-cysteinyldopa (5-S-CD) has been identified as a diagnostic marker of malignant melanoma, but its utility as a biomarker for emerging therapeutic agents remains unknown. We assessed serum 5-S-CD in 12 metastatic melanoma patients (median age, 76 years; six men and six women) who had been treated with nivolumab (Nivo) at Shinshu University Hospital between 2014 and 2016. Serum 5-S-CD and lactate dehydrogenase levels before and at 3-6 weeks of Nivo treatment were obtained and their changes were compared with clinical responses as defined by the Response Evaluation Criteria in Solid Tumors criteria (version 1.1). A decrease of 10 nmol/L or more of serum 5-S-CD was observed only in partial response patients (2/3 cases, 67%), while an increase of 10 nmol/L or more of serum 5-S-CD was witnessed only in progressive disease patients (4/8 cases, 50%). Serum 5-S-CD changes were within ±10 nmol/L in the remaining six patients (partial response, one; stable disease, one; progressive disease, four). The results of the four moderately affected progressive disease patients were suspected to have been influenced by small-sized metastatic lesions, a mixed response that included diminished and enlarged metastatic lesions, prior therapy to Nivo with BRAF inhibitors or radiation, or the development of brain metastasis. Serum 5-S-CD in the early phase of Nivo treatment may be helpful to predict therapeutic response in metastatic melanoma.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Cysteinyldopa; Disease Progression; Female; Humans; L-Lactate Dehydrogenase; Male; Melanoma; Middle Aged; Nivolumab; Prognosis; Response Evaluation Criteria in Solid Tumors; Skin Neoplasms; Young Adult

With the recent development of novel molecular targeted drugs for advanced stage malignant melanoma (MM), including RAF and mitogen-activated protein kinase kinase inhibitors and immune checkpoint blockers, the early detection of relapse is important for managing patients with MM. In this study, we retrospectively analyzed two conventional serum biomarkers, 5-S-cysteinyl-dopa and lactate dehydrogenase, in patients with MM (n = 140) who were treated at a single Japanese institute from June 2007 to June 2015. At the initial hospital visit, serum 5-S-cysteinyl-dopa levels were significantly increased in patients with stages III (n = 38) and IV (n = 20) MM compared with patients with stages 0-II (n = 62) MM. In addition, in patients with stages III and IV MM, serum 5-S-cysteinyl-dopa levels of more than 15.0 nmol/L at initial hospital visit correlated with a poor prognosis. In 11 of 14 patients whose disease progressed during follow up (mostly from stages III-IV), serum 5-S-cysteinyl-dopa levels exceeded the normal limit of 10.0 nmol/L during the clinical detection of distant metastases. These results indicate the usefulness of measuring serum 5-S-cysteinyl-dopa levels at initial hospital visit and during follow up for early and effective therapeutic interventions using newly developed molecular targeted drugs.

Topics: Biomarkers, Tumor; Cysteinyldopa; Dihydroxyphenylalanine; Feasibility Studies; Female; Follow-Up Studies; Humans; Japan; Kaplan-Meier Estimate; L-Lactate Dehydrogenase; Male; Melanoma; Middle Aged; Neoplasm Recurrence, Local; Neoplasm Staging; Prognosis; Retrospective Studies; Skin Neoplasms

TNM staging is mainly used to evaluate the prognosis of melanoma patients. Serum biomarkers such as 5-S-cysteinyldopa (5-S-CD) have occasionally been used but most do not respond until the tumour burden becomes high. Recently, circulating melanoma cells (CMC) have been reported as a possible new biomarker to detect metastasis, monitor treatment response and predict prognosis. The object of this exploratory study was to evaluate the efficacy of CMC to detect metastasis and predict prognosis by cross-sectional and prospective observational analyses, respectively. Altogether 15 patients with stages II-IV melanoma were enrolled and CMC were enumerated by CellSearch system with cut-off values of two cells/7.5 mL. Serum 5-S-CD and lactate dehydrogenase (LDH) were also measured. The sensitivity of CMC and 5-S-CD for the detection of metastasis was 33 and 50%, respectively. The combination of CMC and 5-S-CD showed a sensitivity of 67%, the best performance among CMC, 5-S-CD, LDH and any combination of two of the markers. Additionally, a 30-month prospective observation showed that CMC could segregate patients with poorer prognosis. The median survival time for the patients with <2 CMC and those with ≥2 CMC was 19.5 and 4.5 months, respectively. The limitation of this study is the small sample size. These preliminary results indicate CMC may complement the efficacy of 5-S-CD to detect metastasis and can be a prognostic marker. Although there is still room for improvement to maximise the sensitivity, the CellSearch system is reproducible, standardised and suitable for multi-centre studies.

Topics: Aged; Aged, 80 and over; Biomarkers, Tumor; Cross-Sectional Studies; Cysteinyldopa; Female; Humans; L-Lactate Dehydrogenase; Male; Melanoma; Middle Aged; Neoplastic Cells, Circulating; Prognosis; Prospective Studies; Sensitivity and Specificity; Skin Neoplasms; Survival Rate

Leptin is known to be abnormally expressed in a variety of cancers, and leptin receptors have been reported to be expressed on human melanoma cells. In this study, we evaluated the possibility that the serum levels of leptin receptor could be a tumor marker of malignant melanoma (MM). Serum samples were obtained from 71 patients with MM, and the serum levels of leptin receptor were measured by double-determinant ELISA. Interestingly, serum levels of leptin receptor decreased gradually with the stages of MM, being highest at in situ and lowest at stage IV. There was also a trend of reverse correlation between tumor thickness and serum levels of leptin receptor. To our knowledge, this is the first report investigating the serum levels of leptin receptor in MM, and serum leptin receptor levels may be used as a useful tumor marker of MM.

Topics: Biomarkers, Tumor; Body Mass Index; Case-Control Studies; Cell Line, Tumor; Cysteinyldopa; Disease Progression; Female; Humans; Insulin Resistance; Leptin; Male; Melanoma; Receptors, Leptin; Skin Neoplasms

A new human malignant melanoma cell line, designated DEOC-1, was established from the heel lesion of a 59-year-old man. This cell line has grown well for 84 months. The monolayer, cultured cells are polygonal in shape, appear to be spindle-shaped cells or multipolar cells, and have a tendency to pile up without contact inhibition. The cells are aneuploid, and the modal chromosomal number is in the hyper-triploid range. The cells were transplanted into the subcutis of SCID mice and produced tumors resembling the original tumor. The DEOC-1 cells (1 x 10(6)/5 mL) produced 5-S-cysteinyldopa (5-S-CD). The cells were not sensitive in vitro to any anticancer drug currently used for the treatment of malignant melanoma. Increases in both the protein and the transcriptional levels (mRNA) of multidrug resistance-related genes (multidrug resistance gene 1, multidrug resistance-associated protein 1 and lung resistance-related protein) were observed in DEOC-1 cells. The DEOC-1 cells are well characterized and are a very useful material for basic research of malignant melanoma.

Topics: Animals; Biomarkers, Tumor; Cell Line, Tumor; Cysteinyldopa; Drug Resistance, Multiple; Drug Screening Assays, Antitumor; Humans; Immunohistochemistry; Male; Melanoma; Mice; Mice, SCID; Middle Aged; Neoplasm Transplantation; Polymerase Chain Reaction; Skin Neoplasms; Transplantation, Heterologous

Topics: Adult; Aged; Aged, 80 and over; Cysteinyldopa; Disease Progression; Female; Humans; Kidney Failure, Chronic; Male; Melanoma; Middle Aged; Skin Neoplasms

5-S-L-Cysteinyl-L-dopa is a well-known pigment intermediate and analysis of its serum concentration is well suited for evaluation of treatment and follow-up of stage III and IV malignant melanoma. A simplified analytical method is described using organic extraction followed by clean-up on a boronate gel to capture the compound containing vicinal hydroxyls. Weak acid solution elutes the 5-S-cysteinyldopa suitable for high-performance liquid chromatography (HPLC). The absolute recoveries of cysteinyldopa and its diastereomer 5-S-D-cysteinyl-L-dopa (used as an internal standard) were 81.5 +/- 2.8% and 81.3 +/- 2.7%, respectively, and use of the internal standard for the whole procedure gave an analytical recovery of 101 +/- 0.8%. The limit of quantitation was 1.5 nmol/L and the imprecision of the method was < 5.0% over the analytical range 1.5-500 nmol/L. The method is cheap and easy to perform and compares well with other described techniques. The use of the method is illustrated by results obtained during treatment of a patient with metastatic malignant melanoma.

Topics: Aged; Chemistry, Clinical; Chromatography, High Pressure Liquid; Cysteinyldopa; Humans; Melanoma; Reproducibility of Results; Skin Neoplasms

5-S-Cysteinyldopa (5-S-CD) is a precursor of pheomelanin. S-100B protein is a low molecular weight, acidic, calcium binding, cytoplasmic protein. In this study, the concentration changes of serum 5-S-CD and S-100B protein in melanomas of all stages were examined in parallel and patients were monitored during and after treatment. Serum samples were taken from 478 melanoma patients on 1924 occasions. Of these, 180 cases were regularly monitored. Concentrations of 5-S-CD were determined by high performance liquid chromatography (HPLC), S-100B protein by immunoluminometric assay. The mean/median concentrations of 5-S-CD and S-100B protein in Stage I, II and III patients and in the control group ranged around the normal level. In Stage IV patients, 58.4/50.6% sensitivity, 100% specificity and 100/86.6% positive predictive values were obtained concerning S-100B protein and 5-S-CD, respectively. Recurrence was observed in 57/180 of the regularly monitored patients in Stages I, II and III. In 10/57 (17.5%) of these patients suffering from any type of disease progression increases in both marker levels preceded the detection of metastasis by conventional methods. We can confirm that changes in both marker concentrations correlated with the stages of the patient. The markers are most sensitive in Stage IV patients and also have a high specificity in these patients. In Stage IV melanoma patients, 5-S-CD and S-100B protein levels are independent significant prognostic factors. In almost one fifth of patients both marker levels increased before the detection of metastatic disease with other appropriate, routinely scheduled investigations. This study suggests that serial serum marker measurements in the management and follow-up of melanoma patients should be examined further.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Analysis of Variance; Cysteinyldopa; Female; Humans; Lymphatic Metastasis; Male; Melanoma; Middle Aged; Neoplasm Proteins; Neoplasm Recurrence, Local; Nerve Growth Factors; Prognosis; Prospective Studies; S100 Calcium Binding Protein beta Subunit; S100 Proteins; Skin Neoplasms; Survival Analysis

5-S-Cysteinyldopa (5-S-CD) has been used as a biochemical marker of melanoma progression. In this study, we measured serum levels of 5-S-CD in 2648 samples taken from 218 patients in order to evaluate the usefulness of this parameter in following melanoma progression and prognosis. 5-S-CD levels were significantly elevated above the upper limit of the normal range (10 nmol/l) in stage IV melanoma patients. The sensitivity of elevated serum 5-S-CD levels in detecting distant metastasis was 73%, while the specificity was 98% and the positive predictive value 94%. The sensitivity was improved to 77% when cases of amelanotic melanoma were excluded. Patients without metastases had elevated 5-S-CD values in 5% of the 1480 serum samples. Changes in serum 5-S-CD levels were followed during disease progression until the end stage in 49 patients. In 33% of the patients, elevation of serum 5-S-CD levels preceded clinical detection of visceral metastases, and in 37% elevation of 5-S-CD levels occurred at the same time as visceral metastasis. Patients with elevated 5-S-CD levels before or after surgical treatment had significantly shorter survival times than those with normal levels. These results show that the level of 5-S-CD in the serum is a sensitive and specific marker in predicting distant metastases. Elevated serum levels of 5-S-CD, before or after surgical treatment, is associated with a poor prognosis.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Child; Cysteinyldopa; Disease Progression; Eye Neoplasms; Female; Follow-Up Studies; Humans; Japan; Life Tables; Male; Melanoma; Middle Aged; Neoplasm Metastasis; Neoplasm Staging; Prognosis; Sensitivity and Specificity; Skin Neoplasms; Survival Analysis

5-S-cysteinyldopa is a precursor of pheomelanin. S-100B protein is a low molecular weight, acidic, calcium binding, cytoplasmatic protein. LDH was defined as the most important serum parameter in disseminated melanoma. The aim of the present study was to compare the prognostic values of serum 5-S-Cysteinyldopa, S-100B and LDH concentrations in Stage III-IV melanoma patients. Serum samples were taken from 179 Stage III-IV melanoma patients at diagnosis. Serum 5-S-CD concentrations were determined by HPLC, S-100B protein by immunoluminometric assay while LDH by UV kinetic method. The mean/median concentrations of LDH, S-100B protein and 5-S-CD in Stage III patients ranged around the normal level. In Stage IV, the markers ranked as S100B = 5-S-CD > LDH for sensitivity, S-100B > LDH > 5-S-CD for specificity and LDH = S100B = 5-S-CD for positive predictive value, respectively. Furthermore, mean marker concentrations of patients with progressive disease differed significantly from nonprogresssive cases (when staging categories have been disregarded). Survival analysis indicated, that the initially elevated LDH and S-100B level in Stage IV disease predicts comparably short survival. Results of our study suggest that these serum marker values correlate well with Stages and disease progression. In Stage IV melanoma, the markers had appropriate sensitivity, high specificity as well as important positive predictive value. Among the studied serum markers S-100B protein and LDH proved to be similarly reliable in respect to the clinical outcome.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Cysteinyldopa; Female; Humans; L-Lactate Dehydrogenase; Male; Melanoma; Middle Aged; Neoplasm Staging; Nerve Growth Factors; Prognosis; S100 Calcium Binding Protein beta Subunit; S100 Proteins; Sensitivity and Specificity; Skin Neoplasms; Survival Rate

In a series of 92 patients with malignant melanoma, clinical stage III or IV, both 5-S-cysteinyldopa (5SCD) and 6-hydroxy-5-methoxyindole-2-carboxylic acid (6H5MI2C) were measured in urine during chemotherapy. A total of 434 urine specimens were analysed. The sensitivity of 5SCD for the detection of stage III-IV melanoma was 83%, while the corresponding sensitivity of 6H5MI2C was 52%. Fifty per cent of patients with one metastatic site had increased 5SCD excretion, while all patients with four or more metastatic sites had increased excretion. A significant correlation was found between 5SCD decrease and clinical regression (P<0.001) and between 5SCD increase and clinical progression (P<0.001). Corresponding correlations were not found for 6H5MI2C. Increments in 5SCD excretion (median 269 micromol/mol creatinine) were seen for 83% of the occasions when clinical progression was recorded, and decrements in 5SCD excretion (median 145 micromol/mol creatinine) were seen for 85% of the occasions when clinical regression was seen. During clinical 'stable disease' increases in 5SCD excretion were seen in 59% and decreases in 41%. The median value of 5SCD changes for stable disease was 7.0 micromol/mol creatinine, indicating a chemical marker stability in many cases. We recommend the use of 5SCD in urine as a valuable, reliable and simple biochemical marker to use in the clinical follow-up of melanoma patients with advanced disease.

Topics: Adult; Aged; Aged, 80 and over; Antineoplastic Combined Chemotherapy Protocols; Biomarkers, Tumor; Cysteinyldopa; Disease Progression; Female; Follow-Up Studies; Humans; Indoles; Male; Melanoma; Middle Aged; Skin Neoplasms

5-S-cysteinyldopa (5-S-DC) is a precursor of melanin. Its serum and urinary level can reflect melanoma progression. In this study we examined the concentration changes of 5-S-CD in melanomas of different clinical stages and in patients with different symptoms of melanoma, during and after treatment.. Serum samples were taken from 252 melanoma patients on 765 occasions, from June 1996 to July 1998. Levels of 5-S-CD were determined by HPLC.. The value of 5-S-CD in patients with primary melanoma and in patients without symptoms ranged around the normal level. There was a significant difference between the values of patients with or without symptoms. There was also a significant difference between the 5-S-CD values at clinical Stage I and Stage III, as well as at clinical Stage II and Stage III, respectively. Analysing the values of patients with symptoms we found a significant difference between the mean values of primary tumour and stage III, between values in lymph node metastasis and stage III, between values in lung metastasis and stage III. The tumour burden was found to correlate with a rising marker level. In 7% of the symptomatic patients that had a marker level under the upper limit, amelanotic primary tumour was detected.. According to the high marker level in lung and liver metastases, the marker might be useful in monitoring both patients with disease free ocular melanomas, to detect liver metastasis and high-risk patients after surgical removal of the primary tumour to reveal lung metastases.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Biomarkers; Biomarkers, Tumor; Cysteinyldopa; Humans; Liver Neoplasms; Lung Neoplasms; Melanoma; Middle Aged; Neoplasm Staging; Skin Neoplasms

5-S-Cysteinyldopa (5-SCD) in plasma and urine was determined by means of a newly developed method. This method incorporates optimized conditions for blood collection and storage, as well as a new extraction and separation technique, required for the strong oxidation and light sensitive 5-SCD. The new aspects of the method are the following: immediate centrifugation and freezing of the samples after blood collection, fully automatical solid-phase extraction (SPE) with phenylboronic acid (PBA) cartridges and immediate HPLC injection of the eluate, nearly complete exclusion of light and air-oxygen during extraction, constant sample cooling, use of the more suitable internal standard 5-S-D-cysteinyldopa and easy, sensitive and selective HPLC conditions (RP18-column with isocratic separation and electrochemical detection). The method has a linear range from 0.25 to 50 microg l(-1) and 25 to 5000 microg l(-1) for plasma and urine samples, respectively, a limit of detection of 0.17 microg l(-1), intra-assay variabilities from 1.7 to 3.6%, inter-assay variabilities from 4.0 to 18.3% and an average relative recovery of 103.5% for plasma and 105.4% for urine samples. In our study the measured 5-SCD concentrations of patients with melanomas at various stages correlated better with their clinical pictures than described in literature up to date. The results were obtained in comparison to patients with other skin tumors and in comparison to healthy control persons.

Topics: Automation; Biomarkers, Tumor; Calibration; Case-Control Studies; Chromatography, High Pressure Liquid; Cysteinyldopa; Humans; Melanoma; Prognosis; Reference Standards; Reproducibility of Results; Sensitivity and Specificity; Skin Neoplasms

Serum S-100 protein is widely used as a marker of melanoma and since 5-S-cysteinyldopa (5-S-CD) is a precursor of melanin its serum and urinary levels can reflect melanoma progression. In this study we examined the concentration changes of serum S-100 protein and 5-S-CD in 252 melanoma patients of different clinical stages. Serum samples were taken from 252 melanoma patients at 860 times, from June 1996 to July 1998. The serum S-100 protein was measured by the immunoluminometric assay, levels of 5-S-CD was determined by HPLC. The value of S-100 protein in patients with primary melanoma (0.11m mg/l) and in patients without symptoms (0.15 m mg/l) ranged around the normal level (0.01 0.12 m mg/l). There was a significant difference between the values of patients with or without symptoms. There was a similarly significant difference between the S-100 values of clinical Stage I (0.11 mg/l) and Stage III (2.91 mg/l) as well as between those of clinical Stage II (0.47 mg/l) and Stage III (2.91 mg/l), respectively. Analyzing the values of patients with symptoms we observed significant difference between the S-100 protein values of patients with primary tumor and those with solitary or multiple distant metastases. In case of 5-S-CD significant difference was found between clinical Stage I and III as well as clinical Stage II and III. Furthermore, there was a significant difference between the mean marker values of patients with primary tumor, lymph node, lung metastasis and clinical stage III.

Topics: Biomarkers, Tumor; Cysteinyldopa; Disease Progression; Disease-Free Survival; Humans; Liver Neoplasms; Lung Neoplasms; Lymph Nodes; Melanins; Melanoma; Models, Biological; Neoplasm Metastasis; S100 Proteins; Skin Neoplasms

Glutathione (GSH) and cysteine (CysH) have both been implicated in the biogenesis of the pheomelanin precursor 5-S-cysteinyldopa (5-S-CD). However, recent studies have shown that only CysH is transported across the membrane of isolated melanosomes, and that the positive regulation of CysH in pigment cells leads to an increased production of 5-S-CD. In the present study, the question was examined as to whether melanin precursors and tyrosinase could be coregulated by cellular thiols. To address this issue, the levels of CysH and GSH were varied in normal melanocytes and melanoma cells using buthionine sulfoximine (BSO), an inhibitor of GSH biosynthesis. Treatment with 50-100 microM BSO decreased GSH levels to less than 10% of control, and increased CysH levels between two- and five-fold in both cell types. Concomitant with this, an increase in the ratio of 5-S-CD to DOPA and a decrease in the pigment content of the cells were observed. The decrease in cell pigmentation was associated with strong decreases in tyrosine hydroxylase activity and 14C-melanin production. Only melanoma cells showed a modified tyrosinase isozyme pattern on Western immunoblots in response to BSO, while the mRNA expression of tyrosinase and TRP-1 were unchanged in both cell types. These results suggest that the balance between CysH and GSH, which is partly determined by the rate of utilization of CysH for GSH biosynthesis, regulates not only the levels of 5-S-CD and DOPA but also the melanogenic activity of pigment cells. Since DOPA functions as a cofactor in the monophenolase reaction of tyrosinase, it is proposed that the ratio of 5-S-CD to DOPA may be an important factor in the regulation of tyrosinase activity in situ.

Topics: Buthionine Sulfoximine; Cells, Cultured; Cysteine; Cysteinyldopa; Dihydroxyphenylalanine; Glutathione; Humans; Infant, Newborn; Male; Melanins; Melanocytes; Melanoma; Melanosomes; Monophenol Monooxygenase; Neoplasm Proteins; Skin Neoplasms; Tumor Cells, Cultured; Tyrosine

5-S-Cysteinyldopa (5SCD) and 6-hydroxy-5-methoxyindole-2-carboxylic acid (6H5MI2C) are formed during biosynthesis of melanins. They are used as indicators of pigment formation and markers of melanoma progression in adults and could possibly be used as markers of activity, growth and even malignant transformation in large pigmented naevi in children. We investigated the urinary excretion of these pigment precursor metabolites from 136 children, 5 to 15 years of age. The mean 5SCD excretion was 38.1 mumol/mol creatinine. A significant age-related decrease from a mean of 60.4 mumol/mol creatinine at 5 years of age to 28.0 mumol/mol creatinine at age 15 was found. In a reference group (29 adults, 20-33 years of age) the mean excretion was 48.9 mumol/mol creatinine. The mean excretion of 6H5MI2C was 42.8 mumol/mol creatinine at 5 years of age and 26.1 mumol/mol creatinine at the age of 15. The mean value for the young adults was 33.4 mumol/mol creatinine. No correlation between the mean excretion of 5SCD and 6H5MI2C was demonstrated. We suggest an upper reference level of 90 mumol/mol creatinine for the excretion of 5SCD in the age group 5-11 years and of 60 mumol/mol creatinine in the age group 13-15 years. Corresponding figures for the indole 6H5MI2C are 70 and 60 mumol/mol creatinine. The establishment of reference values in children will make it possible to use 5SCD and 6H5MI2C measurements as diagnostic tools, indicating growth or malignant transformation in giant melanocytic naevi during childhood.

Topics: Adolescent; Adult; Age Factors; Biomarkers, Tumor; Child; Child, Preschool; Cysteinyldopa; Female; Humans; Indoles; Male; Melanins; Nevus, Pigmented; Sex Factors; Skin Neoplasms

Serum 5-S-cysteinyl dopa (5-S-CD), circulating intercellular adhesion molecule-1 (cICAM-1) and soluble interleukin-2 receptor (sIL-2R) have each been reported as useful markers for melanoma progression. To assess the clinical relevance of these three markers, we simultaneously assayed their serum levels in 30 Japanese melanoma patients. Pre-surgical serum levels of 5-S-CD, cICAM-1 and sIL-2R were elevated in six, 13 and five patients respectively. These abnormal values returned to normal after curative surgery in most of the patients, suggesting a direct relationship to the presence of the primary tumour. Pre-surgical values of these three markers, either individually or in combination, did not predict the development of subsequent metastases. The sequential measurements of the three markers in eight patients who had relapse after surgery showed that serum 5-S-CD is the most useful marker for disease progression, although it is dependent on the melanin-producing ability of individual recurrent tumours. sIL-2R seemed to reflect in vivo activation of the host immune system and was a good indicator for predicting occult metastasis in selected cases. Circulating ICAM-1 levels were less relevant to the clinical disease course in our cases, although they tended to increase strikingly after liver metastasis. Our results in this limited number of cases show that the significance of the three markers varied with the individual and suggest that the simultaneous measurement of these markers may facilitate the early detection of metastases and proper therapeutic intervention.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Cysteinyldopa; Female; Humans; Intercellular Adhesion Molecule-1; Male; Melanoma; Middle Aged; Neoplasm Recurrence, Local; Receptors, Interleukin-2; Reference Values; Skin Neoplasms

The effects of insulin on melanogenesis were examined in human Swift melanoma cells. When these cells were grown in a chemically defined culture medium containing insulin (5 microg/ml), they showed a low pigmentation in association with a high activity of glutathione peroxidase (GPO) and a low activity of tyrosinase. In Eagle's minimum essential medium supplemented with foetal calf serum (EMEM-FCS), the Swift cells showed an intense pigmentation in association with a low GPO activity and a high tyrosinase activity. Modulation of GPO activity with sodium selenite had no effect on melanogenesis variables. In contrast, addition of insulin (5 microg/ml) to the EMEM medium led to a marked decrease in tyrosinase activity (p<0.001) and to a concomitant reduction in the levels of 5-S-cysteinyldopa (p <0.01). These results indicate that insulin inhibits the formation of 5-S-cysteinyldopa and that of melanin via the inhibition of tyrosinase activity.

Topics: Calcium; Chromatography, High Pressure Liquid; Cysteinyldopa; Glutathione Peroxidase; Glutathione Reductase; Humans; Insulin; Melanins; Melanoma; Monophenol Monooxygenase; Skin Neoplasms; Sodium Selenite; Tumor Cells, Cultured

The melanin metabolite 5-S-cysteinyldopa (5-S-CD) has been reported to be helpful in detecting occult melanoma metastases and as a prognostic marker in B16 melanoma-bearing mice. The goal of our study was to analyze the significance of the serum 5-S-CD level for the biochemical detection of metastases in human malignant melanoma (MM) and for monitoring the progression or the immunochemotherapeutically induced regression of MM. From 11 patients with metastatic MM observed between 1991 and 1995, serum samples were collected before and after each cycle of immunotherapy or immunochemotherapy. Samples were analyzed for 5-S-CD by automated high performance liquid chromatography with electrochemical detection. Cycles of immunochemotherapy consisted of human interleukin 2 and IFN-alpha (four patients) or of human interleukin 2, IFN-alpha, and dacarbazine (seven patients). Serum value of 5-S-CD in our normal controls was 1.9 +/- 0.6 ng/ml. All patients with metastatic MM showed 5-S-CD serum levels above the upper normal limit of 3.2 ng/ml (10 nM) and ranged from 2.3-fold (4.3 +/- 3.9 ng/ml) of the normal control values in early stages of metastases to more than 50-fold (94.3 +/- 220.3 ng/ml) of the normal control values in advanced stages of the disease. In 28 of 41 (68%) immunochemotherapeutical cycles, a decrease of 5-S-CD was seen during therapy, and in 13 cycles (31.7%), an increase was seen. Patients with more than 68% decreasing cycles (defined as responders; n = 5) showed significantly longer survival times (P = 0.008) than patients with less than 68% decreasing cycles (nonresponders; n = 6). High levels of 5-S-CD were also observed in metastasizing amelanotic melanoma. Serum 5-S-CD is a useful marker for monitoring the clinical course of MM patients, for discriminating between responders and nonresponders to immunochemotherapy, and as a prognostic factor concerning survival time and death risk.

Topics: Adult; Aged; Antineoplastic Agents; Biomarkers, Tumor; Cysteinyldopa; Dacarbazine; Disease Progression; Female; Humans; Immunotherapy; Interferon-alpha; Interleukin-2; Male; Melanoma; Middle Aged; Prognosis; Skin Neoplasms

The cellular concentration of reduced glutathione (GSH) modulates the sensitivity of human melanoma cells to alkylating drugs in vitro. To investigate whether the membrane-associated enzyme gamma-glutamyl transpeptidase (gamma-GTP) involved in GSH breakdown was expressed in melanoma cells, the enzymatic activity of gamma-GTP as well as the secretion of GSH were measured in human melanoma cells from four different cell lines (Me8, JUSO, GLL19, Swift). All the cells showed low gamma-GTP activities (0-1 mU/mg protein) and released GSH in culture supernatants at significant rates. After incubation for 24 h in growth medium containing 0.1 mmol/L cystine, the levels of GSH in supernatants ranged from 56 to 111 nmol GSH/mg protein. The GSH metabolism of melanoma cells was also evaluated by measuring the levels of the melanogenesis intermediate 5-S-cysteinyldopa under different experimental conditions. The results of these experiments suggest that melanoma cells have a low ability to metabolize the tripeptide GSH, which appears to be responsible for GSH secretion and accumulation in culture supernatants.

Topics: Cell Division; Cysteinyldopa; Fibroblasts; gamma-Glutamyltransferase; Glutathione; Humans; Melanoma; Monophenol Monooxygenase; Skin; Skin Neoplasms; Tumor Cells, Cultured

Elevated levels of the phaeomelanin metabolite 5-S-cysteinyldopa and the eumelanin metabolite 6-hydroxy-5-methoxyindole-2-carboxylic acid in urine and serum have been shown in previous studies to correlate with disseminated malignant melanoma. Immunohistochemical detection of S100B protein is an acknowledged method for the diagnosis of malignant melanoma, and it has been suggested that rising serum levels of S100B protein are associated with the survival rate of patients with malignant melanoma. In the present study serum levels of S100B protein and urinary concentrations of 5-S-cysteinyldopa and 6-hydroxy-5-methoxyindole-2-carboxylic acid were measured in 91 patients with histopathologically verified malignant melanoma. At the time of sampling 13 patients were in clinical stage I, 13 in stage II and 65 in stage III. The urinary levels of the melanin metabolites were determined by automated high performance liquid chromatography, and the serum levels of S100B protein by an immunoradiometric assay with two monoclonal antibodies. The overall survival rate was most strongly associated with the serum levels of S100B protein (P < 0.001), but there was also a significant correlation to urinary levels of 5-S-cysteinyldopa (P < 0.001). A corresponding association with urinary levels of 6-hydroxy-5-methoxyindole-2-carboxylic acid was found in only a very few patients with extremely high urinary concentrations. A statistically significant increase in relative hazard was found for S100B protein levels exceeding 0.6 microgram/l (P < 0.001), and predictably for patients in clinical stage III (P < 0.001). An analysis of S100B protein levels in patients in clinical stage III showed a significant correlation to survival (P = 0.005). Our study suggests that of the three biochemical tumour markers, S100B and to a lesser extent 5-S-cysteinyldopa have the greatest potential to be used as predictors of survival prognosis in patients with malignant melanoma.

Topics: Adult; Aged; Aged, 80 and over; Antibodies, Monoclonal; Autoantigens; Biomarkers, Tumor; Calcium-Binding Proteins; Cysteinyldopa; Disease-Free Survival; Female; Humans; Indoles; Male; Melanoma; Middle Aged; Neoplasm Invasiveness; Nerve Growth Factors; Prognosis; Radioimmunoassay; Regression Analysis; S100 Calcium Binding Protein beta Subunit; S100 Proteins; Skin Neoplasms; Survival Rate; Time Factors

The serum concentrations of 5-S-cysteinyldopa (5-S-CD) in pediatric patients with giant pigmented nevi (GPN) were investigated and compared with those of pediatric patients with small- or medium-sized congenital nevi (CN), or non-melanocytic benign skin tumor (control group). Serum 5-S-CD levels in the GPN group (N = 21), particularly in patients less than 5 years old, were significantly higher (highest concentration 38.4, mean +/- S.D. 16.6 +/- 9.4 nmol/L) than those in the CN (N = 22) or control groups (N = 26). Serum 5-S-CD levels in the CN group were not significantly different from those in the control group. There was a significant correlation between the serum 5-S-CD level and size (surface area) of the GNP. There was a significant inverse correlation between the serum 5-S-CD level and age in the CN and control groups, but not in the GPN group. Serum 5-S-CD levels were transiently elevated immediately after treatment of patients with GPN with a combination of skin abrasion and cryotherapy with solid carbon dioxide. These results suggest that serum 5-S-CD levels in the GPN group reflected the 5-S-CD derived from GPN, particularly in patients less than 5 years old. This indicates that melanogenesis may be accelerated in infant patients in the GPN group.

Topics: Adolescent; Age Factors; Carbon Dioxide; Child; Child, Preschool; Cryotherapy; Cysteinyldopa; Dermabrasion; Female; Humans; Infant; Male; Nevus; Nevus, Pigmented; Skin Neoplasms

Two cases of metastatic malignant melanoma of the lower limb who were treated successfully with hyperthermic isolated limb perfusion are reported. One patient was infused with cisdiammine (1.1-cyclobutanedicarboxylate) platinum (II) (carboplatin, Paraplatin, Bristol-Myers Squibb Company, New Jersey, USA), and the other was infused with human natural beta-interferon (Feron, Toray, Tokyo, Japan), via the external iliac artery. The first case showed a remarkable suppression of the growth of multiple metastatic melanoma nodules associated with numerous melanophage infiltrations, as shown histopathologically after the operation. The patient's serum level of 5-S-cysteinyl dopa decreased for the two months following the treatment. In the second case, new formation of metastatic melanoma nodules was completely suppressed for up to 12 months following the operation. Analysis of immunological parameters showed that the number of peripheral CD8+ lymphocytes gradually and constantly increased after the operation, while that of CD4+ lymphocytes transiently increased and then returned to the pre-operative level. Natural killer activity transiently decreased to a slight degree 4 days after the operation and then returned to the pre-operative level 21 days after the operation. Side effects, such as nausea, vomiting and leg discomfort, were seen in the patient (Case 1) treated with carboplatin, but were completely reversible. These results suggest that hyperthermic isolated limb perfusion with concomitant infusion of carboplatin or beta-interferon is effective in suppressing the growth of metastatic malignant melanomas of the lower limb.

Topics: Aged; Antineoplastic Agents; Carboplatin; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Chemotherapy, Cancer, Regional Perfusion; Cysteinyldopa; Female; Humans; Hyperthermia, Induced; Interferon-beta; Killer Cells, Natural; Leg; Lymphatic Metastasis; Melanoma; Middle Aged; Nausea; Remission Induction; Skin Neoplasms; Vomiting

A 36-year-old Japanese female, in whom malignant melanoma was detected from symptoms of brain metastasis, died 7 months later. The serum levels of 5-S-Cysteinyldopa (5-S-CD) and intercellular adhesion molecule-1 (ICAM-1) correlated with the disease progression. These values may be useful markers of disease progression for malignant melanoma, because the material is easily, frequently, and non-invasively obtained at regular intervals.

Topics: Adult; Biomarkers, Tumor; Brain Neoplasms; Cysteinyldopa; Disease Progression; Fatal Outcome; Female; Humans; Intercellular Adhesion Molecule-1; Melanoma; Radionuclide Imaging; Skin Neoplasms; Tomography, X-Ray Computed

Urinary excretion of 5-S-cysteinyldopa (5-S-CD) has been used as a biochemical marker of melanoma progression. Melanomas produce not only 5-S-CD but also 5,6-dihydroxyindole-2-carboxylic acid (5,6DHI2C) as major intermediates in melanin formation. 5,6DHI2C is then metabolized to the two O-methyl derivatives, 5H6MI2C and 6H5MI2C. The aim of this study was to determine which marker in serum and urine most sensitively reflected the progression of melanoma.. Serum and 24-hour urine samples were collected and assayed serially by high-performance liquid chromatography every 1 to 4 months in 28 patients with primary or recurrent melanomas, for up to 48 months.. Serum concentration and urinary excretion of 5-S-CD and 6H5MI2C in patients with melanoma without metastases were close to those obtained from normal subjects. Metastases developed in 9 of the 28 patients. In seven of these nine patients, serum or urinary 5-S-CD values were elevated before or at the time of clinical detection of visceral metastases. However, serum 5-S-CD was elevated significantly earlier and reflected melanoma progression better than the physical examination and/or laboratory tests, such as scintigraphy and echography. Serum 6H5MI2C values exceeded the normal range shortly before death in three patients, and urinary 6H5MI2C did not increase at any stage in most patients, therefore these metabolites did not reflect progression of disease.. Among the four markers, serum 5-S-CD appears to be the best biochemical marker for the detection of progression of melanotic melanoma, a value of more than 10 nmol/l suggesting the presence of metastasis.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Chromatography, High Pressure Liquid; Cysteinyldopa; Female; Humans; Indoles; Male; Melanoma; Middle Aged; Neoplasm Metastasis; Skin Neoplasms

This study reports our success in developing a murine monoclonal antibody (mAb) against phaeomelanin and its major precursor, 5-S-cysteinyldopa (5-S-CD). A competitive enzyme-linked immunosorbent assay was developed and demonstrated that the new mAb, designated as 6D4 (IgG1, kappa), reacted with both 5-S-CD and phaeomelanin, but not with eumelanin. The concentrations required for 50% inhibition of 5-S-CD and phaeomelanin were 65 ng/ml and 95 ng/ml respectively. The minimal amount of 5-S-CD and phaeomelanin which could be detected by mAb 6D4 was approximately 5 and 6 ng/ml, respectively. The immunohistochemical assay indicated that the antigenic epitope(s) recognized by mAb 6D4 was localized in the cytoplasm of certain types of melanocytic tumours, such as superficial spreading melanoma.

Topics: Animals; Antibodies, Monoclonal; Antibody Specificity; Binding, Competitive; Cysteinyldopa; Enzyme-Linked Immunosorbent Assay; Humans; Hybridomas; Immunoglobulin G; Melanins; Melanoma; Mice; Mice, Inbred BALB C; Neoplasms; Nevus, Pigmented; Rats; Skin Neoplasms; Tumor Cells, Cultured

1) The urinary 5-S-CD contents in malignant melanoma subjects (n = 135) and non-melanoma subjects (n = 204) were measured by HPLC. These results suggest that, as a biochemical marker, periodic measurement of urinary 5-S-CD is quite useful for evaluating the determinations of stage classification (UICC, 1987), and the detection of metastases, the therapeutic efficacy of operation or immunochemotherapy against malignant melanoma. 2) Quantitative analyses of 5-S-CD values in tissues from primary malignant melanoma lesions (n = 24) and pigmentary tumors other than melanomas (n = 136) showed 80.6-821.4 ng/mg and N.D.-55.0 ng/mg respectively. In view of the above findings, it was suggested that the pigmentary tumors can be diagnosed as malignant melanoma if the 5-S-CD value in the tissues is higher than 100 ng/mg.

Topics: Biomarkers, Tumor; Cysteinyldopa; Humans; Melanoma; Skin Neoplasms

We previously reported that serum 5-S-cysteinyldopa (5-S-CD) tended to elevate earlier and reflect melanoma progression better than urinary 5-S-CD. In patients without metastatic melanomas, serum concentration and urinary excretion of 5-S-CD and 6-hydroxy-5-methoxy-indole-2-carboxylic acid (6H5MI2C) were within the upper limits of normal controls. In this report, we presented more precisely the changes in these melanin-related markers and clinical courses of four melanoma patients. Serum and 24-hour urine samples were serially collected and assayed every 1 to 4 months. Three of them developed stage IV malignant melanomas and died of metastatic disease. 6H5MI2C in serum and urine did not reflect the progression of disease. Among the 4 parameters considered, 5-S-CD in serum appeared to be the best biochemical marker for melanoma progression. Serum 5-S-CD over the upper limit of 10 nmol/L was suggested as a serious sign of the progression of melanoma.

Topics: Aged; Biomarkers, Tumor; Cysteinyldopa; Female; Humans; Male; Melanoma; Middle Aged; Skin Neoplasms

This article introduces a rapid high-performance liquid chromatographic assay to measure urinary pheomelanin and eumelanin metabolites, 5-S-cysteinyldopa and indoles, 5(6)-hydroxy-6(5)-methoxyindole-2-carboxylic acid.. Our high-performance liquid chromatographic study clearly showed (1) urine of melanoma patients with positive metastasis revealed significant amounts of 5-S-cysteinyldopa and indoles (5,6-dihydroxyindole-2-carboxylic acid plus 6-hydroxy-5-methoxyindole-2-carboxylic acid) above 1 mumol/d and 2 mumol/d, respectively; and (2) in patients with metastasis-free melanoma these melanin metabolites might be excreted into the urine but always less than the two values cited above.. As there is a discrepancy regarding the specificity of 5-S-cysteinyldopa as a marker for estimation of melanoma metastasis, high-performance liquid chromatographic measurement of urinary indoles will provide an additional assay in the detection of melanoma metastasis from an early stage. Both melanoma markers were increased in the urine of patients with metastatic melanoma.

Topics: Adult; Biomarkers, Tumor; Cysteinyldopa; Humans; Indoles; Male; Melanoma; Middle Aged; Skin Neoplasms

To investigate the UV effect on epidermal melanocytes, 21 volunteers and 11 patients with dysplastic nevus syndrome (DNS) received UVB irradiation three times weekly during 17 days. Skin biopsies were taken before and three weeks after the last irradiation (on day 37) from exposed and covered buttock skin. The epidermal melanocyte population density was estimated in dopa-stained split skin preparations. The biopsies taken on day 37 revealed that repeated UVB irradiation induces an increase in the number of melanocytes not only in exposed but also in covered skin. This increased mitotic activity might be a link between sun exposure and melanoma development in covered skin. The size of the proliferative response was inversely correlated to the basal melanocyte number. The larger population increase in skin with few melanocytes might amplify the propagation of DNA damage and increase the likelihood of tumor development. The pigment metabolite 5-S-cysteinyldopa (5-S-CD) was measured in urine before the irradiation and twice weekly until day 38. No correlation was found between the basal 5-S-CD excretion and the size or activity of the melanocyte organ, suggesting that the basal 5-S-CD excretion is mainly of non-melanocytic origin. Despite numerous nevi, DNS-patients did not differ from controls in their 5-S-CD excretion. The normal upper range for the tumor maker 5-S-CD is therefore valid in these melanoma-prone subjects. During the irradiation, subjects with a low tanning ability developed a more pronounced erythema and excreted more 5-S-CD than those with a good tanning ability. This suggests that the UVB-induced 5-S-CD excretion is rather due to melanocyte damage than to an increased melanin synthesis. To investigate the influence of sun exposure on the development of nevi and melanoma (CMM), the distribution over the body surface of CMM, common nevi (CN) greater than or equal to 2 mm and dysplastic nevi (DN) was registered in 121 melanoma patients and 310 controls. Four times as many nevi were found in a sun-exposed area than in a comparable sun-protected area, demonstrating that sun exposure plays an important role in nevus development. Subjects with DNA had a larger difference in nevus counts between the two areas than subjects without DN, indicating a different UV-dose and/or a higher sensitivity to the "nevogenic" effect of UV-light than subjects without DN.(ABSTRACT TRUNCATED AT 400 WORDS)

Topics: Adult; Animals; Cysteinyldopa; Female; Humans; Male; Melanocytes; Melanoma; Middle Aged; Neoplasms, Radiation-Induced; Nevus; Skin Neoplasms; Ultraviolet Rays

Urinary 5-S-cysteinyldopa (5-S-CD) has been used as a biochemical marker of melanoma metastasis. A method was developed for determining the eumelanin-related metabolites 5(6)-hydroxy-6(5)-methyoxyindole-2-carboxylic acids (5H6MI2C and 6H5MI2C) in small volumes of serum. We compared these indoles and 5-S-CD regarding the correlation of their production in melanoma, circulation in blood, and excretion in urine, with the weight of highly pigmented, B16 mouse melanoma. An excellent correlation was found between the serum concentration of 5H6MI2C + 6H5MI2C (r = 0.92) and 5-S-CD (r = 0.89) and tumor weight. However, the urinary excretion of 5H6MI2C + 6H5MI2C and 5-S-CD did not show any significant correlation. These results suggest that 5H6MI2C + 6H5MI2C and 5-S-CD in serum may better reflect melanoma progression than those in urine. Furthermore, comparison of the contents of these melanin-related metabolites between highly pigmented and less pigmented B16 melanomas suggests that 5-S-CD may be accumulated in pigmented melanoma by virtue of binding to melanin and that catechol-O-methyltransferase (COMT) may play a regulatory role in pigmentation.

Topics: Animals; Chromatography, High Pressure Liquid; Cysteinyldopa; Indoles; Male; Melanins; Melanoma; Mice; Mice, Inbred C57BL; Skin Neoplasms; Tumor Cells, Cultured

We describe a 52-year-old man with a pedunculated pigmented eccrine poroma mimicking a nodular malignant melanoma in the occipital region. The tumor was once resected but soon recurred. Histologically, the tumor mass extended from the epidermis downwards into the dermis and contained melanin granules in some areas. The tumor cells were uniformly cuboidal in appearance and had round, deeply basophilic nuclei. Initially, the urinary excretion level of 5-S-cysteinyldopa (5-S-CD) was high, but, after resection of the tumor, the level of 5-S-CD returned to normal.

Topics: Adenoma, Sweat Gland; Cysteinyldopa; Diagnosis, Differential; Humans; Male; Melanoma; Middle Aged; Neoplasm Recurrence, Local; Scalp; Skin Neoplasms; Solitary Pulmonary Nodule

On the basis of clinical and histopathological studies on 17 patients who had been diagnosed as having pigmented Spitz nevus (PSN), pigmented spindle cell nevus (PSCN) was surmised to be a type of pigmented Spitz nevus. In order to distinguish pigmented spindle cell nevus and pigmented Spitz nevus from early melanoma, 5 PSCN cases and 12 PSN cases were analyzed by the fine-needle aspiration fluorescence method, touch fluorescence method and measurement of the 5-S-CD level in the lesion. With the touch fluorescence method, fluorescent tumor cells were detected in one case of PSN. With the fine-needle aspiration fluorescence method, fluorescent tumor cells were detected in one PSCN case and 2 PSN cases. In comparison with fluorescent melanoma cells, the detected fluorescent tumor cells were smaller in size and number and resembled melanocytes. The 5-S-CD level in the lesion was 50 ng/mg or less in all cases, whereas the level in melanoma is known to be a high 100 ng/mg or more. In the final analysis, measurement of the 5-S-CD level in the lesion was concluded to have the greatest utility for differential diagnosis of pigmented spindle cell nevus and pigmented Spitz nevus from early melanoma.

Topics: Adolescent; Adult; Biomarkers, Tumor; Child; Child, Preschool; Cysteinyldopa; Diagnosis, Differential; Female; Histocytochemistry; Humans; Melanoma; Microscopy, Fluorescence; Nevus, Pigmented; Skin Neoplasms

To develop a biochemical diagnosis for malignant melanomas, 5-S-cysteinyldopa in the tissues of malignant melanomas and other pigmented tumors were quantitatively analysed. The mean 5-S-CD value from primary melanotic melanoma lesions was 202.4 ng/mg (80.6-821.4) and those of metastatic melanoma lesions was 214.0 ng/mg (96.2-520.0). There was no significant differences in these 5-S-CD values between the types of primary lesions. On the other hand, the 5-S-CD values from pigmented tumors other than melanomas, such as Spitz nevus, dysplastic nevus, pigmented basal cell epithelioma, and melanoepithelioma Bloch II were less than 55 ng/mg. From these results it is thought that pigmented tumors can be diagnosed as malignant melanomas, if the 5-S-CD values in the tissues are over 100 ng/mg. The mean 5-S-CD values of pigmented macules of SSM was 734.5 ng/mg (320.0-1666.3) and those of ALM was 1.8 ng/mg (0.7-3.6). These data suggest that the biochemical diagnoses of SSM in situ can be based on 5-S-CD values, the diagnoses of ALM in situ can not.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Cysteinyldopa; Dihydroxyphenylalanine; Female; Humans; Male; Melanoma; Middle Aged; Skin Neoplasms

Methods for making diagnoses of early lesions of malignant melanomas using a touch-fluorescence method from the cut surface of the lesions and quantitative analysis of 5-S-CD in the tissues were developed. 1) The touch-fluorescence method and analyses of 5-S-CD values in the tissues were extremely useful in making definitive diagnoses of early lesions of nodular melanoma (NM). 2) Only quantitative analyses of 5-S-CD values in the tissues were useful in making definitive diagnoses of early lesions of superficial spreading melanoma (SSM). 3) It is impossible to definitively diagnose lentigo maligna using either way mentioned above. From these results, it appears that quantitative analysis of 5-S-CD values in the tissue was the most useful method for making definitive diagnoses in case of early lesions of NM and SSM.

Topics: Adolescent; Adult; Aged; Cysteinyldopa; Dihydroxyphenylalanine; Female; Fluorescence; Humans; Male; Melanoma; Microscopy, Fluorescence; Middle Aged; Predictive Value of Tests; Skin Neoplasms

Two patients with metastasizing melanoma and diffuse melanosis have previously been reported to excrete large quantities of trichochromes in the urine. The present study describes 2 further melanoma patients with diffuse melanosis and trichochromuria. The hair of one of the patients which had been red in childhood and turned brown in adult age returned to red with the appearance of melanosis. Normal excretion of a methylated melanocytic metabolite, 6-hydroxy-5-methoxyindole-2-carboxylic acid, was observed in this patient, possibly indicating exhaustion of the methylating system. The other patients excreted large quantities of 6-hydroxy-5-methoxyindole-2-carboxylic acid. Both patients showed highly increased excretion of 5-S-cysteinyldopa. Both patients with melanosis exhibited fine electrone-dense granules in lysosomes of dermal histiocytes. The findings support the concept that trichochromes or similar pigments in dermal histiocytes are responsible for diffuse melanosis in melanoma patients.

Topics: Adult; Aged; Cysteinyldopa; Female; Humans; Indoles; Male; Melanoma; Melanosis; Microscopy, Electron; Skin Neoplasms; Thiazines

Topics: Cysteinyldopa; Dihydroxyphenylalanine; Humans; Indoles; Melanoma; Skin Neoplasms

Topics: Cysteinyldopa; Humans; Melanins; Melanoma; Skin Neoplasms

The effect of anti-Parkinson therapy on the urinary excretion of 5-S-cysteinyldopa (5SCD), a catechol metabolite of dihydroxyphenylalanine (DOPA) and marker for the melanocyte, was studied by means of high performance liquid chromatography. 5SCD was normal in Parkinson patients treated with anticholinergics. DOPA administration increased 5SCD excretion. Carbidopa and DOPA together elevated 5SCD markedly in a dose-dependent manner to values higher than seen in some patients with metastatic malignant melanoma. The effect of anti-Parkinson therapy should be considered when using 5SCD as a tumor marker or when a Parkinson patient has a melanoma.

Topics: Adult; Aged; Carbidopa; Cysteinyldopa; Dihydroxyphenylalanine; Humans; Melanoma; Middle Aged; Parkinson Disease; Skin Neoplasms

In case of uncharacteristic patient complaints or slightly pathologic BSR or liver enzyme levels in patients, suffering from melanoma, the determination of 5-S-cysteinyldopa in the 24-h-urine may be of great diagnostic value. If the complaints and the pathologic values are early symptoms and signs of generalized metastases being clinically not yet manifest, the excretion of 5-S-cysteinyldopa in the urine is often increased. In such cases, a thorough and possibly invasive search for metastases is indicated.

Topics: Cysteinyldopa; Dihydroxyphenylalanine; Humans; Isomerism; Melanoma; Neoplasm Metastasis; Skin Neoplasms

Topics: Adult; Aged; Cysteinyldopa; Dihydroxyphenylalanine; Female; Humans; Male; Melanoma; Microscopy, Fluorescence; Middle Aged; Skin Neoplasms

Cysteinyldopas and Dopa in the urine and tissues of Japanese melanoma patients were investigated quantitatively by means of high-performance liquid chromatography. Cysteinyldopa isomers were detected in the urine of eumelanic Japanese patients. The amount (X +/- SD%) of each isomer of cysteinyldopa in the urine was 80.26 +/- 4.66% in 5-S-cysteinyldopa, 9.39 +/- 1.64% in 2-S-cysteinyldopa, 7.07 +/- 3.33% in 2, 5-S, S-dicysteinyldopa, and 3.28 +/- 1.43% in 6-S-cysteinyldopa. The amount of cysteinyldopa in melanoma tissues was 26-314 times more than that of Dopa. The amount (X +/- SD%) of cysteinyldopa in the tissues was 80.34 +/- 1.75% in 5-S-cysteinyldopa, 11.06 +/- 1.91% in 2-S-cysteinyldopa, 6.27 +/- 1.43% in 2, 5-S, S-dicysteinyldopa, and 2.34 +/- 0.61% in 6-S-cysteinyldopa. The fact that the percentages of each isomer of cysteinyldopa in the urine and in the tissues were approximately constant suggests that the cysteinyldopas secreted from melanoma cells were excreted into the urine without being metabolized.

Topics: Cysteinyldopa; Dihydroxyphenylalanine; Humans; Isomerism; Japan; Lymphatic Metastasis; Melanoma; Skin Neoplasms

Histochemical findings of primary and metastatic amelanotic melanomas were shown by the formaldehyde-induced fluorescence method (Falck and Hillarp). All or some of the amelanotic melanoma cells were discovered to emit green specific fluorescence. Results of the determination of 5-S-cysteinyldopa and DOPA in amelanotic melanoma tissues indicated that the specific fluorescence emitted by these cells is primarily due to the presence of 5-S-cysteinyldopa. The values of 5-S-cysteinyldopa in these tissues were lower than those in melanotic melanoma, but were approximately the same as those in pigmented nevus. When unpigmented tumors were histopathologically revealed to be malignant, amelanotic melanoma could be definitely diagnosed by the fluorescence method of Falck and Hillarp and the biochemical analysis of 5-S-cysteinyldopa in the tissues.

Topics: Adult; Aged; Carcinoma, Basal Cell; Cysteinyldopa; Dihydroxyphenylalanine; Humans; Male; Melanoma; Microscopy, Fluorescence; Nevus, Pigmented; Skin Neoplasms

Topics: Catechol Oxidase; Cysteinyldopa; Dihydroxyphenylalanine; Ditiocarb; gamma-Glutamyltransferase; Humans; In Vitro Techniques; Iodoacetamide; Melanoma; Monophenol Monooxygenase; Skin; Skin Neoplasms

The effect of dopa, cysteine, and glutathione on 5-S-cysteinyldopa (5-S-CD) genesis in melanoma cells cultured in normal and tyrosine- and cysteine-free media has been studied. In normal media only melanotic melanoma cells have been found to secrete 5-S-CD into the medium. In the presence of dopa and cysteine, both, media incubated with and without cells have been found to produce 5-S-CD. In the presence of dopa and glutathione, however, cell-free media did not show the presence of 5-S-CD. In contrast melanoma cell-cultured media has been found to contain large quantities of this amino acid. The optimum condition for maximum production of 5-S-CD via glutathione-dependent pathway has been found to be at the dopa concentration of 10(-5) M when glutathione is present at the concentration of 10(-5) M in the culture medium. Thus dopa concentration with regards to glutathione is 1:1 on the molar basis which is twice the dopa concentration required in in vitro noncellular tyrosinase system. It is suggested that higher dopa requirement in our melanoma cell culture system reflects the co-existence of eu- and pheomelanin synthesis taking place according to their genetically predetermined proportions.

Topics: Animals; Cells, Cultured; Cricetinae; Culture Media; Cysteine; Cysteinyldopa; Dihydroxyphenylalanine; Drug Synergism; Fibroblasts; Glutathione; Humans; In Vitro Techniques; Melanoma; Mice; Neoplasms, Experimental; Skin Neoplasms; Stimulation, Chemical; Tyrosine

Topics: Aged; Cysteinyldopa; Dihydroxyphenylalanine; Humans; Isomerism; Male; Melanoma; Skin Neoplasms

Topics: Cysteinyldopa; Dacarbazine; Dihydroxyphenylalanine; Female; Humans; Lymphoma; Melanoma; Middle Aged; Skin Neoplasms

Topics: Adolescent; Adult; Aged; Child; Cysteinyldopa; Dihydroxyphenylalanine; Female; Humans; Male; Melanoma; Middle Aged; Skin Neoplasms

In a total of 1,828 determinations, urinary excretion of 5-S-Cysteinyldopa was studied over a period of three years in 384 patients treated for melanoma or with metastases of malignant melanoma. By serial investigations the excretion of 5-S-Cysteinyldopa was compared to the course of the disease. In the case of small and circumscribed metastases which could be eliminated by surgical treatment, the excretion of 5-S-Cysteinyldopa remained normal. When the disease became generalized, an increase of the urinary excretion of 5-S-Cysteinyldopa prior to the clinical manifestation of the metastases was observed in only four out of 26 cases. In the remaining cases, the increase of 5-S-Cysteinyldopa coincided with the manifestation of metastases, or the excretion of the substance became pathological when the metastases were already conspicuous. In five patients, the urinary excretion of 5-S-Cysteinyldopa remained normal inspite of widespread disease. Therefore, its diagnostic value seems to be similar to that of the "common" laboratory investigations the results of which are only pathological when the disease has already become generalized. Our investigations demonstrate that serial investigations of the urinary excretion of 5-S-Cysteinyldopa only rarely indicate melanoma metastases prior to their clinical manifestation. In cases of early metastasing melanoma, all common laboratory investigations are of limited value. BSR and GGT levels which become pathological very early in the course of the disease are so sensitive that slightly pathological levels may be ambiguous. In these cases, however, pathological levels of 5-S-Cysteinyldopa most probably will indicate a widespread disease.

Topics: BCG Vaccine; Cysteinyldopa; Dihydroxyphenylalanine; Female; Humans; Male; Melanoma; Melphalan; Neoplasm Metastasis; Skin Neoplasms

Of 16 cell lines derived from 12 human melanomas obtained from 11 patients, all were established as permanent cell lines: 7 from primary tumors and 9 from metastatic tumors. Study of the early subcultures and established cell lines showed that melanocytes passed through a phase of dedifferentiation during which they took on a fibroblast-like appearance and were hypodiploid and nontumorigenic in nude (thymus-deficient) mice. Phenotypic modulation in vitro was shown to be dependent on the culture medium. The lines varied considerably in karyologic and phenotypic expression (as assessed by morphologic appearance and 5-S-cysteinyldopa production). Fibroblast-like, epithelioid, nonpigmented, achromic, and pigmented cells were obtained from the same tumor. Heterotransplantation into nude mice revealed wide variations in tumorigenicity: The latency of the tumors, their size, and infrequent metastases bore no relationship to the phenotypic modulation of the melanocytes as expressed in vitro. Melanogenesis is therefore not related to malignancy; they are two independent characteristics.

Topics: Animals; Cell Differentiation; Cell Line; Culture Media; Cysteinyldopa; Female; Humans; Male; Melanocytes; Melanoma; Mice; Mice, Nude; Microscopy, Electron; Neoplasm Metastasis; Neoplasm Transplantation; Neoplasms, Experimental; Phenotype; Skin Neoplasms; Transplantation, Heterologous

Determinations of the urinary excretion of 5-S-cysteinyldopa were performed in 571 patients previously treated by surgery for melanoma or melanoma metastasis. 90% of the 161 patients with metastases showed values exceeding 0.15 mg/24 h, and 9% of the 410 patients without metastases had such values. The increase in 5-S-cysteinyldopa excretion was generally more pronounced in men with metastases than in women, 98% of the men and 77% of the women with metastases showing values exceeding 0.15 mg/24 h. High levels of 5-S-cysteinyldopa are of grave prognostic significan4% died within one month, and only 3% survived for more than a year. In Sweden, determination of 5-S-cysteinyldopa in patients operated on for melanoma gives maximum information in the winter (October--March), when sun exposure does not influence the excretion levels.

Topics: Adult; Aged; Cysteinyldopa; Dihydroxyphenylalanine; Female; Hair Color; Humans; Lymphatic Metastasis; Male; Melanoma; Middle Aged; Neoplasm Metastasis; Sex Factors; Skin Neoplasms; Sunlight

Topics: Adult; Aged; Cysteinyldopa; Dihydroxyphenylalanine; Female; Humans; Male; Melanoma; Middle Aged; Skin Neoplasms

In an attempt to improve the diagnosis of melanotic tumors, we have compared the diagnosis obtained by histological examination of 216 skin tumors and their metastases with that obtained by using a conjunction of four techniques: tissue culture, cytoenzymology, in situ electron microscopy and 5-S-cysteinyldopa (5-S-CD) assay. In 46 cases the final diagnosis as determined by one or more of these tests differed from the initial histological diagnosis, but was confirmed by repeat histological examination. We conclude that this method presents a valuable new approach to the differential diagnosis of human malignant melanoma.

Topics: Adolescent; Adult; Aged; Ascitic Fluid; Culture Techniques; Cysteinyldopa; Diagnosis, Differential; Female; Histocytochemistry; Humans; Lymphatic Metastasis; Male; Melanoma; Methods; Microscopy, Electron; Middle Aged; Nevus, Pigmented; Pleural Effusion; Skin Neoplasms

Free dopa and 5-S-cysteinyldopa were extracted from two human melanomas. Subsequent hydrolysis of carefully washed melanoma tissue released dopa and 5-S-cysteinyldopa, indication the presence of these catechol amino acids in proteins. Cysteinyldopa-containing proteins may represent the antigens previously demonstrated in human melanomas.

Topics: Cysteinyldopa; Dihydroxyphenylalanine; Humans; Melanoma; Skin Neoplasms

Determination of urine cysteinyldopa excretion is the most sensitive chemical test for the detection of melanoma metastases and has been successfully applied during the Scandinavian winter, when sun irradiation is low. The value of this determination, under Australian conditions of greater sun irradiation, has been assessed by comparing the cysteinyldopa excretion of patients with that of normal subjects exposed to sunlight. Cysteinyldopa is an intermediate in the biosynthesis of the red-brown phaeomelanin. Of 20 patients without known secondary melanoma, the cysteinyldopa concentration of "spot" urines ranged from 0 to 190 (mean 48) microgram/ml; of six known to have local metastases, 0 to 80 (mean 19) microgram/ml; and of four known to have extensive metastases 80 to 1350 (mean 330) microgram/ml. The effect of sun irradiation alone was assessed in nine healthy subjects followed one to 11 weeks before and after recorded periods of sun exposure. The cysteinyldopa concentrations of 24-hour urines ranged from 40 to 3100 microgram/ml. Increases occurred three to 10 days following sun exposure and were greatest following multiple small exposures in an individual with "Celtic" complexion. It is concluded that measurement of cysteinyldopa concentration would be of value in the follow-up of melanoma patients in Australia only if patients could be persuaded to live under conditions free of all direct sun-irradiation.

Topics: Adolescent; Adult; Australia; Cysteinyldopa; Dihydroxyphenylalanine; Humans; Melanoma; Neoplasm Metastasis; Skin Neoplasms; Sunlight

The urine of patients with melanoma metastases and increased urinary excretion of 5-S-cysteinyldopa was examined for trichochromes. Five of 16 patients showed urinary excretion of trichochromes B and C. None excreted trichochromes E or F. All patients showing trichochrome excretion had very large amounts of cysteinyldopa in their urine.

Topics: Adult; Aged; Cysteinyldopa; Female; Hair Color; Humans; Male; Melanins; Melanoma; Middle Aged; Neoplasm Metastasis; Skin Neoplasms

Topics: Cysteinyldopa; Dihydroxyphenylalanine; Humans; Melanoma; Neoplasm Metastasis; Skin Neoplasms

A newly discovered amino acid, 5-S-cysteinyldopa, present in the urine of healthy subjects is excreted in pathological amounts in many patients suffering from melanoma metastases. Increased excretion of 5-S-cysteinyldopa may be observed before metastases become clinically evident. Determination of 5-S-cysteinyldopa is superior to determination of dopa+dopamine in the diagnosis of melanoma metastases.

Topics: Adult; Aged; Cysteinyldopa; Dihydroxyphenylalanine; Eye Neoplasms; Female; Humans; Male; Melanoma; Middle Aged; Neoplasm Metastasis; Skin Neoplasms

Previous studies have demonstrated a specific cytoplasmic fluorescence in human melanocytes, as well as in pigmented nevi and in malignant melanomas, when the formaldehyde histofluorescence method for visualization of certain catechol and indole derivatives was used. In malignant melanoma two fluorogenic substances, dopa and cysteinyldopa, were found previously. In human melanocytes and benign nevi cells the fluorogenic catechols have so far not been characterized, since chemical analyses are difficult to perform on skin, due to the small amounts of catechols present. However, using split thickness skin quantitative determinations are possible by sensitive fluorometric methods. The chemical analyses of cysteinyldopa showed that in human adult skin most or all was located in the superficial layers. The only specific fluorescence in the thin skin was found in dendritic melanocytes. The findings leave little doubt that cysteinyldopa is stored in melanocytes although the possibility of a concomitant occurrence of other thioethers is not excluded. Nevi and giant nevi were also similarly studied and we found considerable amounts of cysteinyldopa in the nevi. It seems as if the cysteinyldopa is stored in the fluorescent nevi cells. There was no consistent difference in the content of the catechol derivatives between intradermal and compound nevi.

Topics: Adolescent; Child; Child, Preschool; Cysteinyldopa; Dihydroxyphenylalanine; Female; Humans; Male; Melanocytes; Nevus; Nevus, Pigmented; Skin Neoplasms