BMP4/Thrombospondin-1 loop paracrinically inhibits tumor angiogenesis and suppresses the growth of solid tumors.

Bone morphogenetic protein 4 (BMP4) has potential as an anticancer agent. Recent studies have suggested that BMP4 inhibits the survival of cancer stem cells (CSCs) of neural and colon cancers. Here, we showed that BMP4 paracrinically inhibited tumor angiogenesis via the induction of Thrombospondin-1 (TSP1), and consequently suppressed tumor growth in vivo. Although HeLa (human cervical cancer), HCI-H460-LNM35 (highly metastatic human lung cancer) and B16 (murine melanoma) cells did not respond to the BMP4 treatment in vitro, the growth of xeno- and allografts of these cells was suppressed via reductions in tumor angiogenesis after intraperitoneal treatment with BMP4. When we assessed the mRNA expression of major angiogenesis-related factors in grafted tumors, we found that the expression of TSP1 was significantly upregulated by BMP4 administration. We then confirmed that BMP4 was less effective in suppressing the tumor growth of TSP1-knockdown cancer cells. Furthermore, we found that BMP4 reduced vascular endothelial growth factor (VEGF) expression in vivo in a TSP1-dependent manner, which indicates that BMP4 interfered with the stabilization of tumor angiogenesis. In conclusion, the BMP4/TSP1 loop paracrinically suppressed tumor angiogenesis in the tumor microenvironment, which subsequently reduced the growth of tumors. BMP4 may become an antitumor agent and open a new field of antiangiogenic therapy.

Absorbed and effective doses from cone beam volumetric imaging for implant planning.

OBJECTIVES: Volumetric CT using a cone beam has been developed by several manufacturers for dentomaxillofacial imaging. The purpose of this study was to measure doses for implant planning with cone beam volumetric imaging (CBVI) in comparison with conventional multidetector CT (MDCT). METHODS: The two CBVI systems used were a 3D Accuitomo (J. Morita), including an image-intensifier type (II) and a flat-panel type (FPD), and a CB MercuRay (Hitachi). The 3D Accuitomo operated at 80 kV, 5 mA and 18 s. The CB MercuRay operated at 120 kV, 15 mA, 9.8 s. The MDCT used was a HiSpeed QX/i (GE), operated at 120 kV, 100 mA and 0.7 s, and its scan length was 77 mm for both jaws. Measurement of the absorbed tissue and organ doses was performed with an Alderson phantom, embedding the radiophotoluminescence glass dosemeter into the organs/tissues. The values obtained were converted into the absorbed dose. The effective dose as defined by the International Commission on Radiological Protection was then calculated. RESULTS: The absorbed doses of the 3D Accuitomo of the organs in the primary beam ranged from 1-5 mGy, and were several to ten times lower than other doses. The effective dose of the 3D Accuitomo ranged from 18 muSv to 66 muSv, and was an order of magnitude smaller than the others. In conclusion, these results show that the dose in the 3D Accuitomo is lower than the CB MercuRay and much less than MDCT.

Cisplatin treatment increases survival and expansion of a highly tumorigenic side-population fraction by upregulating VEGF/Flt1 autocrine signaling.

The cellular and molecular mechanisms of tumor progression following chemotherapy are largely unknown. Here, we demonstrate that cisplatin (CDDP) treatment upregulates VEGF and Flt1 expression leading to the survival and expansion of a highly tumorigenic fraction of side-population (SP) cells in osteosarcoma (HOS), neuroblastoma (SK-N-BE2) and rhabdomyosarcoma (RH-4) cell lines. In all three lines, we show that CDDP treatment increases levels of VEGF and Flt1 expression, and induces enhanced clonogenic capacity and increased expression of the 'stemness'-associated genes Nanog, Bmi-1 and Oct-4 in the SP fraction. In HOS, these changes are associated with the transformation of a non-tumorigenic osteosarcoma SP fraction to a highly tumorigenic phenotype. Inhibition of Flt1 led to complete reduction of tumorigenicity in the HOS SP fraction, and reduction of clonogenic capacity and expression of stemness genes in the SK-N-BE(2) and RH-4 SP fractions. Treatment with U0126, a specific inhibitor of MAPK/ERK1,2 completely downregulates CDDP-induced VEGF and Flt1 expression and induction/expansion of SP fraction in all three cell lines, indicating that these effects are mediated through MAPK/ERK1,2 signaling. In conclusion, we report a novel mechanism of CDDP-induced tumor progression, whereby the activation of VEGF/Flt1 autocrine signaling leads to the survival and expansion of a highly tumorigenic SP fraction.

Differentiation-associated expression and intracellular localization of cyclin-dependent kinase inhibitor p27KIP1 and c-Jun co-activator JAB1 in neuroblastoma.

Neuroblastoma is a unique pediatric cancer, which spontaneously regress in some infants and undergo maturation in older children. The cyclin-dependent kinase inhibitor p27KIP1 negatively control cell cycle progression and its expression is reported to be associated with differentiation and prognosis of some human cancers. To examine whether p27KIP1 is involved in differentiation of neuroblastomas, expression and localization of p27KIP1 in 30 cases of neuroblastic tumors were determined with immunohistochemistry. p27KIP1 was expressed in all cases, but staining intensity and intracellular localization varied in association with tumor differentiation. Primitive small round neuroblasts showed negative or only weak nuclear staining, while differentiating tumor cells displayed a novel, intense cytoplasmic positivity besides the nuclear staining, and mature ganglion cells showed intense positive reaction confined to the nucleus. A neuroblastoma cell line TGW was also immunostained positively for p27KIP1 in the cytoplasm after differentiation induction, and western blot analysis revealed an increase of p27KIP1 in these cells, corroborating the in vivo observations. JAB1, which is thought to bind p27KIP1 and transport it from the nucleus to the cytoplasm for proteasome/ubiquitin-mediated degradation, was found to be localized both in the cytoplasm and the nucleus in undifferentiated and differentiating tumors whereas located predominantly in the nucleus of differentiated tumor cells. These data indicate that the cytoplasmic localization of p27KIP1 in the process of differentiation is due to upregulation of p27KIP1 synthesis and subsequent degradation and suggest a role of p27KIP1 in differentiation of neuroblastoma.

Microtubular aggregates within the rough endoplasmic reticulum of embryonal rhabdomyosarcoma cells: a case report.

This study presents a case of embryonal rhabdomyosarcoma (ERMS) of the forearm soft tissue in a 12-year-old female, in which microtubular aggregates in rough endoplasmic reticulum (rER) were noted ultrastructurally. Histologically, tumor cells consisted of typical rhabdomyoblastoid cells with abundant eosinophilic cytoplasm and relatively immature, small tumor cells. Ultrastructurally, two different types of tumor cells were also identified by light microscopy. More than half the tumor cells possessed the characteristic features of rhabdomyoblastic differentiation, such as abundant thick and thin filaments with Z-bands. The other tumor cells were less differentiated cells in which microtubular aggregates (MA) in rER were observed. MA in rER have been described in several nonepithelial tumors, including malignant melanoma, osteosarcoma, extraskeletal myxoid chondrosarcoma, and chordoma. ERMS is another example of mesenchymal tumor in which MA in rER are observed by electron microscopy. Considering the differential diagnosis among mesenchymal tumors, it is important to know that MA can be also observed in ERMS.

Epidemiology of infant Kawasaki disease with a report of the youngest neonatal case ever reported in Japan.

Our objectives were to describe epidemiological pictures of infant Kawasaki disease (KD) and to report the youngest patient ever reported in Japan. A Japanese database of 105755 KD patients reported in the 25 year period since 1970 was analyzed. Of all the cases registered, there were only six cases aged 30 days or younger and 1768 cases (1.67%) aged 90 days or younger. We reported a typical case of KD (female, the onset at 20 days old) who was the youngest patient ever reported in Japan. The epidemiology of KD in early infants does not contradict the infection theory of the etiology; the rare incidence in the neonatal period can be explained by the protective effects of the passive immunity transferred from the mother and by exposure to the unknown infectious agents.

[Food poisoning caused by enteroinvasive Escherichia coli (O164:H-)--a case in which the causative agent was identified].

Food poisoning due to "Godofu (Sasayuki tofu)" as a main causative foodstuff which broke out on July 14, 1988. There were 670 out of 918 persons who ingested this food who became ill (incidence 73.0%). The main symptoms were diarrhea (93.4%), fever (77.5%), abdominal pain (64.5%), and vomiting (19.9%). A high degree of fever and watery diarrhea were characteristic of this poisoning. The average latent period was 35 hours with a range of one to 156. The O164:H- strains of enteroinvasive Escherichia coli (EIEC) were detected from 22 of the 32 fecal samples collected from the patients, five of ten samples collected from workers engaged in tofu making, and one sample of left-over Godofu. The virulence of EIEC strains isolated from the patients, workers, and left-over food was confirmed by invasion into HeLa and HEp-2 cells, Sereny test, and ELISA test to detect invasive plasmid-derived protein of the organism (conducted at Tokyo Metropolitan Research Laboratory of Public Health). These EIEC strains were sensitive (less than or equal to 0.19 to 6.25 micrograms/ml) to GM, ABPC, CBPC, CER, CET, NA, PB, MINO, TC and CP as well as KM and OFLX which were used for treatment. However, their susceptibility to FOM varied to some extent (6.25 to 25.0 micrograms/ml) and one strain isolated from a tofu worker was resistant to MINO, TC, FOM and CP (25 to greater than or equal to 100 micrograms/ml). Since investigation revealed that Godofu was left at room temperature about 29 degrees C until ingested, we did a experiment to check the bacterial growth in Godofu under similar conditions at the time of outbreak.(ABSTRACT TRUNCATED AT 250 WORDS)

[Case of simultaneous triple cancers of the duodenal papilla, gallbladder and stomach].

The patient was a 76-year-old man with simultaneous triple cancer arising in the duodenal papilla, gallbladder and stomach, whose complaint was epigastralgia. Endoscopic and radiologic studies revealed early gastric cancer (IIc) at the posterior wall of the antrum; ERCP disclosed obstruction with "apple core" appearance in the distal bile duct. Pancreaticoduodenectomy, cholecystectomy and distal gastrectomy with lymph node dissection were performed. Histopathological examination of the resected specimens revealed papillo-tubular adenocarcinoma in the duodenal papilla, well differetiated tubular adenocarcinoma in the gallbladder, and moderately differentiated tubular adenocarcinoma in the gastric mucosa.

Radiation-induced chromosome aberrations in mouse spermatocytes and oocytes.

ICR/Swiss males and females were exposed to 300 R whole-body gamma irradiation. The frequency of aberrations in metaphase I chromosomes recovered from oocytes cultured in vitro was compared with those recovered from spermatocytes irradiated in pachytene and diplotene. The ability of oocytes collected 1 day postirradiation to mature in vitro was not affected, but significantly fewer oocytes cultured 5 days after irradiation matured. The frequency of aberrations in oocytes did not differ from spermatocytes irradiated during diplotene, but significantly more chromosome aberrations were found when pachytene spermatocytes were irradiated. Some variation in the relative frequencies of aberrations was also observed.