PubMed HealthSearch

Biomedical subjects

K Nakashima

Publications and source records attributed to K Nakashima.

At least 19 recordsLinked to original sources

Preparation of a fluorescent derivative of benzoylecgonine, and preliminary studies of its application to the analysis of urine.

A sensitive high-performance liquid chromatographic method using 3-bromomethyl-6,7-dimethoxy-1-methyl-2(1H)-quinoxalinone (Br-DMEQ) as a fluorescent labeling reagent is described for the determination of benzoylecgonine (BE) and ecgonine (EC). The Br-DMEQ derivatives of BE and EC were separated on a C18 column and detected at 455 nm with excitation at 370 nm. The detection limits of the proposed method were 18.7 fmol for BE and 12.5 pmol for EC at a signal-to-noise ratio of 3. Relative standard deviations of five replicate measurements were 1.94% (10 pmol) and 2.98% (50 pmol) for BE and 6.3% (250 pmol) and 5.62% (1.25 pmol) for EC. This method was applied to the determination of BE in human urine. BE was extracted from urine by solvent extraction with chloroform-isopropyl alcohol (9:1, v/v) solution. Levels of 2.5.10(-8) M BE in urine (25 pmol/ml) could be determined.

Chromatography, High Pressure Liquid

Cross-contamination potential with dental equipment.

Some types of reused dental equipment, especially handpieces and their attachments for drilling and cleaning teeth, might be responsible for cross-contamination if patient material were to lodge temporarily in difficult-to-disinfect internal mechanisms. This possibility is worrisome with respect to transmission of hepatitis B and human immunodeficiency viruses (HBV, HIV). Previous cross-contamination studies have relied on laboratory experiments with bacteria or dye tracers. To assess possible risk more thoroughly, we tested 30 new prophylaxis angles and 12 new high-speed handpieces to see whether they would take up and expel contaminants in laboratory and clinical trials. In treatments of three patients, including two infected with HIV, human-specific DNA (beta-globin, HLA DQ alpha) and HIV proviral DNA were detected inside or coming back from the devices. Similarly, when handpieces were operated in contact with blood pooled from HBV-infected patients, HBV DNA was detected in samples taken from inside the equipment and from their attached air/water hoses. When we used bacteriophage phi X174 as a model virus in laboratory tests, many infective viral particles were recovered from internal mechanisms of handpieces, their connecting air/water hoses, and from water spray expelled when the equipment was reused. We recommend that reused high-speed, air-driven handpieces and prophylaxis angles should be cleaned and heat-treated between patients. Further studies are needed to determine ways of eliminating the risks associated with exhaust hoses and air/water input lines.

Bacteriophages

Sexual transmission of hepatitis C virus among female prostitutes and patients with sexually transmitted diseases in Fukuoka, Kyushu, Japan.

The authors investigated the prevalence of antibody to hepatitis C virus (anti-HCV) in 404 female prostitutes, 428 clinic patients with a history of at least one episode of sexually transmitted disease, and 8,944 blood donors who served as the controls. All subjects were Japanese, and all studies were carried out in Fukuoka, Kyushu, Japan, in 1989. The prevalence of anti-HCV was significantly higher in the prostitutes (6.2%), in the female patients with sexually transmitted diseases (6.1%), and in the male patients with sexually transmitted diseases (2.9%) than in the controls (1.5%). Prevalence of anti-HCV increased with age in prostitutes and in the controls. The prevalence of anti-HCV in those who had been involved in prostitution for 1 year or more (8.1%) was higher than in those who had been involved in prostitution for less than 1 year (1.4%), but the difference was not statistically significant. One of the 152 anti-HCV negative prostitutes seroconverted between 1 and 2 years later. Among the subjects with sexually transmitted diseases, patients with a history of at least one episode of syphilis had a significantly higher prevalence of anti-HCV (4.4%) than the controls. Patients with acute urethritis and cervicitis also showed a high prevalence of anti-HCV (3.6% and 6.7%, respectively). These data support the possibility of sexual transmission of hepatitis C virus.

Adolescent

Double antenna structure of chicken prolactin receptor deduced from the cDNA sequence.

Chicken prolactin receptor (cPRLR) deciphered from the cDNA sequence showed a unique double antenna structure in its extracellular domain. The predicted cPRLR preprotein was composed of 831 amino acids and contained a signal peptide and a transmembrane region. The extracellular domain comprised 438 residues, and was divided into two tandemly repeated, highly homologous units, each of which corresponded to the extracellular domains of mammalian prolactin receptors. Both extracellular units of cPRLR possessed two structural features characteristic of the ligand binding units of cytokine/prolactin receptor family, namely two pairs of cysteine residues and a WSXWS motif. These findings strongly suggest that cPRLR contains two repeated ligand binding units, that is a double antenna structure. The cPRLR gene is expressed in a wide range of tissues of laying hen.

Amino Acid Sequence

Unusually high conservation of untranslated sequences in cDNAs for Trimeresurus flavoviridis phospholipase A2 isozymes.

As a step toward understanding the structure and function of phospholipases A2 (PLA2s), we isolated and sequenced several cDNAs encoding Trimeresurus flavoviridis venom PLA2 isozymes including two [Lys49]PLA2s called basic proteins I and II, [Thr37]PLA2, and PLX'-PLA2. Comparison of the nucleotide sequences of these cDNAs with the previously isolated [Asp49]PLA2 cDNA revealed some interesting findings from the viewpoint of evolution. First, the homologies of the 5' and 3' untranslated regions (98% and 89%, respectively) were much higher than that of the protein-coding regions (67%). The predicted secondary structure showed the characteristic stem-loop structures for both the untranslated regions of the mRNAs, suggesting that these regions play some functional role(s) in translation or stability of mRNAs. Second, base substitutions appeared to have occurred at similar rates for the three positions of codons among these PLA2s. The results are discussed in terms of evolution of PLA2s. Northern blot analysis showed that these PLA2s are specific to venom gland.

Amino Acid Sequence

Structure of the chicken growth hormone-encoding gene and its promoter region.

We have cloned the chicken (c) growth hormone (GH)-encoding gene cGH and analyzed its nucleotide sequence including 500 bp of the 5'-flanking region. The cGH gene consists of five exons and four introns as has been observed in the mammalian GH genes. However, the size of the cGH gene is significantly larger than that of analogous mammalian genes, because of its intron size which expands it to 3.5 kb. The transcription start point was determined to be 56 bp upstream from the start codon by the primer-extension analysis. The promoter region of the cGH gene has no overall homology with the corresponding regions of mammalian genes, but contains a short (24 bp) sequence which is highly homologous to the antisense strand sequence of the proximal binding site for a pituitary-specific transcription factor, GHF-1/Pit-1, in the promoter region of the rat GH gene.

Amino Acid Sequence

Chloroplast DNA of black pine retains a residual inverted repeat lacking rRNA genes: nucleotide sequences of trnQ, trnK, psbA, trnI and trnH and the absence of rps16.

A physical map of black pine (Pinus thunbergii) chloroplast DNA (120 kb) was constructed and two separate portions of its nucleotide sequence were determined. One portion contains trnQ-UUG, ORF510, ORF83, trnK-UUU (ORF515 in the trnK intron), ORF22, psbA, trnI-CAU (on the opposing strand) and trnH-GUG, in that order. Sequence analysis of another portion revealed the presence of a 495 bp inverted repeat containing trnI-CAU and the 3' end of psbA but lacking rRNA genes. The position of trnI-CAU is unique because most chloroplast DNAs have no gene between psbA and trnH (trnI-CAU is usually located further downstream). Black pine chloroplast DNA lacks rps16, which has been found between trnQ and trnK in angiosperm chloroplast DNAs, but possesses ORF510 instead. This ORF is highly homologous to ORF513 found in the corresponding region of liverwort chloroplast DNA and ORF563 located downstream from trnT in Chlamydomonas moewusii chloroplast DNA. A possible pathway for the evolution of black pine chloroplast DNA is discussed.

Amino Acid Sequence

Hepatitis C virus detection is facilitated by the combined use of c100 protein and GOR epitope.

Assay for the antibody to the c100 protein (anti-c100) lacks sensitivity in terms of detection of hepatitis C virus (HCV) in all samples. The author used anti-c100 and antibody to the GOR epitope (anti-GOR) by the enzyme-linked immunosorbent assay to examine 524 patients with chronic liver disease and 682 volunteer blood donors in Fukuoka, Japan. The prevalence of HCV infection, as revealed by the presence of anti-c100 and/or anti-GOR, was 3.9% in 540 volunteer blood donors, 12.7% in 142 volunteers with abnormal liver function, 7.4% in 135 patients with HBsAg-positive liver disease and 89.5% in 389 patients with non-A, non-B (NANB) liver disease. These results show a higher prevalence than demonstrated only by the anti-c100 in NANB liver disease patients (82.5%, P < 0.01). The concurrence of anti-c100 and anti-GOR in subjects with HCV infection was 23.8% in 21 volunteer blood donors, 44.4% in 18 volunteers with abnormal liver function and 61.2% in 348 NANB liver disease patients. The concurrence seems to increase with deterioration of liver function. We concluded that combination assay for anti-c100 and anti-GOR demonstrated a more accurate prevalence of HCV infection than single assay for anti-c100 among NANB liver disease patients, and that the presence of anti-GOR plays a role in liver disease in anti-HCV-positive subjects.

Adolescent

Role of cholesterol-accumulating macrophages on vascular smooth muscle cell proliferation.

The proliferation of vascular smooth muscle cells (VSMC) was stimulated by co-incubation with macrophages. Further stimulation was observed when co-incubated macrophages were supplied with LDL or cholesterol. However, the stimulation of VSMC proliferation did not result from co-incubation with macrophages supplemented with acetylated LDL or delipidated LDL. The addition of anti-PDGF antibody partially abolished the stimulation of VSMC proliferation induced by co-incubation with macrophages supplemented with LDL or cholesterol. A high concentration of prostaglandin E2 inhibited the proliferation of VSMC stimulated by PDGF and plasma-derived serum when they were at the G0/G1 stage. However, the inhibitory effect of prostaglandin E2 on proliferation was not observed when cells were incubated with macrophages supplemented with LDL or cholesterol in spite of the promotion under these conditions of prostaglandin E2 production. These results suggest that cholesterol-accumulating macrophages may exert a regulatory effect on the proliferation of VSMC through the synthesis and secretion of platelet-derived growth factor (PDGF) and prostaglandin E2, besides foam-cell formation.

Animals

Recovery functions of somatosensory evoked potentials in parkinsonian patients.

The aim of this study was to analyze the recovery functions of median nerve somatosensory evoked potentials (SEPs) and to clarify changes in the somatosensory system of patients with Parkinson's disease (PD). The central and frontal SEPs and the nerve potential of the median nerve were examined. The latencies and amplitudes of SEPs produced by a single shock in patients with PD were normal. The recovery functions of central SEPs showed a low degree of suppression in patients with PD, although the recovery curve of the nerve potential in PD patients was almost normal. The change in recovery curves of SEPs in patients with sensory complaints were more noticeable than that in patients without sensory disturbance. A low degree of suppression of central SEPs might play a role in the sensory complaints of PD patients.

Adult

Glycated hemoglobin of fractionated erythrocytes, glycated albumin, and plasma fructosamine during pregnancy.

OBJECTIVE: To evaluate glucose metabolism during pregnancy, we measured plasma fructosamine, glycated albumin, and the stable glycated hemoglobin of the light and dense erythrocytes. STUDY DESIGN: The abnormal glucose tolerance group comprised patients with gestational diabetes and those with one abnormal value on a 75 gm oral glucose tolerance test. Erythrocyte fractionation was performed by capillary centrifugation. RESULTS: In normal pregnancy glycated hemoglobin of the light erythrocytes was reduced in the second and third trimesters (3.42% +/- 0.62% [mean +/- SD] [n = 306] in the first trimester, 2.15% +/- 0.48% [n = 353] in the second, and 2.06% +/- 0.58% [n = 300] in the third), and dense erythrocytes were higher in the third trimester (first 4.59% +/- 0.46%, second 4.70% +/- 0.49%, third 5.29% +/- 0.73%). Glycated albumin and fructosamine followed a pattern similar to the light erythrocytes. The group with abnormal glucose tolerance had significantly higher levels of glycated hemoglobin of the light erythrocytes in the first and third trimesters and glycated hemoglobin of the dense erythrocytes and glycated albumin in all trimesters. CONCLUSION: The biphasic change in nonfractionated glycated hemoglobin is the sum of the lower glycated hemoglobin of the light erythrocytes and the higher glycated hemoglobin of the dense erythrocytes in late pregnancy. The stable glycated hemoglobin of fractionated erythrocytes and the glycated albumin accurately reflect maternal glucose metabolism during pregnancy.

Adult

The effects of breastfeeding and presence of antibody to p40tax protein of human T cell lymphotropic virus type-I on mother to child transmission.

We examined the effects of various factors, including duration of breastfeeding, the status of mother's anti-p40tax, and titre of mother's anti-human T cell lymphototropic virus type-I (HTLV-I) on mother to child transmission of HTLV-I in 76 HTLV-I carrier mothers and 175 of their children. The overall prevalence of anti-HTLV-I among children was 16.0%. The prevalence of anti-HTLV-I among children breastfed for over 3 months was significantly higher (27.6%) than that of those breastfed for under 3 months (5.1%; P = 0.012). Of the 78 bottle-fed children, 10 (12.8%) were positive for anti-HTLV-I. In the children breastfed for over 3 months, the prevalence of anti-HTLV-I among 37 children of anti-p40tax positive mothers was 37.8% and that of 21 children of anti-p40tax negative mothers was 9.5%, a significant difference (P = 0.044). These data suggest that about 13% of bottle-fed children born to carrier mothers are infected with HTLV-I by routes other than breast milk, and that the mother's anti-p40tax can serve as a marker of infectivity of HTLV-I in the case of breastfeeding for over 3 months.

Adolescent

Hepatic venous hemoglobin oxygen saturation predicts liver dysfunction after hepatectomy.

A fiberoptic flow-directed catheter inserted into the hepatic vein continuously measures hepatic venous oxygen hemoglobin saturation (ShvO2). This study determined whether intraoperatively measured ShvO2 could predict postoperative serum activities of aminotransferases and patient outcome in 83 patients undergoing hepatectomy. The duration of intraoperative ShvO2 less than or equal to 10, 20, 30, 40, and 50% was calculated in each case. Significant increases in postoperative serum aminotransferases were associated with more than 1, 11, 31, 51, and 181 min of duration of ShvO2 less than or equal to 10, 20, 30, 40, and 50%, respectively. The incidence of postoperative liver failure significantly increased when the duration of ShvO2 less than or equal to 20, 30, and 40% exceeded 11, 31, and 51 min, respectively. The mortality from liver failure was significantly higher when the duration of ShvO2 less than or equal to 30 and 40% exceeded 31 and 51 min, respectively. Therefore, intraoperative monitoring of ShvO2 may predict not only the increase in postoperative serum aminotransferases but also patient outcome in terms of postoperative liver failure after hepatectomy.

Adult

The effect of dobutamine on hepatic blood flow and oxygen supply-uptake ratio during enflurane nitrous oxide anesthesia in humans undergoing liver resection.

Liver surgery is often accompanied by hepatic hypoperfusion and hypoxia, and it is controversial whether catecholamines increase hepatic blood flow and oxygen supply. The effects of 3 micrograms.kg-1.min-1 dobutamine on hepatic circulation and oxygen balance were examined in patients anesthetized with enflurane, nitrous oxide, and oxygen for liver surgery. Dobutamine did not cause a significant increase in hepatic arterial blood flow. However, total hepatic blood flow and portal venous blood flow were increased, resulting in an increase in hepatic oxygen delivery (HDO2). The increase in HDO2 was not associated with an improvement of hepatic oxygen supply-uptake ratio, since hepatic oxygen uptake (HVO2) also increased. After hepatectomy, the increases in portal venous blood flow and HDO2 were not accompanied by an increase in HVO2. The stimulation of hepatocellular oxygen metabolism by dobutamine and depressed responsiveness of adrenoceptors on hepatocytes in which metabolism was already augmented are the likely explanation for the different reactions before and after hepatectomy.

Adult

Spectral analysis of heart rate variability during isoflurane anesthesia.

The autonomic nervous system is an important neural control system for maintaining cardiovascular stability in humans. Analysis of heart rate variations may provide important clinical information on the influence of anesthesia on the autonomic nervous system and the central nervous system. Therefore, the effects of 1.0, 1.5, and 2.0 minimum alveolar concentrations of isoflurane anesthesia on beat-to-beat heart rate variations were studied in ten patients (ASA Physical Status 1). Spectral analysis was used to determine the intensity of the variations. For each power spectrum, the frequency components were identified as follows: 1) the parasympathetically mediated respiratory component (0.15-0.4 Hz) and 2) both parasympathetically and sympathetically mediated components (0.04-0.15 Hz). The latter was subdivided into the low-frequency component (0.04-0.09 Hz) of vasomotor origin and the mid-frequency component (0.09-0.15 Hz) of baroreceptor origin. Marked reductions in the power of heart rate variations, at all frequencies, were found during isoflurane anesthesia, indicating isoflurane decreased total autonomic nervous system activity. Isoflurane decreased the high-frequency and mid-frequency components in a concentration-dependent manner. The low-frequency component increased transiently at 1.5 minimum alveolar concentrations concomitant with the burst suppression in the electroencephalogram. The ratio of mid-frequency to high-frequency components did not change significantly during isoflurane anesthesia compared with the awake period. These frequency characteristics of heart rate variations during isoflurane anesthesia suggest there are dose-related decreases in autonomic nervous system activity in both the vagus and the cardiac sympathetic nerves.

Adult

Methylglyoxal bis(butylamidinohydrazone) exhibits antitumor effect on human malignant melanoma cells but reduces the antitumor action of cisplatin.

The antitumor effect of a polyamine biosynthetic pathway inhibitor methylglyloxal bis(butylamidinohydrazone) (MGBB) on human malignant melanoma (HMG) cells and its combination effect with cisplatin were investigated. The growth of cultured HMG cells was inhibited in a dose dependent manner by either MGBB or cisplatin; complete inhibition of cell proliferation was attained with 5 micrograms/ml of MGBB or 50 micrograms/ml of cisplatin. Pretreatment of HMG cells with MGBB diminished the antitumor action of cisplatin. The cultured HMG cells were inoculated in nude mice and aliquots of the resulting solid tumors (HMG tumor) were transplanted. The growth of transplanted HMG tumors in mice was inhibited markedly by cisplatin (3.8 mg/kg) and moderately by MGBB (10 or 20 mg/kg). The in vivo antitumor effect of cisplatin was also reduced by combined treatment with MGBB.

Animals

Clarification of the structural and functional features of the osmoregulated kdp operon of Escherichia coli.

Expression of the Escherichia coli kdpABC operon, which is responsible for a high-affinity potassium-uptake system, is regulated in response to a change in the medium osmolarity. In this study, we clarified the structure and function of the kdpABC promoter including its regulatory sequence at the molecular level. The canonical -35 and -10 regions determined for the promoter were not fully functional, i.e. in addition to them, a cis-acting sequence located upstream of the -35 region was essential for full activation of the promoter. This upstream sequence was demonstrated to be the target site for the trans-acting activator, KdpE.

Bacterial Proteins