Removal of a cavernous hemangioma in the orbital apex via the endoscopic transnasal approach: a case report.

INTRODUCTION: The aim of this study is to describe the case of a cavernous hemangioma extending from the orbital apex to the pterygopalatine fossa that was completely removed via an endoscopic transnasal approach. CASE REPORT: We report the case of a 48-year-old man who presented with right hemianopsia of the left eye. MRI revealed a 1.5 x 1.1 cm mass lesion extending from the infero-medial part of the left orbital apex to the pterygopalatine fossa. Removal of the lesion was performed via the endoscopic transnasal approach. Using this approach, a wide operative view of the entire extent of the lesion from the optic canal to the orbital apex and the pterygopalatine fossa was obtained, and complete removal of the lesion was performed safely. The pathological diagnosis was cavernous hemangioma. CONCLUSION: The endoscopic transnasal approach is a safe, effective, and less invasive therapeutic modality for the removal of lesions extending from the infero-medial part of the left orbital apex to the pterygopalatine fossa. With appropriate patient selection, this approach improves access and visualization, and it enables performance of operative procedures with much less risk than the conventional microscopic transcranial or transfacial approaches.

Interleukin-1 receptor-related protein ST2 suppresses the initial stage of bleomycin-induced lung injury.

Acute lung injury has a range of causes, and occasionally leads to lethal respiratory failure. Despite advances in treatment, acute lung injury continues to have a high mortality rate, and thus a new therapeutic approach is needed. ST2 is an interleukin (IL)-1 receptor-related protein, and its expression is induced by various inflammatory responses. Recently, ST2 has been speculated to exert anti-inflammatory effects; therefore, we investigated the role of the ST2 in the murine model of acute lung injury. To elucidate the function of ST2 in vivo, mice that transiently overexpressed ST2 protein were prepared using the hydrodynamic gene transfer method, and lung injury was induced by intratracheal administration of bleomycin. In bleomycin-treated ST2-overexpressing mice, the increase of neutrophils in the bronchoalveolar lavage fluid (BALF) was markedly suppressed. Additionally, the levels of tumour necrosis factor-alpha and IL-6, as well as the concentration of albumin, in BALF were reduced compared with those of controls. Furthermore, the pulmonary architecture in ST2-overexpressing mice remained almost normal, and the survival rate was significantly improved. From these results, we concluded that ST2 has the potential to suppress the initial stage of acute lung injury, and therefore it may be a useful reagent for the treatment of acute lung injury.

Lack of B-RAF mutations in head and neck squamous cell carcinoma.

B-RAF is one of the most commonly mutated oncogenes in human cancer. However, the mutation status of B-RAF has not been established completely in HNSCC. We have analysed the mutation status of the kinase domain of the B-RAF gene (exons 11 and 15) in 91 Japanese HNSCC patients as well as 12 HNSCC cell lines. DNA was extracted and amplified by PCR. Mutations were then analysed by SSCP mutation detection method. Since V600EB-RAF constitutes 90 % of the mutations identified in B-RAF in human cancers, we also used MASA analysis to specifically detect this mutation in exon 15 of B-RAF. Using both methods, no mutation was found in both exon 11 and 15 in all patients and cell lines. Mu tations are absent or rare in the kinase domain of B-RAF in Japanese HNSCC. However, more studies are still needed to determine its usefulness as a target for molecular therapy in these patients.

Cooperation of six and eya in activation of their target genes through nuclear translocation of Eya.

Drosophila sine oculis and eyes absent genes synergize in compound-eye formation. The murine homologues of these genes, Six and Eya, respectively, show overlapping expression patterns during development. We hypothesized that Six and Eya proteins cooperate to regulate their target genes. Cotransfection assays were performed with various combinations of Six and Eya to assess their effects on a potential natural target, myogenin promoter, and on a synthetic promoter, the thymidine kinase gene promoter fused to multimerized Six4 binding sites. A clear synergistic activation of these promoters was observed in certain combinations of Six and Eya. To investigate the molecular basis for the cooperation, we first examined the intracellular distribution of Six and Eya proteins in transfected COS7 cells. Coexpression of Six2, Six4, or Six5 induced nuclear translocation of Eya1, Eya2, and Eya3, which were otherwise distributed in the cytoplasm. In contrast, coexpression of Six3 did not result in nuclear localization of any Eya proteins. Six and Eya proteins were coimmunoprecipitated from nuclear extracts prepared from cotransfected COS7 cells and from rat liver. Six domain and homeodomain, two evolutionarily conserved domains among various Six proteins, were necessary and sufficient for the nuclear translocation of Eya. In contrast, the Eya domain, a conserved domain among Eya proteins, was not sufficient for the translocation. A specific interaction between the Six domain and homeodomain of Six4 and Eya2 was observed by yeast two-hybrid analysis. Our results suggest that transcription regulation of certain target genes by Six proteins requires cooperative interaction with Eya proteins: complex formation through direct interaction and nuclear translocation of Eya proteins. This implies that the synergistic action of Six and Eya is conserved in the mouse and is mediated through cooperative activation of their target genes.

Continuous infusion of 5-fluorouracil with versus without low-dose, consecutive administration of cisplatin in advanced colorectal cancer. A prospective randomized phase II study.

Recently, the treatment of advanced gastric cancer by continuous infusion of 5-fluorouracil (5-FU) with low-dose cisplatin (CDDP) has improved efficacy without severe toxicities. The possible effectiveness of 5-FU+low-dose CDDP for colorectal cancer (CRC) is intriguing. One hundred fifty-five patients with far-advanced CRC including at least one measurable lesion were enrolled in a prospective randomized clinical trial funded by the Japanese Foundation for Multidisciplinary Treatment of Cancer. These patients were assigned to the two arms to assess the value of low-dose CDDP when added to a continuous intravenous infusion of 5-FU at a dose of 300 mg/m(2)/24 hrs in a one-week cycle consisting of 5 days of treatment and 2 days of rest for at least 12 weeks. CD-DP was given intravenously at a dose of 3 mg/m(2) on days 1-5 and days 8-12, and then at a dose of 7 mg/m(2) twice a week. Three patients were excluded from the trial. The response rate in the 5-FU+low-dose CDDP arm (n=75) was significantly higher than that in the 5-FU arm (n=77) (25.3% vs. 11.7%; P = 0.037). There was no significant difference in the median overall survival time between the 5-FU+low-dose CDDP arm and the 5-FU arm (479 and 491 days, respectively). Grades 3/4 toxicities occurred infrequently in both arms. The quality of life was almost the same between the arms. Low-dose CDDP improved the response rate while keeping toxicities within clinically acceptable limits. However, this combined treatment did not confer a survival advantage over treatment with continuous infusion of 5-FU alone for patients with far-advanced CRC; that might be attributable to the short CDDP administration setting of 12 weeks.

Heterogeneity of circulating carcinoembryonic antigen analyzed by sandwich-enzyme immunoassays with different specificities.

Nine monoclonal antibodies against carcinoembryonic antigen (CEA) were prepared and used to investigate the immunological and physicochemical heterogeneity of circulating CEA. Two of these, M221-73 and M272-11, recognized "CEA-distinctive" epitopes and they gave sandwich-enzyme immunoassays far less reactive with nonspecific cross-reacting antigen (NCA) and nonspecific cross-reacting antigen-2 (NCA-2). Sandwich-enzyme immunoassays selective for NCA-2 and NCA were also established using suitable monoclonal antibodies as competitive inhibitors against enzyme-labeled antibodies. The studies on the serum CEA levels determined by the "CEA-specific" assay indicated that CEA molecules recognized by M221-73 and M272-11 were generally found in the sera of both cancer patients and normal adults. CEA and related substances in sera were further analyzed by the sandwich-enzyme immunoassays after adsorption on M272-11-coupled immunosorbents and by gel filtration on an Ultrogel AcA-34 column. These studies revealed the presence of a CEA variant detected by the "NCA-2-selective" assay. The variant seemed to be closely related to NCA-2 because it lacked CEA-distinctive epitopes and had an apparent molecular weight similar to that of NCA-2. This variant appeared in the sera of some cancer patients and normal adults.

A comparative case-control analysis of stomach cancer and atrophic gastritis.

We conducted a comparative case-control analysis of stomach cancer and atrophic gastritis involving 427 cases with stomach cancer, 1414 cases with atrophic gastritis, and 3014 control subjects based on a questionnaire survey conducted for the subjects who received gastroscopic examination at Aichi Cancer Center Hospital from April 1985 to March 1989. The risk of atrophic gastritis in both males and females was not associated with any environmental factors. The risk of stomach cancer compared with the control subjects was positively associated with an intake of salted fish guts or cod roe [relative risk (RR) = 1.52, 95% confidence interval (CI) = 1.08-2.15] and smoking (RR for 20 or more cigarettes per day = 2.84; 95% CI = 1.79-4.51) and inversely associated with Western-style breakfast (RR = 0.68; 95% CI = 0.48-0.96) in males. Additionally, the risk of stomach cancer was inversely associated with a daily intake of raw vegetables (RR = 0.56; 95% CI = 0.34-0.94) in males when compared with the patients with atrophic gastritis as controls. Several environmental factors, such as intake of green-yellow vegetables, fruit, and meat, and a family history of stomach cancer, were only associated with intestinal types of cancer in females, whereas a clear difference between diffuse and intestinal types was not observed in males. The results of the present study suggest that risk factors for stomach cancer may be different from those for premalignant lesions.

Distinct mutational spectrum of the p53 gene in lung cancers from Chinese women in Hong Kong.

Accumulating evidence suggests that the p53 gene is a good target for molecular epidemiological studies to search for risk factors in carcinogenic events. The lung cancer incidence for females in Hong Kong is unusually high, ranking among the highest in the world despite a low percentage with a history of smoking. To gain insights into possible etiological risk factors responsible for this high incidence, we examined p53 mutations in 35 lung cancer specimens from Chinese females living in Hong Kong and compared them with 35 matched cases from Japanese women as well as previously reported p53 mutations in the world literature. p53 mutations in exons 5-8 were present in 20 and 31% of the Hong Kong and Japanese cases, respectively. Notably, single-base deletions within runs of identical bases were observed in 3 (43%) of the 7 mutations in the Hong Kong cases, in contrast to the absence of such mutations in the controls and the extreme scarcity in the literature, suggesting that distinct environmental and/or genetic factor(s) might be involved. Although the frequent occurrence of characteristic single-base deletions could be a reflection of mutator mutations leading to inefficient mismatch repair of slipped strand mispairings, none of the lung cancer specimens exhibited such microsatellite instabilities.

Nucleotide sequence of a complementary DNA for human ST2.

Human ST2 cDNA, a homologue of murine ST2 that is only expressed in growth-stimulated BALB/c-3T3 cells and a member of the primary response gene family induced by growth factors, was isolated from the cDNA library of an activated human helper T cell line, 5C10. Human ST2 has 67.6% identity in a 327 amino acid overlap to murine ST2. Furthermore, as in the case of murine ST2, human ST2 encodes a protein remarkably similar in sequence to the extracellular portion of human interleukin 1 receptor, both types 1 and 2. The expression of ST2 in human lymphocytes could trigger further investigations into its physiological role in humans.

Identification of the product of the murine ST2 gene.

The murine ST2 gene is expressed in growth-stimulated BALB/c-3T3 cells. This gene encodes a protein that is similar to the extracellular portions of the interleukin-1 receptors (types 1 and 2). In this study, we prepared a polyclonal antibody against the recombinant ST2 protein produced in Escherichia coli. This antibody detected recombinant ST2 protein in the culture fluid of COS7 cells transfected with a mammalian expression vector (pEF-BOS) carrying ST2 cDNA. Using this antibody, we could detect the ST2 protein in the culture fluid of growth-stimulated BALB/c-3T3 cells, and in the medium of continuously growing cells, but not in that of growth-arrested cells. ST2 proteins produced in COS7 cells and BALB/c-3T3 cells were N-glycosylated as predicted from nine putative N-glycosylation sites in its deduced amino-acid sequence.

Molecular cloning of the murine ST2 gene. Characterization and chromosomal mapping.

The genomic locus of the murine ST2 gene was isolated based on homology with a murine ST2 complementary DNA sequence and its complete nucleotide sequence was determined. The locus is composed of eight exons and seven introns and is approx. 9 kilobase pairs in size. Two Sp1 binding sites are present in the 5' flanking region. The murine ST2 gene, which was expressed only in the growth-stimulated BALB/c-3T3 cells, was mapped to mouse chromosome one, very tightly linked to the interleukin 1 receptor-type 1 locus.

Major prognostic factors of adult patients with advanced T-cell lymphoma/leukemia.

Eighty-one adult patients with advanced T-cell lymphoma/leukemia including 54 with adult T-cell leukemia/lymphoma (ATL), who were treated between 1981 and 1983 with vincristine, cyclophosphamide, prednisolone, and doxorubicin (VEPA) or VEPA plus methotrexate (VEPA-M) in randomized fashion, were evaluated for pretreatment characteristics. The overall complete response (CR) and the 4-year survival rates were 39.5% and 19.4%, respectively, and 69% of 32 CR patients had relapses, indicating the need for development of new effective regimens for the disease. In a multiple logistic regression analysis, only three factors, leukemic manifestation, poor performance status (PS), and a high lactate dehydrogenase (LDH) level, were significantly associated with the poor response rate. In a Cox proportional hazards model analysis, shortened survival was again significantly associated with poor PS and a high LDH level, but not with a clinical diagnosis of ATL. The two factors, PS and LDH level, that were found to be significantly associated with both CR and survival rates, were used to construct a model containing six categories of patients at increasing risk for poor response and shortened survival. These categories divided the patients into three groups with respective CR and 4-year survival rates of 75% and 53% for low-risk, 45% and 15% for moderate-risk, and 15% and 0% for high-risk. The results indicate that PS and LDH levels were the most important in predicting the response and survival of an adult patient with advanced T-cell lymphoma/leukemia. The prognosis of patients with usual peripheral T-cell lymphoma, excluding ATL, was comparable with that of advanced B-cell lymphoma. These results have important implications for the design of new prospective therapeutic trials.

Chemotherapeutic results and prognostic factors of patients with advanced non-Hodgkin's lymphoma treated with VEPA or VEPA-M.

One hundred sixty-three patients with advanced non-Hodgkin's lymphoma including adult T cell leukemia/lymphoma (ATL) were treated from 1981 to 1983 with VEPA (vincristine, cyclophosphamide, prednisolone, and doxorubicin) or VEPA-M (VEPA plus methotrexate) in randomized fashion after stratification by surface marker. The complete response (CR) rate and the 4-year survival rate of patients treated with VEPA-M was 62.2% and 36.9%, respectively, while for those treated with VEPA the rates were 51.9% and 26.6, respectively. The difference was not statistically significant, but pretreatment characteristics predictive for response and survival were interesting. Three factors, leukemic change, poor performance status (PS), and T cell marker, were negatively associated with both CR and survival rates, and high-grade pathology was adversely associated with survival rate in a multivariate analysis. These prognostic factors are somewhat different from those in Western lymphomas. This may be reflection of major differences in patients' characteristics between Japanese and Western lymphomas: in this study, there was a high incidence of T cell lymphoma/leukemia (50%) including ATL (33%), leukemic manifestation (34%), poor PS (34%), and a low incidence of follicular lymphoma (9%). The statistically significant three factors for both CR and survival rates were used to construct a model containing eight categories of patients at increasing risk for poor response and shortened survival. These categories were divided into four groups, with respective CR and 4-year survival rates of 91% and 73%, 67% and 35%, 27% and 7%, and 10% and 5%. Ninety-three patients in whom CR was induced by VEPA or VEPA-M therapy were evaluated for prognostic factors predictive for disease-free survival. A shorter period (less than 28 days) required to achieve CR, a clinical diagnosis of ATL, and a lower hemoglobin level were found to affect disease-free survival adversely. These results have important implications for both the design of prospective randomized therapeutic trials and the determination of optimal therapy for individual patients.

Changes in DNA copy number in primary gastric carcinomas by comparative genomic hybridization.

We examined 33 primary gastric carcinomas using comparative genomic hybridization to detect changes in the DNA copy number and the chromosomal location of these changes. Ninety-four percent (31 of 33) showed 1 or more DNA copy number changes, such as increases at 2p23-p25 (observed in 21% of the total cases), 3q26.3-q27 (24%), 7p15 (24%), 9p22-pter (18%), and 13q22-q34 (21%) and decreases at 1p34.2-p36.2 (18%) and Y (52%). Histological examination indicated that increases at 3q26.1-q26.3 and 7p15 and decreases at 1p36.1-p36. 2 and Y were commonly observed in both differentiated and undifferentiated types. Increases at 3q27, 6q23-q25, and 7cen-p14 and decreases at 1p34.2-p35 and 17p12 were predominantly observed in the differentiated type, and increases at 2p23-pter, 9p22-pter, and 13q31-qter and a decrease at 6p21.3 were predominantly observed in the undifferentiated type. In addition, clinical staging of tumors showed that increases at 2p23-p25, 7p14-p21, 7q31-q32, and 9p22-pter and a decrease at Y were observed in early-stage tumors, whereas increases at 9q32-q33 and 15q26 were observed only in late-stage tumors. Many of the abnormalities detected in this study were not previously reported in gastric carcinomas. Our comparative genomic hybridization results indicate the presence of genetic alterations that may play some important role in the development and progression of gastric carcinomas.

A 63 kDa protein is secreted from BALB/c-3T3 cells entering the G1 phase from the G0 state.

A 63 kDa protein is detectable in the culture fluid of mouse BALB/c-3T3 cells traversing from the G0 state to the G1 phase, whereas it is undetectable in the culture fluid of quiescent or growing BALB/c-3T3 cells. Secretion of the protein is maximal at 10 h after serum addition. G0-specific ts mutant cells (rat tsJT60) also secrete the 63 kDa protein only when the quiescent cells are stimulated by serum addition at permissive temperature. These facts indicate that the 63 kDa protein is secreted only from cells traversing from the G0 state to the G1 phase.

Murine mRNA for the beta-subunit of integrin is increased in BALB/c-3T3 cells entering the G1 phase from the G0 state.

The amount of murine mRNA for the beta-subunit of integrin is enriched 6-fold when BALB/c-3T3 cells traverse from the G0 state to the G1 phase, whereas it remains at the basal level when the cells are growing continuously. The peak of its appearance is at 10 h after serum stimulation. The increase in integrin mRNA at a specific point in cell proliferation may be correlated with growth-signal transduction.

A putative protein of a growth specific cDNA from BALB/c-3T3 cells is highly similar to the extracellular portion of mouse interleukin 1 receptor.

A cDNA clone, which represents a species of mRNA that is expressed in growth-stimulated BALB/c-3T3 cells but not in resting cells, was found to encode a protein remarkably similar in sequence to the members of the immunoglobulin superfamily, especially to the extracellular portion of the mouse interleukin 1 receptor. The immunoglobulin superfamily is believed to be involved in cell adhesion and cell-to-cell interaction. The evidence that a member of this family is induced in the course of the initiation of cell proliferation is intriguing.

Murine ST2 gene is a member of the primary response gene family induced by growth factors.

The murine ST2 gene, which encodes a protein remarkably similar to the extracellular portion of murine interleukin 1 receptor types 1 and 2, is expressed in growth-stimulated BALB/c-3T3 cells in the presence of 50 micrograms/ml of cycloheximide. The treatment with 1,000 U/ml of purified native murine beta-interferon superinduced, rather than suppressed, the ST2 mRNA expression as in the cases of c-myc and JE mRNAs. These results suggested that the murine ST2 gene belongs to the family of primary response genes induced by growth factors. Furthermore, a longer ST2-related mRNA was found in BALB/c-3T3 cells that were stimulated to proliferate in the presence of cycloheximide.

The existence of a growth-specific DNA binding factor for the promoter region of mouse ST2 gene.

A comparison of the 5'-flanking regions of human and mouse ST2 genes revealed the presence of two highly conserved DNA sequences. The promoter activity assay with a luciferase gene as a reporter showed that the deletion of the upstream conserved region diminished the transcriptional activity in growing BALB/c-3T3 cells. By electrophoretic mobility-shift analysis, the presence of a factor that binds to the positive regulatory region of the mouse ST2 gene was found in growing but not in quiescent BALB/c-3T3 cells. These results suggest the functional importance of this conserved region and the requirement of a binding factor for the expression of the ST2 gene.

Different factors bind to the regulatory region of the Na+,K(+)-ATPase alpha 1-subunit gene during the cell cycle.

Three factors that bind to the positive regulatory region (ARE) of the Na+,K(+)-ATPase alpha 1-subunit gene were shown to be present in growing BALB/c-3T3 cells as shown by the gel retardation assay pattern in which three specific complexes (C1, C2 and C3) were identified. The complexes are similar to those observed in MDCK cell nuclear extracts in which linker substitution mutations in the competitor gave parallel specific effects in both cells. During the process of the cell growth cycle, the relative mobility of C3 was altered, and the amount of C1 decreased in the G0 state. All three complexes (C1, C2 and C3) disappeared and other specific complexes with higher mobilities were alternatively observed at 6 h after serum stimulation and thereafter. The expression of the mRNA for the alpha 1-subunit gene was repressed at G0 and gradually increased after serum stimulation. These results suggest that different sets of factors are responsible for the transcription of the gene at different stages of the cell cycle.