Biomedical subjects

Y Sako

Publications and source records attributed to Y Sako.

At least 19 recordsLinked to original sources

PubMed2005

Evaluation of clinical and serological data from Taenia solium cysticercosis patients in eastern Inner Mongolia Autonomous Region, China.

Neurocysticercosis (NCC) is one of the major causes of neurological disease in China. ELISA and immunoblotting using glycoproteins purified by preparative isoelectric focusing were used to detect human cysticercosis in Tongliao area, Inner Mongolia, China in 1998. Approximately 89% (39 of 44 inpatients and outpatients with suspected NCC at Tongliao City Hospital) were residents of Inner Mongolia. About 53% were male and 77% were of working age (18-59 years), and 32% were farmers. Immunoblotting and ELISA showed a high correlation. Of the 44 patients, 31 positive by cerebral computed tomography (CT) scan were confirmed serologically to have cysticercosis. In the ELISA, patients with no lesions by CT scan had lower OD values, similar to those of normal serum. These findings confirm that both ELISA and immunoblotting assays are sufficiently sensitive to detect asymptomatic or symptomatic cysticercosis patients.

Adolescent↗

PubMed2005

Sulfurihydrogenibium yellowstonense sp. nov., an extremely thermophilic, facultatively heterotrophic, sulfur-oxidizing bacterium from Yellowstone National Park, and emended descriptions of the genus Sulfurihydrogenibium, Sulfurihydrogenibium subterraneum and Sulfurihydrogenibium azorense.

A novel thermophilic, sulfur-oxidizing Gram-negative bacterium, designated strain SS-5T, was isolated from the Calcite Hot Springs in Yellowstone National Park, USA. The cells were motile rods (1.2-2.8 microm long and 0.6-0.8 microm wide). The new isolate was a facultative heterotroph capable of using elemental sulfur or thiosulfate as an electron donor and O2 (1-18 %; optimum 6 %, v/v) as an electron acceptor. Hydrogen did not support growth. The isolate grew autotrophically with CO2. In addition, strain SS-5T utilized various organic carbon sources such as yeast extract, tryptone, sugars, amino acids and organic acids. Growth was observed between 55 and 78 degrees C (optimum 70 degrees C; 3.5 h doubling time), pH 6.0 and 8.0 (optimum pH 7.5), and 0 and 0.6 % (w/v) NaCl (optimum 0 %). The G+C content of the genomic DNA was 32 mol%. Phylogenetic analysis based on the 16S rRNA gene sequence indicated that the isolate was a member of the genus Sulfurihydrogenibium. On the basis of the physiological and molecular characteristics of the new isolate, we propose the name Sulfurihydrogenibium yellowstonense sp. nov. with SS-5T (=JCM 12773T=OCM 840T) as the type strain. In addition, emended descriptions of the genus Sulfurihydrogenibium, Sulfurihydrogenibium subterraneum and Sulfurihydrogenibium azorense are proposed.

Base Composition↗

PubMed2003

Evaluation of tongue inspection and serology for diagnosis of Taenia solium cysticercosis in swine: usefulness of ELISA using purified glycoproteins and recombinant antigen.

Evaluation of serology using glycoproteins (GPs) purified by preparative isoelectric focusing (pH 8.8) and recombinant chimeric antigen (RecTs) of Taenia solium was carried out using (1) blood samples on filter papers from pigs infected with different doses of eggs of T. solium in Mexico, (2) serum samples from pigs found infected naturally in Vietnam and Ecuador and (3) serum samples from pigs suspected to be infected with T. solium by tongue inspection in Tanzania. Antibody responses (IgG) were detectable in experimentally infected pigs confirmed harbouring 16 or more cysts at necropsy from 30 days after egg inoculation. One of three pigs naturally infected and harbouring 2.5 cysts/kg muscle and most of pigs harbouring=5.0 cysts/kg were also seropositive by ELISA. Although pigs may be infected with other taeniid species such as Taenia hydatigena, pigs harbouring this parasite were negative in ELISA. Approximately, 76 and 78% of sera from pigs having nodule(s) in the tongue (positive tongue inspection) were serologically positive by both ELISA and immunoblot, respectively. Furthermore, approximately 34 and 18% of sera from pigs having no nodules in the tongue (negative tongue inspection) were also seropositive by ELISA and immunoblot, respectively. ELISA using the two antigens was more sensitive than immunoblot and reliable for differentiation of pigs infected with cysticerci of T. solium from those either uninfected or infected with other taeniid species. Pigs without nodule by tongue inspection should be checked serologically in endemic areas.

Animals↗

PubMed2002

A phylogenetic hypothesis for the distribution of two genotypes of the pig tapeworm Taenia solium worldwide.

Genetic polymorphism was determined among 13 isolates of Taenia solium from various regions using PCR-amplified sequences of 2 mitochondrial genes: cytochrome c oxidase subunit 1 and cytochrome b. The 2 phylogenies obtained were similar to each other regardless of the genes examined. The isolates from Asia (China, Thailand, Irian Jaya and India) formed a single cluster, whereas the isolates from Latin America (Mexico, Peru, Ecuador, Bolivia and Brazil) combined with those from Africa (Tanzania, Mozambique and Cameroon) to form an additional cluster. These results and historical data of swine domestication, distribution of pigs and colonization suggest that T. solium was introduced recently into Latin America and Africa from different regions of Europe during the colonial age, which started 500 years ago, and that the tapeworm of another origin independently spread in Asian countries.

Animals↗

PubMed2002

Effects of biotin on glucotoxicity or lipotoxicity in rat pancreatic islets.

Biotin (vitamin H) plays an important role as a cofactor in glucose or lipid metabolism. We showed that biotin potentiated glucose-induced insulin release in isolated rat islets, while biotin alone did not affect insulin release. Coculture with biotin in islets for 48 hours significantly enhanced glucose-induced insulin release or islet insulin content. Similarly, preproinsulin or pancreatic/duodenal homeobox-1 (PDX-1) mRNA was also enhanced in islets cultured with biotin for 48 hours. Furthermore, we measured effects of biotin on beta-cell function under glucotoxic or lipotoxic states. In islets cultured with high glucose or palmitate for 48 hours, glucose-induced insulin release or islet insulin content deteriorated. Coculture with biotin significantly restored glucose-induced insulin release or islet insulin content together with the restoration of preproinsulin or PDX-1 mRNA. We conclude that biotin exerts its beneficial effects on beta-cell dysfunction induced by glucose or free fatty acids probably through the enhancement of insulin biosynthesis.

Animals↗

PubMed2002

Dogs as alternative intermediate hosts of Taenia solium in Papua (Irian Jaya), Indonesia confirmed by highly specific ELISA and immunoblot using native and recombinant antigens and mitochondrial DNA analysis.

Serology (ELISA and immunoblot) using native glycoproteins, affinity purified glycoproteins, and a recombinant antigen is known to be highly specific to Taenia solium cysticercosis in humans and pigs. These techniques were applied for dogs in the highly endemic area of cysticercosis in Papua (Irian Jaya), Indonesia. Analysis of dog sera by both ELISA and immunoblot revealed 7 of 64 dogs were highly positive. Examination of two sero-positive dogs revealed cysticerci of T. solium in the brain and heart of these dogs. Mitochondrial DNA analysis confirmed that they were the same as T. solium previously confirmed from pigs and biopsies from local people from Irian Jaya. It is suggested that the life cycle of T. solium may be completed not only between humans and pigs but also between humans and dogs.

Animals↗

PubMed2002

Recent advances in basic and applied science for the control of taeniasis/cysticercosis in Asia.

Detection of seven specific bands by immunoblot (IB) using glycoproteins (GPs) purified by lentil-lectin affinity chromatography has been the gold-standard for neurocysticercosis (NCC) serodiagnosis since 1989. However, due to the presence of contaminants, it was impossible to apply the GPs to ELISA. Our group at Asahikawa Medical College (AMC) succeeded in purifying the GPs by preparative isoelectric focusing; these higher quality GPs were suitable for ELISA. Based on the results of both IB and ELISA testing, developed at AMC for a field survey in Irian Jaya, it became evident that that area had pandemic NCC. We found many NCC patients, pigs full of cysts, and one dog infected with two cysts: these findings were based on serology. Recently, we conducted another survey to detect of the worm carriers of T. solium. Three of the 38 local people were positive by copro-antigen specific to Taenia species; these three patients expelled segments of Taenia spp and these were confirmed as those of T. solium by mitochondrial DNA analysis. When viable eggs of any taeniid species could be obtained, they can be developed into metacestodes in NOD-scid mice; it then becomes possible to analyze morphological dynamics, metacestode antigenicity, the efficacy of new metacestocidal drugs, and mitochondrial DNA. Mitochondrial DNA analysis of the specimens obtained in Irian Jaya was compared with that of other isolates worldwide. T. solium is now divided into two genotypes: the Asian type, and the Africa-American type. Some aspects of the pathological differences between the Asian and Africa-American types and the antigenic components of these two types are discussed.

Animals↗

PubMed2001

Single-molecule analysis of chemotactic signaling in Dictyostelium cells.

Single-molecule imaging techniques were used to reveal the binding of individual cyclic adenosine 3',5'-monophosphate molecules to heterotrimeric guanine nucleotide-binding protein coupled receptors on the surface of living Dictyostelium discoideum cells. The binding sites were uniformly distributed and diffused rapidly in the plane of the membrane. The probabilities of individual association and dissociation events were greater for receptors at the anterior end of the cell. Agonist-induced receptor phosphorylation had little effect on any of the monitored properties, whereas G protein coupling influenced the binding kinetics. These observations illustrate the dynamic properties of receptors involved in gradient sensing and suggest that these may be polarized in chemotactic cells.

Animals↗

PubMed2001

Effects of bezafibrate on beta-cell function of rat pancreatic islets.

Bezafibrate is an activator of peroxisome proliferator-activated receptors (PPAR) alpha. The present study was performed to investigate the effects of bezafibrate and the PPAR alpha activator, 4-Cholro-6-(2.3-xylidino)-2-pyrimidin-ylthio acetic acid (WY14643), on the beta-cell function of rat pancreatic islets in vitro. In islets cultured with 300 microM bezafibrate or WY14643 for 8 h, a low glucose concentration induced insulin release and increased the levels of mRNA for PPAR alpha, acyl CoA oxidase, carnitine palmitoyl transferase-1, pyruvate dehydrogenase E1 alpha or pyruvate carboxylase. In contrast, after a 48-h culture period, a high glucose concentration induced insulin release and islet insulin content, but decreased the levels of mRNA for glucose transporter-2 (GLUT-2), preproinsulin or pancreatic/duodenal homeobox-1. Diazoxide, the KATP channel opener, restored these responses. We conclude that bezafibrate enhances insulin release through the activation of PPAR alpha gene expression during a short culture period, whereas it may contribute to beta-cell dysfunction through the mechanism of "excessive stimulation" during longer culture periods.

Animals↗

PubMed2001

Effects of free fatty acids on beta-cell functions: a possible involvement of peroxisome proliferator-activated receptors alpha or pancreatic/duodenal homeobox.

It is well known that acute administration of fatty acids enhances insulin release from beta cells, although chronic exposure to fatty acids inhibits insulin release (lipotoxicity). The mechanism for these reciprocal effects of fatty acids on insulin release remains to be elucidated. The present study was performed to investigate the effects of fatty acids on gene expression related to glucose metabolism or insulin biosynthesis. In islets cultured with palmitate for 8 hours, glucose-induced insulin release was enhanced together with increment of pyruvate carboxylase (PC) mRNA or peroxisome proliferator-activated receptors (PPAR)alpha. In contrast, by extending the culture period up to 48 hours, glucose-induced insulin release or islet insulin content was significantly impaired by the coexistence of palmitate. Concomitantly, PC, PPARalpha, GLUT-2, glucokinase (GK), preproinsulin, or pancreatic/duodenal homeobox-1 (PDX-1) mRNA were significantly suppressed in those islets cultured for 48 hours with palmitate. These data may imply that during short-term culture period palmitate promotes PPARalpha gene expression, which enhances PC mRNA expression leading to the enhancement of insulin release, whereas during long-term culture period, palmitate rather inhibits PPARalpha mRNA, which reduces PC mRNA expression. Furthermore, palmitate reduces GLUT-2, GK, or preproinsulin mRNA expression probably through the inhibition of PDX-1 mRNA.

Animals↗

PubMed2001

Recognition of tRNA by aminoacyl-tRNA synthetase from hyperthermophilic archaea, Aeropyrum pernix K1.

To study the recognition sites of tRNA for archaeal aminoacyl-tRNA synthetase, several aminoacyl-tRNA synthetase genes from hyperthermophilic archaeon, Aeropyrum pernix K1 were cloned and expressed. All the expressed enzymes showed extreme thermostability. Expressed threonyl-tRNA synthetase threonylated not only archaeal (A. pernix and Haloferax volcanii) threonine tRNAs but also Escherichia coli threonine tRNA. However, threonyl-tRNA synthetase from H. volcanii did not threonylate E. coli threonine tRNA.

Desulfurococcaceae↗

PubMed2001

The role of cyclic AMP in the pathogenesis of glucose desensitization in rat pancreatic islets.

Adenosine-3',5'-cyclic monophosphate (cyclic AMP) promotes exocytosis of insulin in pancreatic beta cells. This study was performed to investigate the role of cyclic AMP in the pathogenesis of glucose desensitization in rat pancreatic islets. In islets cultured with high glucose for 48 hours, 27 mmol/L glucose-induced insulin release was markedly impaired, while 3.3 mmol/L glucose-or arginine-induced insulin release was enhanced, indicating glucose desensitization. Islet cyclic AMP content was 190% enhanced in high glucose-culture islets for 48 hours. In islets cultured with dibutyryl-cyclic AMP (dbcAMP) or 3-isobutyl methy-xanthine (IBMX), islet insulin content or 27 mmol/L glucose-induced insulin release was deteriorated. In contrast, 3.3 mmol/L glucose- or arginine-induced insulin release was increased, which was similar to glucose-desensitized islets. Wash-out of dbc AMP for the last 24 hours of the 48-hour culture period restored impaired high glucose-induced insulin release in the same manner as wash-out of high glucose. Diazoxide, the KATP channel opener, also restored impaired high glucose-induced insulin release from dbcAMP-cultured islets. The data suggest that enhancement of cyclic AMP in high glucose-culture islets may be one of the pathogenesis of glucose desensitization.

1-Methyl-3-isobutylxanthine↗

PubMed2001

Pyrobaculum oguniense sp. nov., a novel facultatively aerobic and hyperthermophilic archaeon growing at up to 97 degrees C.

A novel hyperthermophilic, heterotrophic, rod-shaped archaeon was isolated from a terrestrial hot spring at Oguni-cho, Kumamoto Prefecture, Japan. The new isolate, strain TE7T, grew under aerobic, microaerobic and anaerobic conditions. Isolate TE7T grew optimally at 90-94 degrees C and pH 7.0-7.5 (adjusted at 25 degrees C) under atmospheric air with vigorous shaking. Strain TE7T cells were motile rods 2-10 microm in length and covered with a surface-layer lattice. Cell yields at 90 degrees C under aerobic conditions were twice that under anaerobic conditions. Under aerobic conditions, growth was inhibited by elemental sulfur, but thiosulfate stimulated growth. Under anaerobic conditions, no growth was observed in the presence of nitrate and nitrite, but elemental sulfur, thiosulfate, L-cystine and oxidized glutathione stimulated growth. The 16S rDNA sequence of TE7T exhibited a close relationship to the sequences of Pyrobaculum aerophilum and Thermoproteus neutrophilus, which belong to the cluster of the genus Pyrobaculum. DNA-DNA hybridization analysis showed a low level of DNA similarity between TE7T and previously described Pyrobaculum species. As TE7T is phenotypically and phylogenetically different from the other members of this genus, it is described as a new species named Pyrobaculum oguniense (type strain TE7T = JCM 10595T = DSM 13380T).

Aerobiosis↗

PubMed2001

Comparison of the antigenicity of protoscoleces and microvesicles of Echinococcus multilocularis prepared from rats.

Rats are known to be relatively resistant to infection with Echinococcus multilocularis. However, when rats are inoculated with the parasite tissues, E. multilocularis proliferates slowly at first but after 6 months the cysts increase in size considerably and contain large numbers of protoscoleces. As rats survive for 18 months or longer, approximately 100 ml of packed protoscoleces can be produced from each rat. A comparison of the antigenicity of the protoscoleces and microvesicles by immunoblot methods showed that both Em18 and Em16 are shared components between both protoscoleces and microvesicles, although the latter have some additional antigenic components. In antigens prepared from protoscoleces, the banding patterns around Em18 were much simpler than those from microvesicles. Therefore, for serodiagnosis of E. multilocularis, antigens should be carefully prepared from protoscoleces rather than microvesicles from the rat.

Animals↗

PubMed2001

Differential serodiagnosis of cystic and alveolar echinococcosis using native and recombinant antigens in Japan.

Our group at Asahikawa Medical College has established differential serodiagnosis for zoonotic larval cestodiases such as alveolar echinococcosis (AE), cystic echinococcosis (CE) and neurocysticercosis (NCC) using purified specific antigens. In this brief review, we introduce (a) four imported CE cases in Japan, easily identified serologically, (b) most recent advances in serology for differentiation of AE and monitoring of prognosis of AE in Japan. It includes application of affinity purified Em18 and prototype of a recombinant Em18 antigen. Serology using affinity purified Em18 antigens is showing much higher sensitivity for detection of AE cases which are usually undetectable by the ongoing serology for AE authorized in Hokkaido, Japan. As serology for AE, CE or NCC is still not popular in the majority of Asian countries, we expect that this review paper stimulates researchers who are interested in serology or serodiagnosis for these larval cestodiases including AE, CE and NCC.

Animals↗

PubMed2001

NOD-scid mouse as an experimental animal model for cysticercosis.

The major three species of human taeniid cestodes, Taenia solium, T. saginata and T. saginata asiatica (= T. asiatica) which require humans as the definitive host are still not rare in developing countries. Among these, T. solium is the most serious with medical and economic importance. Neurocysticercosis (NCC) in humans is now recognized as the major cause of neurologic disease in the world. As these human taeniid cestodes obligatory require domestic animals such as swine, cattle and swine as the major intermediate host animals respectively, it is not easy to analyze the basic research in these domestic animals. In this brief review, we introduce experimental animal model for these three species in order to obtain fully developed metacestode stage in severe combined immunodeficiency (scid) mice. Non-obese diabetic scid (NOD-scid) mice are expected to be a satisfactory animal model and to have advantages for analysis by several view points of developmental biology with gene expression throughout development, antigenic homology of cyst fluid of these three species, evaluation of drug efficacy or metacestocidal drug designs, confirmation of unknown taeniid gravid segments for identification based on the morphology and DNA analysis of metacestodes. The animal model is not only available for human Taenia spp but can also be applied to other taeniid cestodes of economic importance or in veterinary parasitology.

Animals↗

PubMed2001

Molecular variation of Taenia solium in the world.

Complete nucleotide sequences of the mitochondrial cytochrome b (Cytb) and cytochrome c oxidase subunit I (CO I) genes from various isolates of Taenia solium were examined. Eleven isolates were analyzed; two isolates from China, two isolates from Indonesia, one isolate each from India, Thailand, Mexico, Ecuador, Peru, Mozambique and Tanzania. In both genes, two isolates from Indonesia shared the same sequences. Similarly, the isolate from Mexico shared same sequences with that from Peru, and the isolate from Mozambique shared same sequences with that from Tanzania. Phylogenetic trees inferred from different mitochondrial genes yielded almost the same topology. Both the UPGMA and NJ-trees were also very similar. These trees indicate that T. solium may be diverged to 2 genetic groups; isolates from Asia form one group and isolates from Africa and Latin America belong to the other. It seems that T. solium prevalent in Africa and in Latin America shares the related origin and has recently been introduced to each area, perhaps with domestic pigs or human.

Animals↗

PubMed2001

Recent advances in serodiagnosis for cysticercosis.

Neurocysticercosis (NCC) caused by infection with the larval stage of Taenia solium is an important cause of neurological disease worldwide. Up to the present, many studies on characterizing species-specific antigens of T. solium have been done and several high quality antigens for serodiagnosis are available. Hence the research on serodiagnosis has been shifted to the next phase, stable production of diagnostic antigens using molecular techniques. In order to establish an enzyme-linked immunosorbent assay (ELISA) using recombinant proteins, we carried out molecular cloning and identified four diagnostic antigen candidates (Ag1, Ag1V1, Ag2, and Ag2V1). Recombinant proteins, except Ag2V1, were successfully expressed using an Escherichia coli expression system. Immunoblot analysis using NCC patient sera detected recombinant proteins. But as reactivity to rAg1 was too weak, Ag1 was not suitable for the immunodiagnosis antigen. Therefore Ag1V1 and Ag2 were chosen for ELISA antigens and Ag1V1/Ag2 chimeric protein was expressed. Of 49 serum samples from NCC patients confirmed to be seropositive by immunoblot analysis, 44 (89.7%) were positive by ELISA. Serum samples from patients with other parasitic infections did not recognized Ag1V1/Ag2 chimeric protein. Ag1V1/Ag2 chimeric protein obtained in this study is of value for differential immunodiagnosis.

Amino Acid Sequence↗