Overexpression of Cbfa1 in osteoblasts inhibits osteoblast maturation and causes osteopenia with multiple fractures.

Targeted disruption of core binding factor alpha1 (Cbfa1) showed that Cbfa1 is an essential transcription factor in osteoblast differentiation and bone formation. Furthermore, both in vitro and in vivo studies showed that Cbfa1 plays important roles in matrix production and mineralization. However, it remains to be clarified how Cbfa1 controls osteoblast differentiation, bone formation, and bone remodelling. To understand fully the physiological functions of Cbfa1, we generated transgenic mice that overexpressed Cbfa1 in osteoblasts using type I collagen promoter. Unexpectedly, Cbfa1 transgenic mice showed osteopenia with multiple fractures. Cortical bone, which was thin, porous, and enriched with osteopontin, was invaded by osteoclasts, despite the absence of acceleration of osteoclastogenesis. Although the number of neonatal osteoblasts was increased, their function was impaired in matrix production and mineralization. Furthermore, terminally differentiated osteoblasts, which strongly express osteocalcin, and osteocytes were diminished greatly, whereas less mature osteoblasts expressing osteopontin accumulated in adult bone. These data indicate that immature organization of cortical bone, which was caused by the maturational blockage of osteoblasts, led to osteopenia and fragility in transgenic mice, demonstrating that Cbfa1 inhibits osteoblast differentiation at a late stage.

An interaction between human colon carcinoma cells and hepatocytes activates transforming growth factor-beta1 in vitro.

We established in vitro heterotypic co-cultures of a human colon carcinoma cell line, HT-29, and a human hepatocyte line, tPH5CH, using transwells to investigate possible interactions between colon carcinoma cells and hepatocytes during the metastatic process. Co-culture, but not HT-29 conditioned medium, inhibited tPH5CH cell proliferation, and this inhibition was blocked by an anti-TGF-beta1 antibody. Significantly more activated TGF-beta1 was released by co-culture than by each cell line cultured alone, whereas there were no significant differences between the total TGF-beta1 released by the individual cultured cell lines and that released by co-culture. These data indicate that an interaction between human colon carcinoma cells and hepatocytes activates TGF-beta1 in vitro.

Interferon gamma increases the antitumor activity of mitomycin C against human colon cancer cells in vitro and in vivo.

A combined antitumor activity of mitomycin C (MMC) and interferon gamma-1a (IFN-gamma) was evaluated to be synergistic by the 3-(4, 5-dimethylthiazol-2yl)-2,5-diphenyl-2H tetrazolium bromide assay and human tumor xenografts/severe combined immunodeficient (SCID) mouse system using colon cancer cell lines. The experimental metastasis of WiDr cells in SCID mouse was inhibited by MMC and IFN-gamma with their synergism. Intracellular uptake of MMC in WiDr cells in vitro was significantly increased by IFN-gamma, suggesting the mode of synergism of these agents. This model may also partly explain the antitumor activity of combined MMC, 5-fluorouracil and interleukin-2 treatment on hepatic metastasis of colon cancer.

Combined antitumour activity of mitomycin C, 5-fluorouracil and interleukin-2 against human colon cancer cells.

Combined treatment of interleukin-2 (IL-2), 5-fluorouracil (5-FU) and mitomycin C (MMC) is reported to be effective against hepatic metastasis of colon carcinoma. To clarify this modality, we investigated lymphokine activated killer (LAK) activity of human peripheral lymphocytes on colon cancer cell lines by 51Cr release assay. LAK activity against Colo 205 cells by untreated lymphocytes was 12%, which increased to 25% upon the addition of 5-FU, but remained unchanged upon the addition of MMC. The combined antitumour activity of MMC or 5-FU with or without LAK-supernatant was evaluated by MTT assay; antitumour activity of MMC was increased by the addition of LAK-supernatant. The concentration of interferon-gamma (IFN-gamma) and tumour necrosis factor-alpha (TNF-alpha) was increased in LAK-supernatant, and the antitumour activity of combined IFN-gamma and MMC was synergistic. The mechanism of this therapy seemed to be associated with both increased LAK activity by 5-FU and increased antitumour activity of MMC with IFN-gamma.

Nerve growth factor levels in cerebrospinal fluid from patients neurologic disorders.

Nerve growth factor levels were measured in the cerebrospinal fluid from 73 patients with neurologic disorders and non-neurologic acute illnesses by a two-site enzyme immunoassay. Elevated nerve growth factor levels in cerebrospinal fluid were demonstrated in 2 of 7 patients with bacterial meningitis, 7 of 14 patients with viral meningitis or encephalitis, and 1 with multiple sclerosis. Follow-up examinations of the 3 patients (1 with bacterial meningitis, 1 with viral meningitis, and 1 with multiple sclerosis) at convalescent stage showed diminished nerve growth factor levels in cerebrospinal fluid. None of the other patients showed elevation of nerve growth factor levels in cerebrospinal fluid. Nerve growth factor levels in cerebrospinal fluid were not correlated with cell numbers in patients with meningitis or encephalitis. No relationship was observed between nerve growth factor levels and outcome in patients with viral meningitis or encephalitis and bacterial meningitis. Nerve growth factor in cerebrospinal fluid may play a role in neuronal recovery or function as an immunomodulator in children with inflammatory and immune-mediated neurologic disorders.

Forced normalization induced by ethosuximide therapy in a patient with intractable myoclonic epilepsy.

Forced normalization (FN) is a well known phenomenon of acute psychosis accompanying seizure control. An 11-year-old boy with intractable myoclonic epilepsy and severe psychomotor delay experienced FN during medication of ethosuximide. Although his myoclonic seizures were completely controlled, behavioral changes, more of the manic type, became evident. EEG during this phenomenon showed almost normal findings. Generally, withdrawal of anti-epileptic drugs in patients with FN is still controversial and much debated. We could conclude that the physician should judge comprehensively the treatment considering the more favorable situation for the patient and the family.

Systematic visualization of coronary arteries by two-dimensional echocardiography in children and infants: evaluation in Kawasaki's disease and coronary arteriovenous fistulas.

We describe in our report a systematic approach for visualizing in detail coronary artery anatomy by two-dimensional echocardiography (2DE). The approach provides longitudinal and transverse images of the proximal coronaries, as well as transverse images of the anterior descending and right coronary artery tree. The method is delineated by showing certain cases of Kawasaki's disease (MCLS) and coronary arterial fistula. Using these multiple echo planes, the diagnostic accuracy was improved for both right and left coronary arteries.

[Transposition of the great arteries associated with the straddling tricuspid valve diagnosed preoperatively by two-dimensional echocardiography: report of a case].

A case of transposition of the great arteries associated with the straddling tricuspid valve and the hypoplastic right ventricle was presented, who was diagnosed by two-dimensional echocardiography and finally confirmed by operation. The patient, a 2-year-old girl, had cyanosis, clubbed fingers, effort dyspnea and poor growth. The electrocardiogram revealed both atrial and left ventricular hypertrophy. In the M-mode echocardiogram, the interventricular septal echo was recorded at the level of the midportion of the tricuspid valve and posterior tricuspid leaflet opened behind the interventricular septum during diastole. Two-dimensional echocardiography demonstrated a small right ventricle with the tricuspid valve and its orifice which were straddling over the interventricular septum. Contrast echocardiography disclosed that in early diastole the bolus of contrast echoes passed through the tricuspid orifice into the right and left ventricles simultaneously. Left ventricular angiocardiography in the left anterior oblique projection revealed the negative diastolic silhouette of the tricuspid valve straddling the interventricular septum. Operative findings were as follows; the tricuspid orifice was normal in size, but the crest of the interventricular septum traversed the medial 1/3 of the orifice, the posterior 2/3 of the septal leaflet was supported by an anomalous papillary muscle originated from the left ventricular cavity adjacent to the posterior papillary muscle of the mitral valve and the posterior leaflet of the tricuspid valve was supported with the short chordae from the crest of the septum. The diagnosis of the straddling tricuspid valve was accurately made with the use of two-dimensional echocardiography.