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Biomedical subjects

T Nagai

Publications and source records attributed to T Nagai.

At least 37 records · Page 2Linked to original sources

Saline-assisted endoscopic resection of rectal carcinoids: cap aspiration method versus simple snare resection.

BACKGROUND AND STUDY AIMS: Complete resection of small carcinoid tumors of the rectum is difficult during endoscopic polypectomy, because these tumors are often located in the submucosal layer of the rectal wall. To completely remove these tumors, we have used aspiration lumpectomy. In this study, we evaluated its clinical usefulness in comparison with strip biopsy. PATIENTS AND METHODS: We performed strip biopsy (December 1996 through March 2000) or aspiration lumpectomy (April 2000 through February 2003) in a consecutive series of patients with rectal carcinoid tumor. Eight carcinoid tumors in eight patients were resected by strip biopsy, and eight tumors in eight patients were resected by aspiration lumpectomy. RESULTS: The rate of complete removal of carcinoid tumors with aspiration lumpectomy (100 %) was significantly higher (P < 0.05) than that with strip biopsy (37.5 %). CONCLUSIONS: Endoscopic resection with aspiration lumpectomy is a useful and safe method for resection of small carcinoid tumors of the rectum.

Carcinoid Tumor↗

Abnormal alkaline phosphatase isoenzymes detected in the serum of elderly patients.

Alkaline phosphatase (ALP) isoenzyme analysis of 101,832 serum samples was performed by electrophoresis using cellulose acetate membrane, and abnormal bands at the alpha(1) and alpha(2) globulin positions were detected in 23 samples. The physicochemical properties of these abnormal ALP isoenzyme fractions were examined. In brief, the abnormal fractions were heat-sensitive and inhibited by L-phenylalanine, and neither sialic acid in their polysaccharide chains nor the glycosyl-phosphatidylinositol anchor was detected by the enzyme treatment. The physicochemical properties of abnormal ALP isoenzyme fractions detected in Patients 1 - 22 were similar to those of the adult small intestine type. However, the molecular weight of the adult small intestine type abnormal fractions was smaller than that of the normal fractions. These adult small intestine type abnormal bands at the alpha(1)- to alpha(2)-globulin positions, which were identified in the serum of Patients 1 - 22, were detected in elderly patients. Most of them had various basal diseases such as renal insufficiency, fracture, interstitial pneumonia, and chronic pancreatitis. Some of them had severe diseases such as rectal cancer, descending colon cancer, and septic shock. In Patient 23, the polysaccharide chain had sialic acid and was heat sensitive physicochemical properties that were similar to those of the Kasahara ALP variant.

Adult↗

Multivariate spline interpolation as a novel method to optimize pharmaceutical formulations.

One of the difficulties in the quantitative approach for formulation design is the difficulty in understanding the actual relationship between causal factors and individual pharmaceutical responses. In this regard, several techniques were applied to determine the relationship between causal factors and the pharmaceutical responses. The generation of response surfaces using multivariate spline interpolation (MSI) has provided rapid and detailed information. Nevertheless, no application of MSI in the pharmaceutical field has been reported to date, even though it promises potential applications. To overcome the shortcomings of the classical response surface method, we newly developed a multi-objective simultaneous optimization method, in which MSI had been incorporated. The method was applied to the optimization problem of a transdermal hydrogel formulation for ketoprofen containing several chemical enhancers. Results suggested a superior function of the MSI approach.

Administration, Topical↗

Functional changes and motility characteristics of Japanese Black bull spermatozoa separated by Percoll.

Spermatozoa from two Japanese Black bulls (Bull-ATF and Bull-KTG) were separated by centrifugation at 700 x g for 15min in modified TALP with or without 45-90% Percoll. Control washed spermatozoa and those collected from the bottom of 45 and 90% Percoll fractions were examined for viability and membrane integrity (using Hoechst bis-benzimide 33258 or propidium iodide and 6-carboxyfluorescein diacetate (PI-CFDA)), acrosomal status (using fluorescence isothiocyanate (FITC) conjugated Pisum Sativum agglutinin (PSA) and Peanut agglutinin (PNA), Naphthol Yellow S and Erythrosin B (NE) or triple staining (TS)), capacitation status (using chlortetracycline (CTC)), motility characteristics (using a computer-assisted sperm motion analysis system (CASA)) and for in vitro fertility. Percoll-separated spermatozoa showed greater viability and membrane integrity than controls, as determined by supravital staining. Differences were observed in the results regarding viability and acrosomal status of spermatozoa among sperm staining methods. Bull-ATF, which showed significantly greater in vitro fertility than Bull-KTG (P<0.05), showed a significantly higher rate of CTC-B-pattern (capacitated) spermatozoa (P<0.01) than Bull-KTG. The motility characteristics of control washed spermatozoa and those separated by 45-90% Percoll were analyzed by CASA. More motile and progressively motile spermatozoa were observed in the fraction at the bottom of the 90% Percoll solution than in the 45% Percoll fraction or in controls (P<0.01). Moreover, the spermatozoa of Bull-KTG, which showed lower in vitro fertility than Bull-ATF, did not show significant differences in motility from those of Bull-ATF. These results provided basic information about Japanese Black bull spermatozoa, and suggested that spermatozoa with greater motility and viability can be obtained by Percoll separation than without separation. However, Percoll separation did not enhance their in vitro fertility.

Acrosome↗

Effect of AD-5423 on animal models of schizophrenia: phencyclidine-induced behavioral changes in mice.

The antipsychotic efficacy of AD-5423, which has the properties of both a serotonin 5-HT(2) and a dopamine D(2) receptor antagonist, was evaluated using animal models of schizophrenia. Sensitization to phencyclidine (PCP)-induced hyperlocomotion is considered a model of the positive symptoms of schizophrenia, and was significantly antagonized by AD-5423 and haloperidol. The PCP-induced enhancement of immobility induced by the forced swimming test, a model of the negative symptoms of schizophrenia, was attenuated by AD-5423 but not by haloperidol. Since this attenuated effect of AD-5423 was antagonized by DOI, a serotonin 5-HT(2) receptor agonist, it is postulated to be mediated by serotonin 5-HT(2) receptors. These findings suggest that AD-5423 would be clinically effective against both the positive and negative symptoms of schizophrenia.

Animals↗

Recovery of fish stocks in the Seto Inland Sea.

The total amount of fisheries' yield in the Seto Inland Sea in 1999 was 571,000 MT, consisting of 256,000 MT by fishing and 315,000 MT from aquaculture. About 40,000 people engaged in the fishing and aquaculture industries, earned 129 billion yen (1100 million US$) by fishing and 89 billion yen (770 million $US) by aquaculture. The averaged annual catch for the Seto Inland Sea by fishing was 13 MT/km(2). Division into time periods in terms of eutrophication levels can be made: before 1960 when red sea bream were abundant with ecological divergence (before eutrophication), from 1960 to 1990 when the biomass of anchovy was large (during eutrophication), and after 1990 when the jellyfishes were abundant (excessive eutrophication or high N:P ratio). The fish production will decrease in the sea of jellyfishes. Actually, the amount of catch was 462,000 MT in 1982 which decreased 265,000 MT in 1993, corresponding to 43% in twelve years, then keeping the same level. A big reduction was seen in the catches of the spotlined sardine, anchovy, Spanish mackerel, tiger puffer, short-necked clam, sea cucumber and others. The tiger puffer and Spanish mackerel were abundant as predators in the sea of anchovy. The biomass of anchovy was at its maximum in 1986 and decreased to less than one third in 1996. The stocks of tiger puffer and Spanish mackerel greatly decreased because of the higher fishing pressure compared to the anchovy stock. The fishing power of individual fisheries targeting on the tiger puffer and Spanish mackerel increased substantially when fishing vessel and fishing gear improved, resulting in an excessive fishing effort. A large quantity of small immature fishes is usually caught in the Seto Inland Sea, resulting in growth and/or recruitment overfishing for many species. Hence, it is necessary to promote management of the fisheries so as not to reduce the fish stocks, and to allow the Seto Inland Sea to return from being a sea of jellyfishes to a sea of anchovy, with decreased eutrophication levels.

Animals↗

Stage-specific effects of the osmolarity of a culture medium on the development of parthenogenetic diploids in the pig.

The objective of this study was to investigate the effects of osmolarity of culture media on the development of porcine parthenogenetic diploids. Oocyte-cumulus-granulosa cell complexes were collected from ovaries and then in vitro-cultured for 48 h. The mature oocytes were subjected to a single electro-stimulation (El-St; 100 micros, 1500 V/cm), treated with 5.0 microg/ml Cytochalasin B for 4h and then cultured under various conditions as described below. In Experiment 1, the diploids were cultured for 168 h after El-St in modified Whitten's medium with 256 mOsmol (mWM256), mKRB with 309 mOsmol, and mWM with 309 mOsmol (mWM309), in which the osmolarity was adjusted by addition of NaCl or mannitol, or by reduction of distilled water. In Experiment 2, the diploids were cultured in the five media used in Experiment 1 for the first 48 h, and then in mWM256 until 168 h after El-St. In Experiment 3, the diploids were cultured for the first 48 h in mWM with osmolarity adjusted from 256 to 330 mOsmol by addition of NaCl for the first 48 h and then in mWM256 until 168 h after El-St. In Experiment 4, the diploids were cultured in mWM with 290 mOsmol (mWM290) for the first period of 24, 48, or 72 h, and then in mWM256 until 168 h after El-St. In Experiment 5, after diploids were cultured in mWM290 for the first 48 h, the obtained 4-cell diploids were transferred to mWM with osmolarity adjusted from 200 to 310 mOsmol by addition of NaCl, then cultured until 168 h after El-St. All media were supplemented with 0.5mg/ml hyaluronic acid and 4.0mg/ml bovine serum albumin. The results obtained in Experiments 1-5 indicate that the osmolarity of a medium, but not the Na(+)/K(+) ratio, exerts effects on the development of diploids to the blastocyst stage. The change of osmolarity of the culture media after the 4-cell stage increased the rate of expanded blastocyst formation in porcine diploids. The optimal osmolarities of culture medium for the first 48 h after El-St (before the 4-cell stage) were 290 and 280-320 mOsmol, and those for the later period (after the 4-cell stage) were 256 and 220-270 mOsmol, respectively.

Animals↗

Effects of thioredoxin on the preimplantation development of bovine embryos.

Thioredoxin (TRX) is an ubiquitous protein disulfide reductase, which is known to be involved in the implantation development of mouse embryos. In the present study, recombinant human TRX was used to evaluate its effect on the promotion of preimplantation development of bovine embryos derived from in vitro maturation and fertilization. Supplementation of the medium 24h post insemination with TRX significantly (P<0.05) enhanced the frequency of development to the blastocyst stage in 5% O(2) concentration. The optimal concentration was 0.5 microg/ml (P<0.05, compared with 0, 0.1 and 1.0 microg/ml). This effect of TRX was evident only when added around the time of the first cleavage stage (24 h post insemination); no promotion was found with treatment at 6h (one-cell) or 44 h (six- to eight-cell) after insemination. Moreover, it is of interest that even with the best combination of the dose and timing of TRX treatment (0.5 microg/ml, at 24 h post insemination), no promotion of development was observed when embryos were cultured under 20% O(2). However, a preincubation of TRX in the culture medium under 20% oxygen for 24h did not diminish the promoting effect in the subsequent TRX treatment under optimal conditions, thus suggesting that the possible oxidation of TRX alone may not be the reason for the disappearance of the effect under a high oxygen concentration. These results indicate that TRX does improve the development of bovine embryos in vitro, though unlike the general reducing reagents such as beta-mercaptoethanol or cysteamine, TRX may have to exert its effect at specific times and in more physiologic oxygen environments.

Animals↗

In vitro fertility and motility characteristics of frozen-thawed boar epididymal spermatozoa separated by Percoll.

Frozen-thawed epididymal spermatozoa from four boars were separated through a Percoll gradient, and motility characteristics and in vitro fertility were assessed. Percoll-separated spermatozoa had a significantly higher percentage of motile and progressively motile spermatozoa than those that were not separated (P < 0.0001). However, there were no clear differences in other motility parameters between Percoll-separated and un-separated spermatozoa. Furthermore, sperm agglutination was decreased by Percoll separation (P < 0.05). The effects of Percoll separation on in vitro fertility of spermatozoa differed among boars. In addition, there were large differences in fertility between sperm samples in vitro. Sperm samples, which indicate highly motile and progressively motile, did not always show high in vitro fertility. Furthermore, there was no distinct pattern between fertility in vitro and motility parameters. There was no difference in fertility in vitro between Percoll-separated and un-separated spermatozoa from two of the four boars. However, in vitro fertility of Percoll-separated spermatozoa was higher than that of un-separated spermatozoa from the other two boars.

Animals↗

Immunocytochemical evidence that amyloid beta (1-42) impairs endogenous antioxidant systems in vivo.

Amyloid beta, the major constituent of the senile plaques in the brains of patients with Alzheimer's disease, is cytotoxic to neurons and has a central role in the pathogenesis of the disease. We have previously demonstrated that potent antioxidants idebenone and alpha-tocopherol prevent learning and memory impairment in rats which received a continuous intracerebroventricular infusion of amyloid beta, suggesting a role for oxidative stress in amyloid beta-induced learning and memory impairment. To test the hypothesis, in the present study, we investigated alterations in the immunoreactivity of endogenous antioxidant systems such as mitochondrial Mn-superoxide dismutase, glutathione, glutathione peroxidase and glutathione-S-transferase following the continuous intracerebroventricular infusion of amyloid beta for 2 weeks. The infusion of amyloid beta (1-42) resulted in a significant reduction of the immunoreactivity of these antioxidant substances in such brain areas as the hippocampus, parietal cortex, piriform cortex, substantia nigra and thalamus although the same treatment with amyloid beta (40-1) had little effect. The alterations induced by amyloid beta (1-42) were not uniform, but rather specific for each immunoreactive substance in a brain region-dependent manner. These results demonstrate a cytological effect of oxidative stress induced by amyloid beta (1-42) infusion. Furthermore, our findings may indicate a heterogeneous susceptibility to the oxidative stress produced by amyloid beta.

Amyloid beta-Peptides↗

Differential regulation of Th1 responses and CD154 expression in human CD4+ T cells by IFN-alpha.

Like interleukin (IL)-12, interferon (IFN)-alpha has been shown to play an important role in inducing human Th1 responses. Recent studies have shown that human Th1 responses driven by IL-12 are associated with enhanced expression of CD154. The present study examined the effects of IFN-alpha on CD154 expression in human CD4+ T cells, with special attention to the relationship with Th1 responses. Highly purified CD4+ T cells from healthy donors were stimulated with immobilized anti-CD3 with or without IFN-alpha and IL-12 in the complete absence of accessory cells. IFN-alpha suppressed CD154 protein and mRNA expression in CD4+ T cells at the initial phase of activation with immobilized anti-CD3, but enhanced it in the subsequent maturation phase irrespective of the presence of IL-12. By contrast, IFN-alpha by itself did not enhance IFN-gamma production or mRNA expression in CD4+ T cells in the absence of IL-12 even in the presence of stimulation with anti-CD28, but enhanced it in the presence of IL-12. Accordingly, IFN-alpha enhanced IL-12Rbeta2 mRNA expression in anti-CD3-stimulated CD4+ T cells. Neither IFN-alpha nor IL-12 influenced the stability of CD154 mRNA in anti-CD3-activated CD4+ T cells. These results indicate that IFN-alpha by itself enhances CD154 expression in CD4+ T cells independently of the induction of IFN-gamma mRNA expression. The data also suggest that the optimal induction of human Th1 responses by IFN-alpha might require the presence of IL-12 and that the induction of Th1 responses and CD154 expression in human CD4+ T cells might be regulated through different mechanisms.

Adult↗

Immunohistochemical classification of the localization of laminin in the thickened bronchial epithelial basement membrane of deceased bronchial asthma patients.

To ascertain histological changes in the basal lamina of the bronchial epithelial basement membrane in patients with severe bronchial asthma, an immunohistochemical study was conducted in 43 patients who died of bronchial asthma. Antibodies against laminin, a component of the lamina lucida, were utilized. The results revealed various patterns for immunoreactivity to laminin in the thickened basement membrane layer. We were able to classify these reactivities into four patterns. In Pattern A, laminin reactions branched vertically in relation to the thickened basement membrane layer. In Pattern B, laminin reactions formed lines along the lower margin of the thickened basement membrane layer. In Pattern C, laminin reactions formed lines along the upper margin of the thickened basement membrane layer. Finally, in Pattern D, no laminin reactions were observed. In addition, relationships between immunohistological characteristics of laminin and findings such as epithelial cell shedding, basal cell proliferation and basement membrane layer thickening were investigated. In many Pattern A patients, epithelial cell shedding was observed, but goblet cell hyperplasia and basal cell proliferation were barely detectable. Conversely, in numerous Pattern D patients, epithelial cell shedding was barely seen, but goblet cell hyperplasia and basal cell proliferation were marked. Hence, Patterns A and D were on opposite ends of the spectrum of morphological characteristics associated with severe bronchial asthma. In Patterns B and C, laminin reactions formed lines along the lower and upper margin of the thickened basement membrane layer, respectively. However, no marked differences existed in epithelial cell shedding and basement membrane layer thickening. The present study is thus the first to clarify that laminin reactions in the thickened basement membrane layer vary, and this feature is unique to the bronchi of patients with severe bronchial asthma.

Adult↗

Characterization of monoclonal antibodies against Gnathostoma nipponicum.

Monoclonal antibodies (mAbs) were produced against the proteins of advanced third-stage larvae (AdL3) of Gnathostoma nipponicum. Six mAbs (Gn2C3, Gn2H3, Gn4C3, Gn4E9, GnSH1, and Gn10B7) were obtained as determined by enzyme-linked immunosorbent assay (ELISA). Gn4E9 and GnSH1 seemed to be genus-specific, as they did not cross-react with Anisakis sp., Dirofilaria immitis, Gongylonema pulchrum, Toxocara canis, Trichinella sp., Trichuris vulpis, Metagonimus sp., or Spirometra erinaceieuropaei by ELISA. Immunohistochemistry showed that Gn2C3, Gn4E9, and Gn5H1 reacted strongly with the central esophagus; Gn2H3 reacted with cuticle,muscle, intestine, and the cervical sac; and Gn4C3 and Gn10B7 reacted with cuticle, muscle, esophagus, intestine, and the cervical sac of AdL3. In Western blotting analysis, Gn2C3, Gn4E9, and Gn5H1 reacted to 60-, 53-, 46-, and 41-kDa proteins; Gn4C3 reacted to the AdL3 protein of G. nipponicum (>42 kDa). Moreover, proteins purified using a mAb Gn4E9 immunoprecipitation method (sizes 60-, 53-, 46-, and 41-kDa) were used as antigens in ELISAs. A significant difference (P < 0.01) was shown between mouse sera infected with G. nipponicum and sera infected with Trichnella sp. or not infected. These results provide a rationale for evaluating esophageal proteins for the development of diagnostic methods for detecting G. nipponicum or Gnathostoma sp. infections.

Animals↗

Coumestrol as well as isoflavones in soybean extract prevent bone resorption in ovariectomized rats.

OBJECTIVE: Soybeans contain an abundance of phytoestrogens such as genistein and daidzein, and these compounds are thought to protect against bone loss under estrogen deficient conditions. It is possible, however, that phytoestrogens other than isoflavones may suppress bone resorption. The objective of the present study was to determine whether there are any phytoestrogens other than isoflavones in soybeans that can act as antiosteoporotic agents. METHODS: The isoflavones genistein and daidzein, and an extract of soybeans that contained very low levels of isoflavones were tested for their ability to reduce bone resorption in ovariectomized rats. The extract of soybeans was further analyzed for its main components by thin layer chromatography and gas chromatography-mass spectrometry. RESULTS: The soybean extract, as well as the two isoflavones, were effective to reduce urinary excretion of deoxypyridinoline and pyridinoline, typical markers of bone resorption in ovariectomized rats. However the extract, unlike the isoflavones, increased the uterine weight of ovariectomized rats significantly. Analysis by thin-layer and gas chromatography revealed that the main constituent of this extract exhibited a chromatographic profile corresponding to coumestrol. This substance was positively identified as coumestrol by gas chromatography-mass spectrometry. CONCLUSION: These results suggest that the phytoestrogens including coumestrol and isoflavones in soybeans may exert effective prevention against bone resorption in estrogen deficient conditions.

Amino Acids↗

Ferromagnetic domain structures and nanoclusters in Nd(1/2)Sr(1/2)MnO3.

Magnetic domain structures of Nd(1/2)Sr(1/2)MnO3 were investigated by means of low-temperature Lorentz electron microscopy. On cooling, magnetic domain walls started to appear at 250 K, and they were oriented straight along the [100] and [110] directions. With a further decrease in temperature, the volume of each magnetic domain increased with discontinuous domain-wall jumps. A characteristic granular image was observed at around 140 K, near the charge-ordering transition temperature. We consider that this originated from ferromagnetic nanoclusters that appeared in the antiferromagnetic matrix.

Journal Article↗