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Biomedical subjects

K Fujiwara

Publications and source records attributed to K Fujiwara.

At least 217 records · Page 12Linked to original sources

Enterocolitis associated with dual infection by Clostridium piliforme and feline panleukopenia virus in three kittens.

Dual infection by Clostridium piliforme and feline panleukopenia virus (FPLV) was found in three kittens. In all cases, we found focal necrosis and desquamation of epithelial cells with occasional neutrophil infiltration in the large intestine. Large filamentous bacilli and spores were observed in the epithelium by using the Warthin-Starry method. Electron microscopy revealed the vegetative forms with characteristic peritrichous flagella and spore forms. Immunohistochemically, these bacilli showed a positive reaction with mouse antisera against the RT and MSK C. piliforme strains. Polymerase chain reaction (PCR) using cecum specimens demonstrated the 196-bp band specific to C. piliforme 16S rRNA. All three kittens were also diagnosed as FPLV-infected on the basis of the characteristic mucosal lesions, including intranuclear inclusions and PCR study for the FPLV genomic DNA. The PCR techniques are useful for confirming the C. piliforme and FPLV infection in spontaneous cases.

Animals↗

Perceptibility of body position in anteroposterior direction while standing with eyes closed.

The perception of body position in the anteroposterior direction was investigated by evaluating the reproducibility of the position from a quiet standing posture to forward or backward leaning posture with eyes closed. The subjects were 10 healthy male undergraduates, aged 20 to 28 years. The standing position was represented by the pressure center of the foot, which was shown by the relative distance (%) from the heel to the length of the foot. The reference positions of the pressure center of the foot were set at 10% increments from 20 to 80% of the length of the foot. The subjects attempted to reproduce each reference position 10 times, and the absolute and constant errors of the reproduced position were analyzed. The absolute errors at reference positions of 30 to 60% were distinctly larger than those at the other reference positions. This indicated that the perception of standing positions from 30 to 60% was less accurate. The constant errors at the reference positions of 40 to 60% were significantly positive, which meant that the reproduced position was located farther forward than the reference position.

Adult↗

Starting position of movement and perception of angle of trunk flexion while standing with eyes closed.

The present study attempted to investigate the effect of position on the perception of angle of trunk flexion while standing. For this purpose, the range effect was factored out by setting the constant target angle at 10 degrees, with varied starting positions of trunk flexion. We found that subjects underestimated angle of trunk flexion when the starting position was close to a quiet standing posture, overestimated when close to maximum trunk flexion, and correctly perceived it when at the middle position. Less perceptual distortion was observed at the positions close to maximum trunk flexion in the present study than in our previous one, in which various target angles of trunk flexion were reproduced from a quiet standing posture. The reduced distortion in the present study was believed to have resulted from factoring out the range effect. The flexion angle of the hip joint changed in tandem with that of the trunk, while very little movement was observed in the ankle, knee, and neck joints. Judging from the changing pattern of hip-joint angle, the muscle activity of the erector spinae and biceps femoris increased gradually to 90 degrees trunk flexion. In contrast, the actual increment of muscle activity reached zero or a minimum value at the middle angles as the angle of trunk flexion increased. It was assumed that the abrupt change in kinesthetic information associated with muscle activity exerted a great influence on the perception of trunk flexion.

Hip Joint↗

Expression and characterization of mouse angiotensin II type 1a receptor tagging hemagglutinin epitope in cultured cells.

The octapeptide angiotensin II mediates the physiological actions of the renin-angiotensin system through activation of several angiotensin II receptor (AT) subtypes, in particular AT1 (AT1a and AT1b in the case of rodents). Although we and others have generated mutant mice in which the AT1a gene was disrupted, the function of mouse AT1 remains to be fully elucidated, due to the lack of effective tools involving antibodies against AT1 for detecting biological responses in cellular conditions. To avoid these problems, we constructed the hemagglutinin (HA)-tagged mouse AT1a, and stably introduced this recombinant receptor into human embryonic kidney 293-T cells. Radioligand binding of [(125)I] angiotensin II to AT1a was specific, saturable, and reversible. Scatchard analysis demonstrated that the transfected receptor had a dissociation constant of 1.7 nM with a density of 1.2 x 10(5) sites/cells. Angiotensin II stimulated a rapid increase in cytosolic free calcium, and angiotensin II-induced phosphorylation of extracellular signal-regulated kinases (Erk) was found in a dose-dependent manner. After solubilization, Western blot analysis showed specific interactions between an anti-HA antibody and HA-tagged mouse AT1a. Furthermore, a significant proportion of HA-tagged mouse AT1a was specifically immunoprecipitated with this antibody. In the immunocytochemical and electronmicroscopic studies, treatment of this cell line with angiotensin II resulted in decrease in signals of the surface receptors. Based on these results, the cell line established here provides an excellent tool for studying angiotensin II actions mediated through mouse AT1a, at sub-nanomolar concentrations.

Angiotensin II↗

[Multiple aneurysms of the PICA communicating artery: a case report].

A case with unusual type of aneurysms in the distal posterior inferior cerebellar artery (PICA) is reported here. Though only two cases with a single aneurysm of the PICA communicating artery have been reported previously, the present case is the first one with multiple aneurysms in the PICA communicating artery. A 61-year-old woman with a sudden onset of severe headache, vomiting and unconsciousness was transferred to our hospital. CT scan revealed a hematoma in the fourth, third, and lateral ventricles, and a mild subarachnoid hemorrhage at the posterior fossa. Cerebral angiogram showed the right PICA supplying the hypoplastic left PICA territory through an anastomotic vessel. Two small aneurysms were seen at the tips of hairpin curves of an anastomotic vessel, "the PICA communicating artery". Suboccipital craniotomy was performed, and the ruptured aneurysm was clipped and the unruptured one was wrapped with cotton-sheet. After the operation, her clinical recovery went well and she was discharged on foot.

Aneurysm, Ruptured↗

[A case report of successful mitral valve replacement for congenital mitral stenosis associated with coarctation of the aorta and ventricular septal defect].

A 5-month-old infant with coarctation of the aorta, ventricular septal defect and mitral stenosis known as "Shone's anomaly" is presented. He underwent the repair of coarctation of the aorta by means of the extended aortic arch anastomosis and banding of the pulmonary trunk at 1 month of age and the patch closure of ventricular septal defect and debanding of the pulmonary trunk at 3 months of age in our institution. About 2 months after second surgery, he had been admitted to our institution due to developing tachypnea and he needed the support of mechanical ventilation. The chest X-ray showed pulmonary congestion and the echocardiography revealed only one papillary muscle of mitral valve and pressure gradient about 30 mmHg through mitral valve. Mitral stenosis due to parachute mitral valve was suspected and he was subjected to an emergent surgery. Initially we performed mitral valve repair for parachute mitral valve but echocardiography during the surgery revealed moderate grade of mitral regurgitation and a hemodynamics was not satisfactory. Eventually mitral valve replacement was successfully done with Carbo-Medics mechanical valve (19 mm in diameter) in the position of left atrial wall because his mitral annulus was so small as 10 mm in diameter. The postoperative course was uneventful and the patient has been doing well.

Aortic Coarctation↗

Involvement of growth-related protein in lipopolysaccharide-induced rabbit arthritis: cooperation between growth-related protein and IL-8, and interrelated regulation among TNFalpha, IL-1, IL-1 receptor antagonist, IL-8, and growth-related protein.

We investigated the functional role of a CXC chemokine, growth-related protein (GRO), in the recruitment of neutrophils in lipopolysaccharide (LPS)-induced rabbit arthritis. The amounts of GRO in the synovial fluids (SF) reached the first peak (major) at 2 hours and the second peak (minor) at 9 hours after injection of LPS into the knee joints. Administration of anti-GRO mouse monoclonal antibody inhibited 54% of the peak leukocyte accumulation at 9 hours (neutrophils greater than 95%), which was similar to the inhibition by anti-IL-8 IgG (48%). Co-administration of these inhibitors increased the inhibition up to 70% at 9 hours and also inhibited 65% of the initial phase of leukocyte infiltration at 2 hours (neutrophils greater than 99%), which was not affected by a single administration of each inhibitor. The amounts of GRO in SF at 2 hours were not altered by either anti-TNFalpha mAb or anti-IL-8 IgG, but reduced by rabbit recombinant IL-1 receptor antagonist (rrlL-1Ra) by 39%. The inhibition by rrlL-1 Ra was augmented further to 59% with coadministered anti-TNFalpha mAb. In contrast, the amounts of GRO at 9 hours were reduced by rrlL-1Ra by 67%. There was no additional reduction in the amounts of GRO at 9 hours by either combination of rrlL-1Ra with anti-TNFalpha mAb or anti-IL-8 IgG. Administration of anti-GRO mAb did not alter TNFalpha or IL-8 contents in SF at their peak (2 hours), but reduced the amounts of IL-1beta at 6 hours and IL-1Ra at 9 hours by 42% and 49%, respectively. These results provide evidence for the following: (a) GRO as well as IL-8 are important mediators involved in the recruitment of neutrophils both in the early and the late phase of LPS-induced arthritis, (b) IL-1 produced in the early phase stimulates GRO production, (c) GRO plays a role in the later induction of IL-1beta and IL-1Ra, and (d) induction of GRO is not regulated by IL-8.

Animals↗

[Optimal timing of granisetron administration for the prevention of carboplatin-induced acute emesis].

This prospective pilot study was undertaken to determine whether delayed administration of one of the serotonin-receptor antagonists (granisetron) reduced carboplatin--induced emesis. The subjects were patients who were scheduled to undergo at least three courses of carboplatin based chemotherapy, in whom acute emesis occurred at the first course. Granisetron was administered therapeutically for the first course. Conventional prophylactic granisetron was given 30 min before carboplatin administration for the second course. For the third course, granisetron was administered 5 hr after completion of carboplatin administration. The degrees of appetite loss, nausea and frequency of vomiting on the Day 1 were compared between the three courses. Thirteen patients were evaluated. Acute emesis occurred 8 +/- hr after completion of carboplatin administration in the first courses of these patients. The degree of nausea and frequency of vomiting were reduced by administrating the granisetron five hours after carboplatin when compared with therapeutic granisetron administration or administration 30 min before carboplatin. The degree of appetite loss was significantly lowered by administering the granisetron 5 hr after carboplatin administration. Administration of granisetron 5 hr after carboplatin may thus lessen carboplatin-induced emesis more than conventional granisetron administration. A large-scale comparative study is warranted.

Acute Disease↗

Cervical lesions related to the systemic progression in rheumatoid arthritis.

STUDY DESIGN: Cross-sectional study of cervical involvement in rheumatoid arthritis. OBJECTIVES: To clarify the correlation between the deterioration of cervical lesions and the systemic progression of rheumatoid arthritis. SUMMARY OF BACKGROUND DATA: The natural course of cervical lesions varies. To date, no systemic parameter has been clarified to predict the progression. METHODS: One hundred seventy-three patients with rheumatoid arthritis participated in this study. The authors studied the progression of cervical lesions and investigated the relation between the types of cervical subluxation at the end of study and the following four variables: the serum level of C-reactive protein, the number of joints with erosion, carpal height ratio, and disease subset (least erosive subset, more erosive subset, and mutilating disease subset). RESULTS: Of the 173 patients, 55 already had cervical subluxation before entering the study. During the follow-up period, 44 patients deteriorated radiographically, and 77 (45%) had cervical involvement, including involvement of upper cervical lesions in 65 patients, upper lesions combined with subaxial subluxation in 10, and subaxial subluxation alone in 2. The upper cervical subluxation progressed in the order of anterior atlantoaxial subluxation, atlantoaxial subluxation combined with vertical subluxation, and vertical subluxation alone. Deterioration of upper cervical lesion and occurrence of subaxial subluxation were closely correlated with an elevation of serum C-reactive protein level, an increase in the number of joints with erosion, and a decrease in the carpal height ratio. The incidence of cervical involvement and the extent of deterioration were different among the disease subsets. CONCLUSIONS: The serum level of C-reactive protein, the number of joints with erosion, and the carpal height ratio correlated closely with the extent of the cervical subluxation. The average C-reactive protein values during the follow-up period correlated with progression of the cervical lesions. The classification of rheumatoid disease subset was useful for predicting the terminal feature of the cervical lesions.

Adolescent↗

Activated rat stellate cells express c-met and respond to hepatocyte growth factor to enhance transforming growth factor beta1 expression and DNA synthesis.

Hepatocyte growth factor (HGF) decreases transforming growth factor beta1 (TGFbeta1) levels in the liver and attenuates hepatic fibrosis caused by dimethylnitrosamine in rats. In the liver, HGF is presumed to act predominantly on parenchymal cells, and TGFbeta1 is produced mainly by mesenchymal cells. In hepatic fibrosis, stellate cells play a central role with undergoing activation, which also occurs when the cells are cultured on plastic. Thus, we wondered if HGF could act directly on stellate cells. c-Met was detected in rat stellate cells activated by culture for 10 days, but not in the cells cultured for 3 days. Specific binding of HGF to the activated cells was determined, and Scatchard analysis indicated an apparent Kd of 1.5 nM. c-Met mRNA was detected in freshly isolated stellate cells from rats treated with carbon tetrachloride for 8 weeks, but not in those cells from normal rats. These results indicate that stellate cells express c-met when activated in vitro and in vivo. HGF enhanced TGFbeta1 production and DNA synthesis in the activated cells.

Animals↗

Monoclonal antibody monospecific to glycine for brain immunocytochemistry.

We have developed mouse monoclonal antibodies (AGLY-1-8, all IgG1 subisotype mAbs) against glycine (Gly) conjugated to bovine serum albumin using glutaraldehyde (GA)-NaBH4. Among these, AGLY-4 mAb was found to be the most useful for Gly immunocytochemistry (ICC) in functions of specificity and sensitivity without non-specific immunobinding. AGLY-4 was demonstrated to be monospecific to Gly by an enzyme-linked immunosorbent assay (ELISA) binding test, and not reactive to any of the other amino acids and peptides tested. Using this antibody, indirect immunoperoxidase staining was observed in different regions of the rat brain fixed with GA in combination with borohydride reduction. In contrast, immunoreactivity was quite low in tissues fixed only with GA. Absorption controls indicated that the immunostaining could be completely inhibited by 5 microg/ml of Gly-human serum albumin (HSA) conjugate prepared using GA and NaBH4, which was consistent with the results of an ELISA inhibition test. No cross-reaction occurred with other GA-conjugated amino acids. Dense ICC staining was observed in the rat neurons related to the auditory and vestibular centers, and modest immunostaining was seen in all the structures of the cerebellar cortex except for the Golgi cells which were strongly stained. These results were in complete agreement with the previous methods using polyclonal anti-Gly serum. Also, a new finding was that staining was noticed in certain cells widely distributed in the different brain regions. These results strongly suggest that the monoclonal antibody has a potential for elucidating the precise distribution of Gly-containing cells.

Animals↗

Effects of lysophosphatidic acid on proliferation of stellate cells and hepatocytes in culture.

Lysophosphatidic acid (LPA) is a growth factor-like mediator for fibroblasts or smooth muscle cells produced and released by activated platelets. Platelet activation occurs with hepatic necrosis and subsequent liver regeneration and fibrosis. In the fibrosis, hepatic stellate cells proliferate with phenotypic transformation to myofibroblasts. Thus, effects of LPA on proliferation of hepatocytes and stellate cells were investigated. In cultured rat stellate cells, LPA increased DNA synthesis with enhanced MAP kinase activity. Pertussis toxin (PTX) attenuated this mitogenic action. In contrast, LPA decreased DNA synthesis by cultured rat hepatocytes induced by hepatocyte growth factor (HGF) or epidermal growth factor (EGF) without affecting protein synthesis. Enhanced MAP kinase activity by HGF or EGF was not changed by LPA. This anti-mitogenic action was attenuated by PTX. TGFbeta level in the medium was less than the level effective for inhibiting the DNA synthesis in the presence of LPA. Our results suggest that LPA might affect proliferation of hepatocytes and stellate cells in liver diseases complicating platelet activation.

Animals↗

Involvement of endotoxins or tumor necrosis factor-alpha in macrophage-mediated oxidation of low density lipoprotein.

Oxidation of low-density lipoprotein (LDL) can be mediated in vitro by cultured cells, including macrophages. This cellular oxidation is dependent on the production of free thiols by the cells in the presence of transition metal ions. We now report that the production of thiols by macrophages is greatly enhanced when cells are cultured with lipopolysaccharide (LPS) or tumor necrosis factor-alpha (TNF-alpha). Oxidation of LDL by macrophages is markedly augmented by pre-treatment of the cells with LPS or TNF-alpha. The results suggest that activation by endotoxins or TNF-alpha is a necessary step for macrophages to mediate oxidation of LDL.

Adult↗

Functional transitions in myosin: formation of a critical salt-bridge and transmission of effect to the sensitive tryptophan.

For analyzing the mechanism of energy transduction in the "motor" protein, myosin, it is opportune both to model the structural change in the hydrolytic transition, ATP (myosin-bound) + H2O --> ADP.Pi (myosin-bound) and to check the plausibility of the model by appropriate site-directed mutations in the functional system. Here, we made a series of mutations to investigate the role of the salt-bridge between Glu-470 and Arg-247 (of chicken smooth muscle myosin) that has been inferred from crystallography to be a central feature of the transition [Fisher, A. J., Smith, C. A., Thoden, J. B. , Smith, R., Sutoh, K., Holden, H. M., & Rayment, I. (1995) Biochemistry 34, 8960-8972]. Our results suggest that whether in the normal, or in the inverted, direction an intact salt-bridge is necessary for ATP hydrolysis, but when the salt-bridge is in the inverted direction it does not support actin activation. Normally, fluorescence changes result from adding nucleotides to myosin; these signals are reported by Trp-512 (of chicken smooth muscle myosin). Our results also suggest that structural impairments in the 470-247 region interfere with the transmission of these signals to the responsive Trp.

Animals↗

Contribution of CD14 to endotoxin-induced liver injury may depend on types of macrophage activation in rats.

Activated Kupffer cells and hepatic macrophages can produce massive liver necrosis through microcirculatory disturbance due to sinusoidal fibrin deposition. This mechanism is involved in the development of liver injury after endotoxin administration in rats pretreated with heat-killed Propionibacterium acnes (P.acnes) or undergoing 70% liver resection. The significance of CD14, a receptor for lipopolysaccharide and its binding protein, was evaluated in both models in relation to the activation mechanisms of Kupffer cells and hepatic macrophages. Northern blot analysis revealed that CD14 mRNA expression was increased in the liver of rats following P.acnes administration. In these rats, hepatic macrophages immediately after isolation showed marked increased of CD14 mRNA expression compared to Kupffer cells from normal rats. In contrast, CD14 mRNA expression was minimal in partially resected liver. Interleukin (IL)-18 and IL-2 mRNA expression in the liver and interferon (IFN)-gamma mRNA expression in the spleen were significantly increased in P.acnes-treated rats compared to normal rats, while these increases were absent in partially hepatectomized rats. Thus, CD14 expressed on hepatic macrophages after activation through a cytokine network of IL-18, IFN-gamma, and IL-2 may contribute to endotoxin-induced liver injury in P.acnes-treated rats. In contrast, in partially hepatectomized rats, this network may not operate during Kupffer cell activation, and the liver injury might develop through endotoxin receptors other than CD14 on the cells.

Animals↗