Peripheral lung adenocarcinoma: correlation of thin-section CT findings with histologic prognostic factors and survival.

PURPOSE: To evaluate the prognostic importance of thin-section computed tomographic (CT) findings of peripheral lung adenocarcinomas. MATERIALS AND METHODS: The subjects were 127 patients with adenocarcinomas smaller than 3 cm in largest diameter who underwent at least a lobectomy with hilar and mediastinal lymphadenectomy. The margin characteristics of nodules and the extent of ground-glass opacity (GGO) within the nodules at preoperative thin-section CT were analyzed retrospectively. Regional lymph node metastasis (LNM) and vessel invasion (VI) were histologically examined in surgical specimens. Survival curves were calculated according to the Kaplan-Meier method. RESULTS: The frequencies of LNM (4% [1 of 24]) and VI (13% [three of 24]) in adenocarcinomas with GGO components of more than 50% were significantly lower than those with GGO components of less than 10% (LNM, P <.05; VI, P <.01). The patients with GGO components of more than 50% showed a significantly better prognosis than those with GGO components less than 50% (P <.05). All 17 adenocarcinomas smaller than 2 cm with GGO components of more than 50% were free of LNM and VI, and all these patients are alive without recurrence. Coarse spiculation and thickening of bronchovascular bundles around the tumors were observed more frequently in tumors with LNM or VI than in those without LNM or VI (P <.01). CONCLUSION: Thin-section CT findings of peripheral lung adenocarcinomas correlate well with histologic prognostic factors.

[Interleukin-6 in the cerebrospinal fluid of two patients with herpes zoster meningitis].

Interleukin-6 (IL-6) levels in the cerebrospinal fluid (CSF) and serum were measured in two immuno-competent children with herpes zoster meningitis, who had vesicles, fever, headache and vomiting before admission. The causative agent was identified as varicella zoster virus (VZV) by detecting an increased antibody index in the serum and specific DNA (by polymerase chain reaction) in the CSF. Both patients fully recovered after treatment with acyclovir. The CSF IL-6 levels were high (260.1 pg/ml, 106.1 pg/ml) at the acute stage and thereafter showed a rapid recovery. The serum IL-6 levels were normal. The increased IL-6 level in the CSF may reflect intrathecal inflammatory response following invasion of VZV into the central nervous system.

C-type natriuretic peptide is synthesized and secreted from leukemia cell lines, peripheral blood cells, and peritoneal macrophages.

OBJECTIVE: C-type natriuretic peptide (CNP) is the third member of the natriuretic peptide family. Cultured endothelial cells secrete CNP, and its secretion rate from the endothelial cells is augmented by lipopolysaccharide, interleukin-1beta, and tumor necrosis factor-alpha, which participate in the pathophysiology of inflammation. In this study, we investigated the regulation of CNP secretion from monocytes and macrophages to estimate its contribution to the progression of inflammation. MATERIALS AND METHODS: CNP secretion rates from two human leukemia cell lines (THP-1 and HL-60), human peripheral blood lymphocytes, granulocytes, monocytes, monocyte-derived macrophages, and mouse peritoneal macrophages were measured under conditions with or without stimulation. Immunoreactive CNP levels in the culture media of these cells were measured by a specific radioimmunoassay. RESULTS: The secretion rates of CNP from THP-1 and HL-60 cells were augmented according to the degree of their differentiation into macrophage-like cells under the stimulation with phorbol ester. Peripheral blood monocytes also increased the CNP secretion rate after their differentiation into macrophages. Retinoic acid elicited synergistic effects on the CNP secretion rate from HL-60 cells when administered with lipopolysaccharide, interferon-gamma, interleukin-1beta, tumor necrosis factor-alpha, or phorbol ester. In contrast, the phorbol ester-stimulated CNP secretion rate from THP-1 cells was suppressed with dexamethasone, which inhibits monocyte differentiation into macrophage. CONCLUSIONS: The secretion rate of CNP from monocytes was shown to be regulated based on the degree of their differentiation. This study provides evidence that the monocyte/macrophage system is one of the sources of CNP, especially under inflammatory conditions.

Cardiac fibroblasts are major production and target cells of adrenomedullin in the heart in vitro.

OBJECTIVE: Adrenomedullin (AM) is a potent vasodilator peptide. Plasma AM concentration is increased in patients with various heart diseases, and both myocytes (MCs) and non-myocytes (NMCs) secrete AM and express its receptors. These facts suggest that cardiac cells possess an autocrine/paracrine capability mediated by AM. METHODS: MCs and NMCs were prepared from cardiac ventricles of neonatal rats. AM and endothelin-1 concentrations were measured by radioimmunoassays, and interleukin-6 level by a specific bioassay. Total nitrite/nitrate contents were measured with a fluorescence assay kit. RESULTS: A basal secretion rate of AM from NMCs was 2.8-fold higher than that from MCs. Interleukin-1beta, tumor necrosis factor-alpha and lipopolysaccharide stimulated AM secretion from NMCs but not from MCs. AM stimulated interleukin-6 production in the presence of these cytokines or lipopolysaccharide, which was more prominent in NMCs. In the presence of interleukin-1beta, AM augmented nitric oxide synthesis 2.7-fold in NMCs, but slightly in MCs. NMCs secreted endothelin-1 at a rate nine times higher than MCs, and AM inhibited endothelin-1 secretion from NMCs. CONCLUSION: This in vitro study suggests that AM in the heart is mainly produced in NMCs and exerts its effects through NMCs, especially under inflammatory conditions.

Regulation of adrenomedullin secretion from cultured cells.

Characterization of immunoreactive adrenomedullin (AM) secreted from cultured human vascular smooth muscle cells and 7 other cells indicates that AM is synthesized and secreted from all cultured cells we surveyed. The secretion rate of AM measured ranges from 0.001-6.83 fmol/10(5) cells/h, and endothelial cells, vascular smooth muscle cells and fibroblasts generally secrete AM at high rates. Based on the results of regulation of AM secretion from vascular wall cells, fibroblasts, macrophages and other cells measured in this and previous studies, AM secretion is found to be generally stimulated by inflammatory cytokines, lipopolysaccharide (LPS) and hormones. Especially, vascular smooth muscle cells and fibroblasts elicited uniform and strong stimulatory responses of AM secretion to tumor necrosis factor (TNF), interleukin-1 (IL-1), LPS and glucocorticoid, but endothelial cells did not elicit such prominent responses. AM secretion of monocyte-macrophage was mainly regulated by the degree of differentiation into macrophage and activation by LPS and inflammatory cytokines including interferon-gamma. The other examined cells showed weaker responses to LPS and IL-1. Although cultured cells may have been transformed as compared with those in the tissue, these data indicate that AM is widely synthesized and secreted from most of the cells in the body and functions as a local factor regulating inflammation and related reactions in addition to as a potent vasodilator. The responses of AM secretion to LPS and inflammatory cytokines suggest that fibroblasts, vascular smooth muscle cells and macrophage are the major sources of AM in the septic shock.

[Assessment of solitary hot spots of bone scintigraphy in patients with extraskeletal malignancies].

Bone scintigraphy is widely used to detect bone metastasis owing to its high sensitivity, but solitary focus of increased uptake often causes diagnostic problem because of its low specificity. The purpose of this study was to assess the significance of solitary hot spot detected in patients with extraskeletal malignancies. We reviewed 1,167 consecutive bone scintigraphies of patients with history of lung, breast or prostatic cancer. There was 185 bone scans showing solitary hot spot (lung; 121, breast; 36, prostate; 28). Of the solitary hot spots, 30 (24.8%) in lung cancer, 8 (22.2%) in breast cancer, and 4 (14.3%) in prostatic cancer were a result of metastatic disease. There was no significant difference in the frequency of bone metastasis according to the site of primary tumor. It was relatively higher in the location of pelvis, scapula and thoracic spine. Clinical symptoms, particularly local bone pain, were helpful to diagnose the solitary hot spot.

Hydatidiform mole coexistent with a twin live fetus: a national collaborative study in Japan.

A national collaborative study was conducted in Japan to evaluate the clinical course and the sequelae of patients with hydatidiform mole coexistent with twin live fetus (HMTF). Seventy-two cases of HMTF were diagnosed based on gross appearance and histopathological criteria. In 18 cases, the molar parts were cytogenetically confirmed to be of androgenetic origin (complete mole). The overall incidence of persistent trophoblastic tumour (PTT) in patients with HMTF was 30.6%, and it increased to 50.0% in the 18 patients with proven androgenetic complete mole coexistent with twin live fetus (CHMTF). Among these patients, the mean gestational age at termination of pregnancy or delivery in those who developed PTT (n = 9) and those who did not (n = 9) were 20.6 and 19.4 weeks respectively. The incidence of severe maternal complications was significantly higher in patients who subsequently developed PTT (P < 0.05). The rate of subsequent development of PTT in patients with CHMTF was found to be considerably higher than in a previous study of patients with single complete mole (50 and 12.5% respectively). However, since the risk of malignancy is unchanged with advancement of gestational age, continued pregnancy may be allowed in patients with HMTF provided that severe maternal complications are controlled and fetal karyotype and development are normal.

Phase II study of irinotecan and cisplatin as first-line chemotherapy in advanced or recurrent cervical cancer.

Irinotecan (CPT-11) and cisplatin are singly active against cervical cancer. We evaluated the efficacy and toxicity of CPT-11 plus cisplatin as first-line chemotherapy in patients with advanced or recurrent cervical cancer. Twenty-nine chemotherapy-naive patients with advanced or recurrent cervical cancer were treated with CPT-11 (60 mg/m(2)) on days 1, 8, and 15 by intravenous infusion over 90 min, followed by cisplatin (60 mg/m(2) i.v.) on day 1 over 90 min. The patients' median age was 57 years (range 35-75). Nineteen patients (66%) had advanced primary disease. Six patients with recurrent disease (21%) had been treated with prior radiotherapy. The remaining 4 patients (14%) had residual or recurrent disease after radical surgery. The histologic diagnoses were squamous cell carcinoma in 25 patients (87%), adenocarcinoma in 3, and adenosquamous cell carcinoma in 1. All eligible patients were included in the toxicity and response analysis based on the intent to treat. Two patients (7%) achieved a complete response and 15 (52%) a partial response (overall response rate: 59%, 95% confidence interval; 41-74%). Stable disease was recorded in 6 patients (21%) and progressive disease in 3 patients (10%). In 3 patients, image-guided evaluation of response was judged to be unfeasible at the time of independent extramural review (10%). The median time to response was 32 days (range 16-62 days). The median survival was 27. 7+ months (range, 6.4-52.8+ months). Two dose-limiting side effects were observed: grade 3 (28%) or 4 (45%) neutropenia and grade 3 (7%) or 4 (7%) diarrhea. Other severe toxicities included anemia (45%), thrombocytopenia (3%), nausea/vomiting (31%), and alopecia (7%). The combination of CPT-11 with cisplatin is an active regimen for treatment of advanced or recurrent cervical cancer albeit with a significant degree of myelosuppression.

[Early phase II trial of oral etoposide administered for 21 consecutive days in patients with cervical or ovarian cancer. ETP 21 Study Group--Cervical-Ovarian Cancer Group].

We conducted multi-site early phase II trial or oral etoposide administered for 21 consecutive days in patients with cervical or ovarian cancer in cooperation with 19 institutes. Fifty mg/body of oral etoposide was administered daily for 21 consecutive days. Cycles were repeated every 28 days. In cervical cancer, 24 patients were enrolled and 17 of them were evaluated. The overall response rate including CR and PR was 23.5% (4/17). In ovarian cancer, 18 patients out of 21 enrolled were evaluated. The overall response rate was 16.7% (3/18). The primary toxicity observed was myelosuppression such as leukopenia, neutropenia, hemoglobin decrease and thrombocytopenia. Other adverse effects were anorexia, nausea, vomitting, fatigue, alopecia and stomatitis. From these results we concluded that oral etoposide administered for 21 consecutive days was effective against cervical cancer.

Cationic multilamellar liposome-mediated human interferon-beta gene transfer into cervical cancer cell.

Interferons (IFNs) have antineoplastic activity, but it has been reported that treatment with IFN alone is not effective in many cancers. To enhance the effect of growth inhibition on tumor cells by raising the concentration, we attempted the transfection of cervical cancer cells, HeLa cells, with human interferon-beta (HuIFN-beta) cDNA contained in the expression vector pRSV delivered by cationic multilamellar liposomes, which resulted in the secretion of HuIFN-beta into the medium. The concentration of HuIFN-beta in the medium was 22 IU/ml by 72 hours after transfection of 10 ng DNA, and provoked around 45-fold cell growth inhibitory effect compared with that of exogenously added HuIFN-beta (1000 IU/ml). This strong growth inhibition was considered to be due to the action of HuIFN-beta in a paracrine manner, and a notable fraction of the cell death was apoptotic.

Inductions of immediate early genes (IEGS) and ref-1 by human chorionic gonadotropin in murine Leydig cell line (MA-10).

The effect of human chorionic gonadotropin (hCG) on the expression of immediate early genes (IEGs) including all members of fos and jun family, and c-myc was studied using mouse Leydig cell line (MA-10 cells) by Northern blot analyses. In addition, the induction of ref-1 which enhances DNA binding of fos/jun proteins was also analyzed. HCG induced a rapid and transient expression of c-fos, fosB, c-jun, junB, junD and c-myc with a peak at 30 min to 1 h. In contrast, induction of fra-1 mRNA was delayed with a peak at 3 hr. However, fra-2 mRNA was immediately increased by hCG with a peak at 1 h. The ref-1 mRNA was expressed before the stimulation and its level was not altered by hCG at least for 8 hr. The differential induction of IEGs and continuous expression of ref-1 mRNA suggest an important role of their gene products on the regulation of Leydig cell function by hCG.

Hepatocyte growth factor in human amniotic fluid promotes the migration of fetal small intestinal epithelial cells.

OBJECTIVE: Previously we reported on the abundant existence of hepatocyte growth factor in amniotic fluid. This study was conducted to clarify the effects of hepatocyte growth factor in amniotic fluid on fetal intestinal epithelial cells. STUDY DESIGN: Amniotic fluid samples were obtained from 22 cases at various gestational ages. The effects of amniotic fluid and recombinant human hepatocyte growth factor on proliferation, migration, and morphogenesis of intestine 407 cells (a cell line derived from fetal intestinal epithelial cells) were investigated. RESULTS: The mobility of intestine 407 cells was stimulated by amniotic fluid in proportion to the concentration of hepatocyte growth factor in amniotic fluid with the same effect observed with recombinant human hepatocyte growth factor. This activity was neutralized by addition of antihuman hepatocyte growth factor antibody. Neither increased deoxyribonucleic acid synthesis nor morphogenesis in response to amniotic fluid was identified under the conditions used. CONCLUSION: Amniotic fluid stimulates intestinal epithelial cell migration by way of hepatocyte growth factor in amniotic fluid during development of the fetal intestine.

Increased matrix metalloproteinase-9 activity in human ovarian cancer cells cultured with conditioned medium from human peritoneal tissue.

Ovarian cancer cells disseminate by attachment to the peritoneal mesothelial cell surface of the abdominal cavity. We therefore investigated the influence of conditioned medium (CM) from human peritoneal tissues and mesothelial cells on the secretion of matrix metalloproteinases (MMPs) by ovarian cancer cells. The molecular weights of MMPs stimulating factors derived from human peritoneal tissues and mesothelial cells were estimated using microconcentrators with various cut-off membranes. Human peritoneal tissues were obtained from 12 surgical patients, and mesothelial cells were isolated from three peritoneal specimens. Exposure to CM from peritoneal tissue caused a concentration-dependent increase of the MMP-2 and MMP-9 bands in CM from NOM1 ovarian cancer cells, as shown by zymography. There was a significant difference in the increase of MMP-2 and MMP-9 (2.46-fold and 7.14-fold, respectively, at 0.4 mg/ml protein; P < 0.005). CM from mesothelial cells also significantly increased the secretion of MMP-9 by NOM1 cells. The molecular size of possible MMP-9-stimulating factors secreted by peritoneal tissues and mesothelial cells was above M(r) 100000. Further, CM of peritoneal tissues and mesothelial cells also induced the invasiveness of NOM1 cells. These findings suggest that mesothelial cells may secrete some factors which predominantly induce the MMP-9 production and increase invading cell numbers.

Expression of midkine and pleiotropin in ovarian tumors.

OBJECTIVE: To compare the expression of midkine and pleiotropin in malignant ovarian tumors with that in normal and benign ovarian tissue. METHODS: Total RNA was isolated from 23 samples of normal ovaries, 15 benign ovarian tumors, and 36 malignant ovarian tumors. Midkine and pleiotropin gene expression was examined by using Northern blot analysis. To confirm the localization of midkine expression, in situ hybridization and immunohistochemical analyses were performed. The truncated midkine messenger RNA was examined using polymerase chain reaction with complementary DNA synthesized from total RNA with reverse transcriptase. RESULTS: Expression of midkine gene was observed in 19 of 23 normal ovary samples and in 51 of 53 ovarian tumors (13 of 15 benign, both of the two borderline tumors, and all 36 malignant tumors). Pleiotropin gene was expressed in six normal ovaries and in 24 tumors (nine benign, two borderline, and 13 malignant tumors). The expression of midkine in germ cell tumors was significantly lower than in epithelial tumors, whereas expression in malignant epithelial tumors was significantly higher than in benign ones. In germ cell tumors, two samples with differentiated neural tissues showed high levels of pleiotropin gene expression. In situ hybridization and immunohistochemical analysis showed strong expression of midkine in cancer cells. The truncated midkine messenger RNA was not found in any of the normal, benign, or malignant tissues examined. CONCLUSION: These results suggest an association between midkine and carcinogenesis. Expression of pleiotropin is more restricted, and high levels of its expression may be correlated with neural differentiation.

Levels of hepatocyte growth factor and its messenger ribonucleic acid in uncomplicated pregnancies and those complicated by preeclampsia.

The purpose of this study was to elucidate the possible relationship between hepatocyte growth factor (HGF) expression and the pathogenesis of preeclampsia. The concentration of immunoreactive HGF was measured and the expression of HGF messenger ribonucleic acid (mRNA) assessed in human placentas obtained from two groups: uncomplicated and preeclamptic pregnancies at various gestational weeks. In addition, the localization of HGF mRNA and c-met protein was analyzed using in situ hybridization and immunohistochemical staining, respectively. The expression of HGF mRNA and the concentration of immunoreactive HGF were highest in second trimester and were significantly decreased in preeclamptic placentas compared with the uncomplicated cases in third trimester. HGF mRNA was localized to placental mesenchymal cells, whereas c-met protein was demonstrated on cytotrophoblast. These results provide evidence of an abnormality of HGF expression in the preeclamptic placentas. Such placentas exhibit the abnormally shallow trophoblast invasion of the uterus, and reduced expression of HGF could well account for this morphometric change.

Effects of gonadotropin, estrogen, and progesterone on c-mos gene expression in mouse oocytes in vivo and in vitro.

OBJECTIVE: To analyze the effects of gonadotropin and ovarian steroid hormones on the gene expression of c-mos in mouse oocytes. METHODS: The changes of c-mos messenger RNA (mRNA) levels in oocytes were examined after the administration of pregnant mare's serum gonadotropin (PMSG) in vivo, or after incubation with estrogen and/or progesterone in vitro. Five IU PMSG was injected intraperitoneally to female immature mice, and human chorionic gonadotropin was also injected intraperitoneally 48 hours after the PMSG injection, with or without mating with male mice. The oocytes were collected from follicles or oviducts at 24, 30, 36, 42, 48, 60, 72, and 84 hours after the injection. The RNAs were extracted from 5 oocytes at each time point, and a reverse-transcription polymerase chain reaction using specific primers to c-mos DNA was performed to measure the relative amount of c-mos mRNA. RESULTS: The c-mos mRNA in oocytes at 36 hours after the injection was 2.7 times higher than that at 24 hours. The c-mos mRNA level gradually decreased thereafter, and after ovulation the level was only 1/10 of the peak level. When the oocytes that were retrieved 24 hours after PMSG injection were incubated with 800 ng/ml estradiol 17-beta or 600 ng/ml progesterone for 120 minutes, the c-mos gene expression was significantly suppressed or stimulated, respectively, in comparison with the absence of these substances. CONCLUSION: Although the regulatory mechanism of c-mos gene expression in oocytes is still unclear because the result obtained from the in vitro study, that estrogen suppressed the c-mos gene expression directly, was inconsistent with the result of the in vivo study, that increases of both c-mos mRNA and estrogen occurred simultaneously with PMSG stimulation in the early phase of preovulatory oocytes, our present study revealed that gonadotropin and steroid hormones might affect c-mos gene expression in mouse oocytes indirectly and/or directly.

A randomized trial of cisplatin, vinblastine, and bleomycin versus cyclophosphamide, aclacinomycin, and cisplatin in epithelial ovarian cancer.

After primary cytoreductive surgery, a randomized clinical trial was conducted in women with epithelial ovarian cancer to compare the impact on survival between PVB chemotherapy, consisting of cisplatin, vinblastine and bleomycin, and CAP chemotherapy, consisting of cyclophosphamide, aclacinomycin and cisplatin. There were 148 evaluable patients. One hundred and five patients with stage II, III and IV were analyzed in this study, 49 of them received PVB chemotherapy while the remaining 56 patients received CAP chemotherapy. Sixty-four patients fulfilled the criteria for clinical remission set by the Tokai Ovarian Tumor Study Group [Gynecol Oncol 1993;48:342-348]. The remission rate was 73 and 50% in the PVB and CAP groups, respectively, and showed a significant advantage for the PVB group (p = 0.0139). Moreover, the recurrence rate was 44% in the PVB group and 61% in the CAP group after clinical remission, although there was no significant difference between the two groups. The final survival rate was 32% in the PVB group and 24% in the CAP group. There was a significant difference of survival rate between both groups at 24 months (p = 0.0378) and 48 months (p = 0.0450), but finally no significant difference was found at 96 months (p = 0.0660). Compared to the CAP regimen, the PVB combination has a significantly higher efficacy in remission, but there was no significant difference in the long-term survival rate. Furthermore, multivariate analysis demonstrated that the PVB chemotherapy improved the survival, but it was not significant. The authors conclude that PVB chemotherapy may be more effective than CAP chemotherapy for epithelial ovarian cancer.