Antigen capture assay for detection of bovine leukemia virus proteins by monoclonal antibodies.

A capture monoclonal antibody-based assay has been established for detecting the p24 core protein and the gp51 envelope glycoprotein of bovine leukemia virus (BLV). This assay is rapid, highly sensitive and specific. Viral antigens in test samples were identified using mouse monoclonal antibody-coated or microtiter plates by adding labeled monoclonal antibodies with different epitope specificities. The choice of an appropriate epitope specificity for the specificity of monoclonal antibodies was important for optimal performance of the assay. Results of this assay were in agreement with the syncytia induction assay routinely used for detecting BLV production by cells in vitro. The sensitivity of monoclonal antibody assay was 0.5 ng/ml for p24 and 1.25 ng/ml for gp51, respectively. The specificity was demonstrated by immunoblotting. The assay can be performed in a few hours, is simple, and is comparable with more time-consuming assays with regard to sensitivity and specificity.     

Intra-arterial Autologous Bone Marrow Cell Transplantation in a Patient with Upper-extremity Critical Limb Ischemia

Induction of therapeutic angiogenesis by autologous bone marrow mononuclear cell transplantation has been identified as a potential new option in patients with advanced lower-limb ischemia. There is little evidence of the benefit of intra-arterial cell application in upper-limb critical ischemia. We describe a patient with upper-extremity critical limb ischemia with digital gangrene resulting from hypothenar hammer syndrome successfully treated by intra-arterial autologous bone marrow mononuclear cell transplantation.     

TGF-B3 Dependent Modification of Radiosensitivity in Reporter Cells Exposed to Serum From Whole-Body Low Dose-Rate Irradiated Mice

Prior findings in vitro of a TGF-β3 dependent mechanism induced by low dose-rate irradiation and resulting in increased radioresistance and removal of low dose hyper-radiosensitivity (HRS) was tested in an in vivo model. DBA/2 mice were given whole-body irradiation for 1 h at low dose-rates (LDR) of 0.3 or 0.03 Gy/h. Serum was harvested and added to RPMI (4% mouse serum and 6% bovine serum).This medium was transferred to reporter cells (T-47D breast cancer cells or T98G glioblastoma cells). The response to subsequent challenge irradiation of the reporter cells was measured by the colony assay. While serum from unirradiated control mice had no effect on the radiosensitivity in the reporter cells, serum from mice given 0.3 Gy/h or 0.03 Gy/h for 1 h removed HRS and also increased survival in response to doses up to 5 Gy. The effect lasted for at least 15 months after irradiation. TGF-β3 neutralizer added to the medium containing mouse serum inhibited the effect. Serum from mice given irradiation of 0.3 Gy/h for 1 h and subsequently treated with iNOS inhibitor 1400W did not affect radiosensitivity in reporter cells; neither did serum from the unirradiated progeny of mice given 1h LDR whole-body irradiation.     

Metastatic squamous cell carcinoma of the skin in chronic myeloid leukaemia: complication of hydroxyurea therapy

Hydroxyurea is a ribonucleotide diphosphate reductase inhibitor used in the treatment of patients with myeloproliferative disorders. Hydroxyurea has some dermatological side-effects. It has recently been recognized that hydroxyurea can induce squamous cell and basal cell carcinomas of skin. We present the case of an elderly man with chronic myeloid leukaemia who was treated with hydroxyurea for 4 years, with good control of his disease. However, in addition to the appearance of various skin lesions and cutaneous squamous cell carcinoma after 3 years of therapy, he was found to have a metastatic squamous cell carcinoma after 4 years. Hydroxyurea was discontinued, and he underwent surgery and radiotherapy. The patient subsequently died of ventricular fibrillation. We present this case to draw attention to the association between hydroxyurea and secondary skin cancers and to emphasize the need for dermatological examination before and during the course of hydroxyurea therapy.     

Breast cancer,ovarian gonadoblastoma and cervical cancer in a patient with Peutz-Jeghers Syndrome

BACKGROUND: Peutz-Jeghers Syndrome (PJS) is a rare autosomal dominant disorder characterized by gastrointestinal hamartomatous polyps and mucocutaneous pigmentation. Patients with PJS have increased risk for gastrointestinal, breast, and female genital tract cancers. CASE: Multiple genital tract cancers in a 34-year-old woman with PJS are described. The patient, who was admitted to our department with severe vaginal bleeding, was performed right salpingo-oophorectomy because of pure gonadoblastoma in 1996. In 2003, concomitant to cervical carcinoma, breast cancer was diagnosed. Patient underwent left modified radical mastectomy due to the invasive papillary carcinoma. The patient received six cycles combination chemotherapy and radiation therapy because of stage IIIB cervical cancer. CONCLUSION(S): This is the first case report presenting PJS associated with multiple genital tract tumors including ovarian gonadoblastoma in literature. The clinical significance of these tumors in PJS patients has been reviewed.     

Lymphoepithelioma-Like Carcinoma of the Urinary Bladder: A Case Report and Discussion of Differential Diagnosis

Lymphoepithelioma-like carcinoma (LELC) of the urinary bladder is very rare. It is mandatory to make differential diagnosis among lymphoma, chronic cystitis and LELC because of different therapeutic approach. A bladder tumor was found in a 90-year-old patient suffering from hematuria. After transurethral resection, undifferentiated tumor with prominent lymphoid infiltration was seen on light microscopy. Immunohistochemical examination demonstrated positive staining of tumor cells with cytokeratin (CK), epithelial membrane antigen and CK-20. We presented the case because of its rarity and related literature was reviewed.     

Inhibition of iNOS with S-methylisothiourea was impaired in wound healing in caustic esophageal burn

OBJECTIVE: Stricture formation is a late complication of caustic esophageal burn, which is a common problem in childhood. For this reason, this experimental study was designed to observe the possible effect of nitric oxide on healing and fibrosis formation in caustic esophageal burns. MATERIALS AND METHODS: The rats were divided into five groups. Group A (n=12) received sham burn and treatment with saline injection. Group B (n=34) received caustic burn. Rats in group C (n=31), were given water supplement with 10 g/L L-arginine that was started 24 h preoperatively and continued until postoperative day 4. In group D (n=21), S-methylisothiourea (SMT, specific inducible nitric oxide synthase (iNOS) inhibitor), was injected at a dose of 3 mg/kg i.p. at 30 min before caustic burn, and similar dose was reinjected immediately after caustic burn. SMT 6 mg/kg/day injections continued for 4 days long. In group E (n=22), Nomega-nitro-L-arginine (L-NNA, nonspecific nitric oxide synthase (NOS) inhibitor) was injected at a dose of 15 mg/kg i.p. at 30 min before caustic burn, and similar dose was reinjected immediately after caustic burn. L-NNA 30 mg/kg/day continues for 4 days. RESULTS: Dead rates were significantly higher in group E than in groups A-D. The mean hydroxyproline levels in esophageal tissue were significantly lower in groups A and B than in group D. Histopathologically, tissue damage scores in the esophageal tissue were higher in group D than in groups A-C. CONCLUSIONS: Inhibition of iNOS with SMT was impaired in wound healing due to caustic esophageal burn and provoked collagen accumulation at a later period. Those effects may due to inhibition of antioxidant, immunomodulatory and antifibrotic effects of NO.     

Novel germline mutation in the transmembrane region of RET gene close to Cys634Ser mutation associated with MEN 2A syndrome

Two mutations on the same allele of RET gene were revealed in a family with predisposition to multiple endocrine neoplasia (MEN) type 2A. The first mutation changes codon 634 from cysteine to serine. The second, a novel mutation in codon 641, changes alanine to serine in the transmembrane domain of the RET protein. Two mutations were present in close proximity in both the patients germline and tumor DNA and were absent in DNA isolated from healthy family members and control blood donors. All MEN 2A affected family members suffered from medullary thyroid carcinoma and two of ten patients for pheochromocytoma. No parathyroid gland alterations were observed in patients with two RET gene mutations. Analysis of four genetic polymorphisms in the RET gene showed higher incidence of polymorphisms of exons 11 and 15. The observed allelic imbalance in favor of mutated allele in pheochromocytoma corresponded to higher expression of the RET gene. These observations confirm the multifactorial process leading to development of MEN 2A syndrome.     

Comparison of radioimmunoassay for internal protein of bovine leukemia virus with neutralization test employing VSV-BLV pseudotype

Two methods for the detection of antibodies to the bovine leukemia virus (BLV) in infected animals were compared for their suitability for the early diagnosis of bovine leukemia - pseudotype neutralization test (PNT) employing vesicular stomatitis virus - bovine leukemia virus pseudotypes (VSV-BLV), and radioimmunoassay test (RIA) for major internal viral protein p24 of the BLV. The comparison was made using more than 300 sera from cows of the herds with high incidence of bovine leukemia. In infected animals the presence of antibodies against virus envelope glycoprotein detected by PNT and antibodies against major structural viral protein p24 detected by RIA were found always coincidentally. Both methods were found highly comparable and suitable for early detection of bovine leukemia virus infected animals.