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Biomedical subjects

Y Ohshima

Publications and source records attributed to Y Ohshima.

At least 163 records · Page 9Linked to original sources
PubMed1988

Lectin-mediated cell-attachment and phagocytosis of Staphylococcus saprophyticus strain S1.

Adhesion studies with cryotome sections of human kidney and lung respectively uroepithelial cells together with blocking experiments with competitive carbohydrates suggested that specific attachment of S. saprophyticus strain S 1 to host cells apparently is mediated by lectins. Accordingly, microbial lectin blocking with specific glycoconjugates or lectin dysfunction (after treatment of bacteria with subinhibitory concentrations of tunicamycin) significantly decreased staphylococcal adherence to epithelial cells. Chemiluminescence measurements of human polymorphonuclear leukocyte (PMN) function yielded results suggesting importance of lectin-receptor interaction in phagocytosis, too, since PMN activity was significantly decreased after staphylococcal lectin blocking or dysfunction.

Bacterial Adhesion↗
PubMed1988

Immunostimulating staphylococcal lipoteichoic acid prevents pulmonary tumor colonization in BALB/c-mice.

Immunostimulating and antineoplastic activities of staphylococcal lipoteichoic acid (LTA) were studied in Balb/c-mice. Systemic administration of LTA (1 mg or 2 mg i. p., 7 and 4 days prior to challenge) significantly enhanced chemiluminescence response of peritoneal macrophages (p less than 0.0125) and induced enlargement of the spleen (p less than 0.025) as compared to non-treated controls. In vivo the number of lung colonies was significantly lower (p less than 0.0125) in LTA-treated mice 14 days after challenge with L-1 sarcoma cells.

Adjuvants, Immunologic↗
PubMed1988

Cell surface antigen of encapsulated Staphylococcus epidermidis SE-360 protects mice from homologous infection.

Cell surface antigen was mechanically extracted from encapsulated strain SE-360 of Staphylococcus epidermidis and purified by DEAE-Sephadex A 25 (Cl- form) ion exchange chromatography. This antigen manifested type-specific activity and major sugar constituents were galactose, glucose and N-acetyl-glucosamine at the molar ratio 1.00:9.05:1.65. alpha-D-glucosyl- and N-acetyl-glucosaminyl-residues were closely correlated to the antigenic determinant. In mice, protection against homologous microorganisms could be achieved by active immunization with thus purified antigen. Type-specific opsonin in rabbit anti-SE-360 serum could also be absorbed.

Animals↗
PubMed1988

Intestinal microflora of BALB/c-mice and function of local immune cells.

BALB/c-mice were treated for 7 days with oral nonabsorbable dosages of mezlocillin to achieve digestive tract decontamination. Such a procedure resulted in rapid eradication of most species of aerobic and anaerobic intestinal microflora. Various functions of peritoneal macrophages (e.g. chemiluminescence response, chemotactic motility, bactericidal and cytostatic ability) and lymphocyte proliferation were decreased in decontaminated animals as compared to non-treated controls.

Animals↗
PubMed1988

Short donor site sequences inserted within the intron of beta-globin pre-mRNA serve for splicing in vitro.

We constructed SP6-human beta-globin derivative plasmids that included possible donor site (5' splice site) sequences at a specified position within the first intron. The runoff transcripts from these templates truncated in the second exon were examined for splicing in a nuclear extract from HeLa cells. In addition to the products from the authentic donor site, a corresponding set of novel products from the inserted, alternative donor site was generated. Thus, a short sequence inserted within an intron can be an active donor site signal in the presence of an authentic donor site. The active donor site sequences included a 9-nucleotide consensus sequence, 14- or 16-nucleotide sequences at the human beta-globin first or second donor, and those at simian virus 40 large T antigen or small t antigen donor. These included 3 to 8 nucleotides of an exon and 6 to 8 nucleotides of an intron. The activity of the inserted donor site relative to that of the authentic donor site depended on the donor sequence inserted. The relative activity also strongly depended on the concentrations of both KCl (40 to 100 mM) and MgCl2 (1.6 to 6.4 mM). At the higher KCl concentrations tested, all the inserted, or proximate, donor sites were more efficiently used. Under several conditions, some inserted donor sites were more active than was the authentic donor site. Our system provides an in vitro assay for donor site activity of a sequence to be tested.

Amino Acid Sequence↗
PubMed1987

Recognition of 5' and 3' splice site sequences in pre-mRNA studied with a filter binding technique.

A nuclear extract from HeLa cells was fractionated by DEAE-Sepharose chromatography. The obtained fractions were assayed for binding to an RNA transcript carrying a splice site sequence of 9-16 nucleotides by a filter binding technique. The U1 RNA-rich small nuclear ribonucleoprotein (snRNP) fractions, which showed binding activities for both 5' and 3' splice site RNAs, were studied for the sequence specificity of their binding. Results indicate that the U1-rich snRNP fraction can recognize both 5' and 3' splice site sequences. The U1 RNP, which was highly purified from the snRNP fractions, bound to at least some 5' splice site sequences, but not to a consensus 3' splice site sequence. Therefore, purified U1 RNP can directly recognize a 5' splice site, but not a 3' splice site. The binding activity for the 5' splice sites was lost either by digestion with micrococcal nuclease or by digestion of the 5' end of U1 RNA with RNase H and a complementary oligodeoxynucleotide, indicating the involvement of U1 RNA. Involvement of a protein moiety as well in this binding was suggested by the loss of binding activity upon heating at 60 degrees C. The binding activity to a 3' splice site sequence was not sensitive to digestion by micrococcal nuclease and was removed by protein A-coupled anti-Sm antibody. This activity was found in sucrose gradient fractions of about 8 S.

Base Sequence↗
PubMed1987

Signals for the selection of a splice site in pre-mRNA. Computer analysis of splice junction sequences and like sequences.

To evaluate the importance of the surrounding nucleotide sequence in the selection of a splice site for mRNA, we have carried out computer studies of eukaryotic protein genes whose entire nucleotide sequences were available. A splice site-like sequence that has a significant homology to the consensus splice junction sequences is frequently found within an intron and exon. It is found that the higher the homology of a candidate donor site sequence to the nine-nucleotide consensus sequence, the higher is its probability of being a donor site. For most of the donors, the stability of presumed base-pairing with U1-RNA is higher than that of donor-like sequences, if any, in the adjacent exon and intron. However, homology of a candidate acceptor sequence to the 15-nucleotide consensus is a poor criterion of an acceptor site. The presence of a sequence that could serve as a branch-point 18 to 37 nucleotides before an acceptor does not seem to be critical in distinguishing it from an acceptor-like sequence. For genes of human, rat, mouse and chicken, respectively, nucleotide frequencies around splice junctions of many genes have been calculated. They seem to be different at some positions around a donor site from species to species. The acceptors for these vertebrates have longer pyrimidine-rich regions than the previous consensus sequence. The newly derived nucleotide frequencies were used as the standard to calculate the weighted homology score of a candidate splice site sequence in a gene of the four species. This weighted homology score of the 40 to 60-nucleotide intron-exon sequence is a much better criterion of an acceptor. These results suggest that the most important signal in the selection of a splice resides in the surrounding nucleotide sequence. It is also suggested that the surrounding nucleotide sequence alone is not generally sufficient for the selection.

Animals↗
PubMed1987

Immunostimulating cell surface substance from Staphylococcus epidermidis strain ATCC-31432 prevents metastatic lung colonization in Balb/c-mice.

The antineoplastic activity of cell surface substance from Staphylococcus epidermidis ATCC-31432 (CSS-subfraction 2) in mice and its influence on the activation of human leucocytes was studied. After in vitro incubation with CSS-subfraction 2, human polymorphonuclear leucocytes were evidently stimulated. In vivo (Balb/c-mice) intraperitoneal application of subfraction 2 of CSS induced a considerable splenomegaly and greatly increased IgM levels indicating a strong immuno-stimulating activity. In order to evaluate antineoplastic effects of CSS-subfraction 2, we used sarcoma L-1 cells (Balb/c-mouse origin) which cause heavy tumor colonization of the lung. After single systemic injection of subfraction 2 of CSS, the number of lung tumor-cell colonies drastically decreased. The combination of this immunomodulating therapy with a temporary anticoagulation resulted in a further reduction of tumor colonies in the lungs of Balb/c-mice.

Adjuvants, Immunologic↗
PubMed1987

Neonatal subdural hematoma secondary to birth injury. Clinical analysis of 48 survivors.

In order to evaluate the treatment and prognosis of subdural hematoma in neonates, we analyzed 48 survivors in the 3-year period January 1979 to December 1981. Based on the CT findings, the hematomas were grouped into four types according to location: type I, localized around the posterior interhemispheral fissure (25 cases, 52%); type II, extending from the posterior interhemispheral fissure to the hemispheric convexity (5 cases, 10%); type III, extending from the incisura to the posterior fossa (15 cases, 31%); and type IV, subdural hematoma accompanied by intracerebral hemorrhage (3 cases, 7%). Intracranial pressure was measured via the anterior fontanel in 13 cases. In 10 cases of extensive hemorrhage, the pressure exceeded 200 mm H2O. The age of the patients was from 0 to 7 days. There were 36 mature (75%) and 12 premature (25%) infants. The mothers were primiparous in 27 cases (56%). Fetal presentation was cephalic in 38 cases (79%), in 10 of which (21%) suction delivery was performed, and breech in 11. The fundus oculi was examined in 32 cases. Retinal hemorrhage was noted in 12 cases; it did not correlate with the type of hematoma or the intracranial pressure. Operations were performed in 13 cases; 1 of type I, 4 of type II, 5 of type III, and 3 of type IV. Functional prognoses were found to be as follows: type I, normal 15, abnormal 4, undetermined 6; type II, normal 4, abnormal 1; type III, normal 13, abnormal 1, undetermined 1; type IV, normal 1, abnormal 2 cases.

Birth Injuries↗
PubMed1987

Reverse-transcribed pseudogenes of U1 small nuclear RNA presumably amplified in the rat genome together with the flanking region.

We have examined 25 independent rat genomic clones each of which contains a U1 RNA gene or a pseudogene. We have found five clones whose restriction maps are identical with or overlapping one another. These clones contain sequences which are co-linear with that of U1 RNA except for one or two nucleotides (nt). They also contain 21-23-nt poly(A) stretches immediately after U1 RNA homology. In addition, the U1 RNA-poly(A) regions are surrounded by the same direct repeat sequences. Therefore, they seem to be pseudogenes which have been generated by the reverse transcription of poly(A)-tailed U1 RNA followed by the insertion of the transcript into the genome. Furthermore, conservation of the sequences extends over at least 18 kb of the flanking sequences. This suggests a family of conserved reverse-transcribed pseudogenes, which implies amplification of an original pseudogene. It is also suggested that the target sequence without insertion of a U1 RNA sequence has been amplified. The mechanisms for the amplification and sequence conservation are discussed.

Animals↗
PubMed1987

Platelet aggregation by group B streptococci.

Forty-six strains of group B streptococci (GBS), including various serotypes and non-serotypable strains, were tested for their ability to induce platelet aggregation in human platelet-rich plasma; four strains, all belonging to type III, showed a positive reaction. The characteristics of the reaction were investigated in these four positive strains. Aggregation was dependent on the ratio of bacteria to platelets, being maximal at a ratio of 4.3. Platelet aggregation was inhibited by EDTA (100% inhibition at 3.1 mM), indomethacin (100% inhibition at 10 mM), acetylsalicylic acid (93-100% inhibition at 5.0 mM) and quinacrine (100% inhibition at 0.25 mM). Thus the reaction was cation-dependent and required cyclooxygenase activity. Assays for cytosolic lactate dehydrogenase did not indicate platelet lysis. GBS induced the release of [3H]serotonin, which was maximal (68-78%) at 10 min after the reaction was started. Experiments with gel-filtered platelets suggested that GBS-induced platelet aggregation required both fibrinogen and heat-resistant (56 degrees C, 30 min) serum factors. Type-specific antisera prevented the platelet aggregation activity of heat-killed bacteria, but not of live bacteria. Trypsin digestion of the bacterial cells caused an almost complete loss of the platelet aggregation activity.

Aspirin↗
PubMed1987

Induction of resistance in mice by the capsular polysaccharide antigens of Staphylococcus aureus.

Mice actively immunized with capsular polysaccharides extracted from capsular type strains A, B, C, and D, determined by the serum-soft agar technique, were protected against lethal infection by homologous strains, but no animals survived infection by heterologous substance immunization even with at high doses. Passive protective antibody in rabbit antisera prepared using these strains was absorbed out only by homologous capsular polysaccharide in mice. These results indicated that resistance was specific for capsular polysaccharide. The substance contained mainly neutral sugar, small amounts of hexosamine, methyl-pentose, and phosphate although these amounts varied depending on the capsular types strains.

Animals↗
PubMed1987

Induction and Characterization of Artificial Diploids from the Haploid Yeast Torulaspora delbrueckii.

The yeast Torulaspora delbrueckii, which propagates as a haploid, was made into a diploid by treatment with dimethyl sulfoxide (DMSO) on the regeneration of protoplasts. The diploid state was stably inherited; the cell volume was three times that of the parent strain and the cellular DNA content was two times that of the parental strain. No essential difference was found between diploids induced by DMSO and those formed through intraspecific protoplast fusion. The diploid strains sporulated fairly well, with their cells converting directly into asci. Random spore analysis revealed that diploids induced through protoplast fusion gave rise to auxotrophic segregants (haploids) with the parental genetic marker or to segregants formed by recombination, while diploids induced by DMSO from a doubly auxotrophic parent gave rise to no recombinant, indicating that it was chromosomally homoallelic in nature. The magnesium level in the protoplast regeneration medium was found to be an important factor for inducing diploid formation. At 0.2 mM magnesium diploids appeared even in the absence of DMSO, while at 2 mM magnesium diploids never appeared unless DMSO was added to the regeneration medium. Evidence is provided that the diploids induced by DMSO or a low magnesium level are due to direct diploidization but not protoplast fusion. UV light irradiation of intact cells (without protoplasts), 10% of which survived, also produced diploids among this surviving population. From these results we conclude that the perturbation of protoplast regeneration or of cell division by the treatments mentioned above somehow induced direct diploidization of T. delbrueckii.

Journal Article↗