A new method for strand discrimination in sequence-directed mutagenesis.
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Base Sequence↗
Biomedical subjects
H Ohmori
Publications and source records attributed to H Ohmori.
Intravesical instillation chemotherapy of adriamycin with or without verapamil for the treatment of superficial bladder cancer: the final results of a collaborative randomized trial.
A collaborative randomized clinical trial on the intravesical administration of Adriamycin (ADM) with or without verapamil (VR), a calcium-channel blocker, as chemotherapy of superficial bladder cancer (Ta, T1) was carried out at two universities, Okayama and Kagoshima, and their affiliated hospitals. Arm A consisted of ADM given at 50 mg/50 ml saline, and arm B consisted of ADM given at 50 mg/40 ml saline plus five ampules (25 mg/10 ml saline) of injectable VR. The drugs were instilled into the bladder for 3 consecutive days, and three such courses were given with a 4-day interval between each course for a total of nine instillations. A total of 96 patients (48 in arm A and 48 in arm B) were entered into this study. The two treatment groups showed no significant difference in background factors. Of the 40 evaluated arm-A patients, 24 (60.0%) showed a response (CR + PR), whereas 19 (48.7%) of the 39 patients in arm B responded. The difference between these response rates was not statistically significant. As for adverse reactions to the intravesical chemotherapy, local inflammatory symptoms were observed in half of the patients, although no systemic reaction was observed. No significant difference was found between arm A and arm B, except for urinary turbidity. In conclusion, at the dose employed in the present clinical trial, there was no clear enhancement of the effect of ADM combined with VR in patients with superficial bladder cancer. Further clinical studies are required to determine the optimal doses of ADM and VR for their combination in intravesical chemotherapy.
Administration, Intravesical↗
New familial nephropathy involving glomerular and tubular basement membranes.
We describe two siblings, an 8-year-old boy and a 9-year-old girl, with severe mental retardation, dwarfism, optic atrophy and nephropathy. Laboratory examination showed beta 2-microglobulinuria, decreased creatinine clearance, hypercholesterolaemia and elevated serum levels of muscle enzymes. Renal biopsy from one of the patients demonstrated characteristic ultrastructural changes involving both the glomerular and tubular basement membrane. This group of symptoms and laboratory findings is quite distinct and differs from those of other reported familial nephropathy syndromes. We conclude that this disorder may represent a new syndrome of autosomal recessive inheritance.
Basement Membrane↗
Nucleoside monophosphoramidate hydrolase from rat liver: purification and characterization.
1. Adenosine 5'-phosphoramidate hydrolase of 29 kDa was isolated from rat liver cytosol. 2. It consisted of two subunits of 14 kDa. 3. It hydrolyzed nucleoside 5'-monophosphoramidates into nucleoside 5'-monophosphates and ammonia, while it did not hydrolyze adenylyl phosphoramidate, adenylyl imidodiphosphate and N-phosphorylated compounds like phosphocreatine, N omega-phosphoarginine, 6-phospholysine and 3-phosphohistidine. 4. Divalent cations and cyclic AMP had no effect on the hydrolytic activity.
Animals↗
Predominant suppression of anti-TNP IgE response in mice by monoclonal anti-TNP IgG1 antibody: characterization of its mode of action by in vitro and in vivo studies.
It was found that an antigen-specific IgE response both in vitro and in vivo was strongly suppressed in the presence of IgG1 monoclonal antibody (mAb) against the antigen. Anti-trinitrophenyl (TNP) IgE response was elicited by the co-culture of C3H B-cells and a conalbumin (CA)-specific helper T-cell clone, D10.G4.1, in the presence of 0.1 microgram/ml TNP-CA. Addition of anti-TNP IgG1 monoclonal antibody (mAb) at 1 microgram/ml to the culture resulted in a marked (> 90%) suppression of anti-TNP IgE formation, while anti-TNP IgG1 and IgM responses were affected to a lesser extent (50-60% suppression). Similar observations were made in in vivo experiments. When 100-200 micrograms of anti-TNP IgG1 mAb was injected i.p. into BDF1 mice prior to immunization with TNP-CA, the anti-hapten (TNP) IgE response as well as the IgE response to the carrier (CA) was suppressed by 80-90%, while anti-TNP IgM production was inhibited by less than 50%. Injection of anti-TNP IgM or IgA mAb showed only marginal effects on anti-TNP IgE production. Spleen cells from anti-TNP IgG1 mAb-treated mice cultured in vitro secreted much lower levels of anti-TNP IgE spontaneously than those from untreated mice. In in vitro and in vivo experiments using the F(ab')2 of anti-TNP IgG1 mAb, an IgG1 mAb with an irrelevant specificity and mAb directed to Fc gamma RII, it was shown that the binding of the IgG1 mAb with the antigen and the interaction of its Fc portion with Fc gamma RII are required for the suppressive effects to be exerted.
Animals↗
3-Phosphohistidine/6-phospholysine phosphatase from rat brain as acid phosphatase.
A phosphatase hydrolyzing 3-phosphohistidine and 6-phospholysine was purified from rat brain cytosol to 90% homogeneity on polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. One milligram of protein of the purified phosphatase released inorganic phosphate from 3-phosphohistidine, 6-phospholysine, AMP, GMP, and p-nitrophenyl phosphate at velocities of 6.5, 15.6, 15.0, 6.9, and 8.3 mumol/min, respectively. However, the purified enzyme could not hydrolyze N omega-phosphoarginine and phosphocreatine, which are substrates for phosphoamidase [EC 3.9.1.1]. The molecular masses of the holoenzyme and the subunit were 94 and 50 kDa, respectively, and the sedimentation coefficient of the native enzyme was 6.3 s, indicating that it was a dimeric enzyme of identical subunits. The enzyme functioned well under acidic conditions, and 50% of the activity was inhibited by 30 microM tartrate, 4 microM vanadate, 20 microM molybdate, 4 microM VCl3, or 13 microM MoCl5. These results indicate that the present hydrolase belongs to the acid phosphatase group [EC 3.1.3.2].
Acid Phosphatase↗
A sandwich cup method for the penetration assay of antimicrobial agents through Pseudomonas exopolysaccharides.
We developed new sandwich cup method to assay the penetration of various antimicrobial agents through Pseudomonas exopolysaccharides. Using alginate extracted from mucoid-type Pseudomonas aeruginosa and gellan gum from Pseudomonas elodea, the role of exopolysaccharides as a barrier against drug penetration was examined. The penetration of positively charged hydrophilic drugs such as aminoglycosides and polypeptides was markedly inhibited by the gels tested, but that of beta-lactams, quinolones, and macrolides was not inhibited. The penetration of gentamicin was strongly influenced by the gel concentration, the solution to be used, and the presence of Ca2+. These results suggest that the microenvironment at the infection site could greatly influence drug penetration through biofilms in vivo.
Alginates↗
Over-expression of metallothionein and drug-resistance in bladder cancer.
Metallothionein (MT) in tumor cells has been implicated as one of the factors involved in mechanisms of resistance to anti-cancer drugs, including cis-diaminedichroloplatinum (CDDP) and adriamycin (ADM). The relationship between the expression of MT and chemotherapy with anti-cancer drugs was studied in CDDP- and ADM-resistant human bladder cancer cell lines and tissue samples from clinical cases. In drug-resistant cell lines (T-24/ADM, CI-7/CDDP) established in our laboratory, MT expression was studied by immunohistochemistry using the avidin-biotin peroxidase complex (ABC) method and radioimmunoassay (RIA), using anti-MT antibody. In addition, other potential mechanisms of drug resistance, such as P-glycoprotein expression were examined and the levels of reduced glutathione (GSH), oxidized glutathione (GSSG) and glutathione-S-transferase (GST) determined in these cell lines. The results of these investigations demonstrate that the expression of MT in resistant cell lines increased 2.1- and 2.5-fold when compared with parent cell lines (CI-7, T-24). GSH, GSSG and GST levels were unchanged and P-glycoprotein was not over-expressed. A total of 120 tissue samples from 35 clinical cases of bladder cancer, before and after chemotherapy, were stained for MT which was detected in 10 of the 35 cases before chemotherapy. The incidence of MT expression was significantly higher (p < 0.05) in cases with lower pathological tumor grades.(ABSTRACT TRUNCATED AT 250 WORDS)
ATP Binding Cassette Transporter, Subfamily B, Mem↗
Phorbol ester-induced reversible inactivation of cytotoxic T cell function: correlation with down-regulation of protein kinase C activity.
When an H-2d-specific cytotoxic T lymphocytes (CTL) clone, FC1, was incubated in the presence of 10(-7) M phorbol myristate acetate (PMA) for 10-12 hr, the cytolytic activity of the CTL against H-2d target cells was abrogated, but was reversibly restored to the normal level after subsequent incubation of the cells in PMA-free medium for more than 10 hr. These effects of PMA have been reported (Russell, J.H.: J. Immunol. 133, 907-912 (1984)), but the mode of its action has not been fully investigated. Here, we analyzed the biochemical basis of the PMA-induced loss of cytolytic activity. Cycloheximide completely blocked the restoration of the PMA-suppressed cytolytic activity, suggesting that protein synthesis was required in this process. PMA-treatment did not affect the levels of CD3 and CD8 molecules expressed on the CTL, nor was the level of a CTL-specific serine esterase, BLT esterase, affected by this treatment. However, the target cell-induced release of BLT esterase from the CTL was suppressed if the cells were pretreated with PMA. PMA-treatment of the CTL led to the down-regulation of protein kinase C (PKC) activity by about 50%. On the other hand, staurosporin, an inhibitor of PKC, completely blocked the target cell lysis when added at 10(-6) M. These results suggest that the down-regulation of at least some isoform(s) of PKC is responsible for the PMA-induced loss of the cytotolytic activity of CTL.
Animals↗
Structural analysis of the rhlE gene of Escherichia coli.
The E. coli chromosome is known to carry at least five genes, each of which codes for a "D-E-A-D" box protein that is presumed to possess an ATP-dependent RNA helicase activity. Four of such genes (srmB, deaD, dbpA and rhlB) were already mapped on the E. coli chromosome and their DNA sequences determined. We here report the complete nucleotide sequence of the remaining rhlE gene located at about 17.8 min on the E. coli genetic map. RhlE protein possesses all of the motifs (I to VI) conserved among prokaryotic and eukaryotic "D-E-A-D" proteins and has an arginine-rich carboxyl-terminal region. A null mutant of the rhlE gene was constructed by a new method with a ColE1 plasmid mutant that replicates in RNAse HI-deficient bacterial strains, but not in the wild-type strains. The delta rhlE mutant can grow normally, implying that the rhlE gene product is nonessential for bacterial cell growth.
Base Sequence↗
[Evaluation of analgesic effects of morphine, buprenorphine and pentazocine in rats receiving somatic and visceral stimulation].
The analgesic effects of morphine, buprenorphine and pentazocine examined by behavioral responses to tail-flick (TF) and colorectal distension (CD) were studied in rats. Animals were randomly divided into three groups; morphine (M) groups of 4 mg.kg-1 (n = 7), buprenorphine (B) groups of 0.03 mg.kg-1 (n = 8), pentazocine (P) groups of 3 mg.kg-1 (n = 8). After determinations of TF and CD values, animals were administered each analgesic intraperitoneally, and both procedures were repeated every 5 minutes until 40 minutes. Three drugs produced an almost same degree of increase in pain threshold for TF; % maximum possible effects (%MPEs) were approximately 30%. Similar patterns of increase in pain thresholds for CD were observed in M and B groups; %MPEs were approximately 20%. In contrast, P groups showed a significant increase in thresholds for CD compared to M and B groups; 20 minutes after intraperitoneal administration of P, %MPE was approximately 50%. These results indicate that there are different antinociceptive effects to somatic and visceral stimuli between M, B and P, and suggest that selective opioid agonists modulate responses to different kinds of noxious stimuli.
Abdominal Pain↗
[Differential effects of morphine and non-steroidal anti-inflammatory drugs on somatic and visceral pain in rats].
We studied the differential effects of morphine and non-steroidal anti-inflammatory drug (NSAIDs) on behavioral responses to tail-flick (TF) and colorectal distension (CD) in rats. Animals were randomly divided into three groups; morphine groups (4 mg.kg-1), flurbiprofen axetil groups (30 mg.kg-1) and aspirin DL-lysin groups (100 mg.kg-1). After determining control TF and CD values, animals received each analgesic intraperitoneally, and each procedure was repeated every 10 minutes for 30 or 40 minutes. NSAIDs produced a statistically significant increase in pain threshold for CD, but not for TF. In contrast, almost similar patterns of increase in pain thresholds for CD and TF were observed in morphine groups. The present study demonstrates that NSAIDs have a powerful antinociceptive effect on visceral pain evoked by CD.
Animals↗
[Re-examination of the criteria for clinical evaluation on bacterial prostatitis--analysis of the data of the clinical study of temafloxacin].
The clinical efficacy of temafloxacin (TMFX) was evaluated in the patients with acute prostatitis (AP: 31 cases) and chronic prostatitis (CP: 75 cases), and the criteria for clinical evaluation on bacterial prostatitis by the Japanese UTI Committee were re-examined. The clinical efficacy of TMFX on prostatitis was examined. The number of the evaluable cases by doctors in charge were 30 and 68 for AP and CP, respectively. The efficacy rates were as high as 96.7% for AP and 80.9% for CP. In the patients whose efficacy could be evaluated by the Committee, the efficacy rates were as high as 100% (15/15) for acute bacterial prostatitis (ABP) and 66.7% (18/27) for chronic bacterial prostatitis (CBP). The bacteriological eradication rates were 100% (15/15) for ABP and 81.5% (22/27) for CBP. Adverse drug reactions were observed in 8 of 100 cases (total evaluable cases) but they clinically did not consist any problems. Abnormal laboratory test findings were observed in 9 of 71 cases (total evaluable cases), whose changes were all slight and did not clinically cause any problem. Therefore, TMFX was concluded to be highly useful in the treatment of ABP and CBP. The criteria for clinical evaluation on bacterial prostatitis were re-examined. Concerning the treatment period, comparison of the evaluation at Day 7 with that at Day 14 in ABP revealed that the efficacy was evaluable at Day 7. In the same way, comparison of the evaluation at Day 14 with that at Day 28 in CBP revealed that the efficacy was evaluable at Day 14. Concerning the pathogens, in ABP, the major pathogens were considered to be GNR as well as E. coli. On the other hand, in CBP, GNR and E. faecalis, that were generally authorized to be pathogens, were detected in less than half of the patients, and CNS were detected in the other patients. Among the CNS cases, there were some cases that showed no relationship between improvement of clinical symptoms and bacterial response. Therefore, it is necessary to perform further studies on this matter. In conclusion, regarding the criteria for clinical evaluation on bacterial prostatitis proposed by the Japanese UTI committee, the treatment period for evaluation of efficacy was appropriate, and it is necessary to perform further studies in order to determine the pathogens in bacterial prostatitis.
Acute Disease↗
[Factors determining the level of PaCO2 during laparoscopic cholecystectomy].
Factors responsible for the elevation of PaCO2 were investigated in 30 patients undergoing laparoscopic cholecystectomy using CO2 gas insufflation. Intraperitoneal insufflation of CO2 gas was performed to the level of 15 mmHg of intraperitoneal pressure in each patient and the following measurements were made: body height, body weight, body surface area (BSA), circumference of abdominal wall, obesity index, body mass index (BMI), operating time, total volume of insufflated CO2 gas, and change of peak inspiratory pressure. Rate of elevation of PaCO2 showed the highest correlation with BSA (correlation coefficient -0.691 [P < 0.01]) followed by body weight, body height, and circumference of abdominal wall. These results suggest that during laparoscopic cholecystectomy using CO2 gas insufflation, the degree of CO2 storage capacity has the highest effect on the elevation of PaCO2.
Adult↗
Dihydrostreptomycin modifies adaptation and blocks the mechano-electric transducer in chick cochlear hair cells.
Block of the mechano-electric transduction (MET) channel by dihydrostreptomycin (DHSM) and its effects on adaptation were investigated in dissociated cochlear hair cells of the chick with a whole-cell patch-electrode voltage clamp technique. DHSM reversibly blocked the MET channel in a dose- and voltage-dependent manner. At -50 mV, DHSM blocked the MET channel with a Hill coefficient of approximately 1 and a dissociation constant (Kd) of 2 x 10(-5) M. Rate constants for the DHSM to bind and to unbind to and from the channel were estimated, and could be larger than 5 x 10(7) M-1.s-1 and 1 x 10(3) s-1, respectively. The amplitude of MET current decreased during a constant displacement of the hair bundle. This current decay, the adaptation, disappeared in the DHSM medium. The disappearance and the emergence of adaptation did not have a simple relationship with the block of MET channel by DHSM, but appeared with some delay.
Adaptation, Physiological↗
The secondary antigen-specific IgE response in murine lymphocytes is resistant to blockade by anti-IL4 antibody and an antisense oligodeoxynucleotide for IL4 mRNA.
In order to investigate the role of interleukin 4 (IL4) in the induction of antigen-specific IgE responses, we established culture conditions which allow the induction of anti-trinitrophenyl(TNP) IgE response by the coculture of TNP-keyhole limpet hemocyanin-primed C3H B cells with conalbumin (CA)-specific type 2 helper T (Th2) cell clone, D10.G4.1 in the presence of TNP-CA. A maximum level of anti-TNP IgE was secreted at 1 microgram/ml TNP-CA. By using filter-separated double-chamber culture plates, the physical contact between T cells and B cells was shown to be necessary in this response. Anti-TNP IgE synthesis was not significantly suppressed in the presence of 20-40 micrograms/ml monoclonal anti-IL4 (11B11), nor was the response further enhanced by the addition of recombinant IL4. 11B11 added together with anti-IL5 had also no suppressive effects on the IgE response. This apparent independence on IL4 might be due to the fact that IL4 is transferred from T cells to B cells in a transsynaptical way that would be refractory to the neutralization by 11B11. In order to test this possibility, we synthesized the phosphorothioate analogue of an antisense oligodeoxynucleotide against IL4 mRNA (S-oligo) for inhibiting IL4 production from Th2 cells specifically. S-oligo was effective at 10-20 micrograms/ml in suppressing IL4 production from D10.G4.1 cells by 80-90%. It was demonstrated that S-oligo, either alone or in combination with 11B11, did not significantly suppress anti-TNP IgE response. These results suggest that antigen-specific IgE response in primed B cells does not depend on IL4, but requires cognate interaction with Th2 cells.
Animals↗
Enzyme release assay of human NK cell activity using beta-galactosidase-expressing K562 target cell line.
In the present report, we established a K562 cell line useful for an enzyme release assay of human natural killer (NK) activity. Human myelogenous leukemia cell line, K562, was transfected with a plasmid carrying Escherichia coli beta-galactosidase (beta-gal) gene. A colony that permanently expresses the enzyme activity was isolated, and designated K562/Zneo. Incubation of K562/Zneo cells (1 x 10(4)) with nonadherent human peripheral blood lymphocytes (PBL) resulted in the release of beta-gal activity depending on the incubation time and the number of effector cells. Released beta-gal activity was assayed sensitively by using 4-methylumbelliferyl-beta-D-galactoside, a fluorescent substrate. The cytolytic activity of PBL was augmented significantly when the cells were preincubated with interleukin-2 for 20 h. This enzyme release assay showed a comparable sensitivity to that of 51Cr release assay. Thus, K562/Zneo cell line is thought to be useful for the nonradioactive assay of human NK and lymphokine-activated killer activities.
Cytotoxicity, Immunologic↗
The binding site for fucose-binding proteins of Lotus tetragonolobus is a prognostic marker for transitional cell carcinoma of the human urinary bladder.
BACKGROUND: Carbohydrate signals are important in tumor metastasis. METHODS: Expression of binding sites for fucose-binding proteins (FBP) of Lotus tetragonolobus were immunohistochemically analyzed in patients with transitional cell carcinoma. RESULTS: The survival of patients with invasive bladder cancer was associated with the degree of expression of FBP binding sites. Overall actuarial 5-year survival and cancer-corrected 5-year survival rates were much worse in patients with strong expression of FBP binding sites than in those with no or weak expression (P < 0.001 and P < 0.05, respectively). Strong expression of FBP binding sites in patients with Stage T3-4 disease or disease beyond Stage T2 with lymphatic permeation also was correlated with increased disease progression. Autopsies revealed that FBP binding sites were strongly expressed in all primary tumors with metastasis and in most of the metastatic tissues. CONCLUSION: The degree of expression of FBP binding sites correlates with the increased metastatic potential of bladder cancer and with poor patient survival times.
Adult↗