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Biomedical subjects

M Ueda

Publications and source records attributed to M Ueda.

At least 793 records · Page 44Linked to original sources

Microtubules are required in amoeba chemotaxis for preferential stabilization of appropriate pseudopods.

Amoebae of Physarum polycephalum exhibit chemotactic responses to glucose and to cAMP. The chemotaxing amoebae exhibit alternating locomotive movements: relatively linear locomotion and movements that change the direction of the locomotion. Such locomotive activity is tightly coupled with the changes in the number and the positions of the pseudopods; cells have one pseudopod at the leading edge during their linear locomotion, while they have multiple pseudopods when they are changing the direction of locomotion. Treatment of cells with microtubule-disrupting reagents inhibited the chemotaxis of the cells. To characterize the role of the microtubule system in chemotaxis, we quantitatively analyzed the relationship between the positions of multiple pseudopods of the amoebae and the relative stability of the pseudopods during reorientation. No significant differences were observed in the pseudopod dynamics between the untreated and the treated amoebae. In both cases, one pseudopod at the leading edge continued to expand during linear locomotion. It then split into two to three pseudopods in the reorientation phase, and the positions of the multiple pseudopods were random. Among multiple pseudopods, however, the pseudopods closer to the microneedle tip were selectively stabilized more often than those distant from the tip in the presence of the microtubule system. By contrast, such preferential stabilization of the appropriate pseudopods was completely abolished by microtubule inhibitors. The microtubule-dependent selection of appropriately located pseudopods enables amoebae to turn correctly at the reorientation step.

Animals↗

Development of radioimmunoassay for the novel platelet activating factor receptor antagonist, E6123, and its application to pharmacokinetics in laboratory animals.

A direct radioimmunoassay for the determination of E6123, a novel antagonist of platelet activating factor (PAF) receptor, was developed in order to study the pharmacokinetics at low dose. This procedure used [3H]E6123 as the radioligand and an antiserum obtained from rabbits immunized with the hapten covalently bound to bovine serum albumin. M1B, one of the main metabolites of E6123, exhibited cross-reactivity with antisera. But this metabolite had no effect on measurements of E6123, because the amount of M1B in plasma radioactivity after administration of [14C]E6123 to dogs and monkeys was low. The sensitivity limit of this assay was 25 pg/ml of plasma when 0.1 ml of plasma was used and the assay showed good accuracy and high precision. The validity of the radioimmunoassay was demonstrated by comparative analysis of a number of samples after oral and intravenous administration (1.0 mg/kg) by HPLC-UV method (r = 0.972-0.984, slope = 1.0314-1.2143). The pharmacokinetics of E6123 was studied at a dose of 30 micrograms/kg. After intravenous administration, the plasma concentration-time curves in all species fitted a two-compartment model and the terminal half-lives in guinea pigs, dogs and monkeys (both poor and extensive metabolizers) were 4.77, 1.71, 5.34 and 1.07 h, respectively. After oral administration, the maximum plasma concentrations were obtained within 0.83-3.00 h and the half-life for each animal was almost the same as that after intravenous administration. The mean bioavailabilities of E6123 in guinea pigs, dogs and monkeys (poor and extensive metabolizers) were 106.9, 45.7, 59.1 and 22.8%, respectively.

Animals↗

Pharmacological studies on a new dihydrothienopyridine calcium antagonist, S-312-d. 5th communication: anticonvulsant effects in mice.

S-312, S-312-d, but not S-312-l, L-type calcium channel antagonists, showed anticonvulsant effects on the audiogenic tonic convulsions in DBA/2 mice; and their ED50 values were 18.4 (12.8-27.1) mg/kg, p.o. and 15.0 (10.2-23.7) mg/kg, p.o., respectively, while that of flunarizine was 34.0 (26.0-44.8) mg/kg, p.o. Although moderate anticonvulsant effects of S-312-d in higher doses were observed against the clonic convulsions induced by pentylenetetrazole (85 mg/kg, s.c.) or bemegride (40 mg/kg, s.c.), no effects were observed in convulsions induced by N-methyl-D-aspartate, picrotoxin, or electroshock in Slc:ddY mice. S-312-d may be useful in the therapy of certain types of human epilepsy.

Administration, Oral↗

Evidence that Escherichia coli ubiA product is a functional homolog of yeast COQ2, and the regulation of ubiA gene expression.

The Escherichia coli ubiA gene coding for 4-hydroxy benzoate octaprenyl transferase is thought to be a key enzyme of ubiquinone biosynthesis. Strains with ubiA disrupted were constructed by chromosomal gene replacement with the chloramphenicol resistance gene. The respiration-defective phenotype of the ubiA mutant was complemented by expression of the COQ2 gene encoding the 4-hydroxy benzoate hexaprenyl transferase of Saccharomyces cerevisiae and such strains produced ubiquinone-8. This strongly supports the idea that COQ2 catalyzes the same enzymatic reaction with UbiA and the substrate specificity of COQ2 is broad. Study of the expression of ubiA using an ubiA-lacZ fusion system showed that the ubiA expression was catabolite-repressed by glucose. This repression by glucose was obvious in the arcA mutant. ArcA is the positively acting transcriptional regulator of the oxygen regulated genes. The molecular mass of the protein product of ubiA was 32kD, found using the over-expression of the ubiA gene.

Base Sequence↗

Purification of inulin fructotransferase (DFA I-producing) from Arthrobacter sp. MCI2493 and production of DFA I from inulin by the enzyme.

An extracellular enzyme that produces di-D-fructofuranose-2',1;2,1' dianhydride from inulin was purified from the culture broth of Arthrobacter sp. MCI2493. The molecular weight of the enzyme was 40,000 by gel filtration and SDS polyacrylamide gel electrophoresis. The enzyme had maximum activity at pH 6.0 and 50 degrees C. Using this purified enzyme, 100 g/liter inulin was converted into 60 g/liter of DFA I, nystose, and 1-F-fructofuranosyl-nystose after incubation for 30 h.

Arthrobacter↗

Immunohistochemical and quantitative analysis of cellular and extracellular components of aortic atherosclerosis in WHHL rabbits.

To investigate changes in the major components of atherosclerotic lesions during the progression of this disease, we measured the lesional areas of macrophages, smooth muscle cells, collagen fibers, and extracellular lipid deposits in the aortas of WHHL rabbits. Aortic segments with lesions of various stages were stained for histological and immunohistochemical examination, and the area of each lesional component was measured by a color image analyzer. In the early fatty streaks observed in 3-month-old rabbits, macrophages were predominant in the intima and were also observed in the inner layer of the media. In the transitional lesions (fibro-fatty streaks) found in rabbits at 11 to 15 months of age, an increase in the lesional area of macrophages was prominent compared to other lesional components. Thus, macrophages may play an important role in the progression of aortic atherosclerosis at this stage. In advanced complicated lesions observed in rabbits at 20 to 24 months of age, the area of macrophages and smooth muscle cells did not increase, whereas the area of collagen fibers and extracellular lipid deposits increased. Therefore, both the disruption of foam cells and fibrosis may play an important role in the progression of atherosclerosis at this stage.

Animals↗

[Flare response on bone scintigraphy in metastatic prostate cancer].

Seventy-seven cases of prostate cancer were treated for 5 years at our department and all cases were followed by bone scintigraphy and tumor markers. Of these cases on case of flare response on bone scintigraphy was recognized. A 51-year-old man was hospitalized with chief complaint of lumbago. Serum PAP and gamma-Sm levels were 320 ng/ml and 15 ng/ml, respectively. Prostate biopsy revealed moderately differentiated adenocarcinoma. Bone scintigraphy and CT scan demonstrated multiple bone metastases and lymph nodes involvements. Treatment was started with diethylstilbestrol diphosphate (DES). At one month after the initiation of treatment tumor markers fell down to the normal level and lumbago was diminished, but only serum alkaline phosphatase was elevated and bone scintigraphy showed apparent progression of individual lesions (flare response). The treatment was not altered. At the times after 2, 8, 12 and 36 months successful treatment the bone imaging improved with reduced tracer uptake and no new lesions. The flare response is a healing reaction and is followed apparent improvement. In general, serial bone scintigrams accurately depict the activity of bone metastases in the patients of prostate cancer, but between 1 and 3 months after starting treatment the paradoxical "flare phenomenon" should be taken care.

Adenocarcinoma↗

Isolation of naturally processed peptides from a Toxoplasma gondii-infected human B lymphoma cell line that are recognized by cytotoxic T lymphocytes.

Naturally processed peptides derived from Toxoplasma gondii (T. gondii) were acid extracted from T. gondii-infected cells and detected by cytotoxic T lymphocytes (CTL) derived from peripheral blood lymphocytes of a patient with chronic toxoplasmosis. The CTL lines were obtained by weekly in vitro stimulation with a T. gondii-infected human B cell lymphoma line, ARH, which shares HLA-A2 and -Cw4 determinants with the patient. The lytic activity of these CTL lines against T. gondii-infected ARH and ARH pulsed with fraction 29 of a reversed-phase high-performance liquid chromatography (HPLC) extract from T. gondii-infected ARH was inhibited by an anti-HLA-A, B, C monoclonal antibody (mAb) and an anti-HLA-A2 mAb. Anti-HLA-DR mAb failed to block the lytic activity. Thus, the presentation of peptides by T. gondii-infected cells for CTL is mediated by HLA-A2 molecules. Interestingly, antigen presentation of ARH pulsed with naturally processed HPLC fraction 29 peptides was not inhibited by treatment with brefeldin A. The amino acid sequence of the HLA-A2-bound peptide in fraction 29 was in part consistent with the predictive algorithm of HLA-A2-binding peptide motifs.

Amino Acid Sequence↗

Clinical evaluation of the Endosearch sampler in endometrial cytology-a preliminary report.

The clinical value of the Endosearch endometrial cell sampling device was assessed in 94 cases managed in our outpatient clinic. The Endosearch could be easily inserted into the uterine cavity in 91 (96.8%) of 94 cases without analgesia. Pain and prolonged bleeding accompanied by the insertion of the Endosearch occurred in eight (8.8%) and six (6.6%) of 91 cases, respectively. Satisfactory material for cytologic diagnosis of the endometrial state was obtained in 90(98.9%) of 91 cases, which were comparable to those with the Endocyte. On the other hand, the endometrial tissue samples could be simultaneously obtained with the Endosearch in 78 (85.7%) of 91 cases, and histologic diagnosis was possible in 69(88.5%) of 78 cases. Among them, three of five cases with endometrial hyperplasia and four of five cases with endometrial carcinoma could be correctly diagnosed by the Endosearch histologically. These results suggest that outpatient investigation of the endometrial state by the Endosearch sampler is quite useful not only in cytologic diagnosis but also in histologic evaluation of endometrial lesions.

Aged↗

PCNA expression and nucleolar organizer regions in malignant melanoma and nevus cell nevus.

The histogenesis of malignant melanoma with particular reference to the role of melanocytic nevus is still controversial in oncological pathology. In order to differentiate between the proliferative activity of malignant melanomas and benign melanocytic tumors, immunohistochemical analysis was carried out. We used monoclonal antibody PC10 to identify the Proliferating Cell Nuclear Antigen (PCNA), a highly conserved 36 KD acidic nuclear protein which correlates with cell proliferation, and the AgNOR method to stain the Nucleolar Organizer Regions (NORs) whose number and configurations may reflect protein synthesis activity. 47 cases of primary melanoma, 63 metastatic melanoma lesions, and 23 cases of nevi are included in this study. Spitz nevi showed a higher index of cell proliferation compared with other common benign nevi but a much lower index compared with malignant melanoma cells. The metastatic lesions of malignant melanomas had a higher index of proliferation and activity of cells than the primary lesions. Skin-metastatic lesions had higher indexes of cell proliferation and NOR activity than lymph node-metastatic lesions. These results support the idea that metastatic melanoma cells are derived from a more advanced stage of tumor progression. These data suggest that the combination of PCNA and AgNOR is an useful marker for differentiating benign melanocytic tumors from malignant melanomas.

Humans↗

Intrafamilial outbreak of astrovirus gastroenteritis in São Paulo, Brazil.

This report describes an intrafamilial outbreak of astrovirus-associated gastroenteritis that occurred among 5 children in a family living in the State of São Paul, Brazil. Astrovirus was detected by direct electron microscopy (DEM) and confirmed by solid phase immune electron microscopy (SPIEM) using astrovirus reference serum (T2-6-89). The rise in titre of astrovirus antibody in patients' sera was observed by IEM using acute and convalescent-phase sera. Combined use of the EM techniques to detect virus in stool and antibody rise in sera led us to conclude that this intrafamilial outbreak might have been caused by astrovirus infection. In Brazil, astrovirus may be a more common agent of gastroenteritis than in currently appreciated.

Antibodies, Viral↗

Bacteriological evaluation of a new air turbine handpiece for preventing cross-contamination in dental procedures.

An autoclavable air turbine handpiece, Air Flushing Clean System (AFCS) (Osada Electric Co., Ltd., Tokyo, Japan) was developed for use in dentistry with the objective of reducing cross-contamination. Its potential for bacterial contamination was investigated in vitro using two bacterial strains (Streptococcus mutants ATCC 25175 and Staphylococcus aureus FDA 209 P). In theory, this device should prevent cross-contamination of the internal water and air lines of the handpiece, by maintaining an internal positive pressure even after the turbine is stopped. In the present study, this AFCS device was found to reduce the bacterial contamination within the air turbine handpiece more effectively than the conventional handpiece used according to accepted protocol. The reduction of such contamination by the AFCS is in keeping with the recent objective of the American Dental Association to reduce cross-contamination during dental procedures.

Bacteriological Techniques↗

The localization of basic fibroblast growth factor (FGF-2) in rat submandibular glands.

The immunohistochemical localization of basic fibroblast growth factor (FGF-2) in the submandibular glands of the rat was investigated by use of an antiserum to FGF-2. Nerve fiber bundles with FGF-2-immunoreactivity were found in association with interlobular ducts and blood vessels; they dissociated into single immunoreactive nerve fibers perhaps to terminate in proximity to acinar cells, or to form a reticular fiber network within the tunica adventitia of blood vessels. The FGF-2-immunoreactive neurons were located in the submandibular ganglia, but not in the superior cervical ganglia; hence, at least some of these immunoreactive nerve fibers probably come from the submandibular ganglia and are of parasympathetic origin. Most of the epithelial cells of the intercalated and collecting ducts showed notable FGF-2 immunoreactivity. The characteristic distribution of FGF-2 immunoreactivity in both the neuronal and epithelial tissues of the salivary glands suggests a role of this growth factor in complex physiological processes within the salivary glands.

Animals↗

[Abnormalities and clinical characteristics of growth factors and receptor systems in breast cancer].

Recent progress in molecular biology has revealed that the proliferation of breast cancer is controlled by various growth factors and their receptors. In particular, the amplifications and products of oncogenes which code growth factors and receptors can be different indicators of clinical malignancy from the conventional prognostic factors. We investigated the relation of c-erbB-2 and int-2 gene amplification, expression, ER and EGFR with clinical characteristics. In retrospective study, the cumulative 10-year survival rate of patients with c-erbB-2 and int-2 gene amplification was significantly lower than in patients without amplification. In 72 cases with tumor diameter less than 3.0 cm, which can be an indication of breast preservation surgery, the survival rates of patients in these gene amplified groups were significantly lower than in the non-amplified groups. As for prospective study, the results were the same as those from retrospective study. These data show that the cases with these gene amplification have a high biological malignancy and high risk of recurrence to distant organs, despite the small tumor diameter.

Breast Neoplasms↗

[Anesthetic management for urological surgery of patients with chronic spinal cord injury].

We evaluated retrospectively the anesthetic management and perioperative complications of 47 patients with chronic spinal cord injury for genitourinary procedures. Of the 69 cases, 38 were performed under general anesthesia, 21 cases under spinal anesthesia, one case under epidural anesthesia, four cases under axillary block, and five cases under either sedation or standby. In preoperative laboratory findings, anemia was observed in 41%, hypoproteinemia in 38%, renal insufficiency in 23%, and restrictive pulmonary dysfunction in 69%. Hypotension and bradycardia were seen in 25% and 16% of patients, respectively. Autonomic hyperreflexia was seen in four cases with cervical cord injury: two under spinal anesthesia and two under either sedation or standby. Patients with chronic spinal cord injury have disorders of various organ systems. Careful preoperative evaluation and anesthetic management are required for prevention and prompt treatment of perioperative complications related to the disorders. Autonomic hyperreflexia can be successfully prevented by either general anesthesia or spinal anesthesia. The increased anesthesia safety can be obtained by the successful management of the cardiovascular instability during surgery.

Adolescent↗

Effect of acute lung injury and coexisting disorders on plasma concentrations of atrial natriuretic peptide.

OBJECTIVE: To clarify how plasma atrial natriuretic peptide concentrations vary with the severity of acute lung injury. The influence of coexisting diseases which trigger acute lung injury was also examined. DESIGN: Prospective study. SETTING: Intensive care unit of a university hospital. PATIENTS: Fifty patients who had standard risk factors for acute lung injury including sepsis syndrome, major surgery, prolonged hypotension, aspiration of gastric contents, and burns. Twenty-five of these patients had acute lung injury (group 3) caused by these disorders; the remaining 25 patients had risk factors only (group 2). Ten age-matched normal volunteers were selected as controls (group 1). INTERVENTION: None. MEASUREMENTS AND MAIN RESULTS: Plasma atrial natriuretic peptide concentration was measured in these patients and compared with the severity of acute lung injury. In group 3, a significant increase in the mean plasma atrial natriuretic peptide concentration was observed (188 +/- 78 pg/mL, p < .01) compared with group 2 (54 +/- 28 pg/mL) and the age-matched control group (30 +/- 8 pg/mL). This increase was related to the onset of acute lung injury and returned to control concentrations after recovery. Plasma atrial natriuretic peptide concentrations in group 3 correlated highly with a lung injury score representing the severity of acute lung injury (r2 = .45, p < .01), but did not correlate with other cardiopulmonary variables. CONCLUSION: The results suggest that severity of lung injury, but not other predisposing disorders, may be the key factor leading to the increase in plasma atrial natriuretic peptide concentrations observed in these patients.

Analysis of Variance↗

Classification and surgical treatment of hepatocellular carcinoma (HCC) with bile duct thrombi.

Nine (1.66%) out of 542 cases of HCC treated surgically in our hospital between 1985 and 1992, had macroscopic bile duct thrombi. Three cases presented preoperatively with obstructive jaundice. Two of these received thrombectomy in the hilar bile duct and died of hepatic insufficiency on postoperative days 10 and 66, the other case underwent extended left lobectomy, but also died of renal failure and sepsis 3 months after the operation. In addition, we also treated 6 cases diagnosed at earlier stages than those presenting with obstructive jaundice with both hepatectomy and thrombectomy. In these patients the outcome was as follows: 2 died of recurrent HCC 3 months and 16 months, respectively, after operation, 1 died of apoplexy with no recurrence after 19 months, 1 had a recurrence 5 months after the operation, but is still alive after 7 months, and 2 are still alive 24 months and 60 months after surgery with no recurrence. The outcome is still poor in our series with obstructive jaundice. But in this report, we propose radical surgical treatment for HCC with bile duct thrombi in accordance with our classification, especially for those cases without obstructive jaundice.

Adult↗

[A successful case of PA debanding after percutaneous transluminal angioplasty for restenosis of coarctation complex].

A 11-year-old boy, who had undergone the initial surgical repair for Coarctation complex at 6 months of age was readmitted to our hospital for radical correction. The initial repair at 6 months of age was patch angioplasty of coarctation, PDA division and PA banding. When he was 5 years old, a cardiac catheterization revealed spontaneous closure of VSD, restenosis of coarctation and RV outflow tract stenosis due to PA banding. This time he was treated with 2 steps of procedures. The first step was percutaneous transluminal angioplasty for restenosis of coarctation. Then RV outflow tract repair was performed at the second step. Postoperative course was uneventful.

Angioplasty, Balloon↗