Location of F1 ATPase-like genes on the physical map of the Bacillus subtilis 168 chromosome.
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Biomedical subjects
Publications and source records attributed to T Kudo.
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The present study describes the radiological and histological changes in the canine intervertebral disk after the experimental injection of chymopapain as the chemical reagent, and determines the appropriate dose of the enzyme for treatment of herniated disks. By radiography, narrowing of the disk space was observed within 2 weeks after the injection of chymopapain, and recovered to 74.1% in the 0.1 mg group, 61.1% in the 1.0 mg group and 71.7% in the 10.0 mg group at 12 weeks. The disk space recovery showed a tendency to delay with aging. Microscopically, proteoglycan positive matrix appeared and the nuclear space was reduced in each disk at 2 weeks after chymopapain injection. The nucleus pulposus contained an irregularly-defined mass consisting of clusters of degenerated notochordal cells surrounded by proliferated chondrocytes and collagen matrix. In each disk at 12 weeks after chymopapain injection, the center of the nucleus pulposus was replaced by fibrocartilage tissue. In the disk into which 10.0 mg chymopapain was injected, the nuclear space filled with dense fibrocartilage tissue without a regenerated matrix component and narrowing of the disk were maintained. It is suggested that canine chemonucleolysis with 10.0 mg of chymopapain reduces the interdiskal pressure. This treatment may therefore relieve the signs and symptoms of herniation of the nucleus pulposus, and may effect chemical disk decompression.
In order to meet the present increased demands, we have tried to improve the methods to produce human monoclonal antibodies by using a SCID mouse grafted with human mononuclear cells. Initially, we gave an anti-asialo GM1 antibody to a SCID mouse to suppress the NK activity, as a pretreatment. Then, fifty million human mononuclear cells (MNC) from a healthy volunteer were injected intraperitoneally to the SCID mouse so as to construct a human immune system in the mouse (PBL-SCID mouse). We immunized the mouse with a synthetic peptide (pep 190) conjugated with KLH four times. The spleen cells taken from the immunized PBL-SCID mouse were fused with (mouse x human) heteromyeloma cells. The hybridoma cells were selected in GIT medium containing HAT and IL-6. Among 68 hybridoma-growing wells, we obtained one hybridoma clone (#41-1-4) which secreted a specific antibody to pep 190. The reactivity of this monoclonal antibody was tested by ELISA and the specificity of this antibody was confirmed by an absorption test with different kinds of proteins. This paper is the first report of the successful production of peptide-specific human monoclonal antibody by active in vivo immunization using a PBL-SCID mouse. By this active in vivo immunization system using a PBL-SCID mouse, human monoclonal antibodies for any sort of peptide antigens may easily be made available.
Three cases of successful pregnancies in renal transplant recipients who had undergone transplantation in the Okayama University Medical School Hospital are reported. Two of the women had received an organ from a living relative and one woman received a cadaveric organ graft. These patients, aged 28-37 at the time of the delivery, had received their transplants 2-5 years prior to their conception. The periods of gestation ranged between 35 and 40 weeks. The weight of the babies at birth ranged from 2,380g to 2,500g and the apgar score at 1 min was 8 or 9. None of the infants showed any congenital abnormalities. Lower-segment cesarean section was performed in all of three cases. Serum creatinine levels, an indicator of renal graft function, did not deteriorate during the pregnancy or after delivery. Although further work is needed to solve problems regarding pregnancy in renal transplant recipients, these results encouraged us to meet their hope for a baby.
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Respinomycins are a novel group of anthracycline antibiotics produced by Streptomyces xanthocidicus. Respinomycins A1, A2, B, C and D were isolated by EtOAc extraction, silica gel column chromatography, centrifugal partition chromatography and preparative silica gel thin layer chromatography. Respinomycins A1 and A2 induced the terminal differentiation of human leukemia K-562 cells.
Fifty patients with benign prostatic hypertrophy were treated with chlormadinone acetate (CMA) at the dose of 50 mg/day for 12 months. Subjective symptoms and objective findings were evaluated before, and after 4 and 12 months of treatment. Better improvement was observed in all of the subjective symptoms and in almost all the parameters of objective findings, in proportion to the period of CMA treatment. Generally speaking, 12 months of treatment was more effective than 4 months of treatment. Two patients had impotence (4.0%). Both of them complained of side effects within 4 months after treatment, but they were not severe. In conclusion, long-term treatment of benign prostatic hypertrophy with CMA was useful.
Semiology of postictal aphasia as well as its generating mechanism involving both the epileptogenic zone and language area has not yet been sufficiently elucidated. Therefore, postictal aphasias were studied in 3 patients with localization-related epilepsy. Postictal motor aphasia was observed in a patient (patient 1) with frontal lobe epilepsy whose recovery of language function progressed from loss of language function, to Broca's aphasia, and eventually to poor spontaneous speech. Postictal sensory aphasia was observed in 2 patients (patient 2 and 3) with temporal lobe epilepsy whose recovery of language function progressed from loss of language function, to jargon, and eventually to circumlocutory anomic speech. Both patient 2 and 3 did not show fluent speech. Seizure manifestations, EEG, IMP-SPECT and MRI indicated that epileptogenic zones were in the left frontal lobe including Broca's area in patient 1, the mesial part of the left temporal lobe in patient 2, and the middle and posterior part of the left temporal lobe including Wernicke's area in patient 3. A postictal verbal dichotic listening test showed the reduction of correct responses by right ear in the patient 3. Postictal auditory verbal learning tests showed the impairment of verbal memorization in patient 2 and 3. The impairment of verbal memorization was particularly marked in patient 2. These results suggest that 1) the characteristics of postictal motor and sensory aphasia are clearly elucidated by analyzing the sequence of recovery from postictal language dysfunction, 2) postictal aphasia is generated by the epileptogenic zone involving the language area or the ictal discharges propagating to the language area, and 3) not only the disturbance of language function, but also the impairment of verbal memorization seems to participate in the disturbance of repeating and understanding sentences in the patient with postictal sensory aphasia.
To elucidate the mechanism of general anesthesia, effects of sevoflurane anesthesia on serotonin metabolism in rat brain were studied. Three percent sevoflurane was administered for twenty minutes to Wistar male rats weighing 230-270g under spontaneous respiration. The rats were sacrificed by decapitation and the brains were rapidly removed. They were dissected into nine discrete regions, locus coeruleus, pons plus medulla oblongata, hypothalamus, thalamus, basal ganglia, midbrain, hippocampus, amygdala and cerebral cortex. The contents of serotonin (5-HT) and one of its major metabolites, 5-hydroxyindole-3-acetic acid (5-HIAA) were measured by high performance liquid chromatography with the dual-cell coulometric detector before anesthesia, 20 minutes after the start of anesthesia and at the recovery from anesthesia. Significant increases in 5-HT levels were observed in the pons, hypothalamus, midbrain, amygdala and cerebral cortex by sevoflurane anesthesia as compared with the control group. 5-HIAA levels decreased significantly in the thalamus by sevoflurane anesthesia, while an appreciable increase in 5-HIAA levels was observed in the basal ganglia at the recovery from anesthesia. It is concluded that 5-HT metabolism is significantly suppressed in the pons, hypothalamus, thalamus, midbrain, amygdala and cerebral cortex during sevoflurane anesthesia and this change in 5-HT metabolism may be associated with a mechanism of sevoflurane anesthesia.
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The most characteristic brain lesion of Alzheimer disease is the accumulation of paired helical filaments (PHF) in the affected neurons. Based on solubility in detergents there are two general populations of PHF, the readily soluble (PHF I) and the sparingly soluble (PHF II) types. The major polypeptides of PHF are the microtubule associated protein tau. Tau in PHF is present in abnormally phosphorylated forms. In addition to the PHF, the abnormal tau is also present in unpolymerized form in the AD brain. Small amounts of ubiquitin (%) are associated with PHF II but neither with PHF I nor with the unpolymerized abnormally phosphorylated tau in AD brain. Furthermore, the pretangle neurons can readily be immunolabeled for abnormally phosphorylated tau but not for ubiquitin. The level of tau in neocortex is several-fold higher than in AD aged control cases, but this increase is in the form of the abnormally phosphorylated protein. The microtubule associated proteins from AD brain do not promote the assembly of microtubules in vitro, whereas the in vitro dephosphorylated PHF polypeptides stimulate the binding of GTP to the exchangeable site of tubulin and the assembly of microtubules. In vitro the phosphate groups in PHF are less accessible than those of tau to alkaline phosphatase. It is suggested that a defect in the protein phosphorylation/dephosphorylation system leads to hyperphosphory-lation of tau. The altered tau contributes to a microtubule assembly defect and consequently compromises the axoplasmic flow and leads to neuronal degeneration.
We examined the effects of ketamine (20 mg.kg-1) on plasma ACTH, Comp. B, aldosterone and ir-ANP levels in unstrained rats. We divided rats into two groups, saline group as control (saline group) and ketamine group (ketamine treated rats). Plasma samples were taken before ketamine or saline infusion to determine a pre-treatment value, and samples were also taken at 15, 30, 60 minutes after the infusion. ACTH, aldosterone and ir-ANP were measured by radioimmunoassay. Comp. B was determined by a fluorometric method. We obtained the following results: 1) Plasma ACTH levels increased significantly in the ketamine group, while there was no such a marked increase in the saline group. The slight increase in ACTH in the saline group could have resulted from handling during i.v. infusion or psychic stress; 2) Plasma corticosterone and aldosterone levels increased markedly in both groups. However, the magnitude of the adrenocortical response was significantly greater than in the saline group; 3) Plasma ir-ANP showed no significant change in both groups. These results suggest that ketamine stimulated pituitary-adrenal axis, and the adrenal stimulation produced by ketamine are greater than those induced by the stress of handling or psychic stress.
The contamination of HeLa cells into many culture cells has been suspected. We detected HPV type 18 DNA in two HeLa cell sublines, Hep-2 laryngeal cancer and KB oral cancer cell lines as well as FL cell line derived from normal amniotic membrane which have been extensively used so far in experimental virology. On the other hand, we detected HPV type 18 DNA neither in other epithelial cell lines nor in fibroblastic human embryonic cells. The digestion analysis with several restriction enzymes of the HPV type 18 DNA in HeLa, Hep-2, KB and FL cell lines were almost the same. The molecular size of episomal DNA could not be demonstrated by digestion of these DNAs with XhoI, a non-cut enzyme for HPV type 18 DNA, suggesting all HPV types 18 DNA were integrated. Interestingly enough, the HPV type 18 specific DNA in these cells was cleaved into two fragments with another non-cut enzymes, B g/II and HindIII, suggesting the presence of a single cutting site for each enzymes presumably generated by rearrangements or mutations. These results provide further evidence that the so-called Hep-2, KB and FL cells are all HeLa cell derivatives.
The efficacy of examining antinuclear antibody (ANA) was investigated as a screening test detecting subclinical immune disorders in infertility and sterility. ANA was measured in 116 unexplained infertile or sterile patients. The ANA positive rate was 43.5% in group A (habitual abortion, n = 23), 38.1% in group B (consecutive miscarriages, n = 21), 30.0% in group C (one miscarriage, n = 10), 16.7% in group D (one or more deliveries n = 12) 22.0% in group E (primary sterility, n = 50), and 22.4% in the control group (n = 54). The positive rate for all the infertile patients (group A+B) was 40.9% and significantly higher than that in for the control group (p < 0.05). Ten patients with positive ANA had 12 deliveries and 20 patients with negative ANA had 23 deliveries. The frequency of preeclampsia in the patients with positive ANA was higher than in those with negative ANA (41.7% versus 4.3%, p < 0.05). A higher incidence of premature deliveries was recognized in the patients with positive ANA than in those with negative ANA (41.7% versus 8.7%, p < 0.1). Average birth weight (> 35 week) for patients with and without positive ANA was 2,976g (n = 8) and 3,122g (n = 19) respectively. The former tends to be smaller than the latter (p < 0.1).
An animal model of chronic brain hypoperfusion has been developed by applying coiled clips to the bilateral carotid artery of Mongolian gerbils. The brain tissue damage was neuropathologically studied after 1, 4, 8, and 12 weeks of hypoperfusion. The hippocampus, basal ganglia, and cerebral cortex of the chronically hypoperfused gerbil showed lesions with various severity which are probably due to ischemic episodes. In the cerebral white matter, however, two types of lesions were observed; one similar to those in the gray matter, and the other observed only in the white matter after more than an 8-week duration of brain hypoperfusion. The lesion specific to the white matter showed rarefaction and gliosis without locally associated ischemic changes. This type of the white matter lesion was never found in the gerbil brain before 8 weeks and, significantly, increased in number and size by 12 weeks post operation. The accumulation of the white matter lesions is characteristic in the gerbil with chronic hypoperfusion. The observed white matter-specific lesion resembles the histological changes in aged brain with cerebrovascular diseases.
There was an epidemic of non-A non-B hepatitis in a small area of a town in the central part of Japan, which began with an outbreak of several patients in 1981 and then spread extensively with the result that about one third of the inhabitants showed abnormality in serum liver function tests at the health check performed in 1985. We determined histological diagnoses on that occasion for 167 individuals of the abnormal population and recently assayed antibodies against hepatitis C virus (HCV) for most of their sera left available. Histologically, chronic active hepatitis (CAH) was the major pattern, accounting for 59.3% (99 cases) of the total. Others were chronic persistent hepatitis (CPH) (13.2%), chronic lobular hepatitis (CLH) (16.2%), liver cirrhosis (LC) (6.6%) and fatty liver (4.8%). In the serological studies, the newly developed system to detect antibodies against the viral core protein p 22 was found to be much more sensitive than the conventional system to detect anti C 100-3 antibodies. By using these two methods in combination, we found that 82% were antibody-positive, indicating strong implication of HCV in this epidemic. This was further supported by direct detection of the viral genome in patients' sera by polymerase chain reaction following reverse transcription. We further found a strong correlation between the histological inflammatory activity and the antibody prevalence, since nearly all (97.6%) of the CAH cases were antibody-positive by at least either of the antibody assays, while only about 50% were positive in the less active cases such as CPH and CLH.