PDGFB quantification is a useful tool in the diagnosis of dermatofibrosarcoma protuberans: a study of 10 cases.

BACKGROUND: Genetic aberrations involving the platelet-derived growth factor-beta (PDGFB) gene and the collagen type 1 alpha1 (COL1A1) gene have been implicated in the pathogenesis of dermatofibrosarcoma protuberans (DFSP), a slow-growing and locally infiltrative dermal tumour. AIM: To investigate the genetic rearrangements in 10 patients who presented with a clinical diagnosis of DFSP. METHODS: Total RNA was obtained from frozen sections and sections embedded in paraffin wax, and used for direct sequencing of the cDNA produced from reverse transcription (RT) PCR. The expression of PDFGB mRNA in each of these cases was also examined. RESULTS: Of the 10 samples examined, 9 had the COL1A1/PDGFB fusion transcript by DNA sequencing. The sequenced products showed that there was a fusion between the end of exons 6, 8, 29 (two cases), 38, 25 or 47 (three cases) and the start of exon 2 of the PDGFB gene. Quantitative RT-PCR identified all samples as having significantly higher expression of the PDGFB gene compared with normal skin or dermatofibroma. CONCLUSIONS: Detection of the COL1A1/PDGFB fusion transcript may be important for the diagnosis of DFSP. Furthermore, relative PDGFB gene quantification by real-time PCR may also provide a good diagnostic tool when other methods fail to give conclusive results.

Regulation of high molecular weight-melanoma associated antigen (HMW-MAA) gene expression by promoter DNA methylation in human melanoma cells.

The human high molecular weight-melanoma associated antigen (HMW-MAA) is a membrane-bound chondroitin sulfate proteoglycan that is variably expressed in a high percentage of melanoma cell lines and tumors. Since the mechanism(s) regulating HMW-MAA expression has(ve) not been defined, in this study, we have examined whether promoter DNA methylation regulates the level of HMW-MAA expression. In melanoma cell lines, the level of HMW-MAA mRNA and protein expression is coordinately regulated, implicating a transcriptional control mechanism. Consistent with a role for regulation by DNA methylation, we have found that a dense CpG island flanks the human HMW-MAA gene transcriptional start site. Methylation-specific PCR and sodium bisulfite DNA sequencing analyses indicate that the HMW-MAA promoter is heavily methylated in melanoma cell lines, melanoma lesions and normal lymphocytes that do not express HMW-MAA; in contrast, the HMW-MAA promoter is not methylated in melanoma cell lines and tumors that express this antigen. In addition, HMW-MAA expression is markedly induced in HMW-MAA-negative melanoma cell lines by incubation with the DNA methyltransferase inhibitor 5-aza-2'-deoxycytidine. In summary, our results establish DNA methylation as a key regulator of HMW-MAA expression by human melanoma cells. This information represents a useful background to optimize immunotherapeutic strategies targeting HMW-MAA.

Metastatic melanoma to the palatine tonsil with a favourable prognosis.

Metastasis to the oral cavity from cutaneous melanoma is rare: fewer than 30 cases of metastatic melanoma to the palatine tonsil have been reported. Tonsil metastasis is haematogenously disseminated and therefore usually has a poor prognosis. We present a case of metastatic melanoma to the palatine tonsil occurring 6(1/2) years after removal of the primary cutaneous lesion. The patient has remained disease-free for 18 months since the removal of skin and tonsil metastases. Immunohistopathologically, HLA class II and costimulatory factor B7-2 molecules were concomitantly expressed on melanoma cells: we suggest that the patient was therefore able to develop antimelanoma T-cell activation resulting in prevention of further metastasis, and thus a favourable prognosis.

Detection of minimal residual disease in acute leukemia by Tc-99m MIBI femoral marrow imaging.

To determine the potential of Tc-99m MIBI femoral marrow imaging for detecting minimal residual disease in acute leukemia, MIBI images of 68 patients with acute leukemia and 110 control patients were examined. MIBI accumulation was classified into three patterns: not detectable, mild accumulation, and clearly visualized accumulation. Clearly visualized accumulation was interpreted as abnormal. The mean uptake ratio of the femoral marrow to muscle was calculated. Forty-five patients who were in complete remission (CR) at the time of MIBI imaging had a follow-up study (mean interval, 23 months). Clearly visualized accumulation was demonstrated in 35 patients with acute leukemia: in 7 patients before starting induction chemotherapy, in 12 patients after relapse, and in 16 of the 49 patients in the CR group. Mild accumulation was demonstrated in 14 patients in the CR group and in 13 control group patients. No detectable accumulation was observed in 19 patients in the CR group and in 97 control patients. The marrow and muscle uptake ratio of patients before starting chemotherapy (2.29 +/- 0.26) was greater compared with that in patients after relapse (1.78 +/- 0.44, P < 0.02) and in patients with abnormal accumulation despite complete remission (1.84 +/- 0.36, P < 0.01). The uptake ratio in patients with abnormal accumulation despite CR was higher compared with patients with mild accumulation in CR (1.26 +/- 0.13, P < 0.001) and controls (1.23 +/- 0.10, P < 0.001) who had mild accumulation. Fifteen patients with abnormal accumulation despite CR had a markedly greater relapse rate (66.7% > 10.0%, P < 0.005), a higher mortality rate (46.7% > 6.7%, P < 0.01), and shorter remission time (8.7 +/- 10.2 months < 35.9 +/- 20.1 months, P < 0.001) compared with 30 patients without abnormal accumulation in CR. MIBI femoral marrow imaging may be a useful and simple method for monitoring levels of residual leukemic cells. Clearly visualized MIBI accumulation may be a marker for relapse.

[Drug-induced myocardial disease--adriamycin cardiotoxicity].

Myocardial damage may be induced by several categories of drugs. Adriamycin is one of the most effective chemotherapeutic agents. Unfortunately, it develops a distinctive and life-threatening congestive heart failure by well-characterized, dose-dependent cardiac toxicity. Left ventricular ejection fraction determined using echocardiography or radionuclide angiography does not decrease sensitively and linearly with the adriamycin dose level. Cardiac function is preserved until a critical degree of myocardial damage is reached, after which myocardial performance deteriorates rapidly. Biochemical monitoring may be a novel sensitive and specific method for detecting subclinical myocardial damage. Abnormal cardiac adrenergic neuron activity evaluated with radiolabeled MIBG, impaired myocardial glucose utilization measured with F-18-FDG and plasma brain natriuretic peptide levels increased due to cardiac dysfunction may be particularly sensitive markers of adriamycin cardiotoxicity. It is hoped that prospective clinical study reveals the values of these biochemical markers.

A novel nonsense mutation of the PEPD gene in a Japanese patient with prolidase deficiency.

A nonsense mutation at amino acid residue 184 in the human peptidase D (PEPD) gene caused the production of a truncated polypeptide. Characterizing molecular defects in patients provides clues to elucidate the relationship between the phenotype and the genotype.

A mediastinal hemangioma, associated with perirenal hemangioma and congenital anomaly of the inferior vena cava.

In a 40-year-old man, a mediastinal hemangioma was discovered intially as a compression of the esophagus by upper gastrointestinal endoscopy. Furthermore, perirenal hemangioma and inferior vena cava (IVC) malformation were stimultaneously found. Hemangiomas, which occur in the mediastinal and perirenal area, are extremely uncommon and congenital IVC malformation, like the present case, has not been reported. We review the literature of these vascular abnormalities.

Hepatic subcapsular hematoma after extracorporeal shock wave lithotripsy (ESWL) for pancreatic stones.

We present a patient with complication of huge hepatic subcapsular hematoma after extracorporeal shock wave lithotripsy (ESWL) for pancreatic lithotripsy. The hematoma measured 78-110mm. Angiography showed a subcapsular hematoma, rather than a hematoma in the liver. In the arterial phase, the distal end of the small vessel showed spotty opacification similar to microaneurysma, suggesting that it was an injury caused by separation of the liver and its capsule, caused by the shock waves. The portal vein and hepatic vein were normal. After 8 weeks of conservative therapy, the hematoma was gradually absorbed and the patient was discharged. Eight months after the accident, the hematoma had decreased to 40mm in size. After 20 months, it was completely absorbed. The reported rate of renal subcapsular hematoma after ESWL for renal or ureter stones is 0.1%-0.7%. To date, however, only five cases of hepatic subcapsular hematoma after right renal stone disintegration have been reported. This is the first report of hepatic subcapsular hematoma after ESWL for pancreatic stones.

[A method for improving the accuracy in measurement of 99mTc-GSA liver uptake].

In order to improve the accuracy in the measurement of liver uptake rate of radioactivity at 15 minutes after injection of 99mTc-GSA, the corrected liver uptake rate (LU15VW) for tissue attenuation of gamma ray was measured. On the basis of the results of phantom studies, LU15VW was obtained as a ratio of the liver counts to the calculated counts of the injected dose supposed to homogeneously distribute in the liver of each patient and to decrease in count rate by the tissue attenuation of gamma ray, which was caused by the liver itself and body wall. The values of LU15VW were compared with those of the other 99mTc-GSA indices (LU15, LHL15, and HH15) and of the laboratory tests of liver function in 5 patients with chronic persistent hepatitis (CPH), 25 patients with chronic active hepatitis (CAH) 2A, 8 with CAH 2B, 8 with liver cirrhosis (LC), and 20 with hepatocellular carcinoma. LU15VW showed a good correlation with LU15, LHL15, and HH15 (r = 0.912, 0.864, and -0.869). The relationships between the results of LU15VW and the laboratory tests of liver function such as ICGR15, serum albumin, platelets counts, and hepaplastin test (r = -0.800, 0.684, 0.599, and 0.465) were more excellent as compared with those between the results of the other 99mTc-GSA indices and the laboratory tests. LU15VW was distinctly different in the mean values among the three groups of patients with CAH 2A, CAH 2B, and LC. These results indicated that LU15VW was an useful method for improvement of the accuracy in the measurement of liver uptake rate of 99mTc-GSA.

A case of superficial leiomyosarcoma, and a review of Japanese cases.

We report a case of leiomyosarcoma in a 55-year-old Japanese female. She developed an erythematous nodule, 1.5 cm in diameter, on her right thigh. The totally excised specimen contained tumor cells with variously sized, chromatin-rich nuclei. Gitter staining revealed the so-called encased picture, immunohistopathological staining was positive for smooth muscle actin, and electron microscopy revealed filaments with focal densities. A diagnosis of leiomyosarcoma was made based on these findings, which suggested a muscle cell origin. Normal arrector pili muscles were continuous from the tumor, further suggesting the tumor's origin. The presence of Epstein-Barr virus-encoded RNA (EBER) was examined by in situ hybridization with negative results.

Tc-99m MIBI localization in bone marrow: a marker of bone marrow malignancy.

To determine the potential of Tc-99m MIBI for detecting bone marrow malignancy, MIBI imaging of the femur was evaluated. There was no detectable MIBI activity in 125 of 141 (89%) control patients. Clearly demonstrated focal or tubular MIBI activity indicating intramedullary accumulation was demonstrated in 44 of 45 (98%) patients with proven marrow malignancy: 9 patients with multiple myeloma, 10 patients with malignant lymphoma, 11 patients with acute leukemia, 1 patient with chronic leukemia, and 14 patients with skeletal metastases. No abnormal MIBI activity was observed in the femur in 19 of 22 (86%) patients who had no evidence of malignant involvement in the femoral marrow, in 3 patients with solitary plasmacytomas of the spine, sternum or iliac bone, or in 16 patients with malignant lymphoma. In 12 of 24 patients with acute leukemia in complete remission, no abnormal MIBI accumulation was shown in the femur, but in 12 patients, abnormal accumulation indicating residual leukemic activity was demonstrated. MIBI imaging correlated extremely well with MRI studies; 26 of 28 patients with focal or tubular increased MIBI activity in the femur showed decreased signal on T1-weighted images and a high signal on short inversion recovery images, and 11 patients with no abnormal activity showed a high signal on T1 images. MIBI imaging of the femoral bone marrow may be a new modality for detecting marrow malignancy.

Clinical phenotype of Bart's syndrome seen in a family with dominant dystrophic epidermolysis bullosa.

BACKGROUND: Bart's syndrome is one type of dominant dystrophic epidermolysis bullosa (EB). It is known that, in some familial cases of dominant dystrophic EB, the symptoms differ depending on the individual. We observed the way Bart's syndrome affected four generations in the same family. The proband was a newborn boy who showed congenital localized absence of skin (CLAS) and bullae on the anterior aspects of both legs. Histologically, the bullae were located subepidermally. The CLAS and bullae disappeared within 4 months after birth, leaving scars. His father retained scarring and scaling from the knees down along the anterior aspect of the legs, and the nails of the toes were either lacking or deformed. His paternal grandmother and great-grandmother also presented deformed nails of the toes, although they had not had CLAS or bullae on the legs at birth. The individuals in this family thus showed some heterogeneity depending on the sex: blistering and CLAS were seen on the legs soon after birth in the male family members, but the female members did not share this pattern of symptoms, suggesting that the expression of symptoms may differ depending on the sex of the affected individual.

[Relationship between telomerase activity and clinicopathological findings in gastric cancer].

In recent, telomerase has been drawing the attention, because it plays important role in cell immortalization. Since Kim et al exploited TRAP (telomeric repeat amplification protocol) assay which detected telomerase activity efficiently, telomerase activity of various tissues have been detected. In this study, telomerase activity of gastric cancer of resected or biopsy specimen were detected. And we have examined relationship between telomerase activity and clinicopathological factors. In advanced cancer, telomerase activity is negative in 2 cases and positive in 17 cases. In case with lymph node metastasis, negative in 3 cases and positive in 14 cases. In differentiated type, negative 1 cases and positive 20 cases. In noncancerous gastric mucosa, telomerase activity is positive in only 2 cases of 9 cases with intestinal metaplasia. Previous results were same in biopsy specimen. In summary, telomerase activity was detected in 76.7% of gastric cancer and most of advanced cancer or differentiated type. Some intestinal metaplasia cases with telomerase activity suggested precancerous stage.

[Detection of telomerase activity in intestinal lavage solution from colorectal carcinoma patients].

In terms of colorectal carcinoma, the fecal occult blood test is widely used for mass survey, but has many complicated problems to be overcome. Telomerase activity has been reported in a wide range of malignancies. We have examined telomerase activity of intestinal lavage solution collected from 16 colorectal carcinoma patients and from 10 volunteers (control) by the method of telomeric repeat amplification protocol (TRAP) assay. Patients drunk polyethylene glycol-electrolyte lavage solution (PEG-ELS) before examination. Sample solutions were collected by colonoscope at the beginning of colonoscopy. The telomerase activity from colorectal carcinoma patients were positive 9/16 (56.3%) including 2 cases of early stage. In volunteers, were positive 1/10 (10.0%). This method has, therefore, possibility for a new useful method of diagnosis for colorectal carcinoma.

[Influence of intratumor administration of OK-432 on the tumor selectivity of 5'-DFUR].

The influence of intratumor administration of OK-432 on the tumor-selective antitumor effect of 5'-DFUR was studied in 49 patients with advanced gastric cancer. The patients were divided into 4 groups that received oral 5'-DFUR, intratumor OK-432, oral 5'-DFUR plus intratumor and intracutaneous OK-432, or no therapy before operation. Using surgical specimens, the PyNPase activity and 5-FU content were measured, and the localization of PyNPase was determined immunohistologically. The results were as follows: 1) 5'-DFUR therapy decreased intratumor PyNPase activity whereas administration of OK-432 increased it. 2) PyNPase activity was higher in cancer tissue than in normal tissue for all groups. 3) The 5-FU content of cancer tissue was higher in patients receiving OK-432 plus 5'-DFUR than in patients receiving 5'-DFUR alone. 4) In the resected tumors, PyNPase was mainly localized in the cancer cells of some patients and in the stromal cells of others. Thus, the localization of PyNPase showed two major patterns.

[A case of advanced esophageal cancer made resectable by preoperative combination therapy with 5-FU and CDDP].

The authors experienced a case of advanced esophageal cancer made resectable by combination therapy with 5-FU and CDDP as a neoadjuvant chemotherapy. The patient was a 69-year-old-man suffering from esophageal cancer of A3.N4 (+).Pl0.M0 at stage IV. At this case was diagnosed to be radically unresectable, this form of combination therapy was used. The patient showed PR after 2 courses and the operation could then be conducted. The intraoperative findings revealed fibrous fusion of tumor with the aorta, but no direct invasion. The metastatic lymph nodes were necrotized and reduced. It was evaluated as Grade 3 in accordance with the "Histologic criteria for the effects of anticancer chemotherapy." The postoperative course was favorable without recurrence. This therapy caused no adverse reactions and seems effective as a neoadjuvant chemotherapy for advanced esophageal cancer.

Assessment of adrenergic neuron function altered with progression of heart failure.

UNLABELLED: We used MIBG to evaluate cardiac adrenergic neuron integrity and function in congestive heart failure. METHODS: Rats were treated with adriamycin (2 mg/kg, s.c.) once a week for 7, 8 and 9 wk. In analyzing cardiac adrenergic neuron function, we assessed alterations of uptake-1, exocytotic release and nonexocytotic metabolic release in relation to progression of heart failure. RESULTS: LVEF progressively decreased. Cardiac MIBG accumulation (4 hr postinjection) decreased to 53% of control at 7 wk and markedly decreased to 14% of control at 9 wk, accompanied by massive pleural effusions. Reduction of MIBG accumulation in the lung and spleen, which are adrenergic-rich organs similar to the heart, were less pronounced compared to reduction in the heart. There was no difference in cardiac uptake of 3H-norepinephrine between the control and 8-wk groups. Cardiac uptake of 3H-norepinephrine decreased 91.0% in the control and 90.8% in the 8 wk group by pretreatment of desipramine, indicating no difference in the uptake-1 component. CONCLUSION: Congestive heart failure due to adriamycin cardiomyopathy progressively accelerates exocytotic release of norepinephrine predominantly from cardiac adrenergic neurons, but neuronal uptake function is not disturbed so long as heart failure is not advanced. In the advanced stage, nonexocytotic metabolic release is induced specifically in cardiac adrenergic neurons due to energy depletion and norepinephrine release markedly increases.