Biomedical subjects

Masanobu Maeda

Publications and source records attributed to Masanobu Maeda.

33 records · Page 2Linked to original sources

PubMed2004

Deletion of the kinase domain in death-associated protein kinase attenuates renal tubular cell apoptosis in chronic obstructive uropathy.

Death-associated protein kinase (DAPK) is a Ca2+/calmodulin-dependent serine/threonine kinase that has been implicated as a positive mediator of apoptosis. However, little is known about the involvement of DAPK in the apoptosis associated with several pathological states, except for cancer. Here, DAPK-mutant mice were used in order to examine the role of DAPK in renal cell apoptosis in chronic obstructive uropathy (COU) created by unilateral ureteral ligation. These mice express mutant DAPK with a deletion of 74 amino acids from the catalytic kinase domain. Obstructed kidneys in wild-type and mutant mice were examined for both DAPK protein levels and renal cell apoptosis during the course of COU. Obstructed kidneys in wild-type and mutant mice showed a marked increase in the DAPK and mutant DAPK protein levels, respectively, at day 14 after ureteric ligation. The obstructed kidneys in DAPK-mutant mice displayed a significant attenuation of tubular cell apoptosis, compared with wild-type mice. In contrast, no significant difference in interstitial cell apoptosis was observed between the obstructed kidneys from wild-type and mutant mice. Thus, these results indicate that the part of the kinase domain deleted by the gene targeting is crucial for the execution of tubular cell apoptosis, but is not essential for interstitial cell apoptosis in a COU model in mice.

Animals↗

PubMed2004

Development of vitellogenin assay for endocrine disrupters using medaka (Oryzias latipes).

An enzyme-linked immunosorbent assay (ELISA) was developed for the detection of the egg yolk precursor vitellogenin (VTG) in the liver of medaka (Oryzias latipes) and was employed to establish an in vivo testing system for estrogen and estrogenic compounds using liver homogenates. Results of 3-month-old fish exposed to three reference chemicals (ethynylestradiol, methyltestosterone and flutamide) recommended by Organisation for Economic Co-operation and Development (OECD) for the validation showed the induction and decrease of vitellogenic responses, making the assay using the liver VTG of medaka a possible screening method for not only estrogens but also androgens. In addition, 21-day exposure of male fish to 4-tert-octylphenol and 4-nonylphenol produced concentration-dependent inductions of liver vitellogenin, with the lowest observed effect concentrations of 64.1 microg/L and 22.5 microg/L, respectively, while no significant VTG responses were observed in male and female fish exposed to tributyltin chloride and dibutyl phthalate. This study demonstrates that the VTG assay using liver homogenates from small fish such as medaka can be used as a screening method for environmental estrogens. This is because the sensitivity of the VTG response in medaka may be almost the same as that of other fish using blood samples.

Androgens↗

PubMed2003

Hypotensive effects of neuromedin U microinjected into the cardiovascular-related region of the rat nucleus tractus solitarius.

Neuromedin U (NMU) is a brain-gut peptide with potent contractile effects on the uterus and smooth muscle. Intracerebroventricular injection of NMU reportedly decreased food intake and body-weight gain in the rat. We evaluated the effects of NMU delivered by microinjection into the rat nucleus tractus solitarius (NTS) on cardiovascular responses. At the concentrations used (5, 10 or 50 pmol), the intra-NTS injection of NMU in artificial cerebrospinal fluid produced a significant reduction in both the mean arterial pressure and heart rate. The hypotensive responses were dose dependent. Our findings suggest that NMU may act as a neurotransmitter or neuromodulative substance that causes excitation of neurons in the NTS and that it may play a role in cardiovascular regulation in vivo.

Animals↗

PubMed2003

The current role of preoperative and intraoperative autologous blood donation in pediatric open-heart surgery.

OBJECTIVE: We assessed the current role of preoperative and intraoperative autologous blood donation in pediatric open-heart surgery. METHODS: Group 1 consisted of 51 patients between 5 and 10 years old who underwent preoperative autologous blood donation. Group 2 consisted of 50 age-matched patients without preoperative donation as controls. Intraoperative donation was conducted in both groups prior to cardiopulmonary bypass. We evaluated perioperative blood cell count, blood loss, and the need for homologous blood products. RESULTS: No serious complications occurred in preoperative or intraoperative donation. Total preoperative donation storage was 17.5 +/- 3.4 mL/kg. Intraoperative donation was 21.7 +/- 6.1 mL/kg in Group 1 and 12.8 +/- 4.0 mL/kg in Group 2 (p < 0.001). On admission, serum hemoglobin was lower in Group 1 (12.2 +/- 1.0 g/dL versus 13.6 +/- 1.6 g/dL, p < 0.001) but returned postoperatively to the preoperative value. It hovered at a depressed level in Group 2 (12.2 +/- 1.4 versus 10.2 +/- 1.1 g/dL, p < 0.001). The homologous blood requirement was significantly less in Group 1 than in Group 2 (0% versus 10%, p < 0.05). Postoperative platelet counts showed similar curves, and blood loss was not statistically significantly different between groups. CONCLUSION: Preoperative and intraoperative donations are safe and continue to contribute uniquely to blood conservation, providing important options in comprehensive blood conservation programs in current pediatric open-heart surgery.

Blood Transfusion, Autologous↗

PubMed2003

The antiplatelet aggregation effects of aspirin suppositories.

To confirm that aspirin suppositories are an effective treatment for acute ischemic stroke, we examined the suppressive effects of 200-mg aspirin suppositories on platelet aggregation. Aspirin suppositories suppressed platelet aggregation induced by ADP or collagen, and the suppression continued for 24 h. There was no significant difference in suppression of platelet aggregation between aspirin administered by suppository and orally given aspirin. These results suggest that aspirin suppositories are a useful treatment for acute ischemic stroke.

Adenosine Diphosphate↗

PubMed2003

The Wakayama epileptic rat (WER), a new mutant exhibiting tonic-clonic seizures and absence-like seizures.

A new mutant, the Wakayama epileptic rat (WER), exhibiting both spontaneous absence-like behavior and tonic-clonic convulsions, was identified in a colony of Wistar rats. To determine clear seizure characteristics of this mutant strain, we analyzed the mode of inheritance of the convulsion and observed patterns of electroencephalogram (EEG) during the seizures. F1 progeny were produced between the founder male and normal females of the same colony. Animals were monitored through the inbreeding course to analyze genetic control of epileptic behavior. EEGs were recorded using affected animals in the F3-4 and post F13 generations. After the F2 generation, affected rats spontaneously exhibited both absence-like immobile behavior and tonic-clonic convulsions. The absence-like seizures were characterized by motor arrest and head droop. The tonic-clonic convulsions began with neck and forelimb clonus, wild jumping/running, and opisthotonic posturing, and evolved to tonic, then clonic convulsions. Most convulsion onsets occurred between 25-70 days of age. Mating experiments revealed that 0%(0/18) of the animals in F1, 10%(3/26) in F2, 17%(1/6) in backcross progeny and 86% (100/116) in progeny of crosses between epileptic rats showed tonic-clonic convulsions. Ictal cortical EEGs were characterized by 4-6 (5.1 +/- 0.4, mean +/- SD) Hz spike-and-wave complexes in the absence-like seizures and by low-voltage fast waves in the tonic-clonic convulsions. This new mutant rat spontaneously exhibited both absence-like and tonic-clonic seizures. The tonic-clonic seizure was inherited as an autosomal recessive trait with 86% incidence. Thus, the new mutant rat may become a useful model for studying human inherited epilepsies.

Animals↗

PubMed2003

Effects of 4-nonylphenol on reproduction of Japanese medaka, Oryzias latipes.

We exposed mature Japanese medaka (Oryzias latipes) to 4-nonylphenol (4-NP) to elucidate its reproductive effects and estrogenic responses. For each treatment, eight breeding pairs were exposed for 3 weeks at measured 4-NP concentrations of 24.8, 50.9, 101, and 184 micrograms/L. Egg production of the pairs during the third week of exposure (15-21 d) was significantly decreased in the > or = 101-microgram/L treatment group, and fertility was significantly decreased in the 184-microgram/L group. Induction of testis-ova was observed in male fish at all 4-NP concentrations, whereas abnormality of spermatogenesis was found only in those receiving the 184-microgram/L treatment. The gonadosomatic index of the males in the 184-microgram/L group was also remarkably reduced. Hepatic vitellogenin (VTG) levels were significantly increased in both sexes receiving 4-NP treatments of > or = 50.9 micrograms/L. Overall, the lowest-observed effective concentrations (LOECs) of 4-NP for decreasing reproductive capacity (fecundity and fertility) and eliciting estrogenic responses (induction of testis-ova and VTG) were 101 and < or = 24.8 micrograms/L, respectively. The LOEC for reduced reproduction in medaka was at least four-fold greater than that required to elicit estogenic responses in male fish. Thus, we suggest that the estrogenic responses expressed in male fish are not responsible for the reduction in reproductive capacity.

Animals↗

PubMed2003

Fish full life-cycle testing for the weak estrogen 4-tert-pentylphenol on medaka (Oryzias latipes).

We studied the chronic effects of 4-tert-pentylphenol (4-PP) on reproductive status of medaka (Oryzias latipes) over two generations under continuous exposure, with the goal of verifying the applicability of the fish full life-cycle test (FFLC) for this weak estrogen with this species. The exposure of parental (F0) medaka to 4-PP was begun on embryos within 12 h after fertilization and continued for up to 101 d, with monitoring of embryological development, hatching, posthatch survival, growth, sexual differentiation, reproduction, and hepatic vitellogenin (VTG) levels under flow-through exposure to 4-PP at mean measured concentrations of 51.1, 100, 224, 402, and 931 microg/L. Eggs (F1) spawned from the F0 fish at 99, 100, and 101 d after hatch also were examined for hatchability, survival after hatching, growth, sexual differentiation, and hepatic VTG levels, until 61 d after hatch. In the FFLC with the F0 medaka, the lowest-observed-effect concentration (LOEC) of 4-PP for lethal and sublethal toxicity (as shown by growth inhibition) was 931 microg/L. The LOECs for estrogenic effects (as shown by abnormal sexual differentiation and VTG induction) were 224 and < or = 51.1 microg/L, respectively, and the LOEC for reproductive impairment was 224 microg/L. Therefore, the effective concentrations of 4-PP for abnormal sexual differentiation and reproductive impairment were about four times lower than those for lethal and sublethal toxicity. In the F1 medaka, the LOECs for sublethal toxicity and estrogenic effects were 224 and < or = 51.1 microg/L, respectively. This finding suggests that the continuous exposure to 4-PP over two generations induced these adverse effects at lower concentrations in the F1 generation than those in the F0 generation. Thus, 4-PP has estrogenic effects that reduce the reproductive potential of medaka. The results indicate that the FFLC with medaka is applicable to the evaluation of weak estrogens.

Animals↗

PubMed2003

Effects of 4-nonylphenol and 4-tert-octylphenol on sex differentiation and vitellogenin induction in medaka (Oryzias latipes).

Medaka (Oryzias latipes) were continuously exposed to various concentrations of two alkylphenols, 4-nonylphenol (NP) and 4-tert-octylphenol (OP), from fertilized eggs to 60 d posthatch. The effects on sexual differentiation and hepatic vitellogenin (VTG) induction in medaka were assessed to elucidate the lowest-observed-effect concentrations (LOECs) of NP and OP for these events during early life stages. The LOECs of NP and OP for these events were 11.6 and 11.4 microg/L, respectively. These results suggest that NP and OP may have adverse effects at similar concentrations during early life stage in medaka. Additionally, we investigated whether the abnormal sex differentiation induced by these alkylphenols would be permanent or reversible once the medaka were returned to clean water. The appearance of the secondary sex characteristics reverted from female to male when fish were returned to clean water. However, gonadal histology showed that intersex gonads still existed, even after the fish were transferred to clean water for two months. These results suggest that the induced feminization of secondary sex characteristics in medaka exposed to alkylphenols during the stage of sexual differentiation may not always be permanent, but the gonadal alteration (testis-ova) may continue much longer.

Animals↗

PubMed2002

The expression of Bcl-2 family proteins (Bcl-2, Bcl-x, Bax, Bak and Bim) in human lymphocytes.

Bcl-2 family proteins regulate programmed cell death, and may play an important role in the selection of lymphocytes. We investigated the expression of Bcl-2, Bcl-x, Bax, Bak and Bim in human lymphocytes using flow-cytometry. Bcl-2 was down-regulated in CD4(+)8(+) (DP) thymocytes and CD19(+)38(+) tonsillar lymphocytes (GC B cells). Among DP thymocytes, cells co-expressing CD69 up-regulated Bcl-2, suggesting that the role of Bcl-2 is promoting survival of positively selected DP cells. Unexpectedly, the expression level of Bcl-x was higher in DP cells than in Single Positive (SP) cells and in CD69(+) DP thymocytes it was lower than in CD69(+) DP thymocytes. Expression of Bim was low in DP thymocytes but high in a subset of GC B cells. Bim and Bax were expressed more highly in SP than in DP thymocytes. Among peripheral blood lymphocytes (PBL), CD8(+) T cells expressed an approximately ten-fold higher level of Bcl-x than CD4(+) T cells while both subsets expressed similar levels of Bcl-2. Bak expression was low and Bim expression was absent in PBL. These results suggest that not only Bcl-2 but other members of the Bcl-2 family are involved in T cell development in the thymus and affinity maturation of B cells in the germinal center.

Adult↗

PubMed2002

Effect of 17beta-estradiol on the reproduction of Japanese medaka (Oryzias latipes).

Estrogenic compounds such as 17beta-estradiol (E2) and its analogs are present in the aquatic environment and can adversely affect the reproductive systems of aquatic organisms. Although E2 has been detected at a maximum concentration of 64 ng/l in effluents of sewage treatment works (STWs), few reports address the associated effects on reproduction in fish. Therefore, we exposed adult medaka (Oryzias latipes) to mean measured E2 concentrations of 29.3, 55.7, 116, 227, and 463 ng/l for 21 d and assessed the effects on the egg number and fertility of paired medaka during the exposure period. In addition, we determined the hepatic vitellogenin (Vtg) concentration and histologically assessed the gonads of these fish. The number of egg produced and fertility of the paired medaka exposed to 463 ng/l E2 were significantly less compared with those of the control fish. Males in all treatment groups had developed testis-ova. Males treated with E2 concentrations = 55.7 ng/l contained relating great concentrations of hepatic Vtg. Therefore, although only the greatest E2 concentration tested in our study affected fecundity and fertility, effects of E2 were observed on induction of Vtg and testis-ova in male medaka exposed to environmentally relevant concentrations of E2.

Animals↗

PubMed2002

Involvement of nitric oxide production via kynurenic acid-sensitive glutamate receptors in DOPA-induced depressor responses in the nucleus tractus solitarii of anesthetized rats.

We have proposed the hypothesis that L-3,4-dihydroxyphenylalanine (DOPA) plays a role of neurotransmitter of the primary baroreceptor afferents terminating in the nucleus tractus solitarii (NTS). In the present study, we tried to clarify whether glutamate receptors and/or nitric oxide (NO), important modulators for central cardiovascular regulation, are involved in the DOPA-induced cardiovascular responses in the nucleus. Male Wistar rats were anesthetized with urethane and artificially ventilated. Compounds or antisense oligos (17-mer) for neuronal NO synthase were microinjected into depressor sites of the unilateral nucleus. DOPA 30-300 pmol microinjected into the nucleus dose-dependently induced depressor and bradycardic responses. Prior injection of kynurenic acid (600 pmol) suppressed DOPA (300 pmol)-induced responses by approximately 80%. Prior injection of N(G)-monomethyl-L-arginine 100 nmol, a potent NO synthase inhibitor, reversibly attenuated by approximately 90% DOPA-induced responses, while the D-isomer 100 nmol produced no effect. Furthermore, prior injection of neuronal NO synthase antisense oligos (20 pmol) reversibly reduced by approximately 70% responses to DOPA. Sense or scrambled oligos produced no effect. A NO precursor L-arginine (30 nmol) induced depressor and bradycardic responses, but these responses were not affected by kynurenic acid. These results suggest important roles for glutamate receptors and NO in DOPA induced-depressor and bradycardic responses in the NTS.

Anesthetics, Intravenous↗

PubMed2002

Developmentally regulated expression of survivin in the human thymus.

Survivin is a member of the inhibitor-of-apoptosis (IAP) family. It has been reported to be expressed during development, but not in differentiated normal tissue. However, its expression has been reported to be high in the thymus. To assess the role of survivin in human thymocyte development, we investigated the expression of survivin using reverse-transcriptase-polymerase chain reaction, flow cytometry, and immunohistochemistry in freshly isolated human thymocytes. Survivin was expressed in all thymocyte subsets but its expression level was developmentally regulated. Its expression was low in the double negative (DN) thymocytes, upregulated in double positive (DP) thymocytes, and was highest in the T-cell receptor(high), late DP thymocytes; it was then downregulated in the single positive thymocytes and negative in the peripheral blood T cells. Moreover, there was a positive correlation between the expression of survivin and that of CD69 and Bcl-2 in DP thymocytes. These results suggest that survivin may play an important role in the T-cell development in the human thymus.

Antigens, CD↗

PubMed2002

Effect of ethinylestradiol on the reproduction and induction of vitellogenin and testis-ova in medaka (Oryzias latipes).

Mature medaka (Oryzias latipes) were exposed to ethinylestradiol (EE2) at measured concentrations of 32.6, 63.9, 116, 261, and 488 ng/L for 21 d under flow-through conditions. Effects on reproductive success of the fish as well as on gonadal condition and vitellogenin (VTG) induction were assessed. A significant decrease in fecundity was observed only at the highest EE2 concentration, whereas hepatic VTG concentration in males was increased at concentrations of 63.9 ng/L and greater. In addition, an intersex condition (testis-ova) of the gonad was observed in male medaka exposed to EE2 concentrations of > or = 63.9 ng/L. Overall, these results indicate that the lowest-observed-effect concentrations of EE2 based on reproduction versus induction of both VTG and testis-ova in the medaka were 488 and 63.9 ng/L, respectively. Thus, the physiological and histological measurements were approximately eightfold more responsive to the EE2 exposure than were overt reproductive effects. This suggests that the elevated VTG levels and testis-ova may not be definitely responsible for reproductive impairment of the fish.

Animals↗

PubMedDate not supplied

Death-associated protein kinase localization to human renal tubule cells, and increased expression of chronic obstructive uropathy in rats.

BACKGROUND: Death-associated protein kinase (DAP kinase) is a Ca2+/calmodulin-dependent serine/threonine kinase implicated as a positive apoptosis mediator. However, little is known about DAP kinase involvement with apoptosis in renal diseases. METHODS: In order to determine whether DAP kinase has a role in renal cell apoptosis in kidney diseases, we performed an immunohistochemical study using a monoclonal antibody to DAP kinase. Firstly, by examining the cellular distribution of DAP kinase in normal human renal tissues and cultured proximal tubule cells. We then used western blotting and immunohistochemical analysis to examine directly whether DAP kinase protein levels could be modulated in rat kidneys with chronic obstructive uropathy created by unilateral ureteric ligation. RESULTS: Immunohistochemistry of normal human kidney tissues showed that DAP kinase was exclusively localized in the cytoplasm of renal tubule cells. Expression analysis of DAP kinase using cultured cells confirmed DAP kinase mRNA and protein presence in human renal tubule cells. Immunocytochemical analysis directly visualized DAP kinase in the cytoplasm of the renal tubule cells in culture. Finally, DAP kinase was found up-regulated in renal tubule cells of rat kidneys with chronic obstructive uropathy. CONCLUSIONS: Our study demonstrates that DAP kinase is localized to renal tubule cells, implying a crucial role for DAP kinase in renal tubular cell apoptosis in progressive renal diseases.

Animals↗