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Biomedical subjects

Takeshi Watanabe

Publications and source records attributed to Takeshi Watanabe.

At least 109 records · Page 6Linked to original sources

Characteristics of ureteral bolus in diabetic rats.

Changes in the dynamics of the ureteral bolus in diabetics was investigated by using videomicroscopic imaging. Diabetes mellitus (DM) was induced in 8-week-old male rats by the administration of streptozotocin. The pressure of the renal pelvis (via nephrostomy) and visualization of the dynamics of the ureteral bolus were recorded in three groups (DM, sucrose-fed, and control) 8 weeks after administration. Peristaltic velocity, frequency and bolus length were analyzed on the basis of image processing using indigo carmine dissolved in a saline solution. This was perfused via the nephrostomy. The dilated ureters were macroscopically observed in all diabetic rats. There were significant decreases in the velocity of the bolus in diabetics (4.3+/-0.8 mm/s) compared to sucrose-diuretic (11.0+/-3.2 mm/s) and control (8.3+/-1.4 mm/s) rats. In addition, the length of the bolus in diabetics (9.3+/-1.4 mm) was about twofold longer than those of control rats (4.2+/-0.6 mm). There were also significant decreases in the frequency of renal pelvis contraction in diabetics (19.5+/-1.9 min(-1)) compared to sucrose-diuretic (25.3+/-1.0 min(-1)) and control (29.0+/-2.3 min(-1)) rats. These results indicate that the decrease in the velocity of the bolus and the frequency of renal pelvis contraction result in the disability of urine transport in the upper urinary tract in DM although the volume per bolus increases. Besides hyperglycemia, these changes may predispose diabetics to upper urinary tract infections such as pyelonephritis.

Animals↗

Scaling hypothesis leading to generalized extended self-similarity in turbulence.

A scaling hypothesis leading to generalized extended self-similarity (GESS) for velocity structure functions, valid for intermediate scales in isotropic, homogeneous turbulence, is proposed. By introducing an effective scale ŕ, monotonically depending on the physical scale r, with the use of the large deviation theory, the asymptotic forms of the probability densities for the velocity differences u(r) and for the coarse-grained energy-dissipation rate fluctuations epsilon(r), compatible with this GESS, are proposed. The probability density for epsilon(r) is shown to have the form P(r)(epsilon) approximately equal to epsilon(-1)(ŕ/L)(S(ŕ)[z(ŕ)](epsilon))) with z(ŕ)(epsilon)=ln(epsilon/epsilon(L))/ln(L/ŕ), where L and epsilon(L) are the stirring scale and the coarse-grained energy-dissipation rate over the scale L. The concave function S(ŕ)(z), the spectrum, plays the central role of the present approach. Comparing the results with numerical and experimental data, we explicitly obtain the fluctuation spectra S(ŕ)(z).

Journal Article↗

The role of histamine H(1) receptors in late-phase reaction of allergic conjunctivitis.

The role of histamine H(1) receptors in the late-phase reaction of allergic conjunctivitis was studied using histamine H(1) receptor-deficient mice. To clarify the eosinophil infiltration, which is a reliable indicator of late-phase reaction, eosinophil peroxidase activity in the conjunctiva was measured. Mice were actively immunized with ovalbumin, and conjunctivitis was induced by topical instillation of ovalbumin. A significantly high eosinophil peroxidase level in the conjunctiva was observed in sensitized wild-type mice, whereas sensitized histamine H(1) receptor-deficient mice showed no significant increase in the conjunctival eosinophil peroxidase level. In addition, the elevation of eosinophil peroxidase level observed in sensitized wild-type mice was significantly antagonized by pretreatment with anti-P-selectin antibody. From these findings, it was concluded that eosinophil infiltration into the conjunctival tissue in late-phase reaction of allergic conjunctivitis is mediated by P-selectin stored in endothelial cells via histamine H(1) receptors.

Animals↗

Effects of long-chain fatty alcohol on peripheral nerve conduction and bladder function in diabetic rats.

Diabetic cystopathy as manifested by an enlarged bladder is mainly caused by peripheral neuropathy. Long-chain fatty alcohol, which has been isolated from the Far-Eastern traditional medicinal plant, Hygrophilia erecta, Hochr., has been found to possess some neurotrophic activities on the central neuron. Cyclohexenonic long-chain fatty alcohol (FA) used in this study were synthesized in order to improve the efficiency of the molecules. The effects of this compound on peripheral nerves, however, have not yet been studied. To get more information, we evaluated the effects of this compound on peripheral nerves in streptozotocin-induced diabetic rats in terms of nerve conduction velocity and bladder function. Three experiments were performed 8 weeks after the administration of streptozotocin to 8-week-old rats: (i) motor sciatic nerve conduction (MNCV), (ii) monitoring micturition behavior in the metabolic cage, and (iii) cystometrogram under urethane anesthesia (CMG). Half of the diabetic rats were treated with FA (8 mg/kg/day, i.p.). The difference in MNCV between control rats (49.0 +/- 2.2 m/s) and untreated diabetic rats (42.4 +/- 0.5 m/s) after 8 weeks reached significance (p = 0.0183). FA-administrated diabetic rats showed an improved MNCV (45.8 +/- 1.2 m/s). We also identified a significant improvement of bladder function in these animals. The diabetic rats had a much higher maximal micturition volume per 24 hours (4.9 +/- 0.4 ml) than control animals (1.5 +/- 0.1 ml). However, the diabetic rats treated with FA had a maximal micturition volume of only 3.7 +/- 0.3 ml. Likewise, the diabetic rats had a CMG bladder capacity of 0.90 +/- 0.14 ml while the diabetic rats treated with FA had a capacity of 0.54 +/- 0.07 ml. These results indicate that cyclohexenonic long-chain fatty alcohol has a beneficial effect on peripheral neuropathy and cystopathy in streptozotocin-induced diabetic rats.

Animals↗

Clast5/Stra13 is a negative regulator of B lymphocyte activation.

CD40 is a member of the tumor necrosis factor receptor family and mediates a variety of functions of B cells, including B cell survival, proliferation, immunoglobulin gene class switching, memory B cell formation, and regulation of Fas-mediated apoptosis. To begin to elucidate the molecular mechanism governing such diverse functions of CD40, we have isolated a gene from mouse splenic B cells, termed Clast5, whose expression is strongly repressed during B cell activation. Clast5 is identical with Stra13, a recently identified member of the basic helix-loop-helix family of transcription factors. Clast5/Stra13 is highly expressed in unstimulated, resting B cells and is rapidly downregulated by a variety of stimuli that activate B cells, including CD40 ligand, anti-IgM antibodies, lipopolysaccharides and interleukin-4. Forced expression of Clast5/Stra13 in B cells delayed the cell cycle progression into S phase and strongly suppressed Fas-mediated apoptosis. Moreover, Clast5/Stra13 inhibited the colony formation in fibroblasts. Our results suggest that Clast5/Stra13 functions as a negative regulator of B cell activation by inhibiting cell cycle progression and cell growth.

3T3 Cells↗

Universality of chaotic rare fluctuations in a locally coupled phase map model.

Chaotic fluctuations of the order parameter in a coupled two-dimensional phase map model are numerically investigated. We discuss the system-size N dependence of the statistical properties of rare fluctuations observed in the transition range between the quasiordered chaotic state and the fully developed one. It is found that the normalized probability distribution function (PDF) has a unique functional form irrespective of N. The asymptotic form of the PDF is discussed in connection with the universal distribution for correlated systems proposed by Bramwell et al. [Nature (London) 396, 552 (1998)]. Moreover, it is observed that the power spectrum P(N)(omega) of rare fluctuations asymptotically takes the power-law form P(N)(omega) equivalent to omega(-(1+alpha)) (alpha=0.6 equivalent to 0.7) irrespective of N. This result suggests that the temporal correlation decays as a stretched exponential.

Journal Article↗

The evaluation of putative virulence factors of Helicobacter pylori for gastroduodenal disease by use of a short-term Mongolian gerbil infection model.

Few virulence determinants of Helicobacter pylori have been tested in vivo. We conducted this study to establish an animal model for their screening. Six-week-old male Mongolian gerbils were inoculated with wild-type H. pylori (TN2) or its isogenic mutant with deletion of cagE (TN2DeltacagE), total cag pathogenicity island (TN2DeltacagPAI), HP0499 (TN2DeltaHP499), or HP0638 (TN2DeltaHP638) (n=5 each). The animals were killed 3 weeks later, and the density of bacteria and the degree of inflammation in the stomach were compared. Infection was established in all animals except those inoculated with TN2DeltaHP638. TN2 and TN2DeltaHP499, but not TN2DeltacagE and TN2Deltacag PAI, induced intense inflammation, although the densities of bacteria were similar. The Mongolian gerbil model was useful for the screening of virulence determinants in vivo, which confirmed the importance of cag PAI while questioning that of HP0499.

Animals↗

Directional degradation of beta-chitin by chitinase A1 revealed by a novel reducing end labelling technique.

A novel procedure for labelling the molecular ends of beta-chitin crystals has been established. By introducing a hydrazide derivative of biotin at the reducing end of a chitin chain, followed by a specific interaction between biotin and streptavidin coupled with a colloidal gold particle, the chain directionality of beta-chitin microcrystals could be directly visualized by transmission electron microscopy. This method allowed to certify the parallelism of the chitin chains in the beta-chitin microcrystals, and also to label the reducing tips of beta-chitin microcrystals degraded by Bacillus circulans chitinase A1. With these substrates, the labelling occurred only at their tapered tip, which indicates that the digestion of these crystals proceeded from their reducing end. The generalization of this new labelling method to other polysaccharide crystals is discussed.

Animals↗

Enhanced antinociception by intrathecally-administered morphine in histamine H1 receptor gene knockout mice.

We previously reported that histamine H(1) receptor gene knockout mice (H1KO) showed lower spontaneous nociceptive threshold to pain stimuli when compared to wild-type mice. The objective of the present study was to examine the antinociceptive effect of intrathecally-administered morphine in H1KO mice. The antinociceptive effects of morphine were examined using assays for thermal (tail-flick, hot-plate, paw-withdrawal), mechanical (tail-pressure) and chemical nociception (formalin and capsaicin tests) using H1KO and wild-type mice. In these nociceptive assays, intrathecally-administered morphine produced significant antinociceptive effects in wild-type mice. The antinociceptive effect produced by intrathecally administered morphine was enhanced in the knockout mice. We also examined the effect of an histamine H(1) receptor antagonist, an active (d-) isomer of chlorpheniramine, on morphine-induced antinociception in ICR mice. The intrathecal co-administration of d-chlorpheniramine enhanced the effect of morphine in all nociceptive assays examined. The pharmacological experiments using d-chlorpheniramine further substantiate the evidence for the histamine H(1) receptor-mediated suppression of morphine-induced antinociception. These results suggest that existing H(1) receptors play an inhibitory role in morphine-induced antinociception at the spinal cord level.

Analgesics↗

The tumor-associated antigen, RCAS1, can be expressed in immune-mediated diseases as well as in carcinomas of biliary tract.

BACKGROUND/AIMS: RCAS1, which is recognized by the specific antibody 22-1-1, was first identified as a tumor-associated antigen in gynecological carcinomas. RCAS1 is the ligand of a putative receptor present on lymphocytes, the expression of which is enhanced by lymphocyte activation. RCAS1 inhibits the growth of receptor-bearing cells and induces apoptotic death. Here we examined RCAS1 expression in biliary diseases. METHODS: RCAS1 expression was immunohistochemically examined on tissue samples. Apoptotic death was analyzed by DNA fragmentation detection method. RCAS1 production by cell lines was investigated by flow cytometry, enzyme-linked immunosorbent assay, and reverse transcription-polymerase chain reaction. RESULTS: All cholangiocarcinoma cell lines examined clearly expressed RCAS1 at both the protein and RNA level. Immunohistochemically, RCAS1 was expressed in a high percentage of biliary adenocarcinomas (85.9% of intrahepatic cholangiocarcinomas, 96.4% of extrahepatic cholangiocarcinomas and gallbladder carcinomas). Apoptotic tumor-infiltrating lymphocytes could be found in these specimens. RCAS1 expression was frequently detected also in biliary epithelial cells in cases of immune-mediated cholangitis (74.2% in primary biliary cirrhosis, 66.6% in graft-versus-host disease), although the staining pattern for RCAS1 was different compared with cancer cells. CONCLUSIONS: RCAS1 is highly expressed not only in cancer cells but also in non-tumor bile duct cells subjected to immune attack.

Adenocarcinoma↗

Immune regulation by histamine.

Histamine is a potent bioamine with multiple activities in various pathological and physiological conditions. In addition to its well-characterised effects in the acute inflammatory and allergic responses, histamine regulates several aspects of antigen-specific immune response development. Histamine affects the maturation of dendritic cells and alters their T cell-polarising capacity. Histamine also regulates antigen-specific T helper 1 and T helper 2 cells, as well as related antibody isotype responses. Apparently, diverse effects of histamine on immune regulation are because of differential expression of four types of histamine receptors and their distinct intracellular signals.

Animals↗

Histamine H1 receptors are involved in mouse nasal allergic responses: a demonstration with H1 receptor-deficient mice.

The role of histamine H1 receptors in nasal allergic symptoms (sneezing and nasal rubbing) were studied using histamine H1 receptor-deficient mice. Intranasal instillation of histamine solution resulted in significant increases in sneezing and nasal rubbing in wild-type mice, whereas no increases were observed in histamine H1 receptor-deficient mice. The histamine H1 receptor agonist 2-pyridylethylamine induced sneezing and nasal rubbing in a dose-dependent-manner in wild-type mice, but no such increase was found in histamine H1 receptor-deficient mice. On the other hand, the histamine H2 receptor agonist dimaprit did not increase sneezing and nasal rubbing in wild-type mice. Histamine H1 receptor antagonists such as chlorpheniramine and epinastine significantly inhibited nasal allergic symptoms caused by histamine, but the histamine H2 receptor antagonists cimetidine and famotidine showed no effect. No additional effects were observed by combined use of chlorpheniramine and cimetidine or famotidine compared with cimetidine or famotidine alone. These results suggested that histamine H1 receptors play an important role in nasal allergy symptoms induced by histamine.

Animals↗

New simple method of transabdominal ultrasound to assess the degree of benign prostatic obstruction: size and horizontal shape of the prostate.

BACKGROUND: Transabdominal ultrasound has been the standard clinical tool for evaluating prostatic size; however, the severity of benign prostatic obstruction does not always correspond to the size of the prostate. The purpose of this study was to consider the utility of two parameters, (i) the prostatic volume (Pvol) and (ii) the ratio of the height : width of the maximum horizontal section of prostate (H : W) obtained by transabdominal ultrasound, in screening the degree of benign prostatic obstruction. The results were compared with the linear passive urethral resistance relation (lin-PURR) obtained by pressure-flow studies. METHODS: Fifty-one men with lower urinary tract symptoms were prospectively evaluated in regard to urodynamic parameters, transabdominal ultrasound volume of the prostate, and calculation of lin-PURR. RESULTS: A significant positive correlation was found between Pvol and the degree of benign prostatic obstruction. When the ultrasonographic parameters of greater than 30 mL Pvol and 0.8 H : W were adopted as provisional cut-off points for detecting patients with obstruction, the selected 10 cases had lin-PURR > 3. In addition, there were six cases classified with lin-PURR >or= 3 despite the fact that their Pvol was less than 30 mL. Their H : W values were > 0.8. CONCLUSIONS: Transabdominal ultrasound is useful as a screening test for evaluating the prostate. In addition to the size of the prostate, the parameter of the horizontal shape (H : W ratio) makes an assessment of the degree of benign prostatic obstruction more reliable.

Aged↗

22-Oxacalcitriol prevents progressive glomerulosclerosis without adversely affecting calcium and phosphorus metabolism in subtotally nephrectomized rats.

BACKGROUND: 22-Oxacalcitriol (OCT), an analogue of vitamin D, has been shown to inhibit cell proliferation in cultured mesangial cells. OCT also prevented albuminuria and glomerular injury in an acute model of anti-Thy1 glomerulonephritis. However, potential side effects, including calcaemic actions and tubular dysfunction, of chronic OCT treatment remain unclear. In the present study, we evaluated the effect of OCT in a chronic model of progressive glomerulosclerosis in subtotally nephrectomized (SNX) rats. METHODS: At one week after subtotal nephrectomy, SNX rats were divided into 3 groups having equivalent serum creatinine levels and body weight. OCT (0.08 or 0.4 micro g/kg body weight) was administered intravenously three times per week for 8 weeks to SNX rats. We evaluated effects of OCT on renal function during treatment and on morphologic parameters in glomeruli at 8 weeks. We additionally measured calcium and phosphate levels in serum and urine, and tubular dysfunction markers, including beta(2)-microgloblin (beta(2)m) and N-acetyl-beta-D-glycosaminidase (NAG) levels in urine. RESULTS: OCT treatment significantly suppressed urinary albumin excretion, prevented increases in serum creatinine and serum urea nitrogen, and inhibited glomerular cell number, glomerulosclerosis ratio and glomerular volume in SNX rats at 8 weeks. At that time, OCT-treated groups did not show hypercalcaemia, hypercalciuria or hyperphosphaturia. Furthermore, OCT treatment did not affect beta(2)m or NAG levels in urine, and did not induce histological changes in tubular or interstitial regions. CONCLUSIONS: These findings suggest that OCT may provide a clinically useful agent for preventing the progression of glomerulosclerosis without adversely affecting calcium and phosphorus metabolism or causing subsequent tubular dysfunction.

Animals↗

Comparative study of the reaction mechanism of family 18 chitinases from plants and microbes.

Hydrolytic mechanisms of family 18 chitinases from rice (Oryza sativa L.) and Bacillus circulans WL-12 were comparatively studied by a combination of HPLC analysis of the reaction products and theoretical calculation of reaction time-courses. All of the enzymes tested produced beta-anomers from chitin hexasaccharide [(GlcNAc)(6)], indicating that they catalyze the hydrolysis through a retaining mechanism. The rice chitinases hydrolyzed predominantly the fourth and fifth glycosidic linkages from the nonreducing end of (GlcNAc)(6), whereas B. circulans chitinase A1 hydrolyzed the second linkage from the nonreducing end. In addition, the Bacillus enzyme efficiently catalyzed transglycosylation, producing significant amounts of chitin oligomers larger than the initial substrate, but the rice chitinases did not. The time-courses of (GlcNAc)(6) degradation obtained by HPLC were analyzed by theoretical calculation, and the subsite structures of the rice chitinases were identified to be (-4)(-3)(-2)(-1)(+1)(+2). From the HPLC profile of the reaction products previously reported [Terwisscha van Scheltinga et al. (1995) Biochemistry 34, 15619-15623], family 18 chitinase from rubber tree (Hevea brasiliensis) was estimated to have the same type of subsite structure. Theoretical analysis of the reaction time-course for the Bacillus enzyme revealed that the enzyme has (-2)(-1) (+1)(+2)(+3)(+4)-type subsite structure, which is identical to that of fungal chitinase from Coccidioides immitis [Fukamizo et al. (2001) Biochemistry 40, 2448-2454]. The Bacillus enzyme also resembled the fungal chitinase in its transglycosylation activity. Minor structural differences between plant and microbial enzymes appear to result in such functional variations, even though all of these chitinases are classified into the identical family of glycosyl hydrolases.

Chitinases↗

Videomanofluorometric study in amyotrophic lateral sclerosis.

OBJECTIVE: To elucidate the time-course changes of swallowing function in amyotrophic lateral sclerosis (ALS) by videomanofluorometry. STUDY DESIGN: Videomanofluorometry was conducted on 21 patients with ALS, who were divided into five groups according to type of disease and according to the length of time following the appearance of bulbar symptoms. METHODS: Videomanofluorometry, which is videofluoroscopic and manometric study conducted simultaneously, was performed on patients in the five groups, and swallowing function in each group was evaluated. Some of the patients were followed up by videomanofluorometry performed several times, and their swallowing function changes over time were investigated. RESULTS: A decrease of swallowing pressure first appeared in the oropharynx, then the hypopharynx became involved. Oropharyngeal swallowing pressure had already decreased to approximately 50 mm Hg within 6 months after the appearance of bulbar symptoms; however, hypopharyngeal swallowing pressure was relatively maintained until 1 year after the onset of bulbar symptoms. Most of the patients with ALS maintained normal upper esophageal sphincter relaxation, but upper esophageal sphincter spasm was seen in some patients with ALS. Aspiration was seen in eight cases, five of which showed upper esophageal sphincter spasm. CONCLUSIONS: Patients with ALS gradually face the danger of aspiration as decreases of oropharyngeal and hypopharyngeal swallowing pressure progress. Upper esophageal sphincter spasm occurs in some patients with ALS, and it is an important cause of aspiration. Both videofluoroscopic and manometric evaluation are necessary to assess these conditions, and they are quite useful for follow-up of swallowing function in patients with ALS.

Adult↗

Subtotal nephrectomy stimulates cyclooxygenase 2 expression and prostacyclin synthesis in the rat remnant kidney.

BACKGROUND: Recently, two isoforms of cyclooxygenase (COX) have been identified, a constitutive form (COX-1) and a mitogen-inducible form (COX-2). Several studies have suggested that COX is activated in renal insufficiency, but little is known about the relationship between progression of renal insufficiency and the COX isoforms. METHODS: Five-sixths-nephrectomized (NX) rats were used. 4, 8, and 12 weeks after nephrectomy, the renal cortical prostaglandin contents and the expression levels of the two isoforms of COX were determined by enzyme immunoassay and Western-blotting, respectively. The localization of COX was examined by immunohistochemistry. RESULTS: Renal cortical prostacyclin (PGI2) and COX-2 were significantly upregulated 8 and 12 weeks after NX, while COX-1 remained at the basal level. There was a high correlation between COX-2 and creatinine clearance (r = -0.845). There was also a high correlation between COX-2 and PGI2 (r = 0.816). Immunohistochemistry revealed the expression of COX-2 to be enhanced in the macula densa in NX rats. CONCLUSIONS: Renal cortical COX-2 and prostacyclin were upregulated corresponding to the progression of renal insufficiency in NX rats. These results suggest enhancement of COX-2 expression in the macula densa, perhaps stimulated by a decrease in renal blood flow which upregulates PGI2 synthesis to protect the kidney from ischemia in renal insufficiency.

Animals↗