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Biomedical subjects

M Maeda

Publications and source records attributed to M Maeda.

At least 793 records · Page 44Linked to original sources

Effect of colostomy on the utilisation of dietary nitrogen in the fowl fed on a low protein diet.

1. The effect of the inhibition of urine back-flow into the colon and caeca by colostomy on the utilisation of dietary nitrogen by fowls fed on a low protein diet and receiving free or restricted water supply was investigated. 2. Colostomy caused an increase in water excretion and a resultant increase in water intake to maintain water balance. 3. Colostomy tended to decrease nitrogen balance and nitrogen utilisation (N balance/N intake) to negative values, and these decreases became significant when water was restricted (P less than 0.05). 4. Excretory uric acid, ammonia, urea and total nitrogen were significantly increased after colostomy in water-restricted fowls (P less than 0.05), but such significant effects were not observed, except for ammonia, in fowls given water ad libitum. 5. It is concluded that the back-flow of urine into the caeca plays a significantly useful role in the utilisation of nitrogen in the fowl fed on a low protein diet especially when water intake is restricted.

Animals↗

Occurrence of intraportally-infused urea-15N in the urine of domestic fowl.

1. Measurements were made in situ to determine the occurrence of intraportally infused urea-15N in ureteral urine of the fowl. 2. Of the total amount of infused urea-15N, 15% was excreted intact into the urine (90% of urinary total 15N) whereas 9% remained unchanged in the blood (78% of blood non-protein-15N). 3. The proportions of non-protein-15N in the blood, liver and kidney were 12, 3 and 1%, respectively of the infused 15N. Protein-15N was 3% of that infused in blood and much less in liver and kidney. 4. About 1% of the infused 15N was observed in the urinary uric acid, and 3% of the infused 15N in non-protein N, other than urea, ammonia and glutamine amide N, of blood and liver. 5. No appreciable amounts of 15N were present in ammonia and glutamine amide N of blood, liver or kidney and in uric acid of liver or kidney. 6. The caecal contents contained about 1% of the infused 15N with 15% of this as ammonia-15N. 7. It is concluded that intraportal urea is mostly excreted unchanged into ureteral urine of the fowl.

Animals↗

Dipeptidyl peptidase IV as a differentiation marker of the human endometrial glandular cells.

To investigate the involvement of membrane-bound peptidases in the human endometrial function, we examined the expression of dipeptidyl peptidase (DPP) IV and its enzyme activity. Immunohistological studies revealed that DPP IV was detected on human endometrial glandular cells and endometrial surface epithelium, but not on endometrial stromal cells or decidual cells in the first trimester of pregnancy. DPP IV expression on glandular cells and surface epithelium was weak in the proliferative phase, began to increase gradually in the early secretory phase, and was strong in mid-to late secretory phase and in the first trimester of pregnancy. DPP IV enzyme activity was detected histochemically in glandular cells and surface epithelium in the mid-secretory phase, and became stronger in the late secretory phase, but was rarely detected in the proliferative phase and early secretory phase. During the first trimester of pregnancy DPP IV enzyme activity in glandular cells and surface epithelium was slightly weaker than in the late secretory phase. Endometrial stromal cells and decidual cells, however, had no detectable DPP IV enzyme activity at any time throughout the menstrual cycle or during the first trimester of pregnancy. These findings indicate that DPP IV is a differentiation marker for glandular cells and surface epithelium and that active DPP IV is present in both areas during the peri-implantation period and thereafter.

Adult↗

Expression of aminopeptidase N and neutral endopeptidase on the endometrial stromal cells in endometriosis and adenomyosis.

Indirect immunofluorescence staining revealed that endometrial stromal cells (ESC) in the ectopic endometrium of patients with endometriosis or adenomyosis expressed aminopeptidase N/cluster of differentiation (CD) 13 antigen and neutral endopeptidase/CD10 antigen, both of which are expressed on ESC in the normal endometrium throughout the menstrual cycle. Thus, ESC in the ectopic endometrium resembled ESC in the normal endometrium not only morphologically but also antigenically. Both peptidase antigens may be useful markers for the histological diagnosis of endometriosis and adenomyosis.

Adult↗

Human endometrial stromal cells and decidual cells express cluster of differentiation (CD) 13 antigen/aminopeptidase N and CD10 antigen/neutral endopeptidase.

With specific monoclonal antibodies, we found that human endometrial stromal cells and decidual cells express two function-related surface antigens. Indirect immunofluorescence staining revealed that both endometrial stromal cells and decidual cells during the first trimester of pregnancy expressed cluster of differentiation (CD) 13 antigen and CD10 antigen, which are identical to aminopeptidase N and neutral endopeptidase, respectively. By flow cytometric analysis, CD13 antigen was detected on 82-93% of the examined cells, and CD10 antigen was detected on 75-93% of the examined cells in endometrial stromal cell-enriched preparations. Furthermore, peptidase activity was detected in these cell preparations by an assay based on the hydrolysis of alanine-p-nitroanilide into p-nitroaniline and alanine.

Adult↗

Proof of plasmatic imbibition in rat musculocutaneous grafts: enzymatic proof using peroxidase.

At present, the concept of plasmatic imbibition is generally accepted. That is, observation of weight change or pigmentation methods of the grafts were established to support it. In the present study, we investigated plasmatic imbibition in rat musculocutaneous grafts histologically using peroxidase, which is one of the reductases. Tissue pigmentation by peroxidase became an insoluble sediment that indicated the sites of peroxidase activity. The entire contact surface of the graft with the recipient bed was stained brown within a few minutes after the operation. At 30 minutes, the panniculus carnosus and dermis were stained dark brown diffusely, extending toward the epidermis. This condition continued until the sixth day at least. As a result, peroxidase that was dissolved with the exudate between the graft and recipient bed was imbibed into the musculocutaneous graft.

Absorption↗

448-detector optical recording system: development and application to Aplysia gill-withdrawal reflex.

The spatial resolution of a fast, multichannel, optical recording apparatus has been improved in an attempt to increase the completeness of optical recordings of neuron activity responsible for the Aplysia gill-withdrawal reflex. We developed a new optical apparatus, increasing the number of pixels to 448. Action potential activity from 168 to 192 neurons could be detected. Hence, the number of optically detected neurons increased about 2.5 times in comparison with a previously used 100- or 124-detector optical apparatus. Because optically detected action potential signals were shown to be due to the cell body of a neuron, it was possible to know its position as well as its approximate size. The distribution of the detected active neurons and those neurons that showed an apparent change in spike frequency to the siphon stimulation were determined.

Action Potentials↗

Enhancement of lysosomal enzyme activity by recombinant human tumor necrosis factor and its role in tumor cell killing in vitro.

We investigated the effect of recombinant human tumor necrosis factor (TNF) on the lysosomal enzyme activity of various established cell lines in vitro. Incubation of 1 x 10(6) TNF-sensitive mouse tumorigenic fibroblasts (L-M cells) in the presence of TNF (100 U/ml) for 48 h increased the total (the sum of the enzyme activities in the lysosomes and the cytoplasm) acid phosphatase and beta-glucuronidase activities by 3.7- and 4.2-fold, respectively. The same increase was observed even when 1 U/ml of TNF was added to some cultures and no further augmentation occurred at 10 or 100 U/ml. Measurement of total and free enzyme activities showed that TNF stimulation not only enhanced the total intracellular enzyme activity but also accelerated the conversion into free (cytoplasmic) enzyme activity. Addition of a lysosomotropic agent (methylamine) suppressed both the enhancement of lysosomal enzyme activity and the cytotoxicity of TNF. A similar enhancement of lysosomal enzyme activities was also detected in various TNF-sensitive tumor cell lines, and a strong correlation (acid phosphatase: r = 0.836, beta-glucuronidase: r = 0.910) was observed between the enhancement of enzyme activity and sensitivity to TNF. No such increase was detected in TNF-resistant human diploid cells. These results show that TNF induces the activation and release of lysosomal enzymes in TNF-sensitive cells, and suggest that such events may play an important role in TNF-mediated cytotoxicity.

Acid Phosphatase↗

Mechanism of synergistic cytotoxic effect between tumor necrosis factor and hyperthermia.

We previously reported that recombinant human tumor necrosis factor (rhTNF) and hyperthermia had a synergistic effect against tumors, in vitro and in vivo. We have now investigated the mechanism of this synergy by measuring the lysosomal enzyme activity and hydroxyl radical production of L-M cells treated with rhTNF and/or hyperthermia. A synergistic activation of lysosomal enzyme and the induction of hydroxyl radical production in L-M cells treated with both rhTNF and hyperthermia was observed. A synergistic cytotoxic effect was observed when rhTNF and hyperthermia were combined, and was inhibited by the addition of a reactive oxygen scavenger, dimethyl sulfoxide or bipyridine. The results show that the augmenting effect of hyperthermia on lysosomal enzyme activation and induction of hydroxyl radical production by rhTNF plays an important role in the synergistic cytotoxic effect.

Acid Phosphatase↗

Correlation of HLA types and clinical findings in Japanese patients with hyperthyroid Graves' disease: evidence indicating the existence of four subpopulations.

OBJECTIVES: To re-evaluate the associations of HLA types with Japanese patients having hyperthyroid Graves' disease, HLA types and clinical findings were correlated. DESIGN: Four independent clinical findings (ophthalmopathy, family history, age at onset and size of goitre) and two autoantibody titres, thyrotrophin binding inhibitor immunoglobulin (TBII) and anti-thyroid microsmall antibody (anti-M), were analysed. PATIENTS: Eighty-eight Japanese patients with hyperthyroid Graves' disease and 186 control subjects were assessed. MEASUREMENT: Serological HLA typing was performed on 73 antigens in HLA-A, -B, -C, -DR and -DQ loci. HLA-D and -DP (29 antigens) were determined by the restricted fragment length polymorphism (RFLP) methods. TBII and anti-M were measured by commercially available kits. RESULTS: Patients with potent antibody titres had HLA antigens commonly seen among all the patients with Graves' disease. Interestingly, however, HLA-B35 and -Cw11 were found to relate with negative and/or weak TBII, and HLA-B7 and absence of HLA-Aw19 with negative anti-M. Significant associations were observed between HLA-DRw8 and large goitre and absence of ophthalmopathy, and between HLA-DQw4 and a negative family history of diffuse goitre (corrected P less than 0.05). Several other antigens were also found to be significant. Among these antigens, four pairs of MHC classes I and II were found to relate to the clinical findings independently. HLA-DQw4 and negative -A31 pair was closely related to ophthalmopathy, negative family history and late onset of disease. The HLA-B5 and -Dw12 pair was associated with ophthalmopathy, positive family history and early onset of disease. The HLA-A11 and negative -DPw2 pair was associated with ophthalmopathy, negative family history and early onset of disease. The HLA-Bw46 and -DRw8 pair did not increase in frequency above that seen with HLA-DRw8 alone. These four antigen groups (HLA-DRw8, HLA-DQw4 and negative-A31, HLA-B5 and -Dw12, and HLA-A11 and negative -DPw2) were observed in the majority (68%) of patients with Graves' disease and at a significantly higher incidence than in the control group (P less than 0.05). CONCLUSION: There are four subpopulations of Japanese patients with hyperthyroid Graves' disease. This is one of the reasons why the association of HLA types in Japanese patients is rather weak when they are studied as one group.

Adolescent↗

Occupational or extrinsic stimulation factors and initial signs of progressive systemic sclerosis.

Forty-eight patients with progressive systemic sclerosis (PSS) (man-woman ratio, 7:41) were studied, and detailed questionnaires were used to clarify the relationship between occupational stresses and the initial signs of PSS. All of the patients were right-handed. Raynaud sign was seen in 37 of the 48 patients (77.1%) as the predominant initial sign of PSS, and it first was noticed on the right index and middle fingers. Three sets of extrinsic aggravating factors, vibration stress, fine hand work with mental stress, and exposure to cold, were believed to be related closely to the development of the initial signs because the initial signs appeared after long-term exposure to the extrinsic factors.

Adult↗

Differential expression of aminopeptidase-N on human ovarian granulosa and theca cells.

The expression of aminopeptidase-N and neutral endopeptidase in human ovarian tissue was examined using specific monoclonal antibodies for each of these peptidases and histochemical staining for enzyme activity. Aminopeptidase-N is a membrane-bound metalloprotease catalyzing the removal of N-terminal amino acids from peptides and was detected by immunofluorescence staining on theca interna cells in secondary follicles and on luteinized thecal cells in preovulatory follicles and corpora lutea. However, aminopeptidase-N was not detected on granulosa cells. Peptidase activity was also detected by histochemical staining on theca interna cells and luteinized thecal cells. Luteinized granulosa cells showed peptidase activity, despite the lack of aminopeptidase-N. Neutral endopeptidase was not detected in ovarian granulosa and thecal cells. These observations indicate that aminopeptidase-N can be a useful surface marker for thecal cells.

Adult↗

Human luteal cells express dipeptidyl peptidase IV on the cell surface.

We previously reported that human theca interna cells and small luteal cells express membrane-bound aminopeptidase N, and suggested that membrane-bound peptidases are involved in folliculogenesis and luteal function by regulating extracellular peptide concentrations. In this study, we examined the expression of dipeptidyl peptidase IV (DPP IV), which is a membrane-bound peptidase and has its catalytic domain at extracellular sites, in human granulosa cells, thecal cells of growing, preovulatory, and atretic follicles, as well as corpora lutea. Indirect immunofluorescence staining of ovarian tissues with specific monoclonal antibodies revealed that DPP IV was present in large and small luteal cells in corpora lutea. DPP IV peptidase activity was also detected histochemically in corpora lutea. In growing, preovulatory, and atretic follicles, there was weak immunoreactivity and DPP IV peptidase activity on luteinized theca interna cells, but not on granulosa cells. The expression of DPP IV on the cell surface of large and small luteal cells was confirmed by indirect immunofluorescence staining of freshly isolated luteal cells. These results indicate that DPP IV is a useful surface differentiation marker of human luteal cells and suggest that peptidases are involved in luteal function.

Adult↗

The role of V-ATPase in neuronal and endocrine systems.

Synaptic vesicles have important roles in the neural transmission at nerve terminals: the storage and the controlled exocytosis of neurotransmitters. At least two different factors are responsible for the concentration process: the vacuolar-type H(+)-ATPase (V-ATPase), establishing an electrochemical gradient of protons, and specific transport systems for transmitters. We will discuss our recent progress on the energy-transducing systems in synaptic vesicles: (1) structural aspects of V-ATPase; (2) energy coupling of transport of transmitters; (3) reconstitution of transporters; (4) effects of neurotoxins and neuron blocking agents; (5) function of synaptic-vesicle-like microvesicles from endocrine tissues.

Adenosine Triphosphatases↗

Escherichia coli ATP synthase (F-ATPase): catalytic site and regulation of H+ translocation.

We discuss our recent results on the Escherichia coli F-ATPase, in particular its catalytic site in the beta subunit and regulation of H+ transport by the gamma subunit. Affinity labelling experiments suggest that beta Lys-155 in the glycine-rich sequence is near the gamma-phosphate moiety of ATP bound at the catalytic site. The enzyme loses activity upon introduction of missense mutations in beta Lys-155 or beta Thr-156 and changes catalytic properties upon introduction of other mutations. By analysis of mutations and their pseudo revertants, residues beta Ser-174, beta Glu-192 and beta Val-198 were found to be located near the glycine-rich sequence. The combined approaches of chemical labelling and genetics have been fruitful in visualizing the structure of the catalytic site. Analysis of mutations in the gamma subunit suggests that this subunit has an essential role in coupling catalysis with proton translocation.

Amino Acid Sequence↗

Isolation and characterization of a novel membrane glycoprotein of 85,000 molecular weight from rat liver lysosomes.

We have purified and characterized a novel glycoprotein (r-lamp-3) with an apparent molecular weight (Mr) of 85,000 from membranes of triton-filled lysosomes (tritosomes) by the use of immunoaffinity chromatography on a column of monoclonal antibody-Sepharose 4B. r-lamp-3 accounted for approximately 4% of the total proteins in tritosomal membranes. The isoelectric point (pI) of r-lamp-3 was 4.5 and it was shifted to 6.5 after neuraminidase treatment with its molecular weight decreased by about 7000. Pulse-chase experiments in cultured rat hepatocytes using [35S]methionine showed that r-lamp-3 was initially synthesized as a 77,000 polypeptide and processed to a mature protein with an Mr of 85,000. Upon treatment with endo-beta-N-acetylglucosaminidase H (Endo H), the precursor and mature forms were converted to 55,000 and 73,000 polypeptides, respectively. From the Mr reduction of the precursor form, we estimated the presence of 10--12 N-linked oligosaccharides/r-lamp-3 polypeptide. The data on enzymatic deglycosylation suggested that the mature form of r-lamp-3 contained the same numbers of high mannose-type and complex-type N-linked oligosaccharide chains.

Animals↗

Positron-emitting N-[18F]fluoroalkyl and [18F]fluoropyrrolidinyl analogues of eticlopride as potential in vivo radioligands for dopamine D2 receptors.

N-Fluoroalkyl and 4-fluoropyrrolidinyl eticlopride analogues with high affinity toward central nervous system dopamine D2 receptors in vitro were labelled with positron emitting fluorine-18 (t1/2 = 110 min), and their in vivo biodistribution was investigated in rats. N-[18F]Fluoro-ethyl and -propyl eticlopride derivatives showed poor in vivo selectivity in the rat brain. On the other hand, 4-[18F]fluoropyrrolidinyl eticlopride exhibited almost constant and relatively high striatal concentration. The striatal/cerebellar radioactivity ratio, which corresponds to the ratio of a brain D2 receptor-rich to poor region, gradually increased to 5.2-6.4, 90 min after the injection. The striatal accumulation was selectively inhibited by pre-injection of haloperidol, a dopamine D2 antagonist, without affecting accumulation in other tissues. Thus, the selective striatal accumulation of 4-[18F]fluoropyrrolidinyl eticlopride in striatal tissue appears to be due to the specific binding to dopamine D2 receptors.

Animals↗

Manganese-containing superoxide dismutase in blood and urine during open-heart surgery.

Concentrations of Manganese-containing superoxide dismutase (Mn-SOD) were measured perioperatively by enzyme immunoassay in serial samples of arterial and coronary sinus blood and urine taken from 18 patients undergoing mitral valve surgery. The mean Mn-SOD concentration in the arterial blood samples was 66.2 (SD 16.1 ng/ml) at induction of anesthesia, increased gradually after reperfusion and peaked on the 2nd post-operative day [150 (SD 58.3) ng/ml]. The mean concentration of Mn-SOD in the coronary sinus blood samples was significantly higher than in the arterial samples only at the 6th hour after reperfusion [97 (SD 21.8) ng/ml vs 90.3 (SD 20.9) ng/ml, p < 0.05]. Although concentrations of Mn-SOD in blood did not increase in 8 patients who underwent midline sternotomy for a mediastinal tumor, they increased dramatically in 3 patients who sustained a perioperative myocardial infarction. During open heart surgery the peak values of plasma Mn-SOD concentrations were correlated to that of plasma creatine kinase-MB concentrations (r = 0.5532, n = 18, p < 05) and cardiac ischemic period (r = 0.5186, n = 18, p < 05). Although the meaning of an increase in plasma Mn-SOD concentrations during open heart surgery is not clarified, it may be released from the heart and anywhere also in the body damaged during cardiopulmonary bypass.

Coronary Vessels↗