BMCC1, which is an interacting partner of BCL2, attenuates AKT activity, accompanied by apoptosis.

BNIP2 and Cdc42GAP homology (BCH) motif-containing molecule at the carboxyl-terminal region 1 (BMCC1) gene is highly expressed in patients with favorable neuroblastoma (NB). It encodes a 340-kDa protein with a conserved BCH scaffold domain that may regulate signaling networks and multiple cellular functions, including apoptosis. In this study, we determined the mechanism by which BMCC1 promotes apoptosis in human NB and non-NB cells, as BMCC1 is normally expressed in various organs, particularly in neuronal and epithelial tissues. We demonstrated in this report that BMCC1 was induced by DNA damage, one of the triggers of intrinsic apoptosis. Accordingly, we investigated whether BMCC1 expression impacts intracellular signals in the regulation of apoptosis via its C-terminal region containing BCH scaffold domain. BMCC1 decreased phosphorylation of survival signals on AKT and its upstream kinase PDK1. BMCC1 upregulation was correlated with the activation of forkhead box-O3a (FOXO3a) (a downstream inducer of apoptosis, which is suppressed by AKT) and induction of BCL2 inhibitor BIM, suggesting that BMCC1 negatively regulates phosphorylation pathway of AKT, resulted in apoptosis. In addition, we found that BNIP2 homology region of BMCC1 interacts with BCL2. Intrinsic apoptosis induced by DNA damage was enhanced by BMCC1 overexpression, and was diminished by knockdown of BMCC1. Taken together, we conclude that BMCC1 promotes apoptosis at multiple steps in AKT-mediated survival signal pathway. These steps include physical interaction with BCL2 and attenuation of AKT-dependent inhibition of FOXO3a functions, such as transcriptional induction of BIM and phosphorylation of ataxia telangiectasia-mutated (ATM) after DNA damage. We propose that downregulation of BMCC1 expression, which is frequently observed in unfavorable NB and epithelial-derived cancers, may facilitate tumor development by abrogating DNA damage repair and apoptosis.

Primary hepatoid adenocarcinoma of the uterine cervix.

Hepatoid adenocarcinoma is characterized histologically by neoplastic epithelial cells that resemble hepatocellular carcinoma (HCC) and produce alpha-fetoprotein (AFP). We describe a case of hepatoid adenocarcinoma of the uterus that, unlike any other previously reported case, was strictly confined to the cervix. A cervical biopsy demonstrated poorly differentiated adenocarcinoma, and hysterectomy and bilateral salpingo-oophorectomy were subsequently performed. Histologically, the lesion consisted of solid sheets of hepatoid cells accompanied with areas of endometroid adenocarcinoma. The tumor cells showed strong and diffuse cytoplasmic immunoreactivity with AFP in both medullary and adenocarcinoma components. Metastases to bilateral pelvic lymph nodes were detected 12 months after surgery. Since undergoing total pelvic irradiation, the patient has been alive and in full remission for 22 months. To our knowledge, this is the first report of primary hepatoid adenocarcinoma of the uterine cervix.

Adjuvant chemotherapy in stage I uterine endometrial carcinoma.

OBJECTIVE: We have assessed prognostic factors and the efficacy of adjuvant chemotherapy in stage I uterine endometrial carcinoma. METHODS: 251 primary surgically treated stage I patients were studied. Prognostic factors were evaluated and 5-year and 10-year survival rates were compared in patients with lymph-vascular space invasion to investigate whether adjuvant chemotherapy improves survival. RESULTS: The overall 5-year and 10-year survival rates were 94% and 93%. Multivariate analysis indicates that lymph-vascular space invasion is the most significant prognostic factor in both 5- and 10-year survival rates (P<0.001 at both times) and stage/depth of invasion is significant for the 10-year survival rate (P=0.04). Of 54 patients with lymph-vascular space invasion, statistically significant differences were observed in 10-year survival rate (P=0.02) between patients who had surgery followed by adjuvant chemotherapy (n=23) and patients who had surgery alone (n=31). Toxicities were mild to moderate (30%). CONCLUSIONS: The clinical importance of lymph-vascular space invasion and the efficacy of adjuvant chemotherapy were confirmed. This observation warrants a larger comparative study with adjuvant chemotherapy.

A novel gene (retinovin) expressed selectively in the early stage of chick retinal development.

To understand molecular mechanisms of retinal development, genes expressed selectively only in the early stage of retinal development were isolated by subtractive hybridization based on suppression polymerase chain reaction. The retina has no layered structure in 7-day chick embryos, in contrast with the fully developed multilayered structure of neurons in 15-day embryos. The subtraction between cDNA derived from retinal tissues at these different stages, followed by repeat rounds of 5'-RACE (rapid amplification of cDNA ends) and 3'-RACE, led to isolation of a novel gene with an open reading frame encoding a putative protein with 753 amino acids. Its specific expression in the 7-day embryonic retina was confirmed by Northern blot analysis. The gene, named "retinovin," would be used as a marker for identifying retinal stem cells present at the early stage of retinal development.

Choroidopathy in patients with sarcoidosis observed by simultaneous indocyanine green and fluorescein angiography.

PURPOSE: To examine choroidopathy in patients with sarcoidosis. METHODS: In a prospective clinical study, 10 consecutive patients (20 eyes) with sarcoidosis underwent simultaneous indocyanine green (ICG) and fluorescein angiography (FA) with a double detector of scanning laser ophthalmoscopy. Angiographic findings recorded on videotapes were evaluated, and the relation of ICG angiographic findings with systemic activity and the extent of retinal inflammation was analyzed. RESULTS: Indocyanine green angiographic findings, not evident with fluorescein, were multiple lobular hypofluorescent spots in the posterior pole in 10 eyes of 5 patients, isolated hypofluorescent plaques in 2 eyes of 1 patient, hyperfluorescent spots in 3 eyes of 2 patients, and segmental choroidal vascular wall staining in 6 eyes of 4 patients. The multiple lobular hypofluorescent spots with ICG were observed at a significantly higher rate in eyes with extensive retinal vascular leakage of fluorescein than in eyes with minimal leakage (chi-square test, P = 0.0007). CONCLUSIONS: The patients with sarcoidosis showed choroidal abnormalities that could be revealed by ICG angiography, but not by funduscopy or FA. Simultaneous ICG and FA angiography may be useful for examining choroidal lesions in sarcoidosis.

Acinic cell carcinoma of the palate: case report and immunohistochemical observation.

We report a rare case of acinic cell carcinoma of the palate in a 63-year-old Japanese woman. Clinical examination demonstrated a firm, mobile mass without regional lymph-adenopathy. Histopathologically, the tumor was composed of large, polyhedral or round cells with basophilic granular cytoplasm (serous acinar-like cells) and reticular or clear cytoplasmic cells. These tumor cells were positive for the periodic acid-Schiff reaction, but negative for alcian blue. The tumor nests were separated by thin vascular tissue and incompletely encapsulated. Immunohistochemically, the tumor cells exhibited positive reactivity for alpha-amylase, lactoferrin, secretory component, S100 protein, and epithelial membrane antigen, but were negative for actin, glial fibrillary acidic protein, keratin, and carcinoembryonic antigen. These results suggest that this tumor is well differentiated into serous acinar cells and that the reticular and clear cytoplasmic cells are a modified form of these cells.

Pulmonary lymphangiomyomatosis diagnosed by effusion cytology. A case report.

BACKGROUND: Pulmonary lymphangiomyomatosis (P-LAM) is a rare disease, characterized by an abnormal proliferation of smooth muscle throughout the lung and occurring in females of reproductive age. The typical clinical picture in P-LAM is progressive dyspnea, often punctuated by episodes of chylothorax or pneumothorax, with eventual death from respiratory failure. The definitive diagnosis is usually performed on open lung biopsies. CASE: The cytologic findings in a chylous pleural effusion from a patient with P-LAM are presented. The effusion contained an abundance of globular cell clusters composed of two distinct cell types, inner core spindle cells and surface flat cells. Immunocytochemical examination revealed that the core cells were derived from immature smooth muscle cells and the surface cells from endothelium. Histologic examination of the excised specimen showed the typical findings of P-LAM, and the dilated peripheral lymphatics contained cell clusters similar to those in the pleural effusion. The cell clusters in the lymphatics in the P-LAM lung tissue appeared to have ruptured into the thoracic cavity. CONCLUSION: Cell clusters seem to be pathognomonic of P-LAM, and cytologic examination can obviate the need to perform open lung biopsy to confirm the diagnosis.

[Prognostic significance of histopathological findings (including lymphocyte infiltration) in renal cell carcinoma].

We compared the prognosis with the histopathological findings including intravenous invasion and lymphocytic infiltration inside or adjacent to the primary tumor in 50 renal cell carcinoma patients who underwent radical nephrectomy. We compared the primary tumor to intravenous invasion or metastases in histopathological findings. One-, three- and five-year survival rates for all patients were 91.7, 71.5 and 60.7%, respectively. Significant prognostic factors were tumor size, growth pattern, invasion of fat tissue into peripheral kidneys lymph nodes, distant metastases, intravenous invasion and tumor grade, especially lymph nodes and distant metastases (P < 0.001). Degree of lymphocytic infiltration inside or adjacent to primary tumor was divided into three groups. Five-year survival rates were 86.2% of the patients (n = 15) with apparent lymphocytic infiltration and 48.8% of the patients (n = 18) had few infiltrated lymphocytes. The patients with apparent lymphocytic infiltration showed a trend of better prognosis compared to the patients with few infiltrated lymphocytes (P < 0.1). Tumor grade was higher in 3 of the 28 patients with intravenous invasion and 6 of the 9 patients with distant metastases than in those with primary tumors. However, there was no significant correlation between prognosis and malignant potential of intravenous invasion or distant metastases.

A case of cutaneous malignant fibrous histiocytoma.

A 54-year-old Japanese man with cutaneous malignant fibrous histiocytoma on the back is reported. He not only had a past history of thyroid cancer 1 year prior to the onset of the skin tumor, but also had simultaneous bladder cancer. Despite the early, wide resection, the prognosis was rapid and progressive. Histologically, the primary lesion of the skin tumor was difficult to differentiate from dermatofibrosarcoma protuberans; however, the recurrent and the metastatic lesions changed in appearance.

Airway basophil and mast cell density in patients with bronchial asthma: relationship to bronchial hyperresponsiveness.

We investigated the role of basophils and mast cells in the pathogenesis of bronchial asthma. Eight asthmatics (6 atopic, 2 nonatopic) and 6 control subjects were enrolled in this study. Bronchial responsiveness to acetylcholine (PC20ACh) was measured in asthmatics and endobronchial biopsy from right upper lobe bronchus was performed on the same day. Basophils and mast cells in the airways were identified by immunohistochemistry using a monoclonal antibody against tryptase and anti-IgE. The number of basophils of asthmatics was 52.2 +/- 12.5/mm(2). In contrast, no basophils were found in the airways of control subjects. There was a significant increase of number of mast cells in the asthma group compared to the control group (168.6 +/- 32.6 vs. 22.3 +/- 6.1, p<0.01). There was an inverse correlation between airway basophil and mast cell numbers and PC20ACh (r=-0.82, r=0.72, p<0.05). These findings suggest a possible role for basophils and mast cells in the pathophysiology of asthma.

Positron emission tomography using fluorine-18-fluorodeoxyglucose in malignant lymphoma: a comparison with proliferative activity.

To evaluate the relationship between PET using 18F-fluorodeoxyglucose (FDG) and pathological findings and to compare indices obtained by FDG-PET, FDG-PET was performed in 23 patients with untreated malignant lymphoma. Three indices obtained by FDG-PET, tumor-to-normal contrast ratio (TCR), distribution absorption ratio (DAR), k1k2/(k2 + k3), correlated with proliferative activity which was pathologically estimated both by mitotic count and by proportion of cells in all phases of the cell cycle. The relationship did not significantly change according to which of the three indices was chosen. FDG-PET, which shows the proliferative activity of tumors, is considered to be a useful method for managing tumors.

Prognostic factors for local control in nasopharyngeal cancer (NPC): analysis by multivariate proportional hazard models.

Despite the development of meticulous radiation therapy techniques, the local control of the nasopharyngeal cancer (NPC) remains unsatisfactory. We examined retrospectively the clinicopathologic factors which have impact upon local recurrence-free survival (LRFS) of the 67 patients with NPC who had been irradiated greater than or equal to 40 Gy with curative intent from 1975 through 1988. Three-year LRFS rate was 63%. T stage, histology, the presence of accompanying lymphocytic infiltration, and the properness of radiation therapy had influence upon LRFS with a statistical significance. Multivariate proportional hazard models showed that T stage and pathology retained significance. Radiation therapy properness emerged as a third factor when only the patients irradiated greater than or equal to 60 Gy were included in the analysis. From these results, three groups with low, moderate, and high risk of shortened LRFS could be separated. This risk classification could contribute to the stratification criteria of future study for the improvement of local control by new modalities.

Non-Hodgkin's lymphoma confined to the nasal cavity: its relationship to the polymorphic reticulosis and results of radiation therapy.

From 1975 through 1988, nine patients with locally confined nasal non-Hodgkin's lymphoma (NHL) were treated with radiation therapy in the Department of Radiology, Chiba University Hospital. Immunohistochemical study disclosed that all NHL's have T-lineage. Additionally, unique histological pictures of polymorphism, angiodestruction, and necrosis were seen in most of cases. These three findings are the histological features of polymorphic reticulosis (PMR), which is the main cause of lethal midline granuloma and has recently been shown to belong to T-cell malignancy. Therefore, it is concluded that the nasal T-cell NHL and PMR are really a single disease entity. The predominance of the T-cell lymphoma in the nasal cavity as well as its histological distinctness clearly indicate that the head and neck extranodal NHL cannot be discussed together. Although the disorder was considered to be locally limited at presentation, only 3 of the 9 patients with nasal NHL could be induced into long-term remission with involved field radiotherapy. The distant extranodal spread was the primary cause of failure. Multimodality treatment using intensive chemotherapy might improve the prognosis of nasal NHL.

Expression of functional molecules in non-Hodgkin's lymphoma. Correlation with bone marrow involvement and serum LDH value.

It is well known that non-Hodgkin's lymphoma (NHL) cells express various antigens which are normally involved in a variety of functions. In addition, NHL is diverse in its proliferative capacity. To investigate the relation between these factors and the clinical picture, 45 cases of NHL were studied by immunohistochemistry using snap-frozen materials obtained before therapy. Reactivities with 27 monoclonal antibodies were examined and the results were correlated with clinical findings. The expression of surface mu and ICAM-1 in B-NHLs and CD25 in T-NHLs were significantly associated with bone marrow involvement. B-NHLs without expression of CD21(B2) and T-NHLs with CD25 were seen more frequently in cases with a LDH value of over 500 units/ml. The positivity rate of Ki-67 on B-NHLs was correlated with serum LDH value, NHL histologic classification, and overall survival. These data indicate that immunophenotyping and determination of the proliferative capacity of NHL are of value not only for confirmation of the histopathologic classification of the tumor but also for assessment of clinical behavior.

Study of vimentin expression in non-Hodgkin's lymphoma using paraffin sections.

Human non-Hodgkin's lymphomas were studied by means of an avidin-biotin complex immunoperoxidase method using several monoclonal antibodies against the intermediate filament protein, vimentin. The study cases were 61 B-cell lymphomas (including 2 plasmacytomas) and 30 T-cell lymphomas (including 8 cases of mycosis fungoides). Twelve of the 61 B-cell lymphomas were positive for vimentin, and were composed of extrafollicular-center cells such as immunoblastic and plasmacytoid cells. On the other hand, lymphomas of follicular center cell origin were negative for vimentin. All cases of T-cell lymphoma except for 14 (all of 9 AILD-type lymphomas, all of 4 lymphoblastic lymphomas and one diffuse mixed small/large lymphoma) were positive for vimentin. Although vimentin expression appeared to be influenced by various conditions such as the proportion of T- and B-cell subsets, or B-cell proliferation rate, follicular center cells were constantly negative for vimentin.

Abolishment of c-fos inducibility in ras-transformed mouse osteoblast cell lines.

Proto-oncogene c-fos is induced by many types of cellular stimuli, such as 12-O-tetradecanoylphorbol-13-acetate (TPA), epidermal growth factor (EGF), serum (fetal bovine), calcium ionophore A23187, and dibutyryl cAMP (But2cAMP). In this study, c-fos induction was abolished in ras-transformed mouse osteoblast cells (MC3T3). Transformants of MC3T3 were isolated after transfection with Ki or Ha murine sarcoma virus DNA. All Ki- or Ha-ras transformed MC3T3 clones examined showed exceedingly low levels of c-fos induction by all inducers, as determined by the change in amounts of c-fos mRNA or its product. Induction of other TPA-responsive genes, such as metallothionein, was not altered in some ras-transformed cells; c-myc and c-jun expression was constitutively high in all the ras-transformed clones. Nuclear extracts and gel shift assay showed that the binding activity to c-fos enhancer element (serum response element) was altered in ras-transformed cells. These results indicate that transformation with ras oncogene induces modification of c-fos enhancer binding factors and that this modification is one cause for the decrease in c-fos induction.

In vitro propagation of herbaceous peony (paeonia lactiflora pall.) by a longitudinal shoot-split method.

A procedure for the clonal propagation ofPaeonia lactiflora Pall. cvs. Takinoyosooi and Sarah Bernhardt through shoot tip culture is described. Half strength Murashige and Shoog (1962) medium supplemented with 0.5 mg/l 6-benzylaminopurine plus 1 mg/l gibberellic acid promoted formation and growth of axillary buds. Continuous shoot multiplication was achieved by vertically splitting the shoot axis and subsequent division of elongated axillary shoots every 36 days. High frequency (57-100%) of rooting was obtained on paper-bridge liquid medium supplemented with 1 mg/l indole-3-butyric acid. Half of the rooted plantlets were established on porous soil. Thus, 700 and 300 plants of cv. Takinoyosooi and Sarah Bernhardt could be theoretically obtained from a single bud in one year.

Induction of c-fos proto-oncogene by a chemotactic peptide in human peripheral granulocytes.

The chemotactic peptide, fMet-Leu-Phe (fMLP), induced proto-oncogene c-fos mRNA in purified human peripheral granulocytes. The induction was transient, and was inhibited by pertussis toxin or by an inhibitor of protein kinase C. These results suggest that activation of a guanine nucleotide-binding protein and of protein kinase C is involved in c-fos induction in granulocytes.