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Biomedical subjects

N Sato

Publications and source records attributed to N Sato.

At least 631 records · Page 35Linked to original sources

Development of DNA diagnostic methods for the detection of new fish iridoviral diseases.

A new disease of epidemic proportions caused by fish viruses within the Iridoviridae family inflicts serious damage on red sea breams (Pagrus major) and striped jack (Caranx delicatissimus) populations grown in aquacultures in Japan. A partial segment of the fish iridoviral DNA was directly amplified using the polymerase chain reaction (PCR) with synthetic primers designed from well conserved nucleotide sequences between the frog virus 3 (Ranavirus) and the silkworm iridescent virus type 6. The deduced amino acid sequence from the nucleotide sequence of the PCR fragment demonstrates a high correlation with a partial sequence from the frog virus 3. Using the PCR method with specific primers, we could detect three of four different known types of fish iridoviruses in diseased fishes. To construct more reliable detection methods specific for this viral family, DNA fragments which can specifically hybridize with all of the four known iridoviridae viral DNAs were screened from the genomic library of one iridoviridae strain. The hybridization assay, using a specific fragment which contains regions which are highly homologous with a characterized partial sequence from the frog virus 3, proved to be a reliable diagnostic tool for fish iridoviral diseases.

Animals↗

In vivo anti-tumour effect of 3'-sulphonoquinovosyl 1'-monoacylglyceride isolated from sea urchin (Strongylocentrotus intermedius) intestine.

Extracts from sea urchin intestine were screened for new anti-tumour drugs. Four glycolipids, 3'-sulphonoquinovosyl-1', 2'-diacylglyceride (A-4), 3'-sulphonoquinovosyl-1'-monoacylglyceride (2'-lyso A-4, A-5), NeuGc(alpha)2-6Glc(beta)1-1ceramide (A-6) and HSO3-8NeuGc(alpha)2-6Glc(beta)1-1ceramide (A-7), were isolated from the intestine of sea urchin, Strongylocentrotus intermedius, and characterized by means of proton nuclear magnetic resonance spectroscopy and fast atom bombardment mass spectrometry. When tested for cytotoxic activity against tumour cells in vitro, A-5 showed significant activity, but A-4, -6 and -7 did not. In addition, the hydrophilic derivatives of A-4 or -5 had no cytotoxicity. Furthermore, the anti-tumour effects on nude mice bearing solid tumours of a human lung adenocarcinoma cell line A-549 were evaluated in vivo using A-4 and -5. As a result, A-5 was found to be significantly effective in suppressing the growth of solid tumours, whereas A-4 had no effect. Pathologically, the solid tumours showed haemorrhagic necrosis areas after treatment with A-5. In this study, we have demonstrated the anti-tumour effect of sulphonoquinovosyl-lysoglyceride (A-5), which provides important information that this sulpholipid could be a useful drug for cancer chemotherapy.

Adenocarcinoma↗

A rapid nylon-fiber syringe system to deplete CD14+ cells for positive selection of human blood CD34+ cells. Use of immunomagnetic microspheres.

To achieve a rapid and an efficient purification of CD34+ cells, we devised a nylon-fiber syringe (NF-S) and we manipulated it to deplete adherent cells from normal human blood mononuclear cells (MN cells). The cells processed by NF-S were further purified as the CD34+ fraction, using CD34 monoclonal antibody, immunomagnetic microspheres and chymopapain treatment to detach the microspheres. When steady-state human peripheral blood MN cells were processed by NF-S at 24 degrees C for 5 min, the frequency of monocytes (CD14+ cells) significantly decreased from 22.0 +/- 5.2% to 2.5 +/- 0.4%, with a 73% recovery of CD34+ cells. The subsequent immunomagnetic positive selection achieved preparations of 91 +/- 8% pure CD34+ cells with a 86 +/- 23% yield. The overall yield of CD34+ cells was 44 +/- 11%, and the time required for all procedures was 5 h. There was a tight and an inverse correlation (P < 0.0001) between the frequency of CD14+ cells in the initial cell population and the purity of CD34+ cells in the final preparation. A recommended frequency of CD14+ cells for achieving preparations of over 90% pure CD34+ cells was less than 4.4%. When combining NF-S and immunomagnetic microspheres, efficient bench-top separation of CD34+ cells in steady-state peripheral blood can be done in any laboratory, and fluorescence-activated cell-sorting is not required.

Antigens, CD34↗

Fatal GVHD demonstrating an involvement of respiratory muscle following donor leukocyte transfusion (DLT).

A 41-year-old female patient with AML, who relapsed after an allogeneic BMT from her HLA-identical sister, was treated by a donor leukocyte transfusion (DLT). Thereafter, bone marrow aplasia accompanied by the disappearance of leukemic blasts following the GVHD was observed. The patient died of chronic GVHD with respiratory muscle involvement 19 months after the DLT. Although the DLT was considered helpful in suppressing the proliferation of the leukemic cells, it might also have caused the severe GVHD observed in this case. Efforts to separate the lymphocyte clones responsible for GVL from those for the GVHD thus appear to be necessary for the further development of the therapeutic approach, so-called DLT.

Adult↗

Serum leptin concentration is associated with total body fat mass, but not abdominal fat distribution.

OBJECTIVE: The obese (ob) gene encodes leptin which inhibits appetite and stimulates thermogenesis. Serum leptin concentrations are determined by total body fat mass, but the influence of visceral fat accumulation and other metabolic factors have been clinically determined. METHODS: We determined the correlations between serum leptin concentrations and the total body fat mass, abdominal fat mass, abdominal fat distribution (estimated by ultrasound), and circulating metabolic factors in 104 Japanese healthy subjects (11 men and 93 women). In addition, the effect of food restriction (30 kcal/kg desired body weight/day) for four weeks on serum leptin concentrations were also examined in 30 women. RESULTS: There was a significant correlation between serum concentrations and total body fat mass (r = 0.708, P < 0.0001), the percentage of body fat (r = 0.561, P < 0.001), and the body mass index (BMI, r = 0.630, P < 0.001). Serum leptin concentrations were correlated with abdominal wall preperitoneal and subcutaneous fat pad thickness, but not the abdominal wall fat index (AFI). Serum leptin concentrations were also correlated with serum immunoreactive insulin (IRI), but not glucose, or free fatty acid (FFA) concentrations. The weight loss after food restriction for four weeks significantly (P = 0.016) reduced the serum leptin concentrations with a significant reduction of body fat mass, serum glucose, IRI and FFA concentrations. However, there was no significant correlation of the percentage change in serum leptin concentrations to that in body fat mass after food restriction. CONCLUSION: Serum leptin concentrations are well correlated with total body fat mass in healthy subjects. Differences in abdominal fat distribution do not appear to be related to a difference in the in vivo leptin production from adipose tissue.

Abdomen↗

Suppressive effect of tetraprenylacetone on gastric atrophy induced by short-term administration of N-methyl-N'-nitro-N-nitrosoguanidine in rats.

BACKGROUND: Several studies have been reported on the effects of various therapeutic agents in enhancing or suppressing the carcinogenic activity of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). However, it is still unknown whether a mucosal protective agent could suppress its carcinogenic activity. METHODS: Twenty-five Wistar male rats were divided into four groups: group 1, MNNG alone; group 2, MNNG + tetraprenylacetone; group 3, control; group 4, tetraprenlacetone alone. MNNG 100 mg/mL, was freely given to groups 1 and 2, and tetraprenylacetone (200 mg/kg intraperitoneal) was additionally administered every other day to the rats in groups 2 and 4. The animals were sacrificed at 10 weeks and the gastric mucosa examined. RESULTS: Atrophic changes were observed in the antrum after 8 weeks of oral administration of MNNG. Furthermore, using immunohistological analysis with 5-bromo-2'-deoxyuridine (BrdU), the proliferative zone was found to be enlarged and shifted upward, although the BrdU labelling index of the proliferative zone was unaltered. Intraperitoneal administration of tetraprenylacetone every other day suppressed the MNNG-induced atrophic change and the alterations proliferative markers. Tetraprenylacetone alone did not have an effect either on morphological or proliferative markers. CONCLUSION: These observations suggest that gastric mucosal defensive factors may play critical roles in suppressing atrophic change inducing carcinogenesis by an exogenic carcinogen.

Animals↗

Changes in arrangement and in conformation of molecular components of peripheral T-cell antigen receptor complex after ligand binding: analyses by co-precipitation profiles.

Amounts of co-precipitating CD3 components by anti-T-cell receptor (TCR)V beta or anti-CD4/8 monoclonal antibodies were compared between non-stimulated and stimulated splenic T cells. The amounts of co-precipitating CD3 delta, epsilon and gamma chains with TCR alpha beta and with CD4/8 were not significantly changed after TCR ligation. The apparent amount of CD3 zeta chain co-precipitated with TCR alpha beta increased up to threefold, while the actual amount of co-precipitating CD3 zeta with TCR alpha beta and the total amount of specifically precipitated CD3 zeta are not changed after cross linking of TCR. The apparent amount of CD3 zeta chain co-precipitated with CD4/8 also increased. Unlike co-precipitation with TCR alpha beta, the actual amount of CD3 zeta co-precipitated with CD4/8 increased significantly. This observation suggests a conformational change as well as the relocation of CD3 zeta molecules within the TCR complex after the signal delivery. After TCR ligation, CD3 zeta chains relocate to the vicinity of either CD4 or CD8 molecules. In addition, when cross linking and binding signals are compared, CD3 chains undergo two distinct phases of conformational change. The responses, while the later conformational change caused by the cross linking of TCR does not induce but enhances the proliferative response.

Animals↗

Changes in cerebral blood flow accompanied with reduction of blood pressure treatment in patients with hypertensive intracerebral hemorrhages.

Blood pressure usually is reduced in patients with hypertensive intracerebral hemorrhage for the prevention of the expansion of the hematoma and recurrent hemorrhage in acute stage. However, disturbed autoregulation of cerebral circulation is expected, and decreased cerebral blood flow (CBF) caused by excessive hypotension has been pointed out. There are different mechanisms of action in hypotensives, thereby the influence of hypotension on CBF in patients with the thalamic hemorrhage was investigated using nitroglycerin (TNG), diltiazem hydrochloride (DH) and trimethaphan camsilate (TC). Average CBF in a hemisphere on the hematoma side, the hemisphere without hematoma, and around the hematoma showed a slight decline after administration of TNG or DH. However, CBF declined more, after TC than DH. DH and TNG are preferable in descending order to control blood pressure of patients with intracerebral hemorrhage in the acute stages in view of a smaller decline in CBF.

Adult↗

Isoform-specific monoclonal antibodies against HSP90.

The purpose of this study was to develop monoclonal antibodies (mAbs) that distinguish between the two isoforms of human 90-kDa heat shock protein (HSP90), i.e., HSP90 alpha and beta. Human HSP90 alpha and beta isoforms expressed in Escherichia coli were separately used as antigens for developing the mAbs. Twenty-three and ten mAbs were obtained by immunization of mice with HSP90 alpha and beta, respectively. Among them, ten and three mAbs specifically recognized HSP90 alpha and beta isoforms, respectively, on the criteria of both enzyme-linked immunosorbent and immunoblotting analyses. Immunochemical analysis by use of the mAbs revealed that both of the HSP90 isoforms were present in human cells even under unstressed conditions and that the expression of HSP90 alpha was more strongly induced when the cells were exposed to arsenate. This is the first report of the development of the mAbs discriminating between the two isoforms of HSP90. The mAbs specific for HSP90 isoforms should be useful for the regulational and functional analyses of HSP90 isoforms.

Animals↗

Elevated plasma levels of interleukin-1 receptor antagonist and interleukin-10 in patients with acute myocardial infarction.

The study was undertaken to measure plasma interleukin-1 (IL-1) receptor antagonist and IL-10 concentrations in patients with acute myocardial infarction and to analyze their relationship to the hemodynamics, severity, and prognosis of myocardial infarction in its acute stages. We attempted to define the kinetics of IL-1 receptor antagonist and IL-10 in patients with acute myocardial infarction (n = 34, age 42-91 years, mean 68 years). Plasma IL-1 receptor antagonist and IL-10 levels were measured by enzyme-linked immunosorbent assay. Patients in group A (n = 17) had uncomplicated acute myocardial infarction (Killip class I). Patients in group B (n = 17) had severe acute myocardial infarction (Killip class II, III, or IV). Peak Il-1 receptor antagonist and IL-10 levels in group B were significantly higher (p < 0.05) than those of group A. In group B, the peak IL-1 receptor antagonist levels were significantly correlated with white blood cell counts (r = 0.63, p = 0.006), pulmonary capillary wedge pressure (r = 0.78, p = 0.0002), and cardiac index (r = -0.51, p = 0.04). Peak IL-10 levels were significantly correlated with white blood cell counts (r = 0.60, p = 0.01), the pulmonary wedge pressure (r = 0.73, p = 0.0008), and cardiac index (r = -0.50, p = 0.04). Moreover, a significant correlation was found between the peak IL-1 receptor antagonist and IL-10 levels (r = 0.91, p < 0.0001). The peak IL-1 receptor antagonist levels in nonsurvivors (n = 13) were significantly higher (p < 0.01) than those in survivors (n = 21). The plasma IL-1 receptor antagonist and IL-10 levels were closely correlated with the severity of hemodynamics in acute myocardial infarction and with the clinical status of patients with severe acute myocardial infarction. Results suggest that plasma IL-1 receptor antagonist and IL-10 can serve as prognostic indicators in cases of sever acute myocardial infarction.

Adult↗

Inhibitory effects of metronidazole on anaerobic metabolism of phenylalanine and leucine by Peptostreptococcus anaerobius.

The metabolism of phenylalanine and leucine by Peptostreptococcus anaerobius and the effects of metronidazole on this metabolism were studied. The cells metabolized both phenylalanine and leucine under strict anaerobic conditions and produced hydroxylated products (i.e. phenyllactate from phenylalanine and hydroxyisocaproate from leucine). Other products were phenylpropionate, cinnamate and phenylacetate from phenylalanine; isocaproate, alpha-ketoisocaproate and isovalerate from leucine. Metronidazole inhibited the production of all end-products, with the exception of phenyllactate and hydroxyisocaproate which were increased. Similarly, when the reaction was carried out aerobically, the cells also metabolized both phenylalanine and leucine and all end-products were inhibited again with the exception of phenyllactate and hydroxyisocaproate. Our results suggest that the inhibitory mode of metronidazole is similar to that of oxygen.

Anaerobiosis↗

Molecular mechanism of S-modulin action: binding target and effect of ATP.

S-Modulin is suggested to increase the light sensitivity of rods by inhibiting phosphorylation of light-activated rhodopsin (Rh*) at high Ca2+ concentrations. The inhibition of rhodopsin phosphorylation was almost constant over a wide range of the Rh*/S-modulin ratio (10(-4)-approximately 10[1]). A 125I-labeled cross-linker that had been conjugated with S-modulin interacted with a protein of 60 kDa, a molecular mass close to that of frog rhodopsin kinase. These results suggested that the target molecule of S-modulin is rhodopsin kinase. To investigate the mechanism of the S-modulin action, we measured rhodopsin phosphorylation in the presence and absence of inhibition by S-modulin at various timings of ATP addition. The results suggested the following in situ mechanism of S-modulin action. After light-activation of rhodopsin kinase, the S-modulin/Ca2+ complex binds to the activated kinase and inhibits the phosphorylation of rhodopsin. The complex, however, does not affect the overall kinetics of the phosphorylation. The inhibition of the kinase by S-modulin is reversible in terms of the Ca2+ concentration. On the other hand, the kinase activity decreases as a function of time, probably via autophosphorylation.

Adenosine Triphosphate↗

Cloning of a gene for chloroplast omega6 desaturase of a green alga, Chlamydomonas reinhardtii.

A gene for chloroplast omega6 desaturase, which catalyzes the desaturation of monoenoic to dienoic acids in chloroplasts, was isolated from Chlamydomonas reinhardtii. We first performed reverse transcriptase-polymerase chain reaction with oligonucleotide primers corresponding to regions conserved among plastid omega6 desaturases of higher plants and delta12 desaturases of cyanobacteria, using C. reinhardtii poly(A)+ RNA. An amplified DNA fragment of 0.5 kb, containing a frame for a protein homologous to these desaturases, was used as a probe for screening cDNA and genomic DNA libraries of C. reinhardtii. The cDNA clone of 2.2 kb obtained contained an open reading frame encoding a protein of 424 amino acids with a putative molecular mass of 48.4 kDa, the amino acid sequence of which showed 46-51% homology to those of higher plant plastid omega6 and cyanobacterial delta12 desaturases. Introduction of the cloned genomic counterpart of this cDNA, designated as des6, into a Chlamydomonas mutant with defects in chloroplast omega6 desaturation and in the activities of photosystems I and II (PSI and PSII) complemented the desaturation mutation, indicating that the des6 gene codes for chloroplast omega6 desaturase. The complemented strains did not recover from the photosynthetic lesions, but showed lower PSII activity at 45 degrees C than the desaturation mutant, proving that the photosynthetic lesions in hf-9 are not caused by the desaturation mutation, and that the lowered unsaturation level of chloroplast lipids in the mutant is responsible for the expression at this high temperature of PSII activity, one of the thylakoid membrane functions.

Animals↗

Cellular proliferation and differentiation in rat atrophic gastric mucosa induced by N'-methyl-N'-nitro-N-nitrosoguanidine.

It has been demonstrated that mucosal cell proliferation in atrophic gastritis is significantly accelerated, although the dynamics of the cell proliferation and differentiation have not been well characterized. We sequentially analyzed the markers of proliferation and differentiation of rat atrophic gastritis induced by MNNG. Immunohistochemical staining by anti-BrdU, anti-PCNA, and anti-PDGF-BB antibodies demonstrated that cell proliferation in atrophic gastritis was accelerated not only in the parenchymal tissue but also in the mesenchymal tissue. Expression of a gap junction protein (connexin 32), which is a marker for differentiation of epithelial cells, was reduced during the progression of atrophy. Some apoptotic cells were observed in the lower to middle third of the atrophic mucosa, whereas apoptotic cells were rarely seen in normal mucosa, which suggests that apoptosis in these parts of the mucosa may be related to the occurrence of mucosal atrophy.

Animals↗

Polyarteritis nodosa associated with gastric carcinoma and hepatitis B virus infection.

Several cases of polyarteritis nodosa associated with malignant disorders have been reported, most with bone marrow-related tumors. We report polyarteritis nodosa presenting with a fever of unknown origin and muscle weakness that was complicated by advanced gastric carcinoma and hepatitis B virus-positive cirrhosis. Vasculitis was diagnosed after gastrectomy from histologic findings of arterial vasculitis on the resected gastric carcinoma. Our case is so far the second such report of polyarteritis nodosa associated with gastric cancer.

Hepatitis B↗