Biomedical subjects
Publications and source records attributed to T Kudo.
A new allele of the human diaphorase DIA3 is described. The new allele which is termed DIA5(3) was detected in two Japanese males during the course of electrofocusing analysis of the genetic variants of the human diaphorase DIA3 locus.
The cellulase gene from the alkalophilic Bacillus sp. strain 1139 was cloned in Escherichia coli using pBR322. Plasmid pFK1 was isolated from transformants producing cellulase, and the cloned cellulase gene was found to be in a 4 X 6 kb HindIII fragment. The cellulase gene was subcloned in a functional state on a 2 X 9 kb DNA fragment and its nucleotide sequence was determined. The coding sequence showed an open reading frame encoding 800 amino acids. The pFK1-encoded cellulase had the same enzymic properties as the extracellular cellulase produced by the alkalophilic Bacillus sp. strain 1139, but its Mr was slightly higher.
Most of the cloned penicillinase from alkalophilic Bacillus sp. strain 170 and alkaline phosphatase were released into the culture medium by Escherichia coli strains bearing plasmid pEAP1 or pEAP2 (T. Kudo, C. Kato, and K. Horikoshi, J. Bacteriol. 156:949-951, 1983). We analyzed the basis for excretion of periplasmic enzymes in the cells bearing these plasmids. Several experiments such as subcloning, insertion of a chloramphenicol acetyltransferase cartridge, and DNA sequencing were done. A dormant kil gene in plasmid pMB9 was expressed by a promoter of the inserted DNA fragment of alkalophilic Bacillus sp. strain 170, and as a result, the outer membrane of E. coli became permeable, allowing the proteins to be excreted without cell lysis.
Two plasmids containing the N-acetylneuraminate lyase (NALase) gene (nanA) of Escherichia coli, pNL1 and pNL4, were constructed. Immunoprecipitation analysis indicated that the 35,000-dalton protein encoded in pNL4 was NALase. The synthesis of NALase in E. coli carrying these plasmids was constitutive.
Two genes for cellulases of alkalophilic Bacillus sp. strain N-4 (ATCC 21833) have been sequenced. From the DNA sequences the cellulases encoded in the plasmids pNK1 and pNK2 consist of 488 and 409 amino acids, respectively. The DNA and protein sequences of the pNK1-encoded cellulase are related to those of the pNK2-encoded cellulase. The pNK2-encoded cellulase lacks the direct repeat sequence of a stretch of 60 amino acids near the C-terminal end of the pNK1-encoded cellulase. The duplication of the cellulase genes and the formation of the direct repeat in the pNK1-encoded cellulase occurred at almost the same time.
Postoperative oculomotor palsy is not uncommon in patients with aneurysm, but delayed postoperative peripheral oculomotor palsy due to vasospasm has not been reported. This paper presents such a patient. Oculomotor palsy was successfully treated with volume therapy. The importance of recognizing this entity and initiating appropriate treatment for vasospasm as soon as possible in order to prevent permanent deficit is emphasized.
The effect of long-term pressure overload on myocardial functional and metabolic alterations was investigated in hearts from spontaneously hypertensive rats of 16 weeks (young SHR) and 44 weeks (aged SHR) and age matched normotensive Wistar Kyoto strain rats (young WKY, aged WKY). The hearts were perfused by working heart mode and whole heart ischemia was induced by one-way valve. Following 20 min of ischemia, the hearts were reperfused for 30 min. The heart-body weight ratio in both SHR groups was significantly higher than in the respective age-matched WKY groups. Coronary flow relative to heart weight in both SHR groups was significantly lower than that of the respective age-matched WKY during both preischemic and reperfused periods. There was no significant difference in the recovery rate of cardiac output between young and aged WKY, whereas the young and aged SHR revealed significantly less recovery than their respective age-matched WKY. Tissue creatine phosphate and energy charge in both aged groups were significantly lower than in the young groups. These results indicate that long-term pressure overload increases susceptibility to ischemia and decreases the myocardial reserve presumably resulting from relative ischemia, whereas deterioration was minimal in the normotensive aged rat heart.
A method for assessing inflammatory pain response was developed by modification of the formalin test. Formalin (0.5%, 25 microliters) was injected into the hindpaw of the mouse, and the durations spent in licking or biting response were measured as an indicator of pain response. The response curve was biphasic, having two peaks, from 0 to 5 min (first phase) and from 15 to 20 min (second phase). Morphine, ethylketocyclazocine, ketocyclazocine and pentazocine inhibited the response dose-dependently at the first and the second phases. Aspirin, oxyphenbutazone and dexamethasone inhibited only the second phase. Aminopyrine and mefenamic acid which acted at both central and peripheral sites inhibited both phases; however, the inhibition of the second phase was stronger than that of the first phase. Substance P (SP) antagonist inhibited only the first phase. Bradykinin (BK) inhibitor caused a inhibition of both first and second phases, and pretreatment of compound 48/80 and indomethacin inhibited only the second phase. From these facts, it was suggested that SP and BK played a role in the pain response at the first phase, and histamine, BK and PG were involved at the second phase. Naloxone produced hyperalgesia and bestatin produced analgesia at the second phase; then, it seems that the endogenous opioid system is activated by formalin stimulation and modulates the pain perception. Based on these findings, it is presumed that the pain of the first phase is evoked by the direct stimulation of the nerve fibers, and that of the second phase is due to the inflammatory reaction.
Opiates and opioid peptides were administered in the order of 10(-9)-10(-6) mol peripherally, and their action on pain sensitivity was investigated by the modified formalin test which has two characteristic pain responses (the first and the second phase) in the mouse hindpaw. Opioid peptides (20-500 pmol) had dose-dependent analgesia against both first and second phases, and their action ranked dynorphin greater than [D-Ala2, Met5]-enkephalinamide greater than [Met5]-enkephalin. EKC and morphine (0.4-2.5 nmol) inhibited pain response of the first phase, but produced hyperalgesia in the second phase dose-dependently. Lidocaine hydrochloride had peripheral analgesic action, but was about 500-10000 times weaker than these substances. So, these peripheral analgesic actions have a different mechanism from that of local anesthetic action. N-methyl levallorphan which is thought to be a peripherally selective narcotic antagonist reversed these peripheral analgesic actions at the first and second phases and also prevented the hyperalgesic effects of EKC and morphine at the second phase. Naloxone reversed analgesia at only the first phase. These results suggest that an analgesic mechanism by opioids may exist at the peripheral site as well. Furthermore, it is estimated that a receptor exists which is antagonized by N-methyl levallorphan but not by naloxone and that there is a system of hyperalgesia by EKC and morphine in pain modulation.
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Monoclonal antibody F30 was produced by the fusion of murine myeloma cell line P3-X63-Ag8-653 with spleen cells from a BALB/C mouse immunized with established human pancreatic cancer cell line (PK-1) and the reaction specificity was analyzed. The antigen recognized by monoclonal antibody F30 was different from HLA-associated antigen, beta 2-microglobulin, fetal bovine serum components, ferritin, AFP, or CEA. Monoclonal antibody F30 reacted with all of six pancreatic cancer cell lines established in our laboratory. Cross-reactivity was detected with a colon cancer cell line or an esophagus cancer cell line among various tumor cell lines tested. No reaction was detected with red blood cells, lymphocytes, or lymphoid and myeloid cell lines. By immunoperoxidase staining of frozen sections, the F30-defined antigen was detected not only on pancreatic cancer cell membrane but also on other adenocarcinomas. In addition, the monoclonal antibody F30 had a more wide-spread distribution on normal epithelial cells in the gastrointestinal organs, respiratory system, and urinary system. F30-defined antigen was composed of two protein components with molecular weight of 190 and 160 K. It was indicated that the antigen was an integral protein in the cell membrane since the antigen was not detected in the spent culture medium of antigen-positive cells.
The shape of the human skulls was studied with the aid of a personal computer to establish sex differences in quantitative anatomical terms. On the roentgen cephalograms from 50 adult males and 50 adult females, the lateral profile was transcribed onto an acetate sheet, on which the contour from the nasal apex to the forehead was digitized on a tablet digitizer into a series of dots which were input into the computer system. After this chain of dots was simulated by a spline function, the places most typically reflecting sex difference were determined in the profile based on the radius of curvature computed at each dot. The eminence of glabella and the nasal root, shown to be the places of skull apparently most characteristic of sex, were approximated to circular arcs with the least squares method, the radii of which were expected to serve for sexing. In both places, Student's t-test revealed a significant difference between the male and female groups (p less than 0.01). It was thus demonstrated that in male the eminence of glabella and the nasal concavity develop much more markedly than in female, presenting as a clear skeletal difference between both sexes.
The medical disorders of 269 in-patients with benign prostatic hypertrophy who were over 65 years old and hospitalized between September 1974 and March 1985, were reviewed. The most frequent preoperative disorder was of the cardiovascular system. A comparative examination of the operative group with the non-operative group was made by nine preoperative risk factors, which showed a statistically significant difference at the 0.1% level.
The effect of treatment with intravenously administered Angiotensin II (AT II) on blood flow in normal and malignant tissues was investigated clinically. The time course of the effect of AT II was directly recorded by laser doppler velocimetry (LDV) via a probe placed on the surface of normal and malignant tissues. Intravenous administration of AT II resulted in an approximate 3.5 (1.3-14.0)-fold increase in blood flow in eleven malignant tissues, such as breast cancer with direct extension to the skin and abdominal skin metastasis of gastric adenocarcinoma. On the other hand, the blood flow in normal skin was decreased under AT II-induced hypertension, but a reactive hyperemia-like increase was observed soon after the withdrawal of AT II. These results strongly suggested that intravenously administered AT II can act as an adjuvant to enhance, by varying degrees, drug delivery to tumor tissue in cancer chemotherapy and that the administration of chemotherapeutic agents is undesirable soon after the withdrawal of AT II.
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Changes of endogenous opioid peptide system by pain stimuli were examined using adjuvant-induced arthritic rats. Following the 5th day after adjuvant inoculation, met-enkephalin (ME)-like peptide content in plasma decreased, while after the 12th day, the peptide content increased in adrenal medulla. A significant correlationship was found between pain threshold and plasma ME-like peptide content. Y-20003, a new nonsteroidal agent without inhibition of prostaglandin biosynthesis, showed an analgesic effect which was significantly antagonized by naloxone or Win 44,441-3, suggesting that endogenous opioid peptide system may be involved in the analgesic mechanism of Y-20003.