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Biomedical subjects

H Ueyama

Publications and source records attributed to H Ueyama.

At least 91 records · Page 5Linked to original sources
PubMed1994

Detection of bacteria in phagocyte-smears from septicemia-suspected blood by in situ hybridization using biotinylated probes.

We report herein the detection of intracellular bacteria in phagocyte-smears obtained from septicemia-suspected blood samples by in situ hybridization. This was obtained by using nick-translated biotin-11-dUTP-labeled DNA probes and streptavidin-alkaline phosphatase conjugates for visualization of the hybridized signals. The probes were made from random genomic DNA clones of bacteria which are frequently the causative agents of bacteremia, such as Staphylococcus spp., Pseudomonas aeruginosa, Enterococcus faecalis, Escherichia coli, Klebsiella spp. and Enterobacter spp. When our in situ hybridization method was compared with conventional culture protocols for the ability to detect bacteria from the blood of patients suspected of having septicemia, 30 positive results were obtained in 50 specimens by in situ hybridization methods. In contrast, only 7 positive results were obtained by blood cultures. Thus, even if bacteria cannot be detected by conventional blood cultures and histology, our in situ hybridization method allows for direct observation of bacterial foci in circulating phagocytes and identification of the bacteria. Our investigations suggest that in septicemia, circulating polymorphonuclear neutrophils carry some surviving bacteria as well as metabolized bacterial DNA and RNA for a considerable period of time. Thus, our in situ hybridization method using the phagocyte-smears have diagnostic value for detecting most bacteria which cause septicemia.

Bacteremia↗
PubMed1994

Detection of Staphylococci in mouse phagocytic cells by in situ hybridization using biotinylated DNA probes.

In situ hybridization was used to detect intracellular Staphylococcus aureus and S. epidermidis in mouse phagocytic cells after experimental infection of C3H mice with Staphylococci via abdominal or intravenous injection. Isolated ascites or whole blood were tested by the phagocyte smear technique, using bacteriolytic enzymes to preserve phagocytic cell morphology. The exposed bacterial DNA was visualized as intracellular hybridized signals by use of biotinylated DNA probes and by immunocytochemistry using streptavidin-alkaline phosphatase conjugates as detector molecules. These DNA probes, prepared from randomly cloned genomic DNA fragments of S. aureus and S. epidermidis, were strain-specific and did not cross-hybridize either in situ or on dot-blot hybridization. This technique of in situ hybridization with phagocyte smears is useful for detection and diagnosis of intracellular bacteria regardless of viability.

Animals↗
PubMed1993

Molecular cloning and chromosomal assignment of the gene for human Zn-alpha 2-glycoprotein.

Genomic clones containing the human Zn-alpha 2-glycoprotein gene were isolated. Two of them were for the functional gene overlapped, and the other four were for two different pseudogenes (1 and 2) retaining exon-intron organization. The complete DNA sequence of the functional gene (9.3 kb) and its 5'- and 3'-flanking regions (5.3 and 0.1 kb, respectively) was determined. The gene is composed of four exons; the first exon is for the 5'-untranslated region, the signal sequence, and the first six amino acids; the second, for domain A; the third, for domain B; and the fourth, for domain C and the 3'-untranslated region. The 5'-flanking region contains a TATA box, a CAT box, an octamer sequence, and three possible Spl-binding sites. Ten and three copies of Alu repetitive DNA were identified within the gene and the 5'-flanking region, respectively, and they occupy 30% of the gene. The nucleotide sequences around the exons of pseudogene 1 were also determined; they had high homology (90-91%) with the corresponding region of the functional gene. Southern blot analysis suggested that there are only three genes, including nonfunctional ones, for Zn-alpha 2-glycoprotein in humans. The gene (ZA2G) was mapped to human chromosome band 7q22.1 by fluorescence in situ hybridization.

Amino Acid Sequence↗
PubMed1993

Low plasma apolipoprotein AII levels in human and mouse amyloidosis with mutant transthyretin (Met-30) gene.

We measured the serum apolipoprotein levels in patients with familial amyloidotic polyneuropathy (FAP). The serum apolipoprotein AII levels were much lower than those of the control subjects, while the levels in asymptomatic carriers of the FAP gene were normal. Other plasma apolipoprotein levels, such as apolipoproteins AI, B, CII, CIII, and E, were all within normal ranges. The decrease of apolipoprotein AII in the plasma of FAP patients correlated with the progression of the disease. In a transgenic mice model of FAP carrying human variant transthyretin gene (Met-30), serum apolipoprotein AII levels were decreased in 1.5-year-old mice compared with control mice, while the 3-month-old mice had normal levels. These results suggest that apolipoprotein AII may play an important role in lipid metabolism or amyloid formation in patients with FAP.

Adult↗
PubMed1993

Effect of denervation on overdevelopment of chloroquine-induced autophagic vacuoles in skeletal muscles.

We studied how denervation affects the overdevelopment of autophagic vacuoles in muscles of chloroquine-treated rats. The number of autophagic vacuoles increased significantly in the chloroquine-treated soleus muscles after denervation as compared to similarly treated contralateral, innervated muscles. No vacuoles were present in the denervated and innervated muscles of saline-treated rats. After denervation, the autophagic vacuoles in chloroquine-treated muscle contained numerous glycogen particles and various heterogeneous materials. A biochemical study showed no significant difference in the activities of lysosomal proteases and hydrolases in the chloroquine- and saline-treated muscles after denervation, although these activities were markedly increased in comparison to the same activities in the contralateral, innervated muscles. Chloroquine treatment by itself did not, but denervation with or without chloroquine treatment did enhance the biochemical activities of lysosomal enzymes in the animals. We speculate that denervation induces the marked accumulation of autophagic vacuoles in chloroquine-induced myopathy.

Animals↗
PubMed1993

Postoperative recovery of arterial oxygen saturation determined by pulse oximetry in pediatric patients.

Small children are physiologically subject to arterial oxygen desaturation. However, few reports have referred to the risk factors related to postanesthetic hypoxemia and the duration of hypoxemia. The purpose of this study was to clarify these two aspects. Eighty-five ASA physical status I infants and children were included in the study. They were scheduled for minor surgery. Fifty-six underwent oral endotracheal intubation, and 29 patients breathed from a mask. Anesthesia was maintained with Enflurane or Halothane and nitrous oxide. Arterial oxygen saturation was measured with a pulse oximeter. The measurements were started shortly after patients' arrival in the recovery room, and conducted every 5 min at least for 1 hour. Ten patients had SpO2 values of less than 95%. In all except one, SpO2 decreased within 10 min after arrival in the recovery room. Age, height, and weight of these 10 children were significantly different from the remaining 75, but there were no significant differences in anesthetic duration and postanesthetic awakefulness between the group with postanesthetic hypoxemia and the one without. The importance of monitoring the clinical condition of pediatric patients after general anesthesia is universally acknowledged. Monitoring with the pulse oximeter has proven very useful and shows that, unless oxygen saturation is monitored, all children should receive supplemental oxygen.

Journal Article↗
PubMed1993

Binding of streptavidin to bacteria or fungi and its applications in detecting these microbes.

We have investigated the characteristics and utilities of streptavidin-binding to gram-negative and gram-positive bacteria and Candida spp. The pre-treatment of these microbes with chemical reagents such as CHCl3, NaOH, and Tween 20 have allowed colorimetric visualization under light microscopy or quantitation on nitrocellulose membranes, using streptavidin/biotinylated alkaline phosphatase conjugates. Analysis of this binding was confirmed by western blot. These binding reactions were due to the specific interaction of streptavidin with biotinylated proteins present in the microbes. Competition assays with free biotin or inhibition by an antibiotin antibody confirmed binding to these proteins. With knowledge of these strongly specific interactions, we attempted to reveal the biotinylated proteins within these microbes using clinical specimens. Using phagocyte-smears from blood, urine, and ascites, these intracellular microbes were easily detected by light microscopy. One of the septic blood samples stained by our technique revealed semi-digested microbial signals despite the absence of a signal with routine staining. This detection system, which combines streptavidin as a probe and biotinylated proteins as a microbial marker, is useful in staining for intracellular bacteria or fungi (e.g., microbial infections in phagocyte-smears).

Alkaline Phosphatase↗
PubMed1992

Circulating autoantibody to muscle protein in a patient with paraneoplastic myositis and colon cancer.

A muscle-specific autoantibody was found in a patient with paraneoplastic myositis and colon cancer. By immunoblotting, we found high titers of circulating antibody to a 34-kDa neutral protein in the soluble sarcoplasmic fraction of the rat skeletal muscle. The serum did not react at all with other tissue extracts including the central nervous tissue, liver, or kidney. The possible role of this muscle-specific autoantibody in the pathogenesis of paraneoplastic myositis is discussed.

Adenocarcinoma↗
PubMed1992

[Utility of recombinant human erythropoietin on the anemia of elderly hemodialysis patients].

Recombinant human erythropoietin was administered to 59 patients over 65 years of age receiving maintenance hemodialysis treatment in Kyoto and Shiga district, in order to evaluate its utility on renal anemia. After 6 months of administration, the hematocrit rose from 20.8 +/- 3.5% to 28.0 +/- 3.7% with concomitant improvement of subjective symptoms related to anemia. Twelve patients developed side effects, in 10 of whom elevation of blood pressure was observed. We found no clinically significant abnormalities in the laboratory data. In conclusion, recombinant human erythropoietin is highly useful in the treatment of renal anemia in elderly hemodialysis patients.

Age Factors↗
PubMed1991

Cloning and nucleotide sequence of a human Zn-alpha 2-glycoprotein cDNA and chromosomal assignment of its gene.

A cDNA clone of Zn-alpha 2-glycoprotein (Zn alpha 2gp) was isolated from a human prostate library. The amino acid sequence of prostate Zn alpha 2gp deduced from the nucleotide sequence was identical to the one previously reported on the Zn alpha 2gp protein purified from human blood plasma, except at three positions: the 65th and 222nd amino acid residues were Gln (----Glu) and Glu (----Gln), and there was a two amino acid insertion (Ile-Phe) between the 75th (Glu) and 76th (Met) amino acids. Southern blot analysis of human genomic DNA, however, suggested a single gene encoding Zn alpha 2gp. Using a panel of rodent-human somatic cell hybrids, the Zn alpha 2 gp gene was assigned to human chromosome 7.

Amino Acid Sequence↗
PubMed1991

Alpha-smooth-muscle actin and desmin expressions in human neuroblastoma cell lines.

The neural crest gives rise to a variety of tissues, including peripheral neurons, Schwann cells, melanocytes and ectomesenchymal cells, which include the smooth-muscle cells of large arteries. Cell lines derived from neuroblastoma (a neural-crest tumor) exhibit at least 2 distinct morphological cell types, a neuroblastic phenotype (N-type) and an epithelial-like phenotype (S-type) with characteristics of substrate-adhesiveness. We have analyzed 17 human neuroblastoma cell lines using a panel of monoclonal antibodies (MAbs) against cytoskeletal proteins. Three neuroblastoma cell lines (KP-N-SI, KP-N-YN and SMS-KCN) bound an alpha-smooth-muscle actin antibody. In addition, one of these lines (KP-N-SI) bound anti-desmin MAbs as determined by indirect immunofluorescence. A total of 8 cloned cell lines were obtained from the above parent cell lines. These were composed of either N- or S-type cells and were confirmed to have the same neuroblastoma origin as each parent cell line by chromosomal analysis. Alpha-smooth-muscle actin and desmin were demonstrated in the S-type cloned cells by indirect immunofluorescence, as well as by 2-dimensional Western blot analysis. These results were confirmed by Northern blot analysis using a specific probe (pSH alpha SMA-3' UT) to human alpha-smooth-muscle actin mRNA. These ascertain the presence of alpha-smooth-muscle actin and desmin in neuroblastoma cell lines. These data show that, in addition to giving rise to cells with neural, Schwann-cell and melanocyte markers, neuroblastoma can also give rise to the cells expressing smooth-muscle cell markers.

Actins↗