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Biomedical subjects

R Maeda

Publications and source records attributed to R Maeda.

At least 19 recordsLinked to original sources
PubMed2003

Increase of Th1 type cytokine mRNA expression in peripheral blood lymphocytes of calves experimentally infected with Cryptosporidium parvum.

The expression of the messenger RNA of interleukin-12 (IL-12), interferon-gamma, interleukin-4, interleukin-6, and interleukin-10 was examined by reverse-transcriptase polymerase chain reaction in peripheral blood lymphocytes of calves that were orally inoculated with Cryptosporidium parvum oocysts. In all of the calves, gene expression of interleukin-12, interleukin-6, and interferon-gamma was observed at delivery and this expression was repressed within the next 24h. In calves inoculated with C. parvum, mRNA expression of interleukin-12 and interferon-gamma was noticed on day 3 post-inoculation (p.i.) and increased in the convalescent phase of the infection, whereas in non-inoculated calves no mRNA expression was detectable up to the end of the experiment. No mRNA expression of interleukin-4 or 6 was detected during the experiment. Our observations suggest that systemic Th1 type immune responses are induced in calves infected with C. parvum and may be available for evaluation of the control of the infection.

Animals↗
PubMed2003

Relationship between liver disorders and protection against Eimeria stiedai infection in rabbits immunized with soluble antigens from the bile of infected rabbits.

Soluble antigens exist in the bile of rabbits infected with Eimeria stiedai (E. stiedai) in the acute phase, and rabbits immunized with the antigens show resistance against the infection. In this study, the liver function of rabbits immunized either with the soluble antigens or PBS were examined following the parasite challenge. Rabbits immunized with PBS shed a number of oocysts and showed an increase in r-glutamyltransferase (GGT) activity and a decrease in blood Indocyanine green (ICG) clearance. However, rabbits immunized with the soluble antigens shed a lower number of oocysts and showed a transient increase of alanine-aminotransferase (ALT) activity on Day 8 post-challenge (p.c.). The blood Indocyanine green clearance of the rabbits showed no change throughout the experiment. By histopathological observation of the liver, a number of merozoites were found in the biliary ducts on Day 8 post-challenge in the non-immunized rabbits. In contrast, a number of lymphocytes and neutrophilic leukocytes assembled around the biliary ducts of the immunized rabbits, but few parasites were found there on Day 8 post-challenge. These results suggest that the soluble antigens stimulate local immune reactions, for example around the biliary ducts, resulting in elimination of the parasite's development.

Alanine Transaminase↗
PubMed2003

Eimeria stiedai merozoite 49-kDa soluble antigen induces protection against infection.

A soluble antigen isolated from Eimeria stiedai merozoites with a molecular mass of 49 kDa was detected in the bile of infected rabbits. Rabbits immunized with the antigen shed a lower number of oocysts than did nonimmunized rabbits postchallenge (p.c.). The immunized rabbits showed a marked and transient increase of alanine-aminotransferase (ALT) activity on day 8 p.c. The blood indocyanine green (ICG) clearance and r-glutamyltransferase (GGT) activity showed no change throughout the experiment However, nonimmunized rabbits showed a gradual increase of ALT and GGT in the plasma and a delay of ICG p.c. Many merozoites were observed in the biliary ducts of the nonimmunized rabbits on day 8 p.c. using standard histology. In contrast, in the immunized rabbits, many inflammatory cells were observed around the biliary ducts, but there were few parasites in the tissue. These results suggest that the 49-kDa soluble protein antigen detected in the bile of the infected rabbits was a merozoite-specific antigen, and the immune reaction to the antigen may induce protective effects against the infection.

Alanine Transaminase↗
PubMed2001

Trail antigen in Eimeria stiedai sporozoites associated with a thrombospondin-related motif and the entry of cultured cells.

In order to examine the antigenic similarity and specificity of the trail antigen of Eimeria stiedai and Etp 100, a microneme protein of Eimeria tenella, monoclonal antibodies to the trail antigen of E. stiedai sporozoites were selected by an indirect immunofluorescent antibody method. The monoclonal antibody of one clone, 3D10, reacted with the anterior portion of non-fixed sporozoites. By immunoblotting, the monoclonal antibody was found to react with a 100 kDa antigen of E. stiedai sporozoites, and a 117 kDa antigen of E. tenella sporozoites and merozoites. It was also found to react with a recombinant protein with thrombospondin-/properdin-like motifs homologous to E. tenella microneme protein Etp 100. The monoclonal antibody significantly inhibited the penetration of E. stiedai sporozoites into cultured rabbit hepatobiliary epithelial cells. These results suggest that E. stiedai sporozoites have a trail antigen, located in the anterior region on the outer surface of the sporozoites, which has an epitope with thrombospondin-/properdin-like motifs similar to E. tenella microneme protein Etp 100. This protein may play an important functional role in the process of penetration of host cells.

Amino Acid Sequence↗
PubMed2001

A Novel MAPK phosphatase MKP-7 acts preferentially on JNK/SAPK and p38 alpha and beta MAPKs.

Mitogen-activated protein kinases (MAPKs) are inactivated via dephosphorylation of either the threonine or tyrosine residue or both in the P-loop catalyzed by protein phosphatases which include serine/threonine phosphatases, tyrosine phosphatases, and dual specificity phosphatases. Nine members of the dual specificity phosphatases specific for MAPKs, termed MKPs, have been reported. Each member has its own substrate specificity, tissue distribution, and subcellular localization. In this study, we have cloned and characterized a novel MKP, designated MKP-7. MKP-7 is most similar to hVH5, a member of previously known MKPs, in the primary structure. MKP-7 is predominantly localized in the cytoplasm when expressed in cultured cells, whereas hVH5 is both in the nucleus and the cytoplasm. MKP-7 binds to and inactivates p38 MAPK and JNK/SAPK, but not ERK. Furthermore, we have found that MKPs have the substrate specificity toward the isoforms of the p38 family (alpha, beta, gamma, and delta). MKP-7 binds to and inactivates p38 alpha and -beta, but not gamma or delta. MKP-5 and CL100/MKP-1 also bind to p38 alpha and -beta, but not gamma or delta. Finally, we propose a tentative classification of MKPs based on the sequence characteristics of their MAPK-docking site.

Amino Acid Sequence↗
PubMed2001

Specific IgA antibody response to coproantigens of Cryptosporidium parvum in serum and saliva of calves after experimental infection.

The antibody response to coproantigens of Cryptosporidium parvum was examined in saliva and sera of calves experimentally infected with C. parvum. Coproantigens of C. parvum with approximate molecular masses of 17, 15 and less than 14kDa were found in the feces of infected calves on day 3 or later, and 60 and 23kDa coproantigens observed between days 4 and 9 post-infection, respectively. The antibody reactivity to the coproantigens was mainly attributable to IgA class antibodies in saliva and was detectable during the convalescent phase of infection. A 15kDa protein isolated from the feces of infected calves by immunoaffinity adsorption using a monoclonal anti C. parvum antibody was recognized by IgA antibodies present in the saliva during the convalescent phase of infection. These results suggest that this coproantigen may be released from C. parvum sporozoites and may induce IgA antibody production in the mucosal immune system of infected calves.

Animals↗
PubMed2001

Xmeis1, a protooncogene involved in specifying neural crest cell fate in Xenopus embryos.

Meis1 (Myeloid Ecotropic viral Integration Site 1) is a homeobox gene that was originally isolated as a common site of viral integration in myeloid tumors of the BXH-2 recombinant inbred mice strain. We previously isolated a Xenopus homolog of Meis1 (Xmeis1). Here we show that Xmeis1 may play a significant role in neural crest development. In developing Xenopus embryos, Xmeis1 displays a broad expression pattern, but strong expression is observed in tissue of neural cell fate, such as midbrain, hindbrain, the dorsal portion of the neural tube, and neural crest derived branchial arches. In animal cap explants, overexpression of Xmeis1b, an alternatively spliced form of Xmeis1, induces expression of neural crest marker genes in the absence of mesoderm. Moreover, Xmeis1b induces XGli-3 and XZic3, pre-pattern genes involved at the earliest stages of neural crest development, and like these two genes, can induce ectopic pigmented cell masses when overexpressed in developing embryos. Misexpression of Xmeis1b also induces ectopic expression of neural crest markers along the antero-posterior axis of the neural tube in developing Xenopus embryos. In contrast, Xmeis1a, another splice variant, is much less effective at inducing these effects. These data suggest that Xmeis1b is involved in neural crest cell fate specification during embryogenesis, and can functionally intersect with the Gli/Zic signal transduction pathway.

Alternative Splicing↗
PubMed2001

A 14-3-3 protein homologue is expressed in feline enteroepithelial-stages of Toxoplasma gondii.

Fourteen cDNA clones encoding epitopes of proteins of Toxoplasma gondii feline enteroepithelial-stages parasites were isolated and expressed in Escherichia coli in an effort to determine the antigenecity of the parasites. Sequence analysis showed that four of the cDNA clones had a 930-bp open-reading frame encoding a product showing similarity to the 14-3-3 protein mRNA sequence.(1) Southern hybridization of DIG-labeled positive clone with T. gondii genomic DNA cleaved with EcoRI, BamHI and HindIII resulted in one or two bands in each case. In an immunofluorescence assay, polyclonal and monoclonal antibodies raised against the expressed protein showed strong reactivity with feline enteroepithelial-stages parasites and sporozoites. In a complementation assay in which a plasmid carrying the protein-coding region of the isolated cDNA was introduced into a Saccharomyces cerevisiae mutant, strain DS9-22, the expressed protein showed complementation of the function of the 14-3-3 protein in yeast transformants. These findings suggest that T. gondii parasites produce a protein showing partial homology with members of the 14-3-3 protein family and this protein is expressed in feline enteroepithelial-stages parasites.

14-3-3 Proteins↗
PubMed2001

Identification of a docking groove on ERK and p38 MAP kinases that regulates the specificity of docking interactions.

MAP kinases (MAPKs) form a complex with MAPK kinases (MAPKKs), MAPK-specific phosphatases (MKPs) and various targets including MAPKAPKs. These docking interactions contribute to regulation of the specificity and efficiency of the enzymatic reactions. We have previously identified a docking site on MAPKs, termed the CD (common docking) domain, which is utilized commonly for docking interactions with MAPKKs, MKPs and MAPKAPKs. However, the CD domain alone does not determine the docking specificity. Here we have identified a novel site on p38 and ERK2 MAPKs that regulates the docking specificity towards MAPKAPKs. Remarkably, exchange of two amino acids in this site of ERK2 for corresponding residues of p38 converted the docking specificity for MAPKAPK-3/3pk, which is a dominant target of p38, from the ERK2 type to the p38 type, and vice versa. Furthermore, our detailed analyses with a number of MAPKAPKs and MKPs suggest that a groove in the steric structure of MAPKs, which comprises the CD domain and the site identified here, serves as a common docking region for various MAPK-interacting molecules.

3T3 Cells↗
PubMed2001

Reduction of the infectivity of Toxoplasma gondii and Eimeria stiedai sporozoites by treatment with bovine lactoferricin.

Sporozoites of Toxoplasma gondii preincubated with lactoferricin showed decreased activity in penetration of mouse embryonal cells. Mice inoculated with 10(5) sporozoites preincubated with lactoferricin showed a higher survival rate than those inoculated with the same number of untreated sporozoites. Likewise, sporozoites of Eimeria stiedai preincubated with lactoferricin also showed decreased activity in penetration of rabbit hepatobiliary cells. Rabbits inoculated with 10(5) sporozoites preincubated with lactoferricin shed fewer oocysts than those inoculated with the same number of untreated sporozoites. These results indicate that lactoferricin is effective to reduce the infectivity of sporozoites of Toxoplasma gondii and Eimeria stiedai.

Animals↗
PubMed2001

Identification and the role of soluble antigens detected in bile from Eimeria stiedai-infected rabbits.

Antibodies against Eimeria stiedai sporozoites and merozoites were detected in the sera of rabbits immunized with bile obtained from infected rabbits on the 15th day post-infection. The trails made by gliding sporozoites were also detected by the sera. After penetration into the host cell, an antibody-binding region was observed on the parasitophorous vacuole membranes of the parasites. Rabbits administered a combination of the bile and cholera toxin shed fewer oocysts in the feces after infection than control rabbits. The immunized rabbits developed a high level of IgA antibody against soluble antigens in the bile. By immunoblotting, antigens with molecular masses of 32, 37, and 49 kDa were detected in the bile obtained from infected rabbits on the 15th day postinfection. Absorption treatment with sporozoites reduced or abolished the antibody reactivity to the 32-kDa antigen of merozoites and the bile antigens. However, antibody reactivity to the 37- and 49-kDa antigens still remained. These results indicate that soluble antigens are present in the bile of rabbits in the acute phase of infection, and these may be produced and released by merozoites during the host cell invasion process.

Animals↗
PubMed2001

Isolation of Neospora caninum from the brain of a pregnant sheep.

Neospora caninum was isolated from the brain of a naturally infected pregnant sheep by inoculation of immunodeficient mice with a homogenate of the brain tissue. The ewe showed no clinical signs. Tachyzoites were observed in the tissues of the nu/nu mice injected with the brain tissue homogenate and the diagnosis was confirmed by immunohistochemical staining with anti-N. caninum antibodies and by detecting N. caninum-specific DNA by polymerase chain reaction.

Animals↗
PubMed2001

[Probability of the standardization of control survey in local medical association].

This report discusses about the probability of the standardization of external quality control (control survey) in local medical area such as prefecture size. For example, our control survey by Hiroshima medical association in Hiroshima prefecture is selected and shown on their effort to the standardization of the control survey. This Hiroshima control survey is continuing for 27 years and its purpose is the improvement of the differences between laboratory facilities. For the all standardization of control survey system, tests, devices and reagents, the recent reports of Hiroshima control survey shows that the reference methods by Japan Society of Clinical Chemistry (JSCC) and the enzyme reference materials by Japan Committee of Clinical Laboratory Standardization (JCCLS) are very useful for the improvement of the differences between laboratory facilities in Hiroshima Prefecture.

Chemistry, Clinical↗
PubMed2001

[The introduction of a new clinical laboratory system, so-called the profiles for each diagnosis related group].

We try to establish a new clinical laboratory system, so-called profiles for each diagnosis related group which have features as shown below. 1) The profiles are clinical manuals that are utilized by home doctors and show how to diagnose, follow up, observe complications and judge therapeutic effect. 2) The profiles are constructed on condition that they cover 60% of cases and cases that they cannot handle are supposed to be referred to the major hospitals or the specialized doctors. 3) The profiles are guaranteed to be reformed and maintained according to medical advancement. 4) The target diseases of the profiles are thyroid disease, liver diseases, diabetes mellitus, respiratory disease, infectious disease, renal disease, cardiac disease, hyperlipemia, collagen disease, hematological disease, osteoporosis and gynecological disease. We present the profiles for thyroid disease, hyperlipemia and collagen disease which have been accomplished.

Clinical Laboratory Techniques↗
PubMed2000

Quantitative analysis of microfilarial periodicity of Dirofilaria immitis in cats.

Microfilarial periodicity of Dirofilaria immitis in the venous blood of infected cats was analyzed by a trigonometric model. Cats were infected by subcutaneous transplantation with 120-day-old juvenile D. immitis. Microfilariae in the blood were first observed 98 days after transplantation. Blood was collected at 4h intervals for a 24h period, and examinations were repeated five times in two cats. The calculated periodicity index was 75.1 and 50.3 in these two cats. The estimated hour of peak microfilarial density ranged from 1.00 to 2.84h. Thus, the periodicity of microfilariae of D. immitis in the blood of cats was characterized as nocturnally sub-periodic.

Animals↗
PubMed2000

Active micromixer for microfluidic systems using lead-zirconate-titanate (PZT)-generated ultrasonic vibration.

A micromixer using direct ultrasonic vibration is first reported in this paper. The ultrasonic vibration was induced by a bulk lead-zirconate-titanate (PZT; 5 x 4 x 0.2 mm), which was excited by a 48 kHz square wave at 150 V (peak-to-peak). Liquids were mixed in a chamber (6 x 6 x 0.06 mm) with an oscillating diaphragm driven by the PZT. The oscillating diaphragm was in the size of 6 x 6 x 0.15 mm. Ethanol and water were used to test the mixing effectiveness. The laminar flows of ethanol (115 microL/min) and water (100 microL/min) were mixed effectively when the PZT was excited. The entire process was recorded using a video camera.

Animals↗
PubMed2000

Importance of OGTT for diagnosing diabetes mellitus based on prevalence and incidence of retinopathy.

Study was made on the necessity and importance of the oral glucose tolerance test (OGTT) for diagnosing diabetes mellitus based on prevalence and incidence of diabetic retinopathy. Subjects were 12208 persons undergoing OGTT between 1965 and 1997. The prevalence of retinopathy was significantly elevated with FPG>/=126 and 2-h PG>/=198 mg/dl. The incidence of retinopathy was 15-30/10000 person-years (PY) with FPG<125, but with FPG of 126-139 it was significantly higher (69/10000 PY) and at 140-199 mg/dl it was elevated to 139/10000 PY. Subjects were classified at initial test into FPG<110, 110-125, 126-139, and >/=140 and further into 2-h PG<200 and >/=200 mg/dl for comparison with the incidence of retinopathy. Even with the same FPG, the incidence was two- to threefold higher with 2-h PG>/=200 mg/dl, indicating that 2-h PG was highly associated with the incidence of retinopathy. As for IFG, the prevalence of diabetes as defined by 2-h PG>/=200 in the OGTT increased with elevated FPG, and 33.7% of IFG cases showed 2-h PG>/=200 mg/dl. Based on the prevalence and incidence of retinopathy, we conclude that 126 mg/dl FPG is an appropriate cut-off level, and the OGTT is important for diagnosing mild diabetes mellitus.

Adolescent↗