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Biomedical subjects

M Ota

Publications and source records attributed to M Ota.

At least 253 records · Page 14Linked to original sources
PubMed1990

A simple and rapid method for HLA-DP genotyping by digestion of PCR-amplified DNA with allele-specific restriction endonucleases.

We previously reported a simple and rapid method for HLA-DQA genotyping by digestion of polymerase chain reaction-amplified DQA genes with allele-specific restriction endonucleases. Here we report the application of this method to DP genotyping. The second exon of the HLA-DPB genes was selectively amplified from genomic DNAs of 72 HLA-D homozygous B-cell lines by the polymerase chain reaction method. Amplified DNAs were digested with ApaI, SacI, BstUI, FokI, and RsaI, which can recognize allelic sequence variations in the polymorphic segments of the DPB second exon and then subjected to electrophoresis in polyacrylamide gels. Sixteen different polymorphic patterns of the restriction fragments were found, and twelve were identical to patterns predicted from the known DNA sequences correlating with each HLA-DPw specificity defined by cellular typing. The other four patterns were distinct from those of the known DPw specificities, suggesting the presence of novel DP alleles. This polymerase chain reaction-restriction fragment length polymorphism method provides a simple and rapid technique for accurate definition of HLA-DP types at the nucleotide level, replacing the technically demanding method of primed lymphocyte typing.

Alleles↗
PubMed1990

A simple and rapid method for HLA-DRB and -DQB typing by digestion of PCR-amplified DNA with allele specific restriction endonucleases.

The polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method, which we previously reported as an efficient and convenient typing technique for accurate definition of the HLA-DQA1 and -DPB1 alleles, is now extended and applied to HLA-DRB and -DQB typing. The second exon of the HLA-DRB (B1 and B3 or B4) and DQB (B1 and B2) genes was selectively amplified from genomic DNAs of 70 HLA-homozygous B cell lines by PCR. Amplified DNAs were digested with the restriction endonucleases, which can recognize allelic variations specific for HLA-DR, -DQ, and -Dw allospecificities and then subjected to electrophoresis in polyacrylamide gel. Of DRB genes, FokI, HinfI, HhaI, HphI, KpnI and SacII were selected and the 20 different polymorphic patterns of the restriction fragments thus obtained were found to correlate with each HLA-DR and -Dw type defined by serological and cellular typing. Of the DQB genes, FokI, HaeIII, HhaI, RsaI and Sau3AI produced nine different polymorphic patterns of the restriction fragments, correlating with the HLA-DQ and -Dw types. This PCR-RFLP method provides a simple and rapid technique for accurate definition of the HLA-DR, -DQ and -Dw types at the nucleotide level, eliminating the need for radioisotope as well as allele specific oligonucleotide probes.

Alleles↗
PubMed1990

Antagonism of ethanol intoxication in rats by inhibitors of phenylethanolamine N-methyltransferase.

The probable involvement of brain epinephrine in the expression of the acute sedative and intoxicating effects of ethanol and pentobarbital is demonstrated. Two selective inhibitors of phenylethanolamine N-methyltransferase (PNMT), LY134046 and LY78335, proved to be potent and long-lasting antagonists of ethanol intoxication in rats. Acute antagonism of pentobarbital-induced intoxication was observed with LY134046. The present results are compatible with a role for central epinephrine synthesis in ethanol and pentobarbital-induced sedation and intoxication in rats.

Alcoholic Intoxication↗
PubMed1990

[Quantitation of IgG antibodies to type Ia group B streptococcal type-specific polysaccharides measured by enzyme-linked immunosorbent assay].

The type-specific polysaccharide antigen of the group B streptococcus (GBS) type Ia as extracted and purified according to the procedures of Kane and Karakawa. Using this purified polysaccharide antigen, we made a sensitive and specific assay system of enzyme-linked immunosorbent assay (ELISA) and measured the titres of of the type-specific antibodies in maternal sera and cord blood sera. The titres of antibodies in 78 pregnant women (26 Ia carriers, 18 other types of GBS carriers and 34 non carriers) were compared. A mother of an infant affected by early onset infection of GBS Ia had a titre of antibody 1:10 at delivery, while 2 years later she became a non carrier and had a titre of antibody over 1:160. The titres of antibodies in 27 pair sera of mothers and cords were well correlated.

Antibodies, Bacterial↗
PubMed1990

N-terminal extension of sweet peptides in relation to the structural features of peptide sweeteners.

Sweet aspartyl di- and tripeptide esters were extended toward the N-terminus in relation to the structural features of sweet peptides. The sweet peptides were designed on the basis of the receptor site model. It was found that an extension of the sweet aspartyl dipeptide esters by adding a small D-amino acid residue mostly gave sweet compounds (e.g., D-Ala-L-Asp-D-Ala-OMe), although this significantly decreased their sweetness potencies. Further extension at the N-terminus of the extended sweet tripeptide esters to yield the tetrapeptide esters resulted in a loss of the sweet taste. The N-terminal extension of sweet aspartyl tripeptide esters resulted in faintly sweet or nonsweet tetrapeptide esters. Interestingly, an analogous extension at the N-terminus of the sweet aminomalonyl dipeptide esters gave bitter compounds (e.g., D-Ala-DL-Ama-L-Phe-OMe). These results indicate that the receptor has a small space that can accomodate an additional small D-amino acid residue at the site facing the N-terminus of sweet aspartyl dipeptide esters.

Amino Acid Sequence↗
PubMed1990

[Quantitative evaluation by measuring affected area for cytotoxicity of dental materials].

The agar overlay method was modified and employed in this study to quantitatively evaluate the cytotoxicity of dental materials. The area of fibroblasts affected by the release of toxic substances from tested materials was determined with an image analysis system in order to compare the relative cytotoxicity of amalgams and composite resins. The cytotoxicity of amalgams decreased by 50% or more with aging time. Zinc appeared to be the major contributor to the cytotoxicity of amalgams. The cytotoxicity of chemically cured composite resins was also reduced with hardening time. However, even after 24 hours, the cytotoxicity of all chemically cured composite resins was still apparent. The cytotoxicity of light cured composite resins was dramatically reduced after illumination with the light source. The cytotoxicity of both amalgams and composite resins was strongly dependent upon their hardening process, since aging and curing produced a more stable material which was less likely to release toxic substances into the culture medium. The technique developed in the present study for measuring the area of affected cells to quantitatively evaluate the cytotoxicity of dental materials is simple and reproducible.

Composite Resins↗
PubMed1990

[Transverse strength and acoustic emission characteristics of commercial denture base resins].

Transverse strength and acoustic emission (AE) characteristics were measured by the transverse test in deionized water at 37 degrees C on commercial denture base resins (five heat-cured type resins and one polysulfone). Difference in flexural property of five heat-cured denture base resins was not shown from the transverse deflection according to JIS, but high toughness of polysulfone was recognized in transverse deflection, flexural strength, flexural modulus, flexural rigidity, flexural proof stress, and fracture energy. The five heat-cured denture base resins showed a low AE activity, but the polysulfone resin high AE activity. Significant rates of AE for polysulfone were detected at a kgf of approximately 50-60% the maximum load. The presence of Kaiser effect in its cycle transverse test was confirmed.

Acoustics↗
PubMed1990

Post-hysterectomy extra-uterine endometrial stromal sarcoma: a case report.

Endometrial stromal sarcoma is a relatively rare form of uterine sarcoma. The present paper reports on a patient who was found to have contracted endometrial stromal sarcoma eight years after a hysterectomy. A postoperative specimen revealed this sarcoma to have originated in extra-uterine endometrium of the rectovaginal septum.

Adult↗
PubMed1990

Natural killer cell immunodeficiency in siblings: defective killing in the absence of natural killer cytotoxic factor activity in natural killer and lymphokine-activated killer cytotoxicities.

A immunodeficiency of natural killer cells as effectors for natural killer and lymphokine-activated killer cytotoxicities was first demonstrated in siblings. Two of three male siblings persistently lacked natural killer activity against K562 target cells as assayed by a 51Cr-release assay: percent lysis values were less than 1.0% as compared to the normal lymphocyte values of 43.5% +/- 6.2% (mean +/- SD). Their lymphocytes did not develop natural killer cell activity by changing effector to target ratios, prolonging the incubation time, or stimulating them with interferon-alpha or interleukin 2. Numbers of lymphocytes bearing Leu-7, CD16, or NKH-1 were normal but those of Leu-7-, CD16+ cells were decreased as estimated by flow cytometry. Single cell-in-agarose assays showed normal numbers of natural killer cells capable of binding to a target cell but incapable of killing it. They had depressed levels of lymphokine-activated killer activity, which was totally eliminated by the treatment with OKT3 and complement. This result indicates that the patients' natural killer cells are also defective in the capacity to work as effectors for lymphokine-activated killer activity. The patients' natural killer cells did not produce natural killer cytotoxic factor activity. Antibody-dependent cellular cytotoxicity and cytotoxic T lymphocyte cytotoxicity were normal. These results demonstrate a selective natural killer cell deficiency as effectors for natural killer and lymphokine-activated killer cytotoxicities with a familial tendency, in which there is defective killing with the absence of natural killer cytotoxic factor activity.

Child↗
PubMed1990

A case of death due to dragging by a car: establishment of a homicide because of conscious negligence.

A drunken pedestrian (a 19-year-old male) lying on the street was hit by a car at night and dragged to a distance of about 600 meters. He was taken to a hospital soon but died there from brain injuries. His head and back were strongly and widely grazed against the road surface. Major parts of the left side of his skull and the brain were scraped off. The assailant thought that he had to escape from the place, because he was on probation because of established crime of drunken driving and drug abuse. As he continued to drive even after noticing that his car was dragging the victim, realizing that he might die as a consequence, a case of homicide due to conscious negligence was established.

Accidents, Traffic↗
PubMed1990

[A randomized controlled study of PSK combined immuno-chemotherapy for adenocarcinoma of the lung. The Advanced Lung Cancer Immuno-chemotherapy Study Group].

In a randomized controlled study of chemotherapy (CDDP 100 mg/m2 day 1, VDS 3 mg/m2 day 1, 8, 15) vs. immuno-chemotherapy combined with PSK 3 g/day for adenocarcinoma of the lung (stage III, IV, p. s. 0, 1, 2), response rate for 169 cases with completed extramural review was 14.2%. As for the response rates for 138 complete cases, the chemotherapy group showed 17.9%, and the immuno-chemotherapy group was 16.9%. MST were 330 days and 331 days, respectively. In stage III cases, the response rates were 11.1% in the chemotherapy group and 37.5% in the immuno-chemotherapy group (p = 0.046). MST were 457 days (65.3 weeks) and 576 days (82.3 weeks), respectively. In terms of survival curve, it was suggested that the immuno-chemotherapy group was superior to the chemotherapy group (logrank test p = 0.075), but in stage IV cases, there was nothing outstanding in the immunochemotherapy group.

Adenocarcinoma↗
PubMed1989

Behavioral, hormonal and neurochemical characteristics of aggressive alpha-mice.

The present study examined the behavioral, neurochemical and endocrinological characteristics of aggressive, male alpha-mice. These mice inflict severe bite marks on other male mice in their cage, but are not attacked themselves. The characteristics of the alpha-mice were compared with those of submissive mice, and of control mice taken from cages in which no severe fighting was observed. The behavioral tests used were Porsolt's swim test of behavioral 'despair', a plusmaze test of anxiety, a holeboard test of exploration and locomotor activity, and a test of seizure threshold to bicuculline. The alpha-mice were found to be immobile in the swim test for a shorter time than the submissive and control mice, and the submissive mice for a longer time than the controls. In the holeboard, the alpha-mice spent less time making exploratory head-dips than the other mice. Submissive mice had elevated 5-HIAA levels in the hypothalamus, hippocampus and brainstem, and the alpha mice had reduced concentrations of dopamine in the brainstem. There were no significant differences in plasma corticosterone or testosterone concentrations between the groups. These findings indicate that in alpha-mice, a number of behavioral and neurochemical characteristics appear together with the unusually high aggressiveness towards cage-mates.

Aggression↗
PubMed1989

Autocrine growth factor in defined serum-free medium of human salivary gland adenocarcinoma cell line HSG.

Human salivary gland adenocarcinoma cell line HSG secretes an epidermal growth factor (EGF)-like molecule and contains EGF receptors. Growth of HSG cells is inhibited by glucocorticoid. We have identified that the growth inhibition by glucocorticoid is induced by the reduced secretion of the EGF-like molecule and that addition of anti-human EGF antibody to the culture specifically inhibits the growth of HSG cells, suggesting that autocrine secretion is involved in the growth of HSG cells. To prove that autocrine secretion functions in glucocorticoid-regulated growth of the HSG cell line, we purified the EGF-like molecule from serum-free, defined medium conditioned by the HSG cells and examined the growth-stimulatory effect of the purified molecule. The cultivation of HSG cells in serum-free defined medium, which contains insulin (10 micrograms/ml) and transferrin (10 micrograms/ml) only as proteinaceous components, resulted in establishment of a new cell line (HSG-SF) which had different morphological features from the parental HSG cell line. HSG-SF cells were found to have basically the same responsiveness to glucocorticoid as parental HSG cells. Parental HSG cells secreted high molecular weight EGF-like molecules (Mr 46,000 and 57,000), which were recognized by specific antibody to low molecular weight human EGF (Mr 6,201). From conditioned, serum-free medium of HSG-SF cells, an EGF-like molecule (Mr 46,000) was purified by using an anti-human EGF antibody-coupled Sepharose CL-4B column. This EGF-like molecule induced a maximal increase (36%) in incorporation of [3H]thymidine into DNA of parental HSG cells as well as low molecular weight human EGF. These observations demonstrate that growth of the HSG cell line is regulated by autocrine secretion.

Adenocarcinoma↗
PubMed1989

Effect of isolation on brain monoamines and the behavior of mice in tests of exploration, locomotion, anxiety and behavioral 'despair'.

We have recently found that, besides an increase in aggression, isolation increases social interaction in NIH Swiss mice. In the present study the effect of isolation in other behavioral paradigms and their relation to brain monoamine concentrations were investigated. Mice, isolated for 0-20 days, were tested in the holeboard test of exploration and locomotor activity, the plus-maze test of anxiety, and Porsolt's swim test of behavioral 'despair.' Isolation reduced exploratory head-dipping, increased locomotor activity and increased the preference of mice for the open arms of the plus-maze. The immobility time in the swim test was shortened in mice isolated for 2 or 5 days, suggesting an improved ability to cope with stressful situations. Monoamine assays failed to show significant changes in the noradrenaline, dopamine, HVA, 5-HT or 5-HIAA contents in the amygdala, hypothalamus, hippocampus or brain stem. The results indicate that isolation of NIH Swiss mice for less than three weeks induces several behavioral changes, but is not particularly stressful.

Animals↗
PubMed1989

Availability of immunostaining methods for identification of mixed-up tissue specimens.

The immunostaining method of ABO blood determinants in tissue sections is useful when surgical tissue specimens, submitted for diagnosis, are improperly identified. This article reports two cases in which the surgical specimens became mixed-up and the ABO immunostaining method was used to identify the patients' tissues correctly and prevent unnecessary surgery.

Biopsy↗
PubMed1989

Anaphylactoid reactions to vascular graft material presenting with vasodilation and subsequent disseminated intravascular coagulation.

This report describes five patients who had immediate adverse reactions following placement of a vascular graft. All had unusually persistent decreases in systemic vascular resistance, and four of these patients had bleeding as an early manifestation of this reaction. In two of three patients in whom the graft was replaced, uneventful recovery followed. Both patients in whom the graft was not replaced died. Blood samples from two of the patients demonstrated activation of complement and of the kinin system, whereas control patients did not demonstrate increased levels of activation products from these cascade systems. Recognition of this syndrome is important to patient survival, which appears to depend on rapid replacement of the graft.

Aged↗
PubMed1989

A simple and rapid method for HLA-DQA1 genotyping by digestion of PCR-amplified DNA with allele specific restriction endonucleases.

The second exon of the HLA-DQA1 genes was selectively amplified from genomic DNAs of 72 HLA-homozygous B cell lines by the polymerase chain reaction (PCR). Amplified DNAs were digested with HaeIII, Ddel, ScrFI, FokI and RsaI, which recognize allelic sequence variations in the polymorphic segments of the DQA1 second exon, and then subjected to electrophoresis in polyacrylamide gels. Eight different polymorphic patterns of restriction fragments were obtained, and seven were identical to patterns predicted from the known DNA sequences, correlating with each HLA-DQw type defined by serological typing. The remaining one pattern cannot be explained from the sequence data, suggesting the presence of a novel DQA1 allele at the nucleotide level. This PCR-RFLP method provides a simple and rapid technique for accurate definition of the HLA-DQ types at the nucleotide level, eliminating the need for radioisotope as well as allele specific oligonucleotide probes and can be extended and applied to HLA-DR, -Dw DP typing.

Alleles↗