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Biomedical subjects

K Konishi

Publications and source records attributed to K Konishi.

At least 163 records · Page 9Linked to original sources
PubMed1995

The emergence of free radicals after acoustic trauma and strial blood flow.

The effect of acoustic trauma on cochlear strial circulation was investigated immunohistologically in the guinea pig. Kanamycin was used as a tracer of blood flow. Moreover, histochemical examinations were made to reveal the emergence of free radicals in the cochlea following acoustic trauma. At 5 min (5 min after intense sound exposure 120-125 dB SPL, 3 h) the blood flow in the stria vascularis was greatly diminished. At 2 h the strial blood flow started to recirculate and at 6 h it appeared to have returned to normal. Superoxide anion radicals (O2-) emerged along the luminal membrane of the marginal cells of the stria vascularis at 5 min. O2- disappeared at 30 min, but reappeared at 2 h. The cause of its emergence at 5 min was obscure. However, the strange phenomenon that O2- emerged again at 2 h seemed ascribable to the re-circulation of strial blood flow after sound exposure.

Animals↗
PubMed1995

Natural killer cell response in the inner ear.

Heterologous tumor cells (human chronic myelogenous leukemia K-562 cells) were injected into the perilymph and skin in guinea pigs in order to study the induction of NK cell activity in the inner ear. An in vitro transmission electron microscopic and immunohistochemical study showed that K-562 cells were attacked by guinea pig large granular lymphocytes. K-562 cells injected through the round window membrane were found to be targeted by NK cells emerging from surrounding venules after 7 to 9 days. During this time morhological changes occurred in the organ of Corti and stria vacularis. These findings suggest that the inner ear response to foreign cells induces activation and invasion of NK cells which occur relatively late compared with those in other organs such as skin.

Animals↗
PubMed1995

[A case of gastric cancer responding well to MTX/5-FU (methotrexate/5-fluorouracil) and PMUE (CDDP, MMC, UFT, etoposide) therapies upon lymphnode recurrence].

We encountered a case of non-curatively resected gastric cancer (p1, n4) who responded well to sequential MTX/5-FU therapy and PMUE therapy. A 63-year-old man was admitted to our hospital with complaints of nausea and vomiting. Upper gastrointestinal examination and CT scan revealed Borrmann type 3 gastric cancer with pyloric stenosis and multiple paraaortic lymphnodal metastasis. The patient underwent palliative gastrectomy for extensive gastric cancer (H0, P1, N4, T3, Stage IV b). Histological examination of the resected stomach revealed poorly differentiated adenocarcinoma with paraaortic lymphnodes metastasis (n4) and peritoneal dissemination (p1). Chemotherapy with sequential MTX/5-FU was given 13 times. Ten months after the operation, abdominal pain and back pain required analgesic treatment. Abdominal CT scan revealed increased size of paraaortic lymphnodes, suggesting recurrence. Sequential MTX/5-FU therapy was switched by PMUE therapy. Lymphnode size became smaller and habitual analgesics could be discontinued. Since then he was given MTX/5-FU and PMUE therapies alternately on an ambulant basis. The patient resumed his daily activities at 2 years and 8 months after the operation.

Adenocarcinoma↗
PubMed1995

[A late phase II trial of l-leucovorin and 5-fluorouracil in advanced colorectal cancer. l-Leucovorin and 5-FU Study Group (Japan Western Group)].

A multicentral cooperative study was conducted to evaluate the clinical efficacy and toxicity of l-Leucovorin (l-LV) and 5-fluorouracil (5-FU) in advanced colorectal cancer. The administration schedule was a two-hour intravenous infusion of l-LV (250 mg/m2) and an intravenous bolus injection of 5-FU (600 mg/m2), given one hour after the beginning of the l-LV infusion. Sixty-four patients were treated weekly for six weeks followed by two-weeks rest, and then evaluated for response. Complete response and partial response were obtained in 21 patients (32.8%). The median survival time was 12.8 months. The most prominent side effects were anorexia (57.8%), nausea and vomiting (56.3%), diarrhea (48.4%) and myelosuppression such as leucopenia (54.7%), thrombocytopenia (18.8%) and decreased hemoglobin (40.6%). These side effects, however, were within permissible levels. Severe toxicity was prevented by discontinuance of the treatment. From the present study, l-LV and 5-FU combination therapy seems to be a very promising and useful treatment for patients with advanced colorectal carcinoma.

Adult↗
PubMed1994

Cataloging of the genes expressed in human keratinocytes: analysis of 607 randomly isolated cDNA sequences.

The partial nucleotide sequences of 607 cDNAs randomly isolated from a cDNA library of cultured human epidermal keratinocytes were determined by single pass sequencing. Homology search of the sequences to the non-redundant nucleotide databases revealed that 27% of the cDNAs matched registered human-or non-human genes encoding not only keratinocyte specific genes, but also a variety of functional proteins, the expression of which had not been identified in keratinocytes. Non-matching cDNAs covering 49% of the cDNAs were not homologous even to ESTs from other organs, suggesting that these cDNAs include novel genes expressed in the cells. The large scale sequencing of keratinocyte cDNAs provides a useful molecular source for research into biology and diseases of the skin.

Animals↗
PubMed1994

Identification of cytokine-induced neutrophil chemoattractants (CINC), rat GRO/CINC-2 alpha and CINC-2 beta, produced by granulation tissue in culture: purification, complete amino acid sequences and characterization.

Four basic neutrophil chemotactic factors (chemokines) have been purified from conditioned medium of granulation tissue obtained from carrageenin-induced inflammation in the rat. On the basis of their N-terminal amino acid sequences, one of the chemokines was identical with rat GRO/cytokine-induced neutrophil chemoattractant (CINC) which we reported previously, and another was identical with rat macrophage inflammatory protein-2 (MIP-2). Two other chemokines were novel chemoattractants related to MIP-2. The novel chemokines are referred to as rat GRO/CINC-2 alpha and CINC-2 beta, and consequently CINC and rat MIP-2 are renamed rat GRO/CINC-1 and CINC-3 respectively. The complete amino acid sequences of purified CINC-2 alpha and CINC-3 were determined by analysis of the fragments isolated from proteinase V8-treated CINCs. The cDNA for CINC-2 beta was cloned by reverse transcription/PCR amplification using specific primers starting with total RNA extracted from lipopolysaccharide-stimulated rat macrophages. A comparison of the amino acid sequence encoded by the cDNA with the N-terminal amino acid sequence of purified CINC-2 beta revealed that mature CINC-2 beta is a 68-residue chemoattractant produced by cleavage of a 32-residue signal peptide. The difference in amino acid sequences between CINC-2 alpha and CINC-2 beta consisted of only three C-terminal residues. Rat GRO/CINC-2 alpha is a major chemokine, and the four purified chemokines have similar chemotactic activity, suggesting that they contribute to neutrophil infiltration into inflammatory sites in rats.

Amino Acid Sequence↗
PubMed1994

Analysis of p53 gene mutations and loss of heterozygosity for loci on chromosome 9q in basal cell carcinoma.

To determine the role of the p53 gene in the pathogenesis of basal cell carcinoma (BCC), we screened mutations of the gene in 11 cases of BCCs using the polymerase chain reaction (PCR) and single-stranded conformation polymorphism (SSCP). However, in all the coding exons of the gene analysed, no evidence suggesting the mutations were obtained. On the other hand, in 2 of 5 informative cases of our BCCs (40%) we found loss of heterozygosity (LOH) for loci on chromosome 9q31 which is linked to the Gorlin syndrome, that predisposes to BCC. Therefore, we suggest that a putative tumor suppressor gene on the region of 9q, but not p53 gene, plays a critical role in the pathogenesis of BCC, independent of race.

Aged↗
PubMed1994

Sequencing and characterization of the ntp gene cluster for vacuolar-type Na(+)-translocating ATPase of Enterococcus hirae.

We have previously reported the DNA and amino acid sequences for the three genes (ntpA, ntpB, and ntpK) encoding the A, B, and K (proteolipid) subunits, respectively, of Na(+)-translocating ATPase of a eubacterium Enterococcus hirae (Kakinuma, Y., Kakinuma, S., Takase, K., Konishi, K., Igarashi, K., and Yamato, I. (1993) Biochem. Biophys. Res. Commun. 195, 1063-1069). In this paper we report the entire nucleotide sequence of the ntp gene cluster coding for this multisubunit enzyme. The cluster contained eight other genes; the order of these 11 genes was ntpF, -I, -K, -E, -C, -G, -A, -B, -D, -H, and -J, encoding proteins with predicted molecular weights of 14,255, 75,619, 16,036, 22,699, 38,162, 11,409, 65,766, 51,139, 27,093, 7,164, and 48,869, respectively. The deduced amino acid sequences of these products suggested that NtpI and NtpJ are hydrophobic proteins and others are hydrophilic. The ntpI gene product, which possesses six membrane-spanning segments in its carboxyl-terminal half, resembled the 116-kDa subunit of vacuolar (V)-ATPase in clathrin-coated vesicles. In addition, the NtpE, NtpC, NtpG, and NtpD proteins resembled bovine kidney ATPase E subunit, Saccharomyces cerevisiae Vma6p, Manduca sexta V-ATPase 14-kDa subunit, and Sulfolobus acidocaldarius gamma subunit, respectively, although the similarities between their amino acid sequences were moderate. Other gene products (NtpF and NtpH) did not show significant sequence similarity to other V-ATPase subunits. Since NtpA, NtpB, and NtpK are homologous counterparts of V-ATPase, these findings suggest that the molecular architecture of E. hirae Na(+)-ATPase complex corresponds to the V-type H(+)-ATPase complex distributed in various eukaryotic endomembrane systems. The sequence of the NtpJ product was similar to those of K+ transport systems of S. cerevisiae (Trk1 and Trk2); its meaning will be discussed. This is the first demonstration of a eukaryotic V-ATPase-like Na+ pump in bacteria.

Adenosine Triphosphatases↗
PubMed1994

Rat guanidinoacetate methyltransferase. Effect of site-directed alteration of an aspartic acid residue that is conserved across most mammalian S-adenosylmethionine-dependent methyltransferases.

Most mammalian S-adenosylmethionine (AdoMet)-dependent methyltransferases have a conserved aspartate residue in a sequence oDso (o denotes a hydrophobic amino acid and s denotes a small neutral amino acid). Rat guanidinoacetate methyltransferase has two aspartate residues (Asp-129 and Asp-134) conforming to the motif in close proximity to Tyr-136 that is photoaffinity-labeled by AdoMet (Takata, Y., and Fujioka, M. (1992) Biochemistry 31, 4369-4374). In order to investigate the role of these residues, we prepared variant forms of the enzyme by oligonucleotide-directed mutagenesis. Conversion of Asp-129 to asparagine or alanine resulted in a functional enzyme. Alteration of Asp-134 to glutamate (D134E) and asparagine (D134N) decreased activity, and replacement with alanine (D134A) led to inactivation. Decreases of 3- and 120-fold were found for kcat values of D134E and D134N, respectively. The Km values of D134E for AdoMet and those for guanidinoacetate were increased about 160- and 80-fold over the respective values of wild type. The corresponding increases in D134N were 800- and 50-fold, respectively. Conservative changes of the residues flanking Asp-134 had little effect on activity. Guanidinoacetate methyltransferase obeys an ordered Bi Bi mechanism in which AdoMet binds first. Thus, the large decreases in kcat/Km values for AdoMet indicate that Asp-134 is crucial for binding AdoMet. Spectroscopic studies indicated that the amino acid substitutions of Asp-134 resulted in no significant changes in the secondary and tertiary structures, and urea denaturation experiments showed that the altered enzymes were not destabilized.

Amino Acid Sequence↗
PubMed1994

Serotyping of human rotaviruses in the Tokyo area (1990-1993) by enzyme immunoassay with monoclonal antibodies and by reverse transcription and polymerase chain reaction amplification.

Serotyping of human rotavirus in the Tokyo area was conducted from 1990 to 1993 by enzyme immunoassay with monoclonal antibodies (EIA-MAbs) against VP7 and by reverse transcription and polymerase chain reaction (RT-PCR) amplification of the VP4 and VP7 genes. The results by EIA-MAbs were very similar to those obtained by RT-PCR. Evidence of intraserotypic variations was suggested because strains of undetermined serotypes were detected by either EIA-MAb or RT-PCR. This kind of study is required for vaccine development.

Antibodies, Monoclonal↗
PubMed1994

Interleukin-4 expression in mesangial proliferative glomerulonephritis.

Several cytokines are known to play roles in the pathogenesis of human glomerulonephritis. Recently, some studies proposed a possible relationship between interleukin-4 (IL-4) and glomerulonephritis. To investigate the involvement of IL-4 in human glomerulonephritis, we evaluated renal biopsy specimens using immunohistochemical procedures. Our study revealed that the glomeruli in immunoglobulin A nephropathy and lupus nephritis (World Health Organization grades II and IV) contained IL-4-positive cells, which were CD-3- and CD-4-positive T lymphocytes. CD-8-, CD-14-, CD-19-, or CD-56-positive cells did not react with anti-IL-4. CD-23-positive cells were not observed in the sections used in this study. Interleukin-4-positive cells were not found in the normal control kidney tissues. The degree of the glomerular IL-4-positive cell index correlated well with the glomerular hypercellularity index, but not with any other clinical or histopathologic parameters. Therefore, the present study offers more reliable evidence concerning the association between IL-4 and human glomerulonephritis.

Adult↗
PubMed1994

Red blood cell sorbitol in diabetic children.

When red blood cells (RBC) were incubated with various concentrations of glucose, the RBC sorbitol level increased in a concentration-dependent manner. The elevated RBC sorbitol level was not reduced by further incubation in a glucose-free medium. In both diabetic and non-diabetic children, an increase of RBC sorbitol levels occurred in the oral glucose tolerance test and the return to baseline was delayed in diabetics compared with non-diabetics. In the majority of diabetics (87%), RBC sorbitol levels exceeded the upper limit of the normal range, which was arbitrarily determined as the mean + 2 s.d., in healthy non-diabetic children and adults. A good correlation was observed between RBC sorbitol levels and plasma glucose levels (r = 0.644). In both diabetics and non-diabetics, no correlation was observed between RBC sorbitol levels and age, and in diabetics the RBC sorbitol level was not related to the duration of disease. A good correlation was observed between RBC sorbitol levels, and hemoglobin Alc (HbAlc) or fructosamine levels in diabetic children.

Adolescent↗
PubMed1994

Continuous non-invasive finger blood pressure monitoring in children.

We evaluated the performance of continuous non-invasive finger arterial pressure measurement using the volume-clamp technique (Finapres). This study was designed to compare finger arterial pressure with brachial blood pressure estimated by the auscultatory method in 217 children (90 boys and 127 girls) aged 4-16 years and in 38 adults (aged 18-45 years). Finger and brachial artery pressure readings were obtained consecutively from the ipsilateral side in the supine position. Finger arterial pressure waveforms were recorded in all children except 4 with small and thin fingers. There was good agreement for systolic pressure with only a slight underestimation of 1.9 mmHg and 5.1 mmHg lower for diastolic pressure. This difference most probably reflects inaccuracy of the auscultatory cuff method rather than an error in the Finapres. There was large inter-individual variability in Finapres recordings which might be due to differences in vasomotor tone, as demonstrated by systolic amplification in 5 patients with anorexia. However, Finapres showed a small within-subject variability (3.8 mmHg for systolic and 4.1 mmHg for diastolic pressure) determined in 5 patients during phenylephrine infusion, and as good reproducibility as the auscultatory method. These results suggest that finger arterial pressure measurement in children older than 6 years of age has similar accuracy as that in adults, and that this method is useful for clinical applications in children, especially for the non-invasive evaluation of autonomic control and cardiovascular reflexes involving transient and rapid blood pressure changes.

Adolescent↗
PubMed1994

Development of posture in prone and supine positions during the prenatal period in low risk preterm infants.

The development of posture in the prone and supine positions was studied longitudinally in 10 low risk preterm infants between 31 and 39 weeks' postmenstrual age whose subsequent neurological development at 18 months old was normal. The infants were observed from one week after birth until the expected date of delivery. All postures were analysed according to the procedures described by Prechtl et al. There were large intraindividual and interindividual differences in posture in terms of the duration and incidence of the two positions. There was no age specific preferred posture found in either position. The posture duration in the prone position was longer than that in the supine position, but overall flexed and abducted posture were observed more often in the former. Turning of the head to the right side was also observed more often in the prone than in the supine position.

Female↗