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Biomedical subjects

M Koizumi

Publications and source records attributed to M Koizumi.

At least 19 recordsLinked to original sources

Histochemical demonstration of NADPH-diaphorase activity, a marker for nitric oxide synthase, in neurons of the rat pancreas.

To clarify the role of nitric oxide (NO) in the pancreas, we histochemically investigated NADPH-diaphorase, a marker for NO synthase, in the rat pancreas. NADPH-diaphorase activity was localized in the neurons and endothelium of vessels. The nerve fibers with NADPH-diaphorase activity were observed in both the exocrine and the endocrine pancreas associated chiefly with the vascular systems. Most nerve cell bodies (98.5%) within the pancreatic parenchyma showed strong activity for this enzyme. These results suggest that NO may participate in the hemodynamic control of the pancreas and in the neuronal regulation of pancreatic exocrine and endocrine functions.

Amino Acid Oxidoreductases

Ribozymes designed to inhibit transformation of NIH3T3 cells by the activated c-Ha-ras gene.

We have designed hammerhead ribozymes that cleave c-Ha-ras mRNA mutated at codon 12 (GGU----GUU). Plasmids containing the ribozyme-encoding genes were expressed under the control of the long terminal repeats of Rous sarcoma virus in NIH3T3 cells transfected with the activated c-Ha-ras gene. These ribozymes were found to inhibit formation of foci (by about 50%) by cleaving the oncogene mRNA, rather than by hybridizing to it. Furthermore, when the activated c-Ha-ras gene was cotransfected with the ribozyme-encoding gene, three morphologically flat colonies were found and isolated. We also found that expression of c-Ha-ras was suppressed in cells containing ribozymes.

3T3 Cells

Recognition of bases in Escherichia coli tRNA(Gln) by glutaminyl-tRNA synthetase: a complete identity set.

The fidelity of protein biosynthesis rests largely on the correct aminoacylation of transfer RNAs by their cognate aminoacyl-tRNA synthetases. Previous studies have demonstrated that the interaction of Escherichia coli tRNA(Gln) with glutaminyl-tRNA synthetase (GlnRS) provides an excellent system for studying the basis of this highly specific recognition process. Correct aminoacylation depends on the set of nucleotides (identity elements) in tRNA(Gln) responsible for correct interaction with GlnRS. Specific contacts between tRNA(Gln) and GlnRS include the 2-amino group of guanosines. Therefore, we made a set of tRNA(Gln) variants in which specific guanosines were replaced by inosine using recombinant RNA technology. This resulted in a set of tRNAs that varied by single deletions of the amino group from guanine residues, thus allowing us to test the functional importance of these contacts. In addition, a number of mutants were made by transcription of mutated tRNA genes with base changes at position 10, 16 or 25. In vitro aminoacylation of these mutants showed decreases in the specificity constant (kcat/KM) of up to 300-fold, with kcat being the parameter most affected. These experiments reveal G10 as a new element of glutamine identity. In addition, the interaction of G2, G3 and G10 with GlnRS via the 2-amino group is significant for tRNA discrimination. Based on these results, and on earlier data, we propose a complete set of bases as identity elements for tRNA(Gln).

Amino Acid Sequence

Nucleotide sequence of a gene from Arabidopsis thaliana encoding a myb homologue.

A gene encoding a proto-oncogene, a myb-related gene named Atmyb1, was cloned from Arabidopsis thaliana, and its nucleotide sequence was determined. The Atmyb1 gene contains an intron of 494 bp, and there are no highly homologous sequences present in the A. thaliana genome, but evidence was found that other myb-related genes exist. In the 5' flanking region, we found several typical cis-acting elements found in plant promoters. Sequence comparisons revealed that the ATMYB1 protein has a putative DNA-binding domain with two repeats of tryptophan clusters, which is common in MYB-related proteins in plants, while animal MYB-related proteins contain DNA-binding domains with three repeats of tryptophan clusters. The putative DNA-binding domain of the ATMYB1 protein has higher homology with that of the human c-MYB protein than with those of other plant MYB proteins.

Amino Acid Sequence

Nucleotide sequence of capsid protein gene of rice tungro bacilliform virus.

The sequence of 5,028 nucleotides, including one open reading frame (ORF), of rice tungro bacilliform virus (RTBV) dsDNA was determined. The predicted translational product comprises 1,675 amino acids and has Mr of 194,134 (p194). The amino acid sequences of three tryptic fragments from the 32 k capsid protein of RTBV (p32) were found in the predicted translational product indicating that the ORF codes for the RTBV capsid protein.

Amino Acid Sequence

Expression of vascular adhesion molecules in inflammatory bowel disease.

The expression of the vascular adhesion molecules ELAM-1 (endothelial leukocyte adhesion molecule 1) and VCAM-1 (vascular cell adhesion molecule 1) was evaluated in colonic mucosa of patients with inflammatory bowel disease and normal controls by immunocytochemistry. VCAM-1 was found to be constitutively expressed in lymphoid aggregates in normal colonic mucosa and was not significantly enhanced or altered in distribution in mucosa of patients with inflammatory bowel disease regardless of the activity of the inflammatory process. In contrast, ELAM-1 was not detected by these techniques in normal colonic mucosa (n = 11) or in colonic mucosa of patients with inflammatory bowel disease which was either uninvolved or quiescent (n = 30). However, high levels of ELAM-1 were consistently found on endothelial surfaces in association with active inflammation in affected areas of colonic mucosa in patients with either ulcerative colitis (n = 27) or Crohn's colitis (n = 9). In addition, ELAM-1 appeared to be present within neutrophils which had migrated into crypt abscesses in affected mucosa. Similar analysis was carried out in the cotton-top tamarin (CTT), a primate that experiences an idiopathic chronic diffuse colitis resembling human ulcerative colitis. Although anti-human VCAM-1 antibodies did not react with the CTT, anti-human ELAM-1 stained endothelial surfaces in mucosal biopsies from CTT with active colitis. No ELAM-1 was identified in mucosa of CTT in which colitis activity was quiescent. Thus ELAM-1 is expressed on colonic endothelial surfaces in association with inflammation and may play an important role in facilitating leukocyte migration into sites of active IBD involvement.

Cell Adhesion Molecules

Immunohistochemical demonstration of galaninlike immunoreactive nerves in the human pancreas.

Galanin, the newly discovered 29 amino acid-residue peptide, has been shown to suppress glucose-induced insulin secretion in experimental animals, but its presence and physiological role in the human pancreas have not been established. In this study, the occurrence and distribution of galanin immunoreactivity in the human pancreas was investigated by immunohistochemistry. In addition, the possible coexistence of galanin and vasoactive intestinal peptide immunoreactivity in neural elements of the pancreas was examined. In the human pancreas, galanin immunoreactivity was localized in numerous nerve fibers around glandular acini, ductules and blood vessels, and in a few nerve fibers within islets. Nerve cells with galanin immunoreactivity were frequently noticed. Immunostainings for galanin and for vasoactive intestinal peptide on serial adjacent sections of intrapancreatic ganglia showed the coexistence of the two immunoreactivities in a large proportion (73.3%) of nerve cells. These observations may provide a morphological basis for the possible neurotransmitter or neuromodulator role of galanin in the human pancreas.

Galanin

Potent antitumour activity of (-)-(R)-2-aminomethylpyrrolidine (1,1-cyclobutanedicarboxylato)platinum(II) monohydrate (DWA2114R) against advanced L1210 leukaemia in mice.

The time dependency of the antitumour activity of (-)-(R)-2-aminomethylpyrrolidine(1,1-cyclobutanedicarboxylato++ +)platinum(II) monohydrate (DWA2114R) was examined in mice inoculated i.p. with 10(5) mouse L1210 leukaemia cells. The increase in life span was greater in mice treated with 72 mg kg-1 DWA2114R on the 6th day following tumour inoculation than in mice treated at earlier times. Such superior effects against advanced L1210 were also seen with cis-diammine (1,1-cyclobutanedicarboxylato)platinum(II) (CBDCA) but not seen with the parent compound, cis-diamminedichloroplatinum(II) (CDDP) or other antitumour agents devoid of platinum. After the injection of DWA2114R on day 6, most of the ascites tumour cells accumulated in the S and G2/M phases of the cell cycle and the total cell number markedly decreased from 10(8) to less than 10(6). On the other hand, only a temporary G1 arrest and a less than 50% reduction of the cell number were induced after a similar treatment on day 3. Interestingly, the superiority of DWA2114R for advanced L1210 was lost in athymic nude mice and mice depleted of T cells by anti-thymocyte antisera. In addition, mice cured of advanced L1210 specifically rejected re-inoculated L1210 cells. These results indicate that the superior antitumour activity against advanced L1210 is unique to DWA2114R among the agents tested (except for CBDCA) and is caused by both an increased drug susceptibility of tumour cells and a drug-induced antitumour effect mediated by T cells of the host mice.

Animals

Sulfation of phenolic antibiotics by sulfotransferase obtained from a human intestinal bacterium.

Novel sulfotransferase which was isolated from Eubacterium A-44, a human intestinal bacterium, sulfated phenolic antibiotics, such as amoxicillin, ceaodroxil and cefoperazone. The Km values of sulfotransferase for these antibiotics were 6.9, 4.3 and 22.2 mM, respectively. The Vmax values were 8.3, 3.3 and 1.6 mumol/min/mg protein. The optimal pH of the enzyme was 9.0, a weakly alkaline region. The antibacterial activity of amoxicillin was not altered by enzymic sulfation of the phenolic hydroxyl group.

Animals

A case of eosinophilic granuloma arising in the mandible.

Eosinophilic granuloma is a disease of the reticuloendothelial system and its pathogenesis is not yet completely clear. The histopathological findings are granulomatous proliferation of histiocytes accompanied by diffuse infiltration of eosinophils. We recently experienced a case of eosinophilic granuloma arising monostotically in the body of the mandible in the 76 [symbol: see text] region. There has been no local recurrence 17 months after surgical removal of the lesion.

Adolescent

Cross-ligation and exchange reaction of RNA catalyzed by hairpin ribozymes.

The catalytic domain in the minus strand of the satellite RNA of tobacco ringspot virus (sTobRV(-)) assumes a hairpin-like secondary structure. This ribozyme catalyzes a cross-ligation reaction between substrate RNAs of different lengths. We constructed ribozymes to probe the activities of ligation and RNA fragment exchange.

Base Sequence

[A study on the communication between the pectoral nerve and the extramural nerve branches of the intercostal nerves].

Kumaki et al. (1979) defined the extramural nerve as the rudimentary sensory nerve which appeared on the upper thoracic wall; it branched off the root of the lateral cutaneous nerve of the second, third or fourth intercostal nerve, ran inferomedially adhering to the fascia of the intercostalis externus muscle and ended supplying the membrane covering the adjacent rib. They also stated that the extramural nerve (Rxm) occasionally became a cutaneous nerve which pierced the pectoralis muscles and supplied the skin covering the thoracic wall similar to the lateral cutaneous nerve (Rcl) or the anterior cutaneous nerve (Rca). Further, they proposed that the muscular nerves to the obliquus externus abdominis muscle which are usually situated below the fifth rib might be considered a part of this Rxm series. Although the definition of Rxm is still not widely accepted, Rxm is thought to be a key morphological factor influencing the variations of peripheral nerve arrangement on the thoracic wall. In the student course of gross anatomy dissection at Iwate Medical University School of Medicine during the years 1987-1991, three cases of Rxm communicating with the pectoral nerve and supplying the pectoralis major muscle were observed. Some cases have been reported in which Rcl innervates part of the pectoral muscles. However, the communication between the pectoral nerve and Rxm has not yet been discussed. Therefore, to clarify the morphological significance of the communication between Rxm and the pectoral nerve, the branching pattern and the distribution of the pectoral nerves were extensively investigated and the intramuscular nerve supply of some pectoral nerves, especially the pectoral nerves which communicated with Rxm, was examined in detail under a stereomicroscope. The results are summarized as follows: 1. In the first case, Rxm of the second intercostal nerve originated from Rcl, ran inferomedially adhering to the fascia of the intercostalis externus muscle and pierced the origin of the pectoralis minor muscle at the third intercostal space. Then Rxm turned superolaterally to communicate with a pectoral nerve which originated from the loop composed of the lateral and medial pectoral nerves and passed inferior to the pectoralis minor muscle. After communication, the pectoral nerve with Rxm supplied the caudalmost part of the sternocostal portion of the pectoralis major muscle. In the second case, a similar branch of Rxm of the second intercostal nerve passed inferior to the pectoralis minor muscle.(ABSTRACT TRUNCATED AT 400 WORDS)

Aged

[An aberrant muscular nerve to the latissimus dorsi muscle from the posterior cutaneous nerve of the arm].

In the 1987 student course on gross anatomy dissection of cadavers at Iwate Medical University School of Medicine, an aberrant muscular nerve to the latissimus dorsi muscle arising from the posterior cutaneous nerve of the arm was found bilaterally in a 65-year-old female who had died of acute pneumonia. The case was investigated anatomically. The posterior cutaneous nerve of the arm arose from the radial nerve approximately at the lateral axial border as the first branch independently on the right, and by forming a trunk with the nerve to the long head of the triceps brachii muscle on the left. The aberrant muscular nerve originated from the posterior cutaneous nerve of the arm about 3 cm distally to the point of the branching of the cutaneous nerve from the radial nerve. This aberrant nerve entered the latissimus dorsi muscle near the transitional area from the muscular part to the tendinous part while dividing into three on the right, and without dividing on the left. The side into which the aberrant nerve entered was just the reverse of the side into which the thoracodorsal nerve and artery supplied. The muscular part innervated by the aberrant nerve was bounded by a tendinous intersection for the most part of the latissimus dorsi muscle innervated by the thoracodorsal nerve. The intramuscular nerve distribution of the aberrant nerve was investigated under a binocular dissecting microscope. It was revealed that there was no communication between the ramification of the aberrant nerve and that of the thoracodorsal nerve.(ABSTRACT TRUNCATED AT 250 WORDS)

Aged

[Bilateral absence of the quadrangular space of the axilla].

In the 1988 student course on gross anatomy dissection of cadavers at Iwate Medical University School of Medicine, two cases of the absence of the quadrangular space of the axilla were found bilaterally in a 92-year-old female who had died of heart failure. The cases were investigated anatomically. The tendons of insertion of the latissimus dorsi and the teres major muscles and the tendon of origin of the long head of the triceps brachii muscle were united, forming a conjoint tendon that attached to the infraglenoid tubercle of the scapula and the lower part of the anatomical neck of the humerus adhering to the articular capsule of the shoulder joint. The subscapularis muscle was normal except that a muscular bundle arose from the conjoint tendon and inserted to the lesser tubercle of the humerus and the crest continuing down from the tubercle. A part of the conjoint tendon was covered by the insertion of the subscapularis muscle, and there was no space between the conjoint tendon and the insertion of the subscapularis muscle. The teres major muscle was poorly developed, but the area of origin was rather wide and arose from both the dorsal and costal surfaces of the scapula. The anatomical features of the latissimus dorsi and the long head of the triceps brachii muscles were normal except for the insertion of the former and the origin of the latter. The triangular space of the axilla was found to be surrounded by the conjoint tendon, teres major muscle, and the lateral border of the scapula covered by the subscapularis and the teres minor muscles.(ABSTRACT TRUNCATED AT 250 WORDS)

Aged

[Effect of treatment schedule of an antitumor platinum complex, DWA2114R, on the antitumor activity].

Effects of treatment schedule of (-)-(R)-2-aminomethylpyrrolidine (1,1-cyclobutanedicarboxylato) platinum (II) monohydrate, DWA2114R, on the antitumor activity against murine colon adenocarcinoma (Colon 26) and Lewis lung carcinoma (3LL) were examined. Colon 26 or 3LL tumors were transplanted s.c. into the flank and subsequently DW A2114R was given i.v. by single or multiple injections. The tumor weight was determined on days 14 or 25 and the antitumor effect was evaluated by GIR%. Although the total dose of DWA2114R was identical in both schedules, single injection was superior to multiple. Effect of treatment schedule of cis-dichlorodiammine-platinum (II), CDDP, and cis-diammine (1, 1-cyclobutanedicarboxylato) platinum (II), CBDCA, on the antitumor activities were the same as in case of DWA 2114R, i.e., single injection was superior to multiple.

Animals

[Combination therapy of a new platinum complex, DWA2114R with various antitumor agents against mouse tumors in vivo].

In vivo antitumor activity of (-)-(R)-2-aminomethylpyrrolidine (1, 1-cyclobutanedicarboxylato) platinum (II) monohydrate (DWA2114R) in combination with various antitumor agents was examined using mouse leukemia P388, Meth-A fibrosarcoma and M5076 reticulum cell sarcoma. The combination treatment of DWA2114R with cyclophosphamide, 5-fluorouracil, adriamycin, etoposide or vindesine showed synergistic effects on the prolongation of survival time of mice bearing P388. Among these combinations, combination of DWA2114R with adriamycin or vindesine showed a good synergism. In the combination treatment with vindesine against Meth-A which is not so sensitive to platinum agent, DWA2114R showed a synergistic or additive effect, but cis-diamminedichloroplatinum (II) (CDDP) showed subadditive. Three-drug combination of DWA2114R or CDDP with cyclophosphamide and adriamycin was examined in mice inoculated i.p. or s.c. with M5076. In both models, the combination of DWA2114R was more active than that of CDDP at the dose less than 1/16-fold of the dose of CDDP which is the ratio of two drugs in clinical trial.

Animals