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Biomedical subjects

H Inoue

Publications and source records attributed to H Inoue.

At least 55 records · Page 3Linked to original sources

Stability of p53 protein in rat cells transformed by various viral transforming genes.

To clarify the correlation between metabolic stabilization of p53 protein and cellular transformation, we transformed the normal rat cell line F2408 with various viral transforming genes and examined the expression and stability of p53 protein in these transformed cells by pulse-chase immunoprecipitation experiments. As expected, the level of p53 in the SV40-transformed rat cell line was higher than that in the normal cell line and its stability was also increased. In contrast, in cells transformed with the E7 and E6 genes of human papillomavirus type 16, the level and stability of p53 were similar to those in the normal cells. In cells transformed by the middle T or large T of polyomavirus, v-K-ras, and v-src, the levels and stabilities of p53 were also not elevated, although the level of p53 was increased in activated c-H-ras-transformed cells without an increase in its stability. These results show that increased stability and expression of the p53 protein are not common events in viral transformation of the rat cell line. In addition, we demonstrated with a monoclonal antibody specific for the mutant form of p53 that cellular transformation by viral transforming genes does not involve mutational activation of p53 to an oncogenic form.

Animals

Rat cellular mutants for expression of mRNA from the long terminal repeat of murine retrovirus.

Previously we isolated revertants from a rat cell line transformed by recombinant murine retrovirus containing the v-src gene. These mutant cell lines, R78 and R107, showed low src-kinase activity, but retained wild-type transforming retrovirus, suggesting that a cellular gene involved in viral gene expression was mutated. Southern and Northern hybridization analyses showed that the expression of viral mRNAs from the integrated proviral DNA was reduced in these mutant cells. DNA transfection experiments with various transforming genes and promoters revealed that the mutant cell lines were resistant to transformation by transforming genes expressed under the long terminal repeat (LTR) of Moloney murine leukemia virus (Mo-MuLV). In contrast, these cell lines could be efficiently transformed by the same transforming genes with human metallothionein promoter, polyomavirus promoter-enhancer, and c-H-ras promoter. Transient expression assays using plasmids containing the CAT gene under the LTR of Mo-MuLV also showed that CAT activity expressed under the LTR in these mutant cells was lower than that in the parental cell line, No. 7. These results suggest that cellular mutations of R78 and R107 cells affect specific transcription from the LTR of Mo-MuLV. Studies using various constructs of the LTR CAT indicated that the region responsible for the repression was located in a fragment (-328 to -150) of the LTR containing the 72-bp repeat enhancer. Somatic cell hybridization experiments showed that the mutant phenotype of these mutant cell lines is dominant to that of the parental cell line.

Animals

Enhanced induction of lymphokine-activated killer activity after lentinan administration in patients with gastric carcinoma.

In 15 patients with gastric carcinoma, peripheral blood mononuclear cells (PBM) were obtained serially before and 3, 5 and 7 days after lentinan administration. The generation of lymphokine-activated killer (LAK) activity, induced by in vitro activation of PBM with interleukin 2 (IL 2), was significantly augmented 5 days after a single intravenous dose of 2 mg lentinan, when compared with that before lentinan injection. Natural killer (NK) activity of PBM was also significantly enhanced 7 days after the drug injection. However, the distribution of lymphocyte subsets exhibited no significant change following lentinan administration.

Cell Separation

Hypertrophic cranial pachymeningitis due to Aspergillus flavus.

A 59-year-old woman suffered from occipital headache and bilateral cranial nerve VII, VIII, IX, X, XI and right XII deficit after developing otitis media. Magnetic resonance imaging (MRI) showed a thickening of the dura mater which was enhanced by gadolinium-DTPA (Gd). Aspergillus flavus was identified from the culture of otorrhea. She was treated with miconazole, flucytosin and fluconazole, which resulted in an improvement of the clinical symptoms and a thinning of the Gd-enhanced lesions on MRI. This is the first case of hypertrophic cranial pachymeningitis caused by Asp. flavus infection.

Aspergillosis

A new marker for estimation of bloodstain age by high performance liquid chromatography.

The applicability of a new marker for estimation of bloodstain age by reverse-phase high performance liquid chromatography (HPLC) is described. Using a microBondasphere C18 column with a two step linear gradient of 10.5-46.25% acetonitrile in 0.1% trifluoroacetic acid, an intriguing peak (unidentified) at a retention time of about 5 min was observed on chromatograms from human adult bloodstains and designated as 'X'. The area of this peak, which could be detected in extracts of bloodstains, but not in their fresh whole blood, increased with time. The ratios of the X area to heme area in bloodstains stored at room temperature and 4 degrees C for up to 52 weeks old linearly correlated with stain age by plotting on a double logarithmic scale. In bloodstains exposed to fluorescent light at room temperature, the regression equation calculated from the ratios (Rx) and the ages of stains in weeks (W) is ln(1000.Rx) = 1.1084 + 0.3937.ln(7.W), and the coefficient of correlation (r) is 0.9776 (n = 144, P < 0.001). When stains were stored at 37 degrees C, the ratio transformed into logarithms correlated linearly with stain age. The regression equation describing the relationship in bloodstains exposed to fluorescent light at 37 degrees C is ln(1000.Rx) = 2.4477 + 0.0866.W (r = 0.9826, n = 144, P < 0.001). The results of the present study suggest that the HPLC method may be applicable to the estimation of bloodstain age.

Adult

Complement component C3a or C3a desArg as a new marker for estimation of local vital reactions in incised skin wounds.

The anaphylatoxin C3a or its desArg form (C3a/desArg) generated during complement activation could be detected in the vicinity of incised skin wounds of guinea pigs using immunoblotting methods. The C3a/desArg peptides were detectable immediately after injury in local sites up to 3 mm from the wound edge. In subsequent determinations of up to at least 3-day-old antemortem wounds, the maximum concentration of these peptides was largely localized up to 6 mm from the wound edge at 2 h after injury. In postmortem wounds, however, these peptides were undetectable. When they were released in antemortem wounded tissues they could be detected up to 1 day at 22 degrees C after death. These results suggest that the detection of C3a/desArg in wounds using immunoblotting methods can be useful for distinguishing ante- from postmortem wounds.

Animals

Integrated information-processing system in clinical orthodontics: an approach with use of a computer network system.

A computer network system has been developed in the Orthodontic Clinic, Osaka University Dental Hospital, to improve treatment efficiency and patient service. The system consists of a 32-bit host computer, its peripheral units, and personal computers connected to the host computer by data-transmission circuits, making up a local area network (LAN). It is possible in this system to integrate various types of data, such as the patient's basic information, treatment records, image data, and diagnostic analysis results to construct a relational database. It has been shown that the computer system developed in the orthodontic clinic has various clinical advantages.

Databases, Factual

Autonomic nerve endings in the odontoblast/predentin border and predentin of the canine teeth of dogs.

Autonomic nerve endings in the odontoblast/predentin border and the predentin were observed by electron microscopy in the canine teeth of dogs. Adrenergic nerve endings with a number of very electron-dense granular vesicles and a minority of agranular vesicles of different sizes were clearly observed, but seen rarely, at the odontoblast/predentin border, in the predentin adjacent to the odontoblast processes, and as free endings in the middle part of the predentin. Cholinergic nerve endings were also observed with a number of agranular vesicles of different sizes without any visible exocytosis, in the odontoblast/predentin border, and in the predentin. Near the areas of collagen formation, extensive exocytosis of granular vesicles of different sizes was observed in adrenergic nerve endings located as free nerve endings in the predentin.

Adrenergic Fibers

Importance of atticotomy in chronic otitis media with fixation of ossicles.

Seventy-seven patients with chronic otitis media were reviewed. Of the 77 patients, 42 underwent tympanoplasty with a columella, and 35 underwent myringoplasty and atticotomy. Twenty-five of the latter 35 patients showed less than 10 dB of hearing improvement with a patch test before operation. The ossicular chain was not disconnected in any of the 25 patients, but the mobility was severely restricted. It was interesting that fixation of the malleus was found at surgery in 80% (20/25) of the patients, whereas the stapes was immobilized in only 16% (4/25). Myringoplasty and atticotomy instead of tympanoplasty with a columella was sufficient to achieve hearing improvement in most of such cases. However, conventional atticotomy was not able to achieve improvement in the mobility of the ossicles because the mobility is usually severely restricted at the malleus. Removal of the anterior tympanic spine is the key to recovery of the mobility of the malleus, resulting in hearing improvement. Rate of success was obtained in 24 of 25 ears (96%).

Auditory Threshold

Factors influencing the clinical type and course of myasthenia gravis.

Myasthenia gravis is considered to be an autoimmune disease in which several factors reciprocally influence the clinical type and course. We investigated the relative importance of the following factors: anti-acetylcholine receptor antibody (AChR Ab), HLA, age at onset, autoimmunity, thymic abnormality, duration of treatment, change in AChR Ab titer and immunosuppressive therapy. The pretreatment-AChR Ab titer and HLA were shown to significantly influence the clinical type. On the other hand, the age at onset significantly influenced the clinical course. The finding that with an onset at less than 5-year-old there was a tendency for a good prognosis suggests an association between the immaturity of the muscle and immune systems, and the clinical course.

Adolescent

Role of anisotropy in determining the selective action of antiarrhythmics in atrial flutter in the dog.

OBJECTIVE: The aim was to clarify the electrophysiological and anatomical features of the preferential site of action of antiarrhythmic drugs in the re-entrant circuit of canine atrial flutter. METHODS: Electrophysiological and anatomical findings were correlated in 17 anaesthetised adult mongrel dogs with atrial flutter associated with an intercaval anatomical obstacle, before and after intravenous administration of disopyramide (2 mg.kg-1) and flecainide (2 mg.kg-1). RESULTS: Before drug injection, a rate dependent prolongation of conduction time occurred in the low right atrium where the conduction was slow during atrial flutter. Disopyramide (n = 8 dogs) and flecainide (n = 9 dogs) terminated atrial flutter, with conduction block occurring in this slow conduction area in the low right atrium. Although the degree of drug induced prolongation of refractoriness in this particular area was similar to those in other areas of the right atrium, conduction was depressed to a greater extent in this region. Anatomical study revealed that a thick pectinate muscle that branched from the crista or crista terminalis itself ran perpendicular to the wavefront of the pacing impulse and atrial flutter in this slow conduction area. CONCLUSIONS: These data indicated that slow conduction might be attributed, at least in part, to anisotropic conduction over the thick muscle bundle in the low right atrium, and that antiarrhythmic drugs preferentially produced conduction block in this area. Anisotropic conduction in the low right arium is an anatomical substrate for slow conduction in the re-entrant circuit and for the site preference of antiarrhythmic drugs in the present canine model.

Animals

Modulation of active renin secretion by renin-binding protein (RnBP) in mouse pituitary AtT-20 cells transfected with human renin and RnBP cDNAs.

To investigate the role of renin-binding protein (RnBP) in renin metabolism, RnBP expression plasmid, which was constructed to express human RnBP under the control of mouse mammary tumor virus long terminal repeat, was transfected into mouse pituitary AtT-20 cells together with the expression plasmid encoding human renin. The transfectant secreted prorenin and active renin, whereas RnBP was expressed only in the presence of dexamethasone and without secretion into the medium. The secretion of active renin was stimulated by forskolin, and the stimulation was repressed by dexamethasone. The secretion of prorenin, however, was insensitive to forskolin irrespective of the presence or absence of dexamethasone. Moreover, the forskolin-stimulated release of active renin was hardly repressed by dexamethasone in AtT-20 cells transfected with the renin expression plasmid and a selectable plasmid pMAMneo. Coexistence of RnBP and renin mRNAs in human Wilms' tumor G-401 cells was shown by means of polymerase chain reaction of respective cDNAs from the cells. These results suggest that RnBP modulates the release of active renin in renin-producing cells.

Animals

Site-directed mutagenesis of amino acid residues involved in the glutathione binding of human glutathione S-transferase P1-1.

The four residues of human glutathione S-transferase P1-1 whose counterparts were indicated by X-ray crystallography to reside in the GSH-binding site of pig glutathione S-transferase P1-1 were individually replaced with threonine or alanine by site-directed mutagenesis to obtain mutants R13T, K44T, Q51A, and Q64A. The kinetic parameters, susceptibilities to an inhibitor, S-hexyl-GSH, and affinities for GSH-Sepharose of the latter were compared with those of the wild-type enzyme, and pKa of the thiol group of GSH bound in R13T was shown to be equivalent to that in the wild type. From the results, Lys44, Gln51, and Gln64 were deduced to contribute to the binding of GSH. On the other hand, Arg13 seems to be essential for the enzymatic activity as mainly involved in the construction of a proper structure of the active site.

Amino Acid Sequence

Redundant nerve roots of the cauda equina caused by lumbar spinal canal stenosis.

To investigate pathogenesis of redundant nerve roots of the cauda equina, which were concomitant with severe lumbar spinal canal stenosis, six cadavers were examined anatomically and histopathologically, and quantitative analysis of nerve fibers was performed. In this anatomic study, it was observed that all the redundant nerve roots passed through the constriction of the spinal canal. No significant pathologic change was detected in the spinal ganglia and in the spinal cords except for the posterior column, in which dorsal redundant roots were entering. Redundant nerve roots of unequal length also were observed in the anatomic study. The spatial distribution of redundant nerve roots and the extent of degeneration of nerve fibers in them were established by these histopathologic and quantitative studies. These facts indicated a close causal relationship between redundant nerve roots and constriction of the spinal canal, and that the pathogenesis of redundant nerve roots was a squeezing force acting on the nerve roots at the area of spinal canal constriction.

Cauda Equina

Ubiquitous presence in mammalian cells of enzymatic activity specifically cleaving 8-hydroxyguanine-containing DNA.

Here we report the finding of enzymatic activity that specifically cleaves DNA containing 8-hydroxyguanine (oh8Gua) residues in various mammalian cells. To detect this activity, we used a synthetic double-stranded DNA containing a single oh8Gua at a defined position as the substrate, and analyzed the products of enzymatic digestion by polyacrylamide gel electrophoresis. Two cleavage sites near the oh8Gua residue were detected with partially purified fractions from cow brain and rat liver, and also with preparations from all mammalian tissues examined. These results suggest that enzymatic activity for the removal of oh8Gua from DNA is widely distributed in mammalian cells.

Animals

A case report of inflammatory pseudosarcoma of the urinary bladder.

A case of inflammatory pseudosarcoma of the urinary bladder in a 35-year-old Japanese male is presented. This benign lesion can easily be mistaken for spindle cell sarcoma since it consists of rhabdomyoblast-like elongated strap cells showing infiltrative growth, and whether it is benign or malignant is difficult to determine by microscopic examination. In this case, spindle cell proliferation extended among bundles of the superficial muscle layer. However, no abnormal mitoses, severe nuclear atypia or cellular pleomorphism could be seen, thus indicating inflammatory pseudosarcoma. Although the lesion was not completely resected, no recurrent disease has been clinically observed for two years following transurethral resection. Urologists and surgical pathologists must be able to detect this lesion in order to avoid unnecessary surgical procedures.

Adult

Monoclonal antibody analysis of phosphatidylserine and protein kinase C localizations in developing rat cerebellum.

Understanding the topographical relationships between phosphatidylserine (PS) and protein kinase C (PKC) within neurons can provide clues about the mechanism of translocation and activation of PKC. For this purpose we applied monoclonal antibodies (Abs) of PS and PKC to sections of developing rat cerebellum. The anti-PKC Ab immunohistochemical pattern showed homogeneous staining of Purkinje cells over various postnatal ages, whereas the anti-PS Ab staining showed a heterogeneous localization over these ages. Purkinje cells did not stain well between postnatal day 14 (PND 14) and PND 21, suggesting that the PS was lost from the membrane during preparation of the sections during this period. These data imply that interactions between PS and PKC vary in Purkinje cells during postnatal development.

Aging

[Isolation of vero-cytotoxin-producing Escherichia coli from cattle and serotyping and toxin-typing of the isolated strains].

Two hundred and sixty-six piglets with diarrhea (from 4 farms), 73 healthy pregnant pig (from 2 farms), 27 calves with diarrhea (from 9 farms) and 47 healthy milk cows (from 1 farm) were examined for Vero-cytotoxin-producing Escherichia coli (VTEC), and 52, 11, 15 and 67 strains of VTEC were isolated from 17 piglets, 11 pregnant pigs, 6 calves and 23 milk cows, respectively. All VTEC strains from the piglets produced only VT2vp, while the strains from the healthy pigs did not produce VT2vp, but did VT1 and/or VT2. Most VTEC strains from calves and cows produced VT2vhb and some produced VT2 and VT1. Serotyping of the isolated strains showed that many strains from the piglets belonged either O139:H1, O141:H4 or O141:HUT, but the strains from the pigs were either R-form or O-untypable. Many strains from the calves and cows were serotyped into O116 or O113, but there were several R-form and O-untypable. From these results, it is suggested that VTEC strains, especially from the pregnant pigs, calves with diarrhea and healthy milk cows, which produced the same type of Verotoxins to that produced by human isolates, may become sources of human infections.

Animals