Inhibition of hyaluronan retention by 4-methylumbelliferone suppresses osteosarcoma cells in vitro and lung metastasis in vivo.

BACKGROUND: Hyaluronan (HA) plays crucial roles in the tumourigenicity of many types of malignant tumours. 4-Methylumbelliferone (MU) is an inhibitor of HA synthesis. Several studies have shown its inhibitory effects on malignant tumours; however, none have focused on its effects on osteosarcoma. METHODS: We investigated the effects of MU on HA accumulation and tumourigenicity of highly metastatic murine osteosarcoma cells (LM8) that have HA-rich cell-associated matrix, and human osteosarcoma cell lines (MG-63 and HOS). RESULTS: In vitro, MU inhibited HA retention, thereby reducing the formation of functional cell-associated matrices, and also inhibited cell proliferation, migration, and invasion. Akt phosphorylation was suppressed by MU (1.0 mM). In vivo, although MU showed only a mild inhibitory effect on the growth of the primary tumour, it markedly inhibited (75% reduction) the development of lung metastasis. Hyaluronan retention in the periphery of the primary tumour was markedly suppressed by MU. CONCLUSION: These findings suggested that MU suppressed HA retention and cell-associated matrix formation in osteosarcoma cells, resulting in a reduction of tumourigenicity, including lung metastasis. 4-Methylumbelliferone is a promising therapeutic agent targeting both primary tumours and distant metastasis of osteosarcoma, possibly via suppression of HA retention.

Arterial and cardiopulmonary baroreceptor and chemoreceptor influences and interactions on ear sympathetic nerve discharge in the rabbit.

1. The effects of changing intravascular pressures on integrated ear sympathetic nerve activity (ESNA) were studied in anesthetized artificially ventilated rabbits by inflating aortic and inferior vena caval perivascular balloons under conditions of normal arterial Po2 and during arterial hypoxia. 2. At normal Po2 ESNA was unaffected by arterial and cardiopulmonary baroreflex influences. The small inhibition of ESNA observed during rises in arterial pressure after vagotomy was also present after section of the carotid sinus and aortic nerves, and after cutting both vagi as well. 3. During hypoxia there was marked inhibition of ESNA, which was minimally influenced by vagotomy but abolished by section of the carotid sinus and aortic nerves, suggesting that it was chemoreceptor-mediated. There was a pressure-related rise in ESNA which was abolished by vagotomy and considered to be due to a central nervous chemoreceptor-cardiopulmonary baroreflex interaction.

Quantitative assessment of water diffusion changes in brains of children with neurofibromatosis type I using apparent diffusion coefficient.

We measured diffusion changes in the brains of children with neurofibromatosis type 1 (NF1). Using diffusion-weighted and conventional magnetic resonance (MR) images of 42 children with NF1 (19 girls, 23 boys; 7 months-16 years, mean 6.8 years) and 42 age-matched controls (20 boys, 22 girls; 6 months-17 years, mean, 6.9 years), we calculated the apparent diffusion coefficient (ADC) from the automatically generated ADC maps and placed regions of interest in the pons, middle cerebellar and cerebral peduncles, thalami, globus pallidi and frontal white matter. Evaluating only normal-appearing regions on conventional images, we compared mean ADCs using the unpaired Student t test. Means were not significantly different in frontal white matter but were larger in the other regions in the NF1 (P < 0.01). Although conventional MR showed normal intensity, ADCs of the pons, middle cerebellar and cerebral peduncles, thalami and globus pallidi were significantly larger in the NF1.

The change of vagal activity evoked by spinal cord thermal stimulation in anesthetized rabbits.

Vagal activity decreased significantly during spinal cord warming and increased significantly during spinal cord cooling in anesthetized, immobilized rabbits. The results provide the first direct proff of changes in parsympathetic activity during spinal thermal stimulation.

Patterns of differentiation in various sympathetic efferents induced by hypoxic and by central thermal stimulation in decerebrated rabbits.

The patterns of regional changes of sympathetic efferent activity evoked by thermal stimulation of the spinal cord and by arterial and primary tissue hypoxia were investigated in decerebrated, anesthetized and immobilized rabbits. Decerebration was performed either at the mid- or infracollicular level. The responses of the decerebrated rabbits evoked by spinal thermal stimulation were the same as those of intact rabbits, i.e., splanchnic and cardiac sympathetic activity increased and cutaneous sympathetic activity decreased during warming, while the reverse response was elicited by cooling. It is concluded that the typical thermoregulatory response pattern of the sympathetic nervous system can be produced also after the loss of hypothalamic integration, i.e., by integrative mechanisms in the lower brain stem and the spinal cord. In contrast, the responses of decerebrated rabbits to arterial and primary tissue hypoxia differed from those of intact rabbits in that they consisted in an overall activation in all investigated sympathetic branches. It is confirmed by this result that suprabulbar integration is essential for the generation of the inhibitory components in the differential sympathetic responses to hypoxia, which typically consist in cutaneous and cardiac sympathetic inhibition with splanchnic activation during arterial hypoxia and in cutaneous sympathetic inhibition with cardiac and splanchnic sympathetic activation during primary tissue hypoxia.

Renal sympathetic baroreflex during normoxia and during hypoxia in conscious and in anesthetized rabbits.

The responses of renal sympathetic nerve activity (RSNA) to changes in mean arterial pressure (MAP) during normoxia and hypoxia was studied in conscious rabbits and during anesthesia with pentobarbitone (PB) by determining the RSNA baroreflex curves. In conscious rabbits, the gain in RSNA response was greater and the range of MAP between minimum and maximum levels of RSNA was narrower than in anesthetized rabbits. The renal sympathetic baroreflex was augmented by hypoxia, indicating a central excitatory interaction between the effects of baro- and chemoreceptor stimulation. However, hypoxia produced no significant change in median blood pressure. During anesthesia with PB, resting MAP was decreased, median blood pressure was lowered, and renal sympathetic baroreflexes were less pronounced. Renal sympathetic baroreflex was augmented by hypoxia, and there was a significant increase in median blood pressure. These results provide direct evidence of an inhibitory effect of PB on the response of RSNA to baro- and chemoreceptor stimulation.

[Bone marrow survey by MRI using a table-moving technique].

The bone marrow of the whole body was imaged by MRI in five normal volunteers. The fast short tau inversion recovery (STIR) sequence and sequential imaging using a moving-table system made sagittal and coronal whole body marrow imaging possible within an examination time of 10 minutes. The images obtained showed high resolution and reflected normal red and yellow marrow distribution in all subjects. Additional abnormalities including humeral bone cyst, hepatic cavernous hemangiomas, and maxillary retention cyst were detected. Fast STIR with the table-moving technique was considered a suitable method for rapid bone marrow survey.

Renal and cutaneous vasomotor and respiratory rate adjustments to peripheral cold and warm stimuli and to bacterial endotoxin in conscious rabbits.

In conscious rabbits peripheral cold stimuli decreased respiratory rate and increased cutaneous vasomotor tone while simultaneously renal sympathetic nervous discharge decreased. Peripheral warm stimuli produced the reverse pattern of autonomic effector activity. Injection of a bacterial endotoxin at warm ambient temperature elicited a biphasic fever response. Within the first 60 min cutaneous vasomotor tone increased, simultaneously renal sympathetic activity decreased. Therefore, ear skin vessels dilated and renal sympathetic activity increased by about 100%. Respiratory rate remained depressed during both fever phases. Renal blood flow was investigated in a second series of experiments and showed a negative stimulation but only in the first phase of fever. The results show that renal innervation is involved in the typical thermoregulatory autonomic activity pattern by which temperature homeostasis is preserved. The vasomotor patterns of cold and heat stress developing during fever are compatible with the concept of a changed set-point of the body thermostat. The depressed respiratory rate and the lack of the renal vascular response to the increased nervous activity during the second fever phase and their reversal to normal after acetylsalicylate (ASA) indicate the participation of prostaglandins modifying peripheral and central neurotransmitter mechanisms.

[Normal sternal bone marrow MRI using STIR].

The MR signal of sternal bone marrow was examined in 21 normal volunteers using a sagittal STIR sequence. Craniocaudal phase-encoding with a hand-made positioning device effectively eliminated artifacts due to cardiac pulsation. The sternal bone marrow signal could be classified into three patterns based on the signal distribution. The superior segment of the manubrium showed characteristic age-related changes. This method provided high spatial resolution and excellent bone marrow imaging. Knowledge of a normal marrow signal pattern may be useful for the evaluation of hematological disorders.

Thermally-induced cutaneous sympathetic activity related to blood flow through capillaries and arteriovenous anastomoses.

In the ears of anaesthetized rabbits cutaneous efferent sympathetic nerve activity (SkNA) and blood flow (Q) to capillaries have been measured during various thermal treatments. Warming the spinal cord or skin of the body midside caused a marked decrease in SkNA but capillary Q increased only slightly. Exposure to a warm environment or localized warming of the ear alone induced either a decrease, an increase, or no change in SkNA, but capillary Q always increased markedly. The usual slight increase in capillary Q during spinal warming, was abolished by preventing the usual marked increase in skin temperature. When the spinal cord of the conscious rat was warmed, a marked increase in temperature of the tail (which contains arteriovenous anastomoses, AVA's) indicated dilatation, whereas there was no change in ear temperature (where there are no AVA's). When these results are considered together with recently defined differential influences of reflex and direct effects of temperature on blood flow through cutaneous AVA's and capillaries, it is concluded: (1) That thermally-induced reflex changes in skin blood flow are mediated via sympathetic nervous action on AVA's; (2) Changes in blood flow evoked by direct heating take place through the capillaries, not the AVA's, quite independently of SkNA.

EMG activity in pectoral and femoral muscles during spinal cord cooling in exercising pigeons.

Adult domestic pigeons, with thermodes chronically implanted in the vertebral canal, were trained to walk on a treadmill. In the first series of experiments, EMG activity in a pectoral (M. pectoralis) and a femoral muscle (M. biceps femoris) was measured to determine if shivering could occur during exercise. When the spinal cord was cooled (36.2 +/- 0.5 degrees C) during exercise (0.6 km/h), pectoral muscle EMG activity increased by 80%, while femoral muscle EMG activity did not change significantly. EMG activity remained unchanged during exercise in control experiments where the spinal cord was not cooled. In the second series of experiments, the spinal cord was first cooled (36.1 +/- 0.5 degrees C) for 5 min in resting pigeons and then the treadmill was started. Spinal cord cooling during rest significantly increased pectoral muscle EMG activity but not that of the femoral muscle. Within 1 s after the onset of exercise, EMG activity in the pectoral muscle decreased by 74%. In both series of experiments, shivering was not induced in the femoral muscle. The level of pectoral muscle EMG activity stimulated by spinal cord cooling during exercise in the first series of experiments corresponded to the level to which EMG activity was reduced by exercise during spinal cord cooling in the second series of experiments (192% and 186% in relation to the post-cooling level, respectively). It is concluded that shivering can be induced in the pectoral muscle by spinal cord cooling during exercise in the pigeon.(ABSTRACT TRUNCATED AT 250 WORDS)

[Breath-holding three-dimensional MR imaging: evaluation of phase reordering for motion artifact suppression].

To suppress image degradation due to respiratory corruption during breath holding, a method of reordering of phase encoding was introduced in three-dimensional MR imaging. Without trade-offs in SNR, CNR, and examination time, the method provided a significant reduction of respiratory motion-induced artifacts both in phantom and in vivo experiments. More than twenty seconds of breath holding ensured acceptable image quality regardless of respiratory motion thereafter. Signal intensity change during data acquisition altered the obtained image contrast compared with that of standard sequential linear phase encoding. This technique can be readily implemented on standard two-and three-dimensional pulse sequences and will improve image quality when a patient cannot hold his/her breath during the whole imaging period.

Photocatalytic activity of TiO2 particulate films prepared by depositing TiO2 particles with various sizes.

TiO2 particles of various sizes were prepared by grinding in cyclohexanone, and TiO2 particulate films were obtained by depositing these TiO2 particles with various sizes onto a glass or quartz substrate. The effect of the particle size and thickness on the photocatalytic properties of the films was evaluated via oxidative degradation of gaseous 2-propanol. The initial rate of 2-propanol degradation under UV light irradiation for the films deposited with 30 nm TiO2 particles increased with increasing film thickness up to 600 nm, and reached a saturated value above this film thickness. Photocatalytic activity for the films with thickness below 600nm was larger when smaller TiO2 particles were deposited onto the substrate, due to the increase in the surface area of the particulate films. Furthermore, saturated values of the photocatalytic activity for thick films were smaller for the films deposited with smaller particles, which is mainly attributed to the change in crystal form of the particles during the grinding treatment.

[The effects of flavoxate hydrochloride on the gastrointestinal motility (author's transl)].

The experiments were performed to study the effects of Flavoxate hydrochloride on the gastrointestinal motility and its underlying mechanism. Gastrointestinal motility was inhibited at first and then accelerated markedly after the intravenous injection of Flavoxate hydrochloride (10 mg/kg) in the experiments of anesthetized dogs. For analyzing the underlying mechanism of this response the following experiments were executed. 1) The pendular movements of isolated ileum of rabbits were accelerated with administration of low concentration of Flavoxate hydrochloride less than 10(5) g/ml, while these were reduced with administration of high concentration more than 10(4) g/ml. Because the tetrodotoxin (2.5 times 10(7) g/ml) has no essential effect on these responses, it could be postulated that Flavoxate hydrochloride has direct effect on intestinal smooth muscle itself. 2) Flavoxate hydrochloride (10 mg/kg) induced the differentiated regional response of the sympathetic outflow, that is, the parallel decrease of cutaneous and cardiac sympathetic activities, and the converse increase of splanchnic activity. Therefore, Flavoxate hydrochloride is thought to have an effect on the integrating mechanism of the sympathetic nervous system. This view was also supported by the experiments with neuromuscular preparation of lobster (Panulirus japonicus). It is concluded from these results, that the change of gastrointestinal motility induced by intravenous injection of Flavoxate hydrochloride (10 mg/kg) is influenced both by its effect on the gastrointestinal smooth muscle itself and by its effect via the autonomic nervous system.

Increase in oxygen consumption induced by selective spinal cord cooling in the exercising pigeon.

Six domestic pigeons with chronically implanted spinal thermodes were exercised on a treadmill at neutral ambient temperature. During the exercise the spinal cord was cooled to 34.7 +/- 0.4 degrees C (mean +/- S.E.M.). Oxygen consumption of the pigeons increased from 28.3 +/- 2.1 to 61.2 +/- 3.7 ml X min-1 X kg-1 due to exercise per se, and superimposed cooling of the spinal cord during exercise induced an additional increase in oxygen consumption to 84.9 +/- 4.5 ml X min-1 X kg-1. The result demonstrates that cooling of the spinal cord elicits shivering in exercising pigeons at thermoneutral conditions.

[Detection of recurrent rectal cancer using helical CT with SmartPrep: a new technique for obtaining optimal contrast enhancement].

Seven patients with suspected recurrent rectal cancer underwent Helical CT using SmartPrep. SmartPrep is a software application developed by GE in which the ROI is placed at the desired level of the aorta prior to contrast injection. Using a low-dose continuous scan, the time density curve was obtained instantly, and the optimal timing of scanning was judged from the curve. Four patients who demonstrated intense enhancement at the anastomotic site had local recurrence. No intense enhancement was observed in these areas on conventional delayed CT. Thus, SmartPrep appears to be useful for the accurate diagnosis of recurrent rectal cancer.

[Table-moving contrast-enhanced MR angiography of abdominal aortic aneurysm].

Table-moving contrast-enhanced MR angiography (MRA) was performed in 14 cases of abdominal aortic aneurysm to evaluate its clinical usefulness. In all cases, aneurysms were clearly demonstrated and image quality was clinically acceptable. Findings of reconstructed MRA were highly consistent with those of DSA, and thrombosed areas were confirmed on source images. Main aortic branches including renal arteries, common iliac arteries, and internal and external iliac arteries were readily identified on reconstructed MRA and/or source images. Additional findings such as thoracic aortic aneurysm (n = 1), common iliac aneurysm (n = 6), external iliac aneurysm (n = 1), internal iliac aneurysm (n = 1), femoral arterial obstruction (n = 2), and femoral arterial stenosis (n = 4) were also detected. Although table-moving MRA may have disadvantages like reduced blood signal and limited spatial resolution compared with the conventional contrast-enhanced technique, the images that were obtained provided sufficient contrast and resolution for preoperative evaluation. Because abdominal aortic aneurysm is accompanied by various arterial abnormalities in many of the large arteries, table-moving MRA was considered a suitable technique for comprehensive assessment.