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Biomedical subjects

T Ohshima

Publications and source records attributed to T Ohshima.

At least 325 records · Page 18Linked to original sources
PubMed1989

Variations in protein binding of drugs in plasma and serum.

The free fractions of five drugs were determined in plasma treated with EDTA and in serum. The free fractions of phenytoin, sodium valproate, and phenobarbital in serum were significantly higher than in plasma (P less than 0.05). In contrast, the free fractions of carbamazepine and theophylline were significantly lower in serum than in plasma. Although we observed minor differences in the protein patterns obtained with two-dimensional electrophoresis, these did not have an important influence on protein binding. No significant differences were observed between plasma and serum in the physiological data, except for pH. Binding of these drugs by protein was shown to be pH-dependent. We conclude that serum may be better than plasma for determination of the free fraction, because the free fraction in plasma is affected by anticoagulants such as EDTA salts and heparin.

Anticonvulsants↗
PubMed1989

[Comparison of conventional chemotherapy and short-term intensive chemotherapy in adult acute myeloid leukemia].

We designed a short term intensive chemotherapy (DCMP-85 regimen) that completed 4 courses of postremission therapy without maintenance therapy in an effort to improve remission durations for adult patients with AML. Twenty six patients aged 14-65 yr were entered into this study. The rate of complete remission is 76.9%. A Kaplan-Meier analysis of patients entering remission predicts that 56% of patients will remain in remission at 3 years (median follow-up is 12 mo). Comparison three different conventional chemotherapies (DC(M)P, DCMP Two Step and BH-AC DMP), DCMP-85 has achieved higher CR rate and 3-year leukemia-free survival rate, so far. Therefore, we conclude short term intensive chemotherapy is getting better results.

Adolescent↗
PubMed1989

[An autopsy case of multiple system atrophy with many senile plaques].

A 60 year-old man was admitted to our hospital because of gait disturbance and dizziness. At 57 years of age, he noticed his walking unstable. After then, he had dizziness due to orthostatic hypotension, urinary difficulty, loss of livid, and forgetfulness. Neurological examination revealed he had severe orthostatic hypotension, cerebellar ataxia, dysarthria, hyperreflexia of four limbs, myoclonus of right leg, and atonic bladder. His brain CT showed cerebellar atrophy. Thereafter he had recurrent syncopic attacks. His gait disturbance progressed steadily, so he became bedridden. In his terminal stage, his limbs showed rigidity. About 3 years later he died of pneumonia and sepsis. At autopsy brain weighted 1,230 g. Glossly the putamens was bilaterally shrunken, the color of the substantia nigra and locus ceruleus became pale. Base of the pons and the cerebellum were atrophic. Microscopical examination confirmed the degeneration of striato-nigral and olivo-ponto-cerebellar systems without Lewy body. In the spinal cord there was depletion of neuronal cells in the intermediolateral nuclei and Onufrowitz nuclei. In addition to the conventional neuropathological staining methods, we performed the immunohistochemical studies using monoclonal antibody against synthetic peptide of beta protein which detected senile plaque of every stages with formic acid pretreatment, and compared to the modified Bielschowsky method and Congo red method. Our case showed many very primitive and primitive senile plaque in neocortices and hippocampal region. A few neurofibrally tangle were seen in hippocampus. We supposed our case might combine multiple system atrophy and Alzheimer' pathology.

Alzheimer Disease↗
PubMed1989

[Salivary calculus of the parotid gland: report of a case].

We reported a case of parotid sialolithiasis. A man, aged 72, complained of swelling of the left buccal region accompanied by a sensation of inflammation. On diagnosis of sialolithiasis of the parotid gland, a surgical operation was performed with a small incision in the left cheek region. About one month after the operation, a salivary calculus was discharged spontaneously from the left buccal region. He recovered satisfactory without the formation of a parotid salivary fistula. Morphological observation on the salivary calculus was carried out using the scanning electron microscope (SEM).

Aged↗
PubMed1989

Activation of oncogenes in human oral cancer cells: a novel codon 13 mutation of c-H-ras-1 and concurrent amplifications of c-erbB-1 and c-myc.

By NIH3T3 transfection assay in conjunction with in vitro transient neomycin selection, activated c-H-ras-1 oncogenes were detected in two squamous cell carcinoma cell lines, ZA and HOC-313, newly established from human oral cancer patients. ZA had a point mutational activation at the 13th codon, this activation of c-H-ras-1 being novel in human cancer cells, while HOC-313 appeared to have an activation at the 12th codon. In ZA, 16- to 32-fold amplification of the EGF receptor gene, c-erbB-1 and a few-fold amplification of c-myc were detected. The significance of these findings is discussed in relation to multistep carcinogenesis in human cells.

Aged↗
PubMed1989

Protein binding characteristics of a new bronchodilator, 1-methyl-3-propylxanthine (MPX), in different species.

The protein binding of a new bronchodilator, 1-methyl-3-propylxanthine (MPX), in different animal species sera (human, dog, rabbit, rat and mouse) was investigated in vitro by ultrafiltration method. Interspecies differences in the binding affinity and binding capacity for MPX were observed. The dissociation constant (Kd1) for the high affinity binding site of human serum was approximately 17 times higher than that of rat serum. Those of rat serum and rabbit serum were nearly equal. On the other hand, the binding indices (n1p1/kd1) of human serum and rat serum were comparable. The binding of MPX to various proteins such as human serum albumin (HSA), alpha 1-acid glycoprotein (AGP) and isolated human lipoproteins was also investigated. The binding of MPX to HSA was lower than that of human serum protein but the number of specific binding sites (n1) in human serum was in agreement with that in HSA. The binding curves of MPX to AGP and lipoproteins were almost linear and the binding percentages were less than 7% in both cases. These data suggest that MPX is exclusively bound to albumin and that AGP and lipoproteins are of little importance in the protein binding of MPX. The present data appear to be useful for predicting the role of protein binding on the pharmacokinetics of MPX in different animal species.

Animals↗
PubMed1989

[Anesthetic management of a patient with perforated necrotic trachea after radical esophageal cancer surgery].

We anesthetized a 69-year-old man with perforated necrotic trachea after radical esophageal cancer surgery. We recognized during the reoperation that necrotizing part of the trachea was perforated from the cricoid cartilage to 2cm above the carina. We ventilated the both lungs successfully without leaks by inserting a double-lumen tube into the trachea after tracheostomy. Postoperatively, the carina and the main bronchi were also necrotized and perforated. We ventilated the both lungs without leaks by inserting two single-lumen tubes into the left and right bronchi separately. But, eventually the distal part of the both bronchi was also necrotized and perforated. Finally, the patient died of respiratory failure due to hypoventilation on the 7th day after the re-operation. We should remember this type of emergency complication after radical esophageal cancer surgery.

Aged↗
PubMed1989

Ultrastructural changes in human erythrocytes caused by heat and the localization of ABO blood group activity.

The ultrastructural changes in human erythrocytes caused by heat and the localization of ABO blood group activity have been studied electron microscopically. Osmotically hemolyzed cells were found to show varying degrees of cell matrix loss and a swelling of the membranes. By means of immunoferritin staining, ferritin particles, which indicate the localization of ABO group activity, were found to bind in clusters on small process-like structures on the membranes. Erythrocytes heated to 60 degrees C showed a hemolytic change; the cell matrix was released and small globular substances (ca. 50-100 nm in diameter) aggregated on the cytoplasmic side of the partially swollen cell membranes, a phenomenon that was not noted in erythrocytes that undergo a hypo-osmotic change. Erythrocytes heated to 100 degrees C underwent fragmentation, though the cell matrix was almost preserved and aggregated substances of varying size were noted inside the membranes. ABO group activity was demonstrated almost evenly on the sub-membrane aggregated substances and on the surface of the thermo-changed erythrocytes.

ABO Blood-Group System↗
PubMed1988

Gene cloning and sequence determination of leucine dehydrogenase from Bacillus stearothermophilus and structural comparison with other NAD(P)+-dependent dehydrogenases.

The gene for leucine dehydrogenase (EC 1.4.1.9) from Bacillus stearothermophilus was cloned and expressed in Escherichia coli. The selection for the cloned gene was based upon activity staining of the replica printed E. coli cells. A transformant showing high leucine dehydrogenase activity was found to carry an about 9 kilobase pair plasmid, which contained 4.6 kilobase pairs of B. stearothermophilus DNA. The nucleotide sequence including the 1287 base pair coding region of the leucine dehydrogenase gene was determined by the dideoxy chain termination method. The translated amino acid sequence was confirmed by automated Edman degradation of several peptide fragments produced from the purified enzyme by trypsin digestion. The polypeptide contained 429 amino acid residues corresponding to the subunit (Mr 49,000) of the hexameric enzyme. Comparison of the amino acid sequence of leucine dehydrogenase with those of other pyridine nucleotide dependent oxidoreductases registered in a protein data bank revealed significant sequence similarity, particularly between leucine and glutamate dehydrogenases, in the regions containing the coenzyme binding domain and certain specific residues with catalytic importance.

Amino Acid Oxidoreductases↗
PubMed1988

Yeast KEX2 genes encodes an endopeptidase homologous to subtilisin-like serine proteases.

Yeast Saccharomyces cerevisiae KEX2 gene previously isolated, was characterized as the gene encoding a calcium-dependent endopeptidase required for processing of precursors of alpha-factor and killer toxin. In this study, we report the amino acid sequence of the KEX2 gene product deduced from nucleotide sequencing. Our results indicate that the KEX2 gene contains a 2,442-bp open reading frame encoding a polypeptide of 814 amino acids. The deduced amino acid sequence contains a region extensively homologous to the members of subtilisin-like serine protease family near the N-terminus. A putative membrane-spanning domain near the C-terminus was also detected. These facts indicate that the KEX2-encoded protein may function as a membrane-bound, subtilisin-like serine protease.

Amino Acid Sequence↗
PubMed1988

Cloning and expression in Escherichia coli of two additional amylase genes of a strictly anaerobic thermophile, Dictyoglomus thermophilum, and their nucleotide sequences with extremely low guanine-plus-cytosine contents.

An obligately anaerobic and extremely thermophilic bacterium, Dictyoglomus thermophilum, produces multiple extracellular amylases. In addition to one of the amylase genes, amyA, which we previously cloned and characterized, we have cloned two additional genes, amyB and amyC, coding for amylases of this thermophile, into Escherichia coli and determined their nucleotide sequences. The two amylase genes were expressed under the control of E. coli promoters. Almost all activity was detected in the intracellular fraction in the E. coli cells. The molecular mass and NH2-terminal amino acid sequence of the AmyB enzyme, which was purified from an E. coli transformant containing the amyB gene, confirmed that the reading frame of amyB consisted of 562 amino acids (Mr 67,000). The molecular mass of the AmyC enzyme, estimated by activity staining of a crude extract of E. coli containing amyC, confirmed that AmyC consisted of 498 amino acids (Mr 59,000). The optimal temperatures for AmyB and AmyC activities on soluble starch were 80 degrees C and 70 degrees C, respectively. Both AmyB and AmyC showed a pH optimum of 5.5. AmyB and AmyC showed a different pattern of starch hydrolysis when examined by thin-layer chromatography. Some homology in the amino acid sequences with the functional regions of Taka-amylase A was found in both AmyB and AmyC. The codon usage in the amyA, amyB and amyC genes was highly biased, which reflects the fact that the guanine-plus-cytosine (G + C) content of DNA of D. thermophilum is 29 mol%. The distribution of G and C at each position of the codons was non-random; the G + C content of the first position of codons is significantly high, whereas that of the third position is somewhat low. In addition, codons consisting only of A and T were preferentially used in this thermophile.

Amino Acid Sequence↗
PubMed1988

Inhibitory activity of interleukin 2-activated lymphocytes on human granulocyte precursors (CFU-g).

Interleukin 2-activated lymphocytes (lymphokine-activated killer [LAK] cells) cultured from 2 to 14 days were added to the cultures of granulocyte precursors (CFU-g). The LAK cells inhibited colony formation of granulocyte precursors; LAK cells cultured for five days showed the strongest inhibitory activity on colony formation. The presence of cell-to-cell interaction between LAK cells and bone marrow mononuclear cells (BMNC) in CFU-g assays emphasized the LAK cell-derived colony inhibitory activity (LAK-CIA), but cell-to-cell interaction was not always a requirement for LAK-CIA, since LAK cells were also found to inhibit colony formation without such interaction. This report shows that LAK cells can inhibit in vitro colony formation of granulocyte precursors. We therefore concluded that the observed CIA is caused by soluble factor(s) derived from LAK cells, and that E-rosette-forming cells are manifesting LAK-CIA.

Colony-Forming Units Assay↗
PubMed1988

Human-type blood group activities on chimpanzee erythrocytes with special reference to M and N.

Human-type blood group activities on the red blood cells (RBCs) of three chimpanzees were individually examined with commercial mouse monoclonal antibodies (anti-A, -B, -H, -M, -N, -Lea, and -Leb) as well as lectins (UEA-I and VGA) and conventional polyclonal antisera for the systems ABO, MN, Lewis, Rh-Hr, P, Kell, Kidd, Duffy, and Lutheran. For further analysis of the MN antigens, treatment of the RBCs with sialidase, trypsin, and chymotrypsin were employed. The activities recognized among the three chimpanzees were A, H, M, N, Leb, c, S, k, and Jka. The RBCs of the three individuals possessed the A antigen which showed the same serologic activity as the human A1. Those chimpanzee RBCs showed higher H-activity than the human A1 RBCs. The Lewis b activity was revealed by the absorption-elution method. The RBCs of the three individuals showed a reactivity to the polyclonal anti-M reagents, which was affected by both the sialidase and trypsin treatment. The RBCs of two individuals were agglutinated with the monoclonal anti-N. The receptor was sensitive to sialidase and chymotrypsin. The RBCs of the three individuals, however, did not react with the monoclonal anti-M or with one of the polyclonal anti-N. These results indicate structural differences in the glycophorins and MN antigens between the human and chimpanzee.

ABO Blood-Group System↗
PubMed1988

Immunocytochemical study on the ultrastructural localization of human-type ABO (H)-blood group activities in a macaque (Macaca irus).

The immunocytochemical study on the ultrastructural localization of human-type ABO(H)-activities in a crab-eating macaque (Macaca irus) was carried out by using postembedding and immuno-gold staining method. The tissue specimens examined were the esophagus, stomach (St), small intestine (Si), large intestine, liver, kidney, and pancreas. The specimens from these organs and submandibular gland (Sg) of a human (O-group) were used as staining reaction controls. Primary and secondary antibodies were commercially obtained mouse monoclonal anti-A, -B, -H (IgM), and goat anti-mouse IgM labeled with colloidal gold particles (luminal diameter 20 nm), respectively. The results were as follows: (1) In macaque specimens, only A-activity could be observed as the location of gold particles on the peripheral rim of serous secretory granules (Sg) and of epithelial cells (esophagus), the mucous droplets in epithelial cells and brush border (St, Si), the intracellular secretory canaliculi [ISC (St)] and the zymogen granules and secretory ducts (pancreas). Gold particles could be also noted at the Golgi apparatus and nascent secretory granules. (2) By periodic acid-thiocarbohydrazide-osmium tetroxide (PA-TCH-OS) reaction, hexose-rich neutral mucopolysaccharides were noted on the peripheral rim of serous secretory granules (Sg), the mucous droplets (St, Si), the ISC (St), and the brush border (Si). Such a distributional pattern corresponded well with that of gold particles, indicating that the substances were responsible for ABO(H)-activities.

ABO Blood-Group System↗
PubMed1988

Immunohistochemical study of blood group activities in the alimentary canal in normal and pathologic conditions with reference to the nature of epithelial mucopolysaccharides.

ABO(H)- and Lewis-activities in the human alimentary canal were investigated by the avidin-biotin-peroxidase complex method using commercial mouse monoclonal antibodies in order to clarify (1) the changes apparent in these activities in carcinoma of the digestive tract or premalignant intestinal metaplasia of the stomach and (2) the relationship of the activities to the histochemical properties of epithelial mucins examined by Alcian blue (pH 2.5)/periodic acid-Schiff (PAS) double staining. In intestinal metaplasia, ectopic goblet cells showed various degrees of ABO(H)-activities according to the PAS stainability of mucins and revealed Lea-activity even in a Le(a-b+) group. In carcinoma of the alimentary canal, ABO(H)- and Lewis-activities compatible with the donors' blood groups were located mainly at the PAS-positive cell surface, and in the Golgi bodies and secretions. It was thus concluded that quantitative and/or qualitative changes in blood group activities were closely associated with those of epithelial mucopolysaccharides.

ABO Blood-Group System↗