Replication-competent retrovirus vector-mediated prodrug activator gene therapy in experimental models of human malignant mesothelioma.

Replication-competent retrovirus (RCR) vectors have been shown to achieve significantly enhanced tumor transduction efficiency and therapeutic efficacy in various cancer models. In the present study, we investigated RCR vector-mediated prodrug activator gene therapy for the treatment of malignant mesothelioma, a highly aggressive tumor with poor prognosis. RCR-GFP vector expressing the green fluorescent protein marker gene successfully infected and efficiently replicated in human malignant mesothelioma cell lines, as compared with non-malignant mesothelial cells in vitro. In mice with pre-established subcutaneous tumor xenografts, RCR-GFP vector showed robust spread throughout entire tumor masses after intratumoral administration. Next, RCR-cytosine deaminase (RCR-CD), expressing the yeast CD prodrug activator gene, showed efficient transmission of the prodrug activator gene associated with replicative spread of the virus, resulting in efficient killing of malignant mesothelioma cells in a prodrug 5-fluorocytosine (5FC)-dose dependent manner in vitro. After a single intratumoral injection of RCR-CD followed by intraperitoneal administration of 5FC, RCR vector-mediated prodrug activator gene therapy achieved significant inhibition of subcutaneous tumor growth, and significantly prolonged survival in the disseminated peritoneal model of malignant mesothelioma. These data indicate the potential utility of RCR vector-mediated prodrug activator gene therapy in the treatment of malignant mesothelioma.

A clinical study of patients with pemphigus vulgaris and pemphigus foliaceous: an 11-year retrospective study (1996-2006).

Only a few reports have been published of detailed clinical studies of pemphigus in Japan. The aim of this study was to determine the clinical characteristics of patients with pemphigus vulgaris (PV) and pemphigus foliaceous (PF), who were newly diagnosed in the dermatology department of Kurume University Hospital, Japan, over the past 11 years. The primary site of involvement was the oral mucosa in 21 patients (75%) with PV. At the initial visit, most of the patients with PV had moderate to severe disease. With regard to management, systemic corticosteroids were the mainstay of treatment for patients with PV, and plasmapheresis was the most frequently used adjuvant therapy. Dapsone was the mainstay of treatment for the patients with PF. The patients were investigated for any association with an underlying malignancy; in patients with PV, lung, stomach and uterine cancers (one patient each) were seen.

Serum from normal elderly individuals contains anti-basement membrane zone antibodies.

BACKGROUND: Bullous pemphigoid is an autoimmune bullous disease with circulating anti-basement membrane zone antibodies, and it commonly affects elderly individuals; however, the reasons for the late onset of the disease are unclear. DESIGN: The anti-basement membrane zone antibodies in serum samples from normal elderly subjects were compared with those in serum samples from normal young subjects. PARTICIPANTS: Serum samples from 32 elderly and 28 young normal individuals and 10 patients with bullous pemphigoid were used. INTERVENTIONS: Indirect immunofluorescence against guinea pig esophagus or human salt-split epidermis and immunoblotting against human and guinea pig epidermis were performed. RESULTS: Serum samples from young individuals were devoid of anti-basement membrane zone antibodies against guinea pig esophagus and human salt-split epidermis. Among 32 serum samples from elderly patients, 6 cases (19%) were positive for anti-basement membrane zone antibody for guinea pig esophagus, and in those the titers were 10 in 3 cases and 40, 80, and 320 in the others. One case was positive against human split epidermis at a titer of 10. An immunoblotting analysis showed that the antigenicity of the 230-kd and 180-kd bullous pemphigoid antigen from guinea pig epidermal extract was similar to that of human epidermal extract; however, the molecular weight was slightly different. The 4 cases of elderly serum that recognized guinea pig esophagus basement membrane zone showed positivity with the 230-kd peptide in the guinea pig epidermal extract; however, they were negative with the human epidermal extracts. Direct immunofluorescence observation of these cases showed that deposition of IgG or C3 was not present in cryostat sections from flexor arm surfaces. CONCLUSIONS: The serum samples from elderly subjects possessed a relatively high incidence of anti-basement membrane zone antibodies detectable with guinea pig esophagus as substrate. This observation of a specific immune defect in elderly individuals might explain why they are more susceptible to developing bullous pemphigoid.

Secondary amyloidosis complicating arthropathic psoriasis.

We report a patient with secondary systemic amyloidosis complicating arthropathic psoriasis. At autopsy amyloidosis was observed in the spleen, kidneys, liver, gastro-intestinal tract, lung, heart, pancreas, adrenal glands, thyroid and gall-bladder in addition to the skin by histological, histochemical and ultrastructural studies. The amyloid deposits in these organs were of amyloid A protein type.

A flow cytometric analysis of the DNA content of Paget cells in the epidermis and the tumor of extramammary Paget's disease.

In the present study, we have examined whether or not the Paget cells in cases with dermal invasion or metastasis exhibit a different DNA distribution pattern to that seen in the epidermis in extramammary Paget's disease. The DNA contents of Paget cells from the epidermis and the tumor were determined by flow cytometry in two cases of extramammary Paget's disease with tumor formation, dermal invasion and metastases. The DNA histogram of Paget cells from the epidermis of two cases appeared to be close to the normal ploidy, whereas the histogram from the tumors was aneuploid in both cases. Our data suggest that the behavior of Paget cells in the epidermis is different from that in the tumor.

Enhanced expression of bcl-2 inhibits apoptosis in cultured human keratinocytes.

The molecular mechanisms regulating epidermal differentiation and apoptosis have not been elucidated. Bcl-2, one of the candidate genes for suppressing apoptosis, was originally cloned from the breakpoint of at (14;18) translocation present in many human B cell lymphomas. In this study, the influence of bcl-2 on apoptosis was observed in transfected keratinocytes. After transfection of pEF-BOS vector with/without bcl-2, the expression of coded protein and the viability under starved conditions were examined. The bcl-2-transfected keratinocytes had cytoplasmic positive staining with anti-bcl-2 monoclonal antibodies, however the vector only transfected cells were devoid of the reaction products. The viability of transfected keratinocytes under starved conditions, with a lack of epidermal growth factor and bovine pituitary extract, was maintained in bcl-2 transfected cells; while the vector only transfected cells showed apoptotic cell death. The present result indicates that bcl-2 suppresses apoptotic cell death under starved conditions due to a lack of epidermal growth factor and bovine pituitary extract.

A case of mammary Paget's disease without an underlying carcinoma: microscopic analysis of the DNA content in Paget cells.

A 72-year-old woman had a 7-year history of a scaly red area on the right breast which had enlarged asymptomatically. A biopsy of the nipple was taken, and, following the finding of Paget's disease, a modified radical mastectomy was carried out. On histological examination of the entire breast specimen by serial sections, no evidence of an intraductal adenocarcinoma was found. Fractionation of Paget cells was performed from the epidermis. Stripped skin was treated with EDTA and trypsin, and epidermal cell suspensions were obtained. They were layered onto discontinuous Percoll gradients and centrifuged. Paget cells fell into three fractions with densities of 1.041, 1.058, and 1.078. Electronmicroscopically, the purity of fractionated cells obtained by this method ranged from 55 to 74% with viabilities of from 70 to 90%. Microscopic analysis of the DNA content in these cells was performed. The DNA histogram was close to the normal ploidy. This may explain why the mammary Paget's disease lesions in this case enlarged rather slowly.

The effect of orally-administered retinoid on lectin binding in guinea pig keratinocytes.

Retinoids influence proliferation and differentiation of epithelial cells. The present paper was designed to examine the change of lectin binding in isolated keratinocytes from retinoid-treated guinea pigs. Aromatic retinoid, etretinate, in peanut oil (5 mg/kg/day) was given orally for a period of 14 days. Skin samples were obtained from animals on days 1, 3, 7, and 14 after beginning the administration. Free keratinocytes were obtained by treatment with EDTA and trypsin. They were separated into 3 fractions by centrifugation on a continuous colloidal silica (Percoll) density gradient. The cells in each fraction were stained by biotinyl lectins and avidin-FITC, and the fluorescence intensity was measured by cytofluorometry. The lectins used were Concanavalin A (Con A), soybean agglutinin (SBA), peanut agglutinin (PNA), Dolichos biflorus agglutinin (DBA), and Ricinus communis agglutinin (RCA). The PNA-binding was diminished in both the intermediate and lower density fractions at 7 and 14 days. The binding of SBA was reduced in all fractions from 1 to 7 days. With Con A, RCA and DBA, no changes in intensity were found.

Cell cycle analyses of extramammary Paget's disease.

The cell cycle of Paget cells in extramammary Paget's disease was investigated using microscopic analysis of the DNA content, and using immunohistochemistry for 5-bromodeoxyuridine (BrdU) incorporation. Stripped skin was treated with tetrasodium ethylendiamine tetraacetate dihydrate (EDTA) and trypsin, and epidermal cells suspensions were obtained. Microscopic analysis of the DNA content in the cells was performed. The resulting DNA histogram was close to the normal ploidy. Specimens from the involved skin were incubated with BrdU, and sections were immunostained with anti-BrdU monoclonal antibody. A few nuclei showed positive staining for BrdU. The present study showed that the proliferative potential of Paget cells in extramammary Paget's disease was low. This may explain why lesions of extramammary Paget's disease enlarge rather slowly.

[Impacted bilateral first permanent molars induced by the migrated second permanent molars].

It is a relatively common clinical experience to see an impacted permanent tooth. In most cases, the cause is considered to be the existence of a supernumerary tooth or developmental crowding, but occasionally, the impacted tooth may occur with no apparent cause. The tooth most frequently impacted in the dental arch is a maxillary permanent canine or the third permanent molar, but the occurrence of the first permanent molar is an extremely rare situation. The patient, an eight-year, seven-month-old male was referred to the Pediatric Dentistry Clinic at the Higashi-Nippon-Gakuen University, School of Dentistry, for a detailed dental examination of his impacted lower permanent first molars. The oral findings through dental examination revealed that the bilateral upper first permanent molars were erupted, while the lower ones showed no clinical signs of eruption. Radiographic examination revealed that the lower second permanent premolars placed under the distal root of the second deciduous molars, and the first molars, inclined to the mesial position, placed at the under the second premolars bilaterally. There was nothing particular in his medical and dental history during the "teething" period. The authors determined that the cause of this impacted condition for the first molars seemed to be induced by a migration of an unerupted second permanent premolars. After extraction of the second deciduous molars and second permanent premolars, orthodontic therapy was undertaken in an attempt to move the impacted first molars into the arch.(ABSTRACT TRUNCATED AT 250 WORDS)