Long PCR amplification of Plasmodium falciparum DNA extracted from filter paper blots.
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Animals↗
Biomedical subjects
A Kaneko
Publications and source records attributed to A Kaneko.
Long-term clinical impact of occult hepatitis B virus infection in chronic hepatitis B patients.
BACKGROUND/AIMS: Long-term clinical outcomes of occult hepatitis B virus (HBV) infection were studied. METHODS: Fifteen chronic hepatitis B patients were monitored for a median of 4.4 years (range 0.9-15.3) after hepatitis B surface antigen (HBsAg) seroclearance. Serum HBV DNA was measured by real-time detection polymerase chain reaction. Thirteen patients underwent liver biopsies at the end of follow-up and liver histology was evaluated by Ishak score. Liver HBV DNA was also measured for 12 patients. RESULTS: At the end of follow-up, HBV viremia was absent in 13 (87%) patients, and antibody titers to hepatitis B core antigen showed an inverse correlation with time from HBsAg seroclearance (r=-0.554; P=0.0040). However, all patients retained liver HBV DNA and tested positive for the covalently closed circular HBV DNA replicative intermediate. The hepatic HBV DNA loads had no relation to liver histology. Paired biopsies from 11 patients disclosed that each necroinflammatory score significantly improved after HBsAg seroclearance. Amelioration of liver fibrosis was also evident in eight (73%) patients (P=0.0391 by signed rank test). CONCLUSIONS: A long-standing but strongly suppressed HBV infection may confer histological amelioration after HBsAg seroclearance.
Adult↗
Tubal compliance--changes with age and in tubal malfunction.
OBJECTIVE: It is known that eustachian tubal compliance has influence on the tubal function. We applied the direct measurement method to ascertain whether or not aging and tubal diseases are related to the compliance of the cartilaginous part of the eustachian tube. METHODS: We developed a method for directly measuring tubal compliance by inserting a balloon catheter, which is inflated with water, into the eustachian tube and comparing the change in balloon pressure with the balloon volume. This method is advantageous in that one can directly measure tubal compliance in the cartilaginous part of the eustachian tube. Using this method we examined tubal compliance in 16 normal young volunteer ears, 21 normal elderly volunteer ears, 11 elderly ears with patulous tube and 12 elderly ears with stenotic tube including otitis media with effusion. RESULTS: In adults the tubal compliance increases in value with age. Tubal compliance of the elderly patulous tube was significantly higher than that of the normal elderly one, on the other hand the compliance of the elderly stenotic tube was significantly lower than that of the normal one. CONCLUSION: Tubal compliance was considered to be changed with aging and influence of the patulous and stenotic tubes.
Adult↗
Protective effect of OK-432 on mice against endotoxemia and infection with Pseudomonas aeruginosa and Salmonella enteritidis.
OK-432 has been used clinically as a biological response modifier for cancer therapy. We investigated here the protective effects of OK-432 against endotoxic shock and infectious death caused by Pseudomonas aeruginosa and Salmonella enteritidis in mice and proposed a possible mechanism. Pretreatment of OK-432 reduced the lethality of lipopolysaccharide (LPS)-induced endotoxic shock in D-(+)-galactosamine-sensitized C3H/HeN mice. OK-432 did not affect the TNFalpha production in blood, but it did decrease the susceptibility to TNFalpha. Furthermore, an acceleration of LPS clearance from blood was detected. The pretreatment of OK-432 also decreased the lethality of mice in bacterial infection caused by P. aeruginosa and S. enteritidis. The rapid decrease of the viable bacteria from the circulating blood and in spleen and liver in mice was observed in a manner similar to LPS clearance. These findings indicate that the protective effect of OK-432 against the endotoxemia and bacteremia may depend on an up-regulation of clearance of LPS and bacteria and the augmented resistance to TNFalpha.
Animals↗
Molecular heterogeneity of heat-stable toxin-producing and non-producing Escherichia coli O25 strains isolated in Japan.
Escherichia coli O25 strains that produce heat-stable toxin (ST) have been recently isolated in Japan, and epidemiological study of this type of enterotoxigenic E. coli is required. In this study the heterogeneity of 16 ST-producing and non-producing strains of E. coli O25 was investigated. All eight ST-producing strains were shown to have STIb gene, and seven of them had similar profiles of plasmids, ladder-banding of LPS in SDS-polyacrylamide gel electrophoresis, and chromosomal DNA digestions in pulsed-field gel electrophoresis (PFGE). In contrast, ST-non-producing strains were more heterogeneous in all parameters examined. PFGE of the digested chromosomal DNA with several restriction enzymes was proved to be an effective procedure to compare the closely related strains of E. coli O25.
Bacterial Proteins↗
Persistent Na+ current and Ca2+ current boost graded depolarization of rat retinal amacrine cells in culture.
Retinal amacrine cells are depolarized by the excitatory synaptic input from bipolar cells. When a graded depolarization exceeds the threshold level, trains of action potentials are generated. There have been several reports that both spikes and graded depolarization are sensitive to tetrodotoxin (TTX). In the present study, we investigated the contribution of voltage-gated currents to membrane depolarization by using rat GABAergic amacrine cells in culture recorded by the patch-clamp method. Injection of a negative current induced membrane hyperpolarization, the waveform of which can be well fitted by a single exponential function. Injection of positive current depolarized the cell, and the depolarization exceeded the amplitude expected from the passive properties of the membrane. The boosted depolarization sustained after the current was turned off. Either 1 microM TTX or 2 mM Co2+ suppressed the boosted depolarization, and co-application of TTX and Co2+ blocked it completely. Under the voltage clamp, we identified a transient Na+ current (fast I(Na)), a TTX-sensitive persistent current that reversed the polarity near the equilibrium potential of Na+ (I(NaP)), and three types of Ca2+ currents (I(Ca)), L, N, and the pharmacological agent-resistant type (R type). These findings suggest that the I(NaP) and I(Ca) of amacrine cells boost depolarization evoked by the excitatory synaptic input, and they may aid the spread of electrical signals among dendritic arbors of amacrine cells.
Animals↗
Our recent studies on sensory transduction: from vision to taste.
The transduction in photoreceptors and chemoreceptors has common mechanisms. In photoreceptors, activation of rhodopsin by light triggers the enzymatic cascade mediated by a G-protein, transducin. In olfactory cells activation of the receptor molecule by odorant triggers the enzymatic cascade mediated by a G-protein, Golf. In taste receptor cells biochemical studies have also suggested a metabotropic transduction hypothesis, but we recently identified a cationic channel that was directly gated by a bitter taste substance. By reviewing these recent studies carried out in our laboratory in the last 10 years, the transduction machinery in these sensory receptor cells are summarized.
Chemoreceptor Cells↗
[Convulsion following the combination of single preoperative oral administration of enoxacine and single postoperative intravenous administration of flurbiprofen axetil].
We experienced a case of convulsion following the combination of single oral administration of enoxacine before an emergency operation and single postoperative intravenous administration of flurbiprofen axetil. The patient was an 87-year-old female referred to our hospital for severe abdominal pain. She was diagnosed as having the strangulation ileus, then underwent the emergent operation of partial resection of the necrotic small intestine under general anesthesia. Unfortunately we did not know that she had temporarily received oral enoxacine 200 mg, a new quinolone, administered by the previous doctor on the day before the operation. After the operation, flurbiprofen axetil 50 mg, a nonsteroidal anti-inflammatory drug, was given intravenously in thirty seconds due to postoperative pain. One minute after administration of the drug, she immediately developed a convulsive fit, severe disturbance of consciousness and apnea. We then administered at once, a single dose of diazepam intravenously for convulsion treatment, kept her airway open and controlled her ventilation. Convulsion disappeared in a minute and her condition improved gradually. We suspect that convulsive seizure may have been induced by the drug interaction between single oral dose of enoxacine before the operation and single intravenous dose of flurbiprofen axetil after the operation. We also suspect that the serum concentration of enoxacine was kept high because of metabolic disturbance and renal dysfunction resulting from her old age and dehydration. This case suggests that medication before the emergency operation must be considered in anesthetic management because of the possible side effect such as convulsion induced by the drug interaction between neuquinolones and anti-inflammatory drugs.
Administration, Oral↗
Molecular epidemiology of methicillin-resistant Staphylococcus aureus in a hospital in Chiang Mai.
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Cross Infection↗
[Two-step interferon rebound therapy for refractory chronic hepatitis C].
In almost all patients in whom interferon(IFN) treatment dose not result in persistent negative findings for HCV-RNA, HCV-RNA levels show a rebound after discontinuing the administration. In most patients, HCV-RNA levels after administration are increased compared to pretreatment values. When a rapid increase in HCV-RNA levels causes transient exacerbation of transaminase levels, HCV-RNA levels then rapidly decrease. In two-step interferon rebound therapy(TIRT), IFN is additionally administered when HCV-RNA levels are decreased. We previously reported that TIRT was useful for treating type 1b patients with an HCV-RNA level of 1 Meq/ml or more who did not respond to IFN treatment. In the year 2000, health insurance covered the additional administration of IFN. The use of TIRT for additional administration may further improve treatment response in patients who do not respond to IFN treatment.
Adult↗
Demonstration of genome DNA diversity of strains of urease-positive thermophilic Campylobacter isolated from the natural environment by pulsed-field gel electrophoresis analysis.
Pulsed-field gel electrophoresis (PFGE) analysis was carried out after separate digestion with Apa I, Sal I and Sma I of the genomic DNA from sixteen isolates of urease-positive thermophilic Campylobacter (UPTC) obtained from the natural environment, namely from oysters and mussels, in Northern Ireland. Five NCTC strains previously isolated in England were used for the analysis. Although the eight isolates of UPTC in Northern Ireland and a strain of UPTC in England showed that one or no fragments appeared after digestion with Apa I around 1,900 to 1,640 kb region of the gel, Apa I was shown to cut the genomic DNA from all of the other twelve strains of UPTC and Sal I and Sma I from all of the 21 strains in a distinctly different and distinguishable manner. Consequently, the present study clearly demonstrated that the sixteen isolates of UPTC in Northern Ireland and the five strains in England gave the diversity of the genomic DNA by using PFGE. Some strains of UPTC examined were shown to carry genomes from 1.6 to 1.9 Mb in length, thus the heterogeneous profiles of PFGE and the length of the genomes are thought to occur among the isolates of UPTC in Northern Ireland, as well as among the five representative strains of UPTC from NCTC.
Animals↗
Endorphin and enkephalin ameliorate excessive synovial cell functions in patients with rheumatoid arthritis.
OBJECTIVE: To determine whether endorphin (END) and enkephalin (ENK) modulate excessive synovial cell functions in patients with rheumatoid arthritis (RA). METHODS: Effects of leucine-enkephalin (leu-ENK), methionine-enkephalin (met-ENK), and beta-endorphin (END) on proinflammatory cytokine and matrix metalloproteinase (MMP) production by RA synovial cells were analyzed by immunoblotting, and their mRNA expression by reverse transcription-polymerase chain reaction (RT-PCR) using limiting dilution of complementary DNA. Expression of opioid receptors on RA synovial cells was assessed by immunohistochemical staining, radioreceptor assay, and RT-PCR. RESULTS: Leu-ENK, met-ENK, and END inhibited tumor necrosis factor-alpha and interleukin 1beta production at the level of mRNA expression. ENK and END inhibited MMP-9 production and its enzymatic activity by RA synovial cells. The mu-subtype opioid receptor was expressed in the RA synovial lining and sublining cells. Radioreceptor assay suggested expression of high affinity receptor for END on RA synovial cells. The mu-subtype opioid receptor-specific antagonist, naloxone, reversed the inhibitory effect of the opioid peptides. The opioid peptides inhibited nuclear translocation and phosphorylation of the transcription factor, cyclic AMP responsive element binding protein (CREB) in RA synovial cells. CONCLUSION: Leu-ENK, met-ENK, and END inhibited excessive RA synovial cell functions in vitro. The opioid hormones may have not only antinociceptive action, but also antiinflammatory effects on synovitis itself in RA.
Aged↗
Pulmonary Mycobacterium avium infection in an immunocompetent young adult related to use of home bath with a circulating water system.
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Adult↗
A new member of the HCO3(-) transporter superfamily is an apical anion exchanger of beta-intercalated cells in the kidney.
The kidneys play pivotal roles in acid-base homeostasis, and the acid-secreting (alpha-type) and bicarbonate-secreting (beta-type) intercalated cells in the collecting ducts are major sites for the final modulation of urinary acid secretion. Since the H(+)-ATPase and anion exchanger activities in these two types of intercalated cells exhibit opposite polarities, it has been suggested that the alpha- and beta-intercalated cells are interchangeable via a cell polarity change. Immunohistological studies, however, have failed to confirm that the apical anion exchanger of beta-intercalated cells is the band 3 protein localized to the basolateral membrane of alpha-intercalated cells. In the present study, we show the evidence that a novel member of the anion exchanger and sodium bicarbonate cotransporter superfamily is an apical anion exchanger of beta-intercalated cells. Cloned cDNA from the beta-intercalated cells shows about 30% homology with anion exchanger types 1-3, and functional expression of this protein in COS-7 cells and Xenopus oocytes showed sodium-independent and 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid-insensitive anion exchanger activity. Furthermore, immunohistological studies revealed that this novel anion exchanger is present on the apical membrane of beta-intercalated cells, although some beta-intercalated cells were negative for AE4 staining. We conclude that our newly cloned transporter is an apical anion exchanger of the beta-intercalated cells, whereas our data do not exclude the possibility that there may be another form of anion exchanger in these cells.
4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
Malaria eradication on islands.
BACKGROUND: To be successful, a malaria control programme needs to be tailored to the local epidemiological characteristics. Vanuatu consists of 80 inhabited islands in the Southwest Pacific, with hypoendemic and mesoendemic malaria and suitable conditions for sustained parasite elimination. We aimed to assess whether malaria can be eliminated on isolated islands. METHODS: Weekly mass drug administration of chloroquine, pyrimethamine/sulfadoxine (Fansidar), and primaquine was carried out on the entire population of 718 inhabitants of Aneityum island for 9 weeks in 1991 before the onset of the rainy season. Simultaneously with the administration of drugs, permethrin-impregnated bednets were distributed to the entire population. Larvivorous fish were also introduced into several identified breeding sites of Anopheles farauti. Periodic malariometric monitoring has continued for the past 9 years. Two additional islands of Vanuatu, one with and one without malaria transmission, have been monitored for comparison. FINDINGS: High community involvement as measured by drug compliance (88.3%) and bednet provision (0.94 nets per villager) has resulted in sustained interruption of malaria transmission in Aneityum. The surveys showed complete absence of Plasmodium falciparum after mass drug administration, and P. vivax disappeared from 1996 onwards, with the exception of two instances of imported infections (one mixed infection in 1993 and one P. vivax infection in 1999). INTERPRETATION: Malaria can be eliminated on isolated islands with well-adapted short-term mass drug administration and sustained vector control if there is a high degree of community participation.
Adolescent↗
MgcRacGAP is involved in the control of growth and differentiation of hematopoietic cells.
In a search for key molecules that prevent murine M1 leukemia cells from undergoing interleukin (IL)-6-induced differentiation into macrophages, we isolated an antisense complementary DNA (cDNA) that encodes full-length mouse MgcRac-GTPase-activating protein (GAP) through functional cloning. Forced expression of this antisense cDNA profoundly inhibited IL-6-induced differentiation of M1 cells into macrophage lineages. We also isolated a full-length human MgcRacGAP cDNA, which encodes an additional N-terminal polypeptide of 105 amino acid residues compared with the previously published human MgcRacGAP. In human HL-60 leukemic cells, overexpression of the full-length form of human MgcRacGAP alone induced growth suppression and macrophage differentiation associated with hypervacuolization and de novo expression of the myelomonocytic marker CD14. Analyses using a GAP-inactive mutant and 2 deletion mutants of MgcRacGAP indicated that the GAP activity was dispensable, but the myosin-like domain and the cysteine-rich domain were indispensable for growth suppression and macrophage differentiation. The present results indicated that MgcRacGAP plays key roles in controlling growth and differentiation of hematopoietic cells through mechanisms other than regulating Rac GTPase activity.
Amino Acid Sequence↗
A method to identify cDNAs based on localization of green fluorescent protein fusion products.
We previously established a high-efficiency, retrovirus-mediated expression cloning method. Using this system, we now have developed an expression cloning method (FL-REX; fluorescence localization-based retrovirus-mediated expression cloning) in which cDNAs can be isolated based on the subcellular localization of their protein products. Complementary DNAs generated from mRNA using random hexamers were fused to the cDNA of green fluorescent protein (GFP) in the pMX retrovirus vector. The resulting cDNA-GFP fusion library was transfected into retrovirus-packaging cells, and the derived retroviruses were used to infect NIH 3T3 cells. Infected cells then were screened to identify cDNAs of interest through the subcellular localization of the GFP-fusion products. Using FL-REX, we have identified 25 cDNAs, most of which showed reasonable subcellular localization as GFP-fusion proteins, indicating that FL-REX is useful for identification of proteins that show specific intracellular localization.
3T3 Cells↗
Tetrodotoxin-sensitive persistent current boosts the depolarization of retinal amacrine cells in goldfish.
To light illumination retinal amacrine cells respond with graded depolarization accompanied by a spike discharge. It has been assumed that the graded depolarization is produced solely by the excitatory synaptic inputs from bipolar cells. Here we demonstrate that a tetrodotoxin (TTX)-sensitive persistent current also contributes to the graded depolarization. This current was isolated in amacrine cells of the goldfish retinal slice preparations by the whole-cell patch clamp technique. The activation threshold of the persistent current was about -50 mV, approximately 10 mV more positive than the membrane potential in the dark. Therefore, it is highly likely that the TTX-sensitive persistent current is a booster of the excitatory postsynaptic potential in amacrine cells.
Action Potentials↗