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Biomedical subjects

S Homma

Publications and source records attributed to S Homma.

At least 271 records · Page 15Linked to original sources

Integration of hepatitis B virus DNA into cells of six established human hepatocellular carcinoma cell lines.

The authors successfully established 5 hepatocellular carcinoma cell lines, JHH-1, 2, 4, 5 and 6, derived from hepatitis B surface antigen seronegative patients and one line. JHH-7, from a patient considered to be a hepatitis B virus carrier. In the culture media of any of the JHH cell lines, including JHH-7, hepatitis B surface antigen was not detected by radioimmunoassay. However, in JHH-7, integration of hepatitis B virus DNA was confirmed at two sites on the chromosomes of this line by Southern blot hybridization. In contrast, in other JHH cell lines derived from hepatitis B surface antigen seronegative patients, integration of hepatitis B virus DNA into the chromosomes of cells was not detected. These cell lines will be useful in the investigation of hepatocellular carcinoma development, especially for research into non-A/non-B hepatitis viruses.

Albumins↗

[Establishment and characterization of a human hepatocellular carcinoma cell line JHH-7 producing alpha -fetoprotein and carcinoembryonic antigen--changes in secretion of AFP and CEA from JHH-7 cells after heat treatment].

A human hepatocellular carcinoma cell line, JHH-7, was established from resected liver tumor of a 53 year old male with hepatitis B virus infection. JHH-7 was composed of polygonal epithelial cells and functionally synthesized and secreted human albumin, AFP, CEA and ferritin. No HBsAg was detected in the culture supernatant of JHH-7 cells. Changes of secretion of AFP and CEA from JHH-7 cells after heat treatment was studied using a temperature gradient incubator. Secretion of AFP decreased along with the inhibition of cell proliferation by heat treatment. Secretion of CEA, however, did not decrease even though the cells were damaged.

Biomarkers, Tumor↗

Causes of the urinary concentrating defect in mice with nephrogenic diabetes insipidus.

In a strain of mice called DI +/+ Severe, nephrogenic (or vasopressin-resistant) diabetes insipidus is caused by an inability of the antidiuretic hormone (ADH, or vasopressin) to increase the water permeability of the renal collecting system. That inability, in turn, arises from abnormally high activity of the enzyme cAMP-phosphodiesterase, specifically of the isozyme type III (PDE-III), which hydrolyzes cAMP and prevents the intracellular buildup of this second messenger. Two rather specific inhibitors of PDE-III, rolipram and cilostamide, used either in vitro or in vivo, reverse the deficiencies in DI +/+ Severe mice by increasing intracellular cAMP and water permeability toward or to their normal values. These results have implications for the treatment of nephrogenic diabetes insipidus in human patients.

3',5'-Cyclic-AMP Phosphodiesterases↗

[Analysis for the integrated hepatitis B virus genome in cells of established human hepatocellular carcinoma cell line JHH-7].

Authors have successfully established 7 human hepatocellular carcinoma (HCC) cell lines. In one of these cell lines, JHH-7, derived from HBs antigen positive HCC, hepatitis B virus (HBV) genome was analyzed by Southern blot hybridization. Digesting with Hind III restriction endonuclease, two bands were obtained at 6.0 and 2.5 kb, that is the integration of HBV genome was confirmed at the two sites of chromosomes in this cell line. And analyzing with 32P labeled HBV-DNA fragments for probes, it was indicated that integrated HBV genome was incomplete one containing from X to C region.

Blotting, Southern↗

Hypotonic diuresis following oropharyngeal stimulation with water in humans.

The role of oropharyngeal mechanisms in body water regulation was studied in 12 human males by measuring urine output and osmolality before and after drinking a very small volume of distilled water (0.15 ml/kg b.w.t.). Hypotonic diuresis was resulted only in the subjects (n = 6) who drank only sufficient water to keep their oropharynx moist continually over a 20 min period but not in those who (n = 6) drank the same volume of water within several seconds. Sham verbal instructions on drinking induced no changes in subjects examined. These results suggest that oropharyngeal afferents alone, and neither gastric afferents nor psychosomatic effects, may account for hypotonic diuresis following water intake in man.

Adult↗

Cell death of motoneurons in the chick embryo spinal cord. X. Synapse formation on motoneurons following the reduction of cell death by neuromuscular blockade.

Chronic treatment of chick embryos with neuromuscular blocking agents, such as curare, rescues motoneurons from naturally occurring cell death. In the present study, embryos treated with curare from E6 to E9 had 35% more motoneurons than controls on E10 and 42% more than controls on E16. Previous studies have shown that several aspects of motoneuron differentiation occur normally in curare-treated embryos. We report here that dendrite growth and arborization is also unaltered on E10 and E16 following curare treatment. A quantitative analysis of afferent synapses on motoneurons shows that the packing density of both axosomatic and axodendritic synapses is also normal on E10 in curare-treated embryos, despite the greater number of motoneurons present. This indicates that the interneurons that provide presynaptic input to motoneurons are able to compensate for the increased number of synaptic sites made available by curare treatment. However, by E16 the packing density of synapses is reduced by about half. Because motoneurons and their dendrites continue to grow between E10 and E16, the further increase in synaptic sites made available in curare-treated embryos apparently exceeds the compensatory capacity of presynaptic interneurons on E16. One can conclude from these results that the increased survival of motoneurons in curare-treated embryos is not owing to an increase in afferent synapses. Motoneurons in these embryos continue to survive in the face of either no change (E10) or a reduction (E16) in the number of axodendritic and axosomatic synapses. Therefore, increased motoneuron survival in this situation is very likely regulated primarily by motoneuron-target interactions.

Animals↗

In vitro motor activity of intestinal segments of streptozotocin diabetic rats.

The spontaneous motor activity of the isolated segments of the gut of streptozotocin-diabetic rats was studied in vitro. In normal preparations, the frequency of intraluminal pressure changes was little influenced by raising the intraluminal pressure. In contrast, in diabetic preparations, the frequency tended to increase with a rise of the intraluminal pressure, and the pressure waves were more irregular than in controls. The motor frequency with a pressure load of 3 cm H2O in the duodenum was 33/min in control rats and 21/min in diabetic rats. The motor frequencies in the other intestinal segments were also higher in normal preparations than in diabetic ones, although these differences were statistically not significant. The amplitude of intraluminal pressure changes increased according to the increase of pressure load, both in normal and diabetic preparations. These amplitudes were higher in normal than in diabetic preparations; however, the differences were statistically significant only in the jejunum. Insulin treatment (5 U/day) for one month, one to two months after streptozotocin injection, lowered the plasma glucose level to nearly normal and increased the body weight up to 80% of the normal but did not re-establish the normal motor frequency in the duodenum. Moreover, treatment of diabetic rats with aldose reductase inhibitor, ONO-2235 (per os, 50 mg/kg b.w./day for one month, one to two months after streptozotocin injection) did not re-establish the normal rhythms in duodenum. The pacemaker activity as well as mechanical properties in the intestinal tract may be disturbed in diabetic preparations.

Animals↗

Long-lasting afterdischarge of alpha-motoneurons after muscle vibration or electrical stimulation of group I afferent fibers in the anemically decerebrated cat.

Experiments were performed on anemically decerebrated cats. Motoneuronal activity in the spinal cord was intracellularly or extracellulary recorded and muscle vibration or electrical stimulation of the peripheral nerve was applied for 1 s every 2 s. Such stimulation elicited monosynaptic responses and afterdischarge, a response which lasts after cessation of the stimuli. Stimulus frequencies from 50 to 200 Hz were effective in eliciting the afterdischarge, while frequencies higher or lower than this range of stimulation were ineffective. The interspike intervals of the motoneuron afterdischarge were almost constant and did not correspond to the stimulus frequency. The mean intervals obtained from the motoneurons examined ranged from 70 to 120 ms. Power spectrum analysis of the membrane potential ripples recorded immediately after stimulation revealed increased activity at 38 and 62 Hz components. Since the spike interval of the afterdischarge in this motoneuron was 80 ms, the interval was almost equal to 5- and 3-fold the cyclic time of the increased frequency components. This indicates that the interval of afterdischarge was determined by the specifically increased frequency components of the membrane potential ripples.

Animals↗

Compliance changes of the gastrointestinal tract in streptozotocin-induced diabetic rats.

In order to elucidate diabetic gastrointestinal disorders, we measured the length, diameter, volume, and intraluminal pressure of the isolated segments of the duodenum, jejunum, ileum, and proximal colon during injection of Tyrode's solution into them, as well as wet weight. Wet weight of the stomach and of each intestinal segment of 3 cm length were similar between normal and diabetic preparations, except the duodenum. Wet weight of the diabetic duodenum was significantly heavier than that of normal. However, capacity of all diabetic gastrointestinal segments was significantly larger than that of normal ones, even after corrected for wet weight (ml/g wet weight). During saline injection, normal intestinal segments were more easily distended longitudinally than circumferentially. Contrarily, diabetic ones were distended more circumferentially than normal, as well as longitudinally. Pressure-volume relationships showed that pressure inside of the diabetic gastrointestinal tract increased much more moderately than that of normal one according to volume increase during saline injection. Similarly, tension inside of diabetic intestinal segments increased much more moderately than that of normal ones. Chord and slope compliance of diabetic gastrointestinal tract was generally larger than those of normal one. Histologically, there are no remarkable differences in cross-sectional area between normal and diabetic intestinal segments after usual fixation without intraluminal fixative injection. However, diabetic segments were much more remarkably dilated than normals were, when fixed after fixative injection. Greater compliance or distensibility of the diabetic gastrointestinal tract seemed to be one basic ground for dilatation, atony, larger appearance, transit delay, and motile disorders of the diabetic gastrointestinal tract.

Abdominal Muscles↗

[Cisplatin combination chemotherapy in advanced lung cancer in the aged].

Eleven aged patients over 65 years of age with advanced lung cancer (mean age = 70.8 +/- 1.4, non-small cell:small cell = 9:2, stage III:IV = 5:6) were treated with combination chemotherapy consisting of cisplatin (50 or 80 mg/m2) and vincaloids (vindesine 3 mg/m2 or etoposide 80 mg/m2). To evaluate this cisplatin combination therapy, the aged group was compared with a young group consisting of eleven patients (mean age = 53.3 +/- 1.7, non-small cell:small cell = 9:2, stage III:IV = 5:6) matched for cell type, stage and dose regimen. The mean dose of cisplatin was 58.2 mg/m2 in the aged and 63.6 mg/m2 in the younger group. A notable reduction in tumor size was observed in 9.1% of the aged and 27.3% of the young, while one-year survival rate was 63.6% in the aged and 72.7% in the young. The common side effects were nausea and vomiting, while diarrhea was seen in 18.2% of the aged. Neutropenia, anemia and thrombocytopenia were found in both groups and the time course of myelosuppression in the aged (18.2 +/- 0.8 days) was significantly shorter than that in the younger patients (22.0 +/- 1.4 days, p less than 0.05). With regard to nephrotoxicity, creatinine clearance rate in the aged decreased remarkably from 56.9 to 38.9 ml/min, while there was no significant change in BUN, serum creatinine and urine NAG between the aged and the young. Disorders of electrolytes such as hypokalemia and hyponatremia were seen in 45.5% of the aged. We conclude that advanced lung cancer in the aged was effectively treated with cisplatin combination therapy with tolerable nephrotoxicity and myelosuppression.

Aged↗

Morphological characteristics of human fat-storing cells fractionated and cultured by newly techniques.

Human non-parenchymal cells, especially fat-storing cells (FSCs), were isolated and primarily cultured using 1-2 g of normal human liver tissue obtained in conjunction with tumor biopsies. The human hepatic FSCs were cultured by a modified Howard and Pesch method. Microscopically the cultured human FSCs showed characteristic fat droplets like those in in vivo FSCs. The FSCs from tumor liver with serious fibrosis contained fewer fat droplets, and the fibrous constituents were especially abundant. The intermediate filaments extending longitudinally were characteristic of the cultured FSCs. In the human FSCs observed by the plasma polymerization replica method, the cells adhered and were stretched relatively thin. Particularly, the ends of the processes adhered and stretched like a folding fan to the bottom of the dish.

Adipose Tissue↗

[Combination therapy of hyperthermia and other methods in liver and bile tract cancers--evaluation of these methods using cancer cell lines in vitro].

In order to evaluate the combination therapy for liver and bile tract cancer, the effects of anticancer drugs and hyperthermia were observed using cultured human cancer cell lines. In the case of gall bladder cancer cell line (NOZ), combination of adriamycin and hyperthermia showed more effective inhibition for cell proliferation than MMC + hyperthermia and 5-FU + hyperthermia. Hepatocellular carcinoma cell line (JHH-4) showed remarkable inhibition of cell growth and secretion of albumin by combination treatment of adriamycin and hyperthermia. Morphologically, JHH-4 cells were enlarged and the nucleus was also enlarged with combination adriamycin and hyperthermia by phase contrast microscopy. Cytoskeleton of JHH-4 cells became irregular and intercellular borderline was unclear by plasma polymerization replica method (PPRM). The effects of BRM (OK-432 and TNF) on HCC cell lines was also investigated. OK-432 directly inhibited proliferation of JHH-4 cells. We observed internalization of OK-432 by JHH-4 cells with TEM and 16-mm movie. TNF showed various effects on human HCC cell lines. Proliferation of two cell lines was inhibited, and one tended to be enhanced after the addition of TNF to the medium. Hyperthermia influenced the effects of TNF to HCC cell lines. We think that this paper is a very significant study for improving the therapy for hepato-biliary cancers.

Antineoplastic Agents↗

[The significance of moment analysis of spirogram].

In order to assess the significance of moment analysis of transit times on spirograms, we investigated the characteristics of this analysis and the distribution of mechanical time constants of the lung derived from this method. Assuming the log-normal distribution of time constants, mu is the mean and sigma is the standard deviation of the natural logarithms of the time constants. The normal value for mu was -0.64 +/- 0.41 and for sigma was 1.54 +/- 0.42 (mean +/- 1 S.D.) in 94 normal males. The smoking habits enhanced the response of sigma on age-effect, but did not enhance the response of V25/HT. Compared with the normal group, excessive sigma was observed in the patients with bronchial asthma, pulmonary emphysema and interstitial lung disease. Mu was especially greater in the patients with the former two diseases. In bronchial asthma, mu decreased and forced expiratory volume in 1 second (FEV 1.0) increased after inhalation of salbutamol, but these did not always correlate. Mu correlated with some clinical conditions. Moment analysis was available on the volume-time profile of quiet breathing with good reproducibility. The first moment (alpha 1) and standardized first moment (alpha 1') showed an increase in bronchial asthma and pulmonary emphysema, and decreased with treatment in bronchial asthma. It was concluded that the moment analysis of transit times on spirogram is useful as an index of time-course aspects of expiration.

Adult↗

Retinol transport in cultured fat-storing cells of rat liver. Quantitative analysis by anchored cell analysis and sorting system.

It is difficult to study the mechanism of specific transport of vitamin A in fat-storing cells (FSC) in vivo. In this study, transport of vitamin A added to the medium was quantitatively analyzed in cultured FSCs by means of the spontaneous fluorescence emitted by vitamin A. By density-gradient centrifugation with 38% Percoll, an FSC-rich fraction was separated from normal rat liver cells. The FSCs were observed to retain cytoplasmic fat droplets even on days 3 and 4 of culture. The FSCs containing fat droplets were selected for this experiment by checking their emission of vitamin A fluorescence. To analyze the vitamin A content of isolated cells, we employed a newly developed anchored cell analysis and sorting system (ACAS 470), which provides fluorescence analysis and sorting of adherent cells under the phase contrast microscope by utilizing a laser with its irradiation range narrowed to 1 micron. Vitamin A fluorescence was detectable by this system even in the cultured FSCs. After 24 hours of culture of FSCs in medium with 1 x 10(-6) M vitamin A added, the strength of fluorescence per FSC was 24.3 +/- 11.2 x 10(5)/cell for control, 61.5 +/- 17.6 x 10(5)/cell for retinyl acetate, 26.0 +/- 12.6 x 10(5)/cell for retinyl palmitate, and 59.0 +/- 15.1 x 10(5)/cell for retinol. Thus, retinol and retinyl acetate were transferred to FSCs in significant amounts without the participation of retinol-binding protein. Furthermore, an extended examination was made of the mechanism of the retinol transport observed in this study. Transport was never inhibited by the presence of vitamin E or azide. Retinol may be transferred by passive transport attributable to the concentration gradient rather than by active transport or through cell membrane damage by retinol itself. There was a tendency for inhibition of the transport of retinol into the cells in fetal calf serum. This inhibition may have occurred because the retinol-binding protein or other serum proteins had decreased the concentration of free retinol.

Animals↗

[Examination of the effects of hyperthermia and anticancer agent on cultured human gall bladder cancer cells by DNA fluorimetric assay].

Tumor cells are generally more sensitive to hyperthermia than normal cells. We applied hyperthermia for bile duct cancer therapy. We also studied the effect of hyperthermia and anticancer agents using cultured cancer cells. In this study, we examined the effect of hyperthermia and adriamycin by using newly established cultured human gall bladder cancer cells. In order to calculate cell numbers, we used the new DNA fluorimetric assay with fluorochrome dye Hoechst 33342, developed by Richards et al. For the purpose of confirming the usefulness of this fluorimetric assay, we compared this assay with methylene blue and 3H-thymidine incorporation methods. The following results were obtained: 1) the combination effect of hyperthermia and adriamycin showed an additive effect, 2) the effect of hyperthermia was only intensified according to the extension of heating time, but the combination treatment resulted in a maximum effect at the early stage of heating, 3) DNA fluorimetric assay with Hoechst 33342 dye was a sensitive, easy and rapid method for calculating cell numbers compared with methylene blue and 3H-thymidine incorporation methods, and more useful for studies of cultured cells.

Cell Count↗

[Spinal cord compression by primary epidural non-Hodgkin lymphoma].

A 63-year-old man was admitted because of sudden paraparesis. Myelography revealed a complete block at the level of T-3. The spinal epidural tumor was histologically diagnosed as malignant lymphoma, diffuse large cell type and immunohistochemically B-cell type. A thorough work-up for metastasis including MRI was negative. He was postoperatively treated with combination chemotherapies (VEPA), followed by irradiation to the region. The residual tumor disappeared on chest-CT and the patient's neurological status rapidly improved. He is still in remission for 18 months and has full performance status. This case is a very rare lymphoma originated from spinal-epidural space.

B-Lymphocytes↗