Brief report: polymorphic microsatellite markers for the diagnosis of graft-versus-host disease.
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Biomedical subjects
Publications and source records attributed to K Takai.
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Nuclear DNA ploidy has been shown to have an important prognostic association for patients with adenocarcinoma of the prostate. Flow cytometry and static image analysis are ploidy methods that have been used in prostate carcinoma. Fluorescence in situ hybridization (FISH) using chromosome-specific probes can be used to evaluate the ploidy of interphase nuclei. In this study FISH was compared with flow cytometry and static image analysis in determining ploidy in paraffin-embedded tissue from 34 prostatic adenocarcinomas. Ploidy status using FISH was determined by enumerating centromeres of two chromosomes (8 and 12) by use of directly-labeled alpha-satellite DNA probes in isolated whole nuclei obtained by the Hedley technique. All three methods identified 11 of 34 cases as diploid and 17 of 34 cases as nondiploid (82% concordance). Six cases were discordant; two cases had discrepant results by each method. Ploidy classification as determined by FISH had an 88% concordance with ploidy classification by either flow cytometry or static image analysis. In conclusion, FISH was found to be a sensitive method of ploidy analysis in isolated paraffin-embedded nuclei from prostate adenocarcinomas. When the chromosomes commonly involved in aneuploidy have been identified in prostate adenocarcinoma, FISH has the potential to provide greater sensitivity for aneuploidy detection compared with currently available methods.
In an attempt to know the applicability of non-cultural detection technique for N. gonorrhoeae and C. trachomatis on pharyngeal and rectal swabs, the swabs taken from N. gonorrhoeae- and C. trachomatis-free subjects were tested by Gonozyme, Chlamydiazyme, Ideia Chlamydia, Gen-Probe and Gen Probe Pace 2. As N. gonorrhoeae- and C. trachomatis-free subjects, the castrated males older than age 70, with known prostate cancer treated at least for 6 months with anti-androgens who were seronegative for C. trachomatis and had a history of Ofloxacin treatment in the last 3 months and whose pharyngeal and rectal swabs were negative for N. gonorrhoeae, were subjected for study. The false negative rates were for N. gonorrhoeae from the pharynx by Gonozyme 57.7%, by Gen-Probe 0%, by Gen-Probe Pace 2 0%, from rectum by Gonozyme 0%, by Gen-Probe 0% by Gen-Probe Pace 2 0%, for C. trachomatis from pharynx by Chlamydiazyme 93.8%, by Ideia Chlamydia 8.3%, by Gen-Probe 0%, by Gen-Probe Pace 2 0%, from rectum by Chlamydiazyme 94.4%, by Ideia Chlamydia 70.9%, by Gen-Probe 20.0% and by Gen-Probe Pace 2 0%. To investigate the incidence of pharyngeal and rectal infection by N. gonorrhoeae among the patients with gonococcal genital infection, as well as pharyngeal and rectal infection by C. trachomatis among the patients with chlamydial genital infection, pharyngeal and recal swabs obtained from the patients with genital infection by N. gonorrhoeae or C. trachomatis were tested by Gen-Probe Pace 2.(ABSTRACT TRUNCATED AT 250 WORDS)
The expression of T-cell antigen receptor (TCR) alpha beta was investigated in rat CD4- CD8- thymocytes during thymic reconstitution after the exposure of animals to irradiation or glucocorticoid. The effect of the immunosuppressant FK506 on the expression of TCR alpha beta in rat CD4- CD8- thymocytes was also examined. The percentage of CD4- CD8- thymocytes constituted 2.6% of total thymocytes and that of CD4- CD8- TCR alpha beta high cells constituted 12.6% of CD4- CD8- thymocytes in normal adult Lewis rats. The percentage of CD4- CD8- TCR alpha beta high cells increased during thymic reconstitution after irradiation, and maximally constituted 28.6% of CD4- CD8- thymocytes on day 7. Similar results were obtained during thymic reconstitution after glucocorticoid treatment. In contrast, continuous treatment with FK506 for 7 days markedly decreased not only the percentages of CD4+ CD8- TCR alpha beta high and CD4- CD8+ TCR alpha beta high thymocytes, but also that of CD4- CD8- TCR alpha beta high thymocytes. These results indicate that rat CD4- CD8- thymocytes contain a subpopulation of mature (TCR alpha beta high) cells. The possible implications of the existence of this subpopulation with regard to thymocyte differentiation and maturation are discussed.
To determine the effects of FK506 on the intrathymic maturation of T cells, we analyzed changes in surface antigen expression by regenerating thymocytes after sublethal irradiation. Two-color flow cytofluorometric analysis of the regenerating thymocytes revealed that FK506 had few effects on the maturation of rat thymocytes from CD4-8- to CD4+8+, and from TCR alpha beta- MHC class I- to TCR alpha beta low MHC class I+ (TCR alpha beta, alpha and beta chains of the T cell antigen receptor). However, defects in the maturation of CD4+8+ cells to CD4+8- cells and of TCR alpha beta low MHC class I+ cells to TCR alpha beta high MHC class I+ cells were observed after FK506 administration. Furthermore, three-color analysis also detected a maturational defect of CD4-8+ TCR alpha beta high cells. Immunohistochemical analysis using MoAbs specific for MHC and TCR alpha beta revealed marked atrophy of the thymus medulla, suggesting disturbances in thymocyte regeneration and maturation. These results suggest that the maturation of CD4-8- TCR alpha beta- MHC class I- cells to CD4+8+ TCR alpha beta low MHC class I+ cells may be FK506 resistant, whereas FK506 may interfere with the maturation of CD4+8+ TCR alpha beta low MHC class I+ thymocytes not only to CD4+8- TCR alpha beta high MHC class I+ cells but also to CD4-8+ TCR alpha beta high MHC class I+ cells. We also demonstrated the possibility of enrichment of CD4-8- cells after sublethal irradiation and the existence of small subpopulations of CD4+8- TCR alpha beta- and CD4-8+ TCR alpha beta- cells which might mature to CD4+8+ cells in rat thymus.
We investigated the effects of FK506 and glucocorticoids (GC) on rat thymocytes using flow cytofluorometry. Rats were treated with GC (0.1 mg/body, by single injection), with FK506 (1 mg/kg/day, for 7 days), or with FK506 and GC. GC alone significantly decreased the percentage of CD4+8+ thymocytes and increased the percentages of CD4-8-, CD4+8- and CD4-8+ thymocytes on day 7. FK decreased the percentage of CD4+8- and CD4-8+ thymocytes and increased the percentage of CD4+8+ thymocytes on days 5 to 14. FK and GC induced a significant decrease in the number of CD4+8+ thymocytes greater than that seen with GC alone on day 7. The absolute number of TCR alpha beta high MHC class Ihigh thymocytes after FK and GC was significantly lower than that of the control group, and was slightly lower than that after FK alone on day 14. These results suggest that combined treatment with FK506 and GC acts complexly to decrease rat CD4+8+ thymocytes and prevents thymocyte differentiation and maturation.
We reported a surviving case of 6-year-old boy with pontine hematoma. He complained of headache as an initial symptom and developed progressively Foville's syndrome with impairments of the IX-XII cranial nerves. Although brainstem tumor was suspected initially using CT scan, MRI revealed the existence of hematoma in the ventromedial pons. During the first 4 months of his clinical course, Gd-DTPA did not demonstrate any enhancement in that lesion. However, hemangiomatous lesion was suspected by subsequent serial MRIs with positive Gd-DTPA enhancement. Using conservative treatment including oral corticosteroids, all the neurological deficits disappeared in several months and he did not show any recurrence of clinical signs for 3 years. It was suggested that MRI was very useful in the differential diagnosis and the follow-up of hematoma in the posterior fossa.
We treated eleven cases of primary ureteropelvic junction obstruction with percutaneous endopyeltomy. Endopyelotomy was successful in nine of the eleven cases. Complications during and after endopyelotomy occurred in three cases, two of which had pyelonephritis and the other severe postoperative hemorrhage requiring blood transfusion due to incision of a posterior crossing vessel. In this last case, the kidney was supplied by three arteries. The lower segmental artery passed behind the ureter. We emphasized that angiography should be performed to rule out the presence of a posterior crossing vessel before endopyelotomy, when it is doubtful if an extrinsic cause of ureteropelvic junction obstruction is present.
3'-N-Aminoacyl analogs of puromycin with nonnatural aromatic amino acids were synthesized and their inhibitory activity in E. coli in vitro protein synthesizing system was evaluated. The analogs with L-2-naphthylalanine, L-p-biphenylalanine, L-2-anthrylalanine and trans-L-p-phenylazophenylalanine were found to inhibit the protein synthesis with high efficiency. The inhibition suggests that these nonnatural amino acids are accepted by the active center of the E. coli ribosomal A site. A model for the adaptability of nonnatural aromatic amino acids to the active center is proposed.
KW-2149 is a new derivative of mitomycin C (MMC). The plasma concentrations, distribution, metabolism, and excretion of [3H]-KW-2149 in normal and tumor-bearing mice after i.v. administration of 16.6 mg/kg were investigated. The plasma radioactivity decreased biexponentially after i.v. administration in normal mice. However, the unchanged drug disappeared rapidly, showing a half-life (t1/2) of 9.7 min, which was shorter than MMC's (18 min). The radioactivity was excreted in mouse urine (33%) and feces (58%) within 144 h. High radioactivity was distributed in the gallbladder, liver, kidney, pancreas, and lung at 1 h after i.v. administration to normal mice. The tumor concentration was lower than the plasma or blood concentration. The lowest radioactivity was observed in the brain. The metabolic rate of KW-2149 was very rapid. The methyl sulfide form (M-16), the symmetrical disulfide dimer (M-18), and the albumin conjugate were detected in plasma, which possessed anticellular activity. The specific anticellular activity of these compounds against uterine carcinoma (HeLa S3) was 1/100, 1, and 1/20 respectively, as compared with that of KW-2149.
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Bile duct injury can occur more frequently during laparoscopic cholecystectomy than in open cholecystectomy. Three cases of common bile or hepatic duct injuries occurred in a series of eighty laparoscopic cholecystectomies; In case 1, the common bile duct was misidentified as the cystic duct. In case 2, bile peritonitis occurred on the fourth postoperative day caused by necrosis of the common hepatic duct involving the cautery surrounding it. In case 3, a bile leak occurred due to an incision at the confluence of the cystic and common duct. Dissection of the cystic duct at the infundibulum of the gallbladder, blunt dissection of the Calot's triangle, the handling of clips with special attention for safety were thought to be necessary in order to lower the risk of bile duct injury. Preoperative endoscopic retrograde cholangiography (ERCP) is recommended to avoid bile duct injury.
Subsequent of the discovery that RNA can have site specific cleavage activity, there has been a great deal of interest in the design and testing of trans-acting catalytic RNAs as therapeutic agents. We have chemically synthesized and experimentally analyzed chimeric DNA-RNA ribozymes in which the catalytic center remains as RNA.
The disappearance of thigmonastic mobility of a sensitive plant, Mimosa pudica L., caused by volatile anesthetic agents such as methoxyflurane, chloroform, halothane, enflurane or sevoflurane revealed that the response to anesthetic agents in plants maybe similar to that in animals. In terms of reversible anesthesia, animals seem to equal plants in order of potency of anesthetics, and the minimum immobilizing concentration (MIC; %) in the plant shows a good correlation with MAC in the human (MICplant = 5.8 MAChuman + 0.01, r2 = 0.946). These phenomena suggest the existence of a certain common mechanism in anesthesia between animals and plants.
A 64-year-old man had been found to have primary splenic lymphoma (stage III) seven years after the diagnosis of rheumatoid arthritis (RA). Histological diagnosis of the lymphoma was diffuse, medium sized cell type (LSG) or intermediate lymphocytic lymphoma (ILL). Splenectomy and ten courses of CHOP regimen produced continuing remission. After tow years, he suffered from peripheral neuropathy due to vasculitis of polyarteritis nodosa (PN) type. He was treated with prednisolone (PSL) and cyclophosphamide (CPM) for malignant rheumatoid arthritis. One year later, evaluation for intermittent hematuria revealed bladder cancer and he underwent total cystectomy. He has been treated with small doses of PSL under observation. The high incidence of ILL in lymphomas developing in patients with autoimmune diseases of the thyroid and salivary glands has been reported. This case suggests an association between antecedent RA and splenic lymphoma, the influence of splenectomy and chemotherapy on occurrence of rheumatoid vasculitis, and a causal relationship between CPM and bladder cancer.
When rabbit skeletal muscle myofibrils were treated with a solution containing 0.1 mM Ca2+ and 30 micrograms of leupeptin/ml, alpha-connectin, which forms very thin filaments in myofibrils, was split into beta-connectin and a 1,200-kDa subfragment. A part of beta-connectin located near the junction between beta-connectin and the subfragment seems to have an affinity for calcium ions and to be susceptible to the binding of large amounts of calcium ions. The calcium-binding site on beta-connectin is localized near the N2 line in the I band, and the subfragment is localized adjacent to the Z disk. It is possible that connectin filaments change their elasticity during the contraction-relaxation cycle of skeletal muscle at the physiological concentration of calcium ions. Because postmortem skeletal muscles lose their elasticity and become plastic in association with the calcium-specific splitting of connectin filaments, the splitting is considered to be a factor in meat tenderization during postrigor ageing.
To investigate the effect of milrinone (MIL) on systemic capacitance vessels, we measured the mean circulatory filling pressure (MCP) in anesthetized open chest dogs. We measured hemodynamic parameters (a) at baseline blood volume (BV), (b) immediately after bloodletting (5 ml/kg), and (c) immediately after dextran injection (5 ml/kg). These measurements were taken in a control group (n = 8) and in a MIL group (n = 8), where MIL (100 + 2.5 micrograms/kg/min) was administered i.v. The extra volume (EV) was obtained by extrapolating the straight line that was fit to the MCP-BV plot. MIL significantly decreased the EV, from 22.8 +/- 1.0 to 19.1 +/- 0.4 ml/kg, indicating that MIL dilates the systemic capacitance vessels. MIL shifted the right ventricular output curve to the left and upward and shifted the venous return curve to the left and rotated it clockwise. Thus, venous return was increased by decreasing the resistance to venous return and by improving right ventricular pump function. Next we evaluated the effects of MIL (100 micrograms/kg) on systemic capacitance and resistance vessels from changes seen in the MCP and the total peripheral resistance (TPR), respectively. In the untreated dogs, MIL decreased TPR significantly without affecting MCP. In dogs pretreated with total spinal anesthesia or phenoxybenzamine, MIL significantly decreased both MCP and TPR. This suggests that the intrinsic venodilator action of MIL is modified by the baroreflex in untreated animals. MIL decreased the MCP and TPR elevated previously by norepinephrine. One would, therefore, predict that MIL would dilate the systemic capacitance vessels in patients with congestive heart failure.
In anesthetized open-chest dogs, an intravenous bolus injection of endothelin-1 (ET-1, 400 pmol/kg) caused transient hypotension (initial hypotensive phase; phase 1), followed by a continuous elevation of blood pressure (late hypertensive phase; phase 2). The constriction and dilation of the systemic capacitance and resistance vessels were evaluated from the change in mean circulatory pressure (MCP) and in total peripheral resistance (TPR) in phases 1 and 2. To examine the modification of the action of ET-1 on the blood vessels by the baroceptor reflex or by the endothelium-derived relaxing factor (EDRF) released by ET-1 in phase 1, we performed experiments in dogs under total spinal anesthesia (TSA group), methylene blue-treated dogs (MB group) as well as in the untreated dogs (control group). ET-1 decreased the TPR significantly, and increased the MCP significantly in phase 1 in the control (n = 8) and TSA (n = 8) groups; there was no difference between the groups. ET-1 had no significant effect on TPR but increased the MCP significantly in MB group (n = 8) during phase 1. The percentage increase of MCP in the MB group significantly exceeded that of the control group. ET-1 increased both the TPR and MCP significantly in phase 2 in the control group (n = 8). This study indicated that the vasoconstrictor action of ET-1 on the systemic capacitance vessels in phase 1 did not result from a baroceptor reflex, and that the vasodilator action of ET-1 on the systemic resistance vessels may be at least in part mediated via EDRF released by ET-1. We suggest that the vasoconstrictor action of ET-1 on the systemic capacitance vessels is strong, but the vasodilator action of EDRF on the systemic capacitance vessels is weak.