Search PubMed⌕ Search

Biomedical subjects

S Iwai

Publications and source records attributed to S Iwai.

At least 145 records · Page 8Linked to original sources
PubMed1994

[The optimal period for orally administered fluoropyrimidines as an adjuvant chemotherapy for gastric cancer--a pilot study using 5-FU tablets compared with surgical operation alone].

Long-term oral administration of fluoropyrimidines such as 5-fluorouracil (5-FU) or tegafur is commonly used as an adjuvant chemotherapy for gastric cancer, but the optimal period or optimal total doses of fluoropyrimidines have not been studied. Two hundred cases of macroscopical Stage II and III curatively resected gastric cancer patients were entered in this study, and divided into three groups (6 months group: mitomycin C was given i.v. at day 0 and day 1 and 5-FU tablets were orally administered at a dose of 200 mg/day for 6 months. 12 months group: MMC was given the same as for the 6 months group and 5-FU tablets were administered for 12 months. Surgery alone group: No chemotherapy, operation only). As the result, 185 cases were eligible. There was no significant difference between the 6 months group and the 12 months group among Stage II patients. Although there was also no significant difference between the 2 groups in Stage III patients, the survival curve of 12 months group was always higher than in the 6 months group. When comparing with surgery alone group, 5-year survival of the 12 months group was always higher than in the surgery alone group of Stage III patients; however, the survival rate in the 6 months group was worse than in the surgery alone group at Stage II and III. These results suggest that MMC i.v. and 12 months or over administration of 5-FU tablets is useful for Stage III gastric cancer patients, and that cooperative study is required comparing with surgery alone in Stage II patients.

Administration, Oral↗
PubMed1994

[Severe infection in gastroenterological field; MRSA (methicillin resistant Staphylococcus aureus) enterocolitis].

There has been a rising trend in the number of methicillin-resistant Staphylococcus aureus (MRSA) infections in recent years, and postoperative MRSA enterocolitis has been on the increase. We conducted a nation wide questionnaire on postoperative enterocolitis from January 1988 to June 1990. Among the 169 institutes which replied, there were 126 cases of postoperative MRSA enterocolitis in 53 institutes. Most of the cases based on malignant disease, especially gastric cancer. Their onset began with diarrhea and fever, between the second and fifth postoperative days. 18 patients died, giving a mortality rate of 14.3%. Examination of the recent antibiotic susceptibility demonstrated that MRSA had rapidly acquired multiple drug resistance. It is important that every effort be made to protect against hospital infection.

Aged↗
PubMed1993

Binding of nucleic acids to E. coli RNase HI observed by NMR and CD spectroscopy.

To clarify the mechanism by which the RNA portion of a DNA/RNA hybrid is specifically hydrolyzed by ribonuclease H (RNase H), the binding of a DNA/RNA hybrid, a DNA/DNA duplex, or an RNA/RNA duplex to RNase HI from Escherichia coli was investigated by 1H-15N heteronuclear NMR. Chemical shift changes of backbone amide resonances were monitored while the substrate, a hybrid 9-mer duplex, a DNA/DNA 12-mer duplex, or an RNA/RNA 12-mer duplex was titrated. The amino acid residues affected by the addition of each 12-mer duplex were almost identical to those affected by the substrate hybrid binding, and resided close to the active site of the enzyme. The results reveal that all the duplexes, hybrid-, DNA-, and RNA-duplex, bind to the enzyme. From the linewidth analysis of the resonance peaks, it was found that the exchange rates for the binding were different between the hybrid and the other duplexes. The NMR and CD data suggest that conformational changes occur in the enzyme and the hybrid duplex upon binding.

Base Sequence↗
PubMed1993

Cyclobutane thymine dimers in a ras proto-oncogene hot spot activate the gene by point mutation.

ras proto-oncogenes with a cyclobutane-type thymine photodimer (cis-syn or trans-syn isomer) were constructed by replacement of a portion of the gene with a chemically synthesized fragment. When the genes were transfected by the calcium phosphate method into mouse NIH3T3 cells, they induced focus-formation, indicating that both photoproducts were mutagenic in mammalian cells. Sequence analysis of the ras gene fragments derived from the transformed cells showed that the genes were activated by a point mutation. The mutations detected most frequently were 3'-T-->A for the cis-syn isomer and 5'-T-->A for the trans-syn isomer. In contrast, a different trend of mutations was observed when a primer on a DNA template with a cis-syn dimer was extended in vitro by either DNA polymerase beta or alpha.

3T3 Cells↗
PubMed1993

Hydrogen-bonding contacts in the major groove are required for human immunodeficiency virus type-1 tat protein recognition of TAR RNA.

The binding site for tat on TAR RNA was analysed by preparing a series of model RNA substrates carrying site-specific functional group modifications. The test RNAs were prepared by annealing two short synthetic oligoribonucleotides to form a duplex structure with a U-rich bulge and flanking sequences identical to TAR RNA. Tat binds these duplex RNAs with approximately half the affinity for wild-type TAR RNA. Substitution at positions U23 or U25 by the base analogue, O4-methyl-dT, which is deficient in its ability to hydrogen-bond at the N3 position reduces tat affinity more than 20-fold. Modifications to purines in the stem of TAR RNA that affect hydrogen-bonding ability in either the major or the minor groove of duplex RNA were also tested. Removal of the nitrogen atom at either the N7 position of G26 or at the N7 position of A27 reduces tat affinity 10- to 20-fold. By contrast removal of the exocyclic amino group in the minor groove at position G26, by substitution with inosine, does not affect tat binding significantly. A single methylphosphonate substitution at the phosphate bond between A22 and U23 also leads to a significant loss of tat binding ability, whereas all other methylphosphonate substitutions in the U-rich bulge are not harmful to tat binding. We conclude that tat forms multiple specific hydrogen bonds to a series of dispersed sites displayed in the major groove of the TAR RNA molecule. These include the N3-H of U23, the N7 of G26, the N7 of A26 and the phosphate between A22 and U23.

Base Sequence↗
PubMed1993

Epididymal lymphangiectasis.

We report on a 59-year-old man with epididymal lymphangiectasis. The patient had noticed somewhat intermittent intrascrotal painless swelling. The interstitium of the right caput epididymis exhibited a number of large, dilated lymphatic vessels forming irregular channels among and around the epididymal ducts. The afferent epididymal ducts showed dilatation similar to that of the lymphatic vessels except for focal cuboid epithelial linings. Spermatogenesis in the right testis was preserved. Lymphangiectasis in the epididymis is infrequent and needs to be differentiated from other intrascrotal lesions.

Diagnosis, Differential↗
PubMed1993

HIV gene regulatory proteins tat and rev and their interactions with synthetic RNA.

Synthetic oligoribonucleotides have been prepared and annealed to form model RNA duplexes that mimic the high affinity RNA recognition sites for the HIV-1 tat and rev proteins. The contributions of individual functional groups on the model RNAs to the specificity of binding by their respective proteins were studied by use of oligoribonucleotides containing site-specifically modified nucleotides. Both tat and rev appear to recognise specifically a limited number of functional groups in the major groove of an RNA double helix distorted by virtue of unpaired or non-Watson-Crick paired nucleotides.

Base Sequence↗
PubMed1992

Recognition of the high affinity binding site in rev-response element RNA by the human immunodeficiency virus type-1 rev protein.

The Human Immunodeficiency Virus type-1 rev protein binds with high affinity to a bubble structure located within the rev-response element (RRE) RNA in stemloop II. After this initial interaction, additional rev molecules bind to the RRE RNA in an ordered assembly process which requires a functional bubble structure, since mutations in the bubble sequence that reduce rev affinity block multiple complex formation. We have used synthetic chemistry to characterize the interaction between rev protein and its high affinity binding site. A minimal synthetic duplex RNA (RBC6) carrying the bubble and 12 flanking base pairs is able to bind rev with 1 to 1 stoichiometry and with high affinity. When the bubble structure is inserted into synthetic RNA molecules carrying longer stretches of flanking double-stranded RNA, rev forms additional complexes resembling the multimers observed with the RRE RNA. The ability of rev to bind to RBC6 analogues containing functional group modifications on base and sugar moieties of nucleoside residues was also examined. The results provide strong evidence that the bubble structure contains specific configurations of non-Watson--Crick G:G and G:A base pairs and suggest that high affinity recognition of RRE RNA by rev requires hydrogen bonding to functional groups in the major groove of a distorted RNA structure.

Bacteriophage T7↗
PubMed1992

Photoaffinity labeling of T4 endonuclease V with a substrate containing a phenyldiazirine derivative.

T4 endonuclease V recognizes thymine photodimers in DNA duplexes and, in a two-step reaction, cleaves the glycosyl linkage of the 5'-side thymidine and the phosphodiester linkage. To determine the amino acid residues responsible for binding thymine photodimers, a photoaffinity reagent, 4-(1-azi-2,2,2-trifluoroethyl)-benzoate, was linked to the aminoalkylphosphonate of a thymine photodimer in a 14-mer duplex. The reactive substrate was treated with the enzyme under UV light (365 nm). The nascent enzyme and the modified enzyme were treated with lysyl endopeptidase, and the peptide maps were compared. Three peptides from the C terminus were found to interact with the reactive oligonucleotide to various extents. The three modified peptides were isolated and analyzed by Edman degradation. The amino acid residues Gly-133, Tyr-129, and Thr-89 were partially linked with the reactive substrate and may be involved in the binding of thymine photodimers.

Affinity Labels↗
PubMed1992

[Hospital acquired infection in surgical field and its countermeasure present situation of anaerobes, P. aeruginosa and MRSA infection].

The important organisms of hospital acquired infection in surgical field are anaerobes, P. aeruginosa and MRSA. From 1977, isolation frequency of anaerobes from pus has been increasing remarkably and it became 60% in 1978. The isolation rates of P. aeruginosa increased 20 to 30%. Since 1988, the incidence of isolation of MRSA has increased markedly. Almost of all the coagulase typing of MRSA were type II, and it suggested the possibility of hospital infection. In 1988, high antimicrobial activity against MRSA was observed with vancomycin, imipenem, minocycline and new quinolones, but in 1990 the multiresistant strains against imipenem, minocycline and new quinolones were increased. Among the various antimicrobial agents examined, vancomycin demonstrated the strongest activity to MRSA. The symptomatic infections were postoperative enterocolitis, respiratory infection, biliary tract infection and wound infection. The most severe symptom was observed in MRSA enterocolitis. We conducted a nation wide questionnaire of postoperative enterocolitis from January 1988 to June 1990. 169 institutes replied, and 126 cases were involved in postoperative MRSA enterocolitis in 53 institutes. Eighteen cases died, and mortality rate was 14.3%. In the treatment of MRSA infection vancomycin was the most effective antibiotics. It is the most important to prevent nosocomial infection.

Anti-Bacterial Agents↗
PubMed1991

How does RNase H recognize a DNA.RNA hybrid?

The mechanism of RNase H substrate recognition is proposed from a model of a chemically modified DNA.RNA hybrid Escherichia coli RNase H complex. Site-directed mutagenesis of the enzyme and substrate titration observed by heteronuclear two-dimensional NMR spectra have been carried out. A model complex has been built, based on free structures of the enzyme and the substrate independently determined by x-ray crystallography and NMR distance geometry, respectively. In addition to steric and electrostatic complementarities between the molecular surfaces of the enzyme and the minor groove of the hybrid in the model, putative hydrogen bonds between the polar groups in the enzyme and 2'-oxygens of the RNA strand of the hybrid fix the hybrid close to the active site of the enzyme. The enzymatic activities of the mutant proteins and the changes in NMR spectra during the course of substrate titration are consistent with the present model. Moreover, the specific cleavage of the RNA strand in DNA.RNA hybrids can be explained as well as cleavage modes in modified heteroduplexes. A mechanism of enzymatic action is proposed.

Amino Acid Sequence↗
PubMed1991

Pathological study on beagles after long-term oral administration of cadmium.

Histopathological, histochemical, and electron microscopic examinations were performed on beagles after a long-term oral cadmium (Cd) administration of greater than 8 years. Although renal atrophy was remarkable in groups receiving doses of 50 and 100 mg/kg body weight/day, bone lesions could not be demonstrated by roentgenological and histopathologic examination. It was noticed that concomitant regeneration or recovery and cell death of the epithelium occurred in the proximal convoluted tubules. The cell death was consistent with apoptosis, a special feature of cell death, which was shown to play a major part in the tubular damage of cadmium by electron microscopic examination. Fatty degeneration of the pars recta tubules was seen to show dose-dependence. The intrarenal cadmium was localized predominantly in the cytoplasm of the proximal tubular epithelium by histochemical and ultracentrifugal cell fractionation examinations. Although no remarkable changes were found in the other organs, aggregates of siderophages in the liver and focal hemorrhage in the spleen, known as spontaneous lesions, might be related to Cd intoxication. In conclusion, the present study revealed that no bone lesions occur with Cd administration in adult beagles in spite of long-term administration. An excessive cell death to regeneration or recovery in the proximal tubules might result in the renal cortical atrophy. No remarkable changes were seen in the glomeruli and distal nephrons, which were in good agreement with Cd distribution.

Administration, Oral↗
PubMed1991

[A case of colon cancer with severe anal bleeding caused in a preoperative chemotherapy].

UFT was orally administered at a dose of 600 mg/day as a preoperative chemotherapy for a 44-year old male with colon cancer. On the seventh day of the administration, severe anal bleeding occurred, and an emergent colectomy was performed. The macroscopic findings of the resected tumor were different from the first endoscopic picture, and the histopathological findings showed the significant effects of preoperative chemotherapy.

Adenocarcinoma↗
PubMed1991

[Laboratory interpretive reporting system. A prototypical approach for knowledge representation in acid-base and electrolyte disorders].

It is important to realize the problem-oriented diagnosis and the goal-oriented one to support general physicians as clinical decision makers under so many laboratory tests more than full with using practically. A prototypical model for computer-assisted interpretation of laboratory data was presented. To achieve both function of the problem-oriented inference and the goal-oriented one, knowledge base in this system was structured using the methods of semantic network and qualitative reasoning. Superficial knowledge level characterized by the intuitive reasoning from test results to diseases and deep knowledge level referring to the relations among the pathophysiologic states were stratified, and the heuristics based on the experiences at bed-side were incorporated in this knowledge representation. The pathophysiologic states were distilled from the textbooks and the articles, and were linked together by the relationships of cause and result, such as functional, constraint or conceptual relationship. Diagnostic knowledge on the computer was reconstructed by the causal network representing the pathological process as qualitative chaining between the pathophysiologic states, and was corrected and compiled by the clinical experiences and the experimental facts. Abnormal test data became a trigger of the start of inference, and presumptive pathophysiologic states and diagnoses were fired on the causal network. Consequently, the most important pathophysiologic states might become to be clear to understand the cause of changes of laboratory data.

Acid-Base Imbalance↗
PubMed1990

Synthesis and characterization of a substrate for T4 endonuclease V containing a phosphorodithioate linkage at the thymine dimer site.

A dodecadeoxyribonucleotide containing a cis-syn thymine dimer with a phosphorodithioate linkage was synthesized on a solid support using a dinucleotide coupling unit prepared by UV-irradiation of dithymidine monophosphorodithioate followed by S- and 5'-O-protection and 3'-phosphitylation. A photodimer-containing dodecamer without phosphate modification was also synthesized. The dodecamers were hybridized to the complementary sequence, and the duplexes used as substrates for T4 endonuclease V. This enzyme cleaved the phosphate-modified substrate more slowly than the unmodified duplex with the same dissociation constant.

Base Sequence↗
PubMed1990

Identification of the amino acid residues involved in an active site of Escherichia coli ribonuclease H by site-directed mutagenesis.

The amino acid residues essential for the catalytic activity of ribonuclease H (RNase H) from Escherichia coli (E. coli) were identified by site-directed mutagenesis. It has been proposed by computer analysis that E. coli RNase H has homologous amino acid sequence with the RNase H domains of various retroviral reverse transcriptases (RTs) (Johnson, M. S., McClure, M. A., Feng, D. F., Gray, J., and Doolittle, R. F. (1986) Proc. Natl. Acad. Sci. U. S. A. 83, 7648--7652). Of the eight highly conserved residues examined, Asp10, Glu48, and Asp70 were found to be crucial for RNase H activity. Determination of the kinetic parameters for the mutated enzymes using the chemically synthesized nonanucleotide duplex as a substrate demonstrated that these residues are involved in the catalytic site rather than the substrate-binding site. These residues are fully conserved in the amino acid sequences of not only retroviral RTs but also hepadnaviral, plant viral and retrotransposon RTs. This strongly suggests that they are also involved in the active site of these RTs and RT related enzymes.

Amino Acid Sequence↗