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Biomedical subjects

T Mikami

Publications and source records attributed to T Mikami.

At least 127 records · Page 7Linked to original sources

Novel species specific antigens of trypanosoma congolense and their different localization among life-cycle stages.

Seven monoclonal antibodies (mAbs) were raised against Trypanosoma congolense procyclic form (PCF). Localization of the antigens recognized by the mAbs was determined in bloodstream form (BSF), PCF, epimastigote form (EMF) and metacyclic form (MCF) by confocal laser scanning microscopy (CLSM). Two mAbs (10F9 and 20H12) showed different fluorescent patterns among different life-cycle stages of the parasite. The 10F9 recognized a 76 kDa antigen of all life-cycle stages of the parasite and the antigen localization corresponded with that of a mitochondrion. While the 20H12 recognized 119 and 122 kDa antigens of all the life-cycle stages and the antigen localization corresponded with a flagellum in BSF and MCF, tip of a flagellum in PCF, and part of cytoplasm in EMF. Moreover, the 20H12 did not react to T. brucei gambiense, T. b. rhodesiense and T. evansi antigens in both CLSM and immunoblotting. Therefore, the antigens recognized by the 20H12 seem to be T. congolense specific. Although, further studies will be required for a full characterization of the T. congolense specific 119 and 122 kDa antigens, the mAb 20H12 and the specific antigens may be useful in not only establishment of T. congolense specific diagnosis methods but also studies on molecular mechanisms regulating differentiation of the parasite during life-cycle.

Animals↗

Cloning of full length cDNA, high-level expression and biological characterization of feline IL-12.

A full length cDNA of feline interleukin(IL)-12 p35 and p40 subunits was cloned. By transferring the plasmids containing both the subunit genes to mammalian cells, we expressed biologically active feline IL-12. The expressed feline IL-12 has interferon-gamma-inducing activity against both human and feline peripheral blood mononuclear cells (PBMC) and stimulates cytotoxic T lymphocyte activity against herpes simplex virus-infected human PBMCs. There were two kinds of molecules (p75, p80) in the purified recombinant feline IL-12, and both molecules exhibited biological activity. The difference between p75 and p80 was the degree of the glycosylation of the p35 chain. Moreover, when we modified the cDNA of p35 by changing some codons and deleted the 5' and 3' non-coding regions, the expression level of IL-12 increased about 100 fold.

Amino Acid Sequence↗

Antigens expressed in feline enteroepithelial-stages parasites of Toxoplasma gondii.

In an investigation aimed to identify Toxoplasma gondii antigens expressed in feline enteroepithelial-stages parasites, a cDNA library was constructed and fourteen positive clones were isolated by immunoscreening using sera from cats immunized with feline enteroepithelial-stages parasites. By DNA sequence homology analysis, these fourteen isolated clones were classified into four groups: hypoxanthine-guanine phosphoribosyl transferase (HGPRT) cDNA, heat shock protein 70 (HSP70) cDNA, 14-3-3 protein homologue cDNA, and cDNA encoding an unknown product. In an indirect immunofluorescence antibody test, sera from mice immunized with the recombinant protein encoded by the cDNA for HGPRT, HSP70, 14-3-3 protein, or the unknown product each showed a relatively high level of immunoreactivity with feline enteroepithelial-stages parasites.

Animals↗

Antibody reactivity to Cryptosporidium parvum in saliva of calves after experimental infection.

Antibodies against Cryptosporidium parvum in the saliva and sera of three calves experimentally infected with this parasite were examined by an indirect immunofluorescence antibody test and immunoblotting. Salivary anti-C. parvum IgA antibody appeared on day 12 post-challenge and had a tendency to increase transiently between days 15 and 30 post-challenge. Salivary anti-C. parvum IgG antibody levels showed a gradual increase along with the change in IgA antibody levels during the infection. In contrast, serum anti-C. parvum IgA antibody levels showed only a slight increase between days 15 and 30 post-challenge. Serum anti-C. parvum IgG antibody levels rose on day 12 post-challenge and one calf maintained relatively high level up to the end of the experiment. In immunoblotting, an antigen with a molecular mass of 15 kDa was found to react strongly to salivary IgA antibody and a 27 kDa antigen to react to serum IgG antibody.

Animals↗

Protein analysis of Babesia caballi merozoites by two-dimensional polyacrylamide gel electrophoresis and western blotting.

Babesia caballi merozoites were prepared by combining two improved methods of cultivation and purification of merozoites using Percoll-gradiation, and the protein compositions of merozoites were analyzed by two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting. The relative molecular masses of the major proteins and protein masses separated by electrophoresis were >94, 80-70, 50-45, 34-30, 30-28 and 18 kDa. By Western blotting, twelve proteins or protein groups were recognized by pooled sera from two horses experimentally infected with B. caballi. Among twelve proteins, five new proteins (54, 30-26, 24, and two 18 kDa) were identified, and the 48 kDa protein was revealed to consist of 2 components in the B. caballi merozoite. One protein (54 kDa) of B. caballi was also recognized by the pooled sera from two horses experimentally infected with B. equi.

Animals↗

Immunization with recombinant surface antigens p26 with Freund's adjuvants against Babesia rodhaini infection.

The surface proteins of Babesia rodhaini have previously been shown to induce a high degree of protective immunity. In the present study, one of those proteins, B. rodhaini antigen p26 was expressed in Escherichia coli and in insect cells infected with a recombinant baculovirus. These proteins were recognized by immune serum from a drug-cured BALB/c mouse. While BALB/c mice immunized with both recombinant antigens and Freund's adjuvants showed 40-100% survival rate against challenge infection with B. rodhaini, saponin failed to induce protection, although significant levels of B. rodhaini-specific antibodies were produced in both immunized mice (1:1,000-2,000 by indirect immunofluorescent antibody test). The immunization of IFN-gamma-deficient mice with the recombinant proteins was not protective against B. rodhaini infection, indicating that IFN-gamma is one of the important factors for the survival against lethal B. rodhaini infection.

Animals↗

Contrastive prevalence of feline retrovirus infections between northern and southern Vietnam.

The prevalence of infections with three feline retroviruses; feline leukemia virus (FeLV), feline immunodeficiency virus (FIV) and feline foamy virus (FeFV), was examined in domestic cats (Felis catus) and leopard cats (Felis bengalensis) in southern Vietnam in 1998. We then compared this data with our previous study in northern Vietnam in 1997. None of the cats had FeLV antigens in both the northern and southern areas. In contrast, there is a great distinction in the seropositivity of FIV. Twenty-two percent of domestic cats had FIV antibodies whereas no FIV positive cats were detected in northern area. FIV may have entered southern Vietnam recently and spread rapidly. FeFV infections were found in both areas, suggesting that FeFV might be present in the cat populations in Vietnam from the earliest time.

Animals↗

Morphological changes of Babesia gibsoni grown in canine red blood cell-substituted severe combined immune deficiency mice.

Canine red blood cell-substituted severe combined immune deficiency (Ca-RBC-SCID) mice were prepared for canine Babesia gibsoni infection. The Ca-RBC-SCID mice infected with B. gibsoni developed a high level of parasitemia, and showed clinical symptoms such as anemia and hemoglobinuria, which are similar to those observed in dogs infected with B. gibsoni. The B. gibsoni parasites grown in Ca-RBC-SCID mice showed marked morphological changes, including a significantly larger size of parasites than those in dogs and abundant RBCs containing 4, 8, 16, and 32 parasites. The multiple infection may have resulted from 1 parasite because the posterior end of each parasite in a multiply infected cell was connected. The parasites grown in SCID mice retained their infectivity and virulence to dogs and their morphology was dramatically restored to the original state when they were returned to dogs.

Animals↗

Differences between the silencing-related properties of the extreme carboxyl-terminal regions of thyroid hormone receptors alpha 1 and beta 1.

Human thyroid hormone receptor (TR) is encoded by two distinct genes, TR alpha and TR beta. TR heterodimerizes with retinoid X receptor (RXR) and binds efficiently to the thyroid hormone (T(3)) response element (TRE) of target genes. In the absence of T(3), unliganded TR suppresses the basal promoter activity of positively regulated genes (silencing). Silencing mediator for retinoid and thyroid hormone receptors (SMRT) and nuclear receptor co-repressor (N-CoR) interact with unliganded TR and function as corepressor proteins. Previously, we found beta F451X with carboxyl (C)-terminal 11-amino acid deletion had stronger silencing potency than wild-type TR beta 1 and beta E449X with C-terminal 13-amino acid deletion on a subset of TREs. In the present study, to assess the isoform-specific effects of the C-terminal truncations on TR silencing, we constructed two mutant TR alpha 1s (alpha F397X and alpha E395X) with the same respective C-terminal truncations as beta F451X and beta E449X and analysed their silencing activities. Unlike beta F451X and beta E449X, alpha F397X and alpha E395X showed similarly stronger silencing potency than wild-type TR alpha 1. We further studied the abilities of wild-type and the mutant TR beta 1s and alpha 1s on RXR and co-repressor binding by a two-hybrid interference assay. beta F451X had significantly stronger abilities to bind to RXR and SMRT than did wild-type TR beta 1 and beta E449X. In contrast, wild-type TR alpha 1, alpha F397X and alpha E395X showed similar abilities to bind to RXR and SMRT. beta E449X and alpha E395X, which have identical C-terminal truncation, showed less ability to bind to N-CoR than did wild-type TR beta 1 and beta F451X and wild-type TR alpha 1 and alpha F397X respectively. These results indicate that an identical C-terminal truncation gives rise to different effects on TR beta 1 and alpha1 with respect to silencing potency, RXR binding and SMRT binding. The difference in the silencing potency among wild-type TR beta 1, beta F451X and beta E449X correlated well with the difference in the ability to bind co-repressor SMRT.

Animals↗

[Myelopathy caused by tentorial dural arteriovenous fistula: a rare case report].

A 51-year-old man with myelopathy due to intracranial dural arteriovenous fistula (dural AVF) is reported. At age 46, the patient experienced subarachnoid hemorrhage caused by rupture of the dural AVF and underwent embolization of the lesion at another hospital. At this time, the patient complained of numbness in his legs and showed paraplegic gait disturbance. MRI scan revealed swelling of the cervical spinal cord. Cerebral angiograms demonstrated the recurrence of tentorial dural AVF fed by bilateral meningohypopheseal trunks and the right posterior inferior cerebellar artery. Arteriovenous shunt (AV shunt) flow was drained into the anterior spinal vein. It seemed that the swelling of the spinal cord and myelopathy was caused by venous hypertension of spinal veins. After surgical interruption of the right petrosal vein which connected dAVF with cerebellar veins, AV shunt was obliterated successfully. Postoperative cerebral angiograms showed disappearance of dural AVF. The patient became ambulant and his cervical spinal cord appeared normal on the postoperative MRI scan. Surgical interruption of the draining vein was simple, effective and essential treatment.

Arteriovenous Fistula↗

[Vaccine application of recombinant herpesviruses].

Recently, genetic engineering using recombinant DNA techniques has been applied to design new viral vaccines in order to reduce some problems which the present viral vaccines have. Up to now, many viruses have been investigated for development of recombinant attenuated vaccines or live viral vectors for delivery of foreign genes coding immunogenic antigens. In this article, we introduced the new vaccine strategy using genetically engineered herpesviruses.

Alphaherpesvirinae↗

[Recent advances in echocardiographic diagnosis of ischemic heart disease].

Before the early 1990s, the diagnostic usefulness of echocardiography for ischemic heart disease had been relatively limited compared with that for other cardiac diseases such as valvular disease, congenital anomalies and cardiomyopathies. The principal role of echocardiography was to assess persistent regional wall motion abnormality as well as to detect complications of myocardial infarction. However, recent technological advances have created many newer applications of echocardiography in this field. One of the most important advances was seen in the field of stress echocardiography. Dobutamine stress echocardiography has become an established method of diagnosing transient myocardial ischemia due to coronary stenosis and assessing the myocardial viability of a persistently akinetic segment. More recently, several new contrast agents have been developed or will be available in the near future to visualize the blood stream within the left heart cavity and myocardial blood flow. Simultaneously, new ultrasound technologies including harmonic imaging and gated intermittent imaging have enhanced the selective visualization of contrast agents and will contribute to noninvasive imaging of coronary microcirculation. Harmonic imaging has also been shown to improve quality of B-mode image without a contrast agent and will play an important role in the clinical recognition of wall motion abnormality in patients with ischemic heart disease. Recent advances in three-dimensional technology have enabled accurate measurements of left ventricular volume and ejection fraction without any geometrical assumption, which may be especially important for the evaluation of ischemic patients who often have a deformed left ventricular cavity due to remodeling.

Echocardiography↗

[Acute surgical and endovascular therapy for stroke: especially patients with brain infarction].

From January 1994 to December 1997, 845 patients with stroke were admitted to Hakodate Municipal Hospital. They consisted of 514 patients with brain infarction, 206 with brain hemorrhage, 121 with subarachnoid hemorrhage and 4 with intracranial hemorrhage from arteriovenous malformation. The clinical categories of brain infarction were as follows; atherothrombotic recognized in 158 patients, cardioembolic in 114, lacunar in 217 and other categories in 25. With regard to the cures of brain infarction in the acute phase, direct percutaneous transluminal angioplasty (direct PTA) was carried out on three patients with atherothrombotic infarction, immediate PTA on two, superselective fibrinolytic therapy on two, and STA-MCA anastomosis on three. In all, ten atherothrombotic patients (6.3%) were treated by acute surgical or endovascular therapy. On the other hand, superselective fibrinolytic therapy was carried out on 35 patients (30.7%) with cardioembolic infarction. There were no patients in the lacunar infarction group who were given acute surgical treatment. Neurological improvement after 24 hours was recognized in 4 patients (40%) of 10 with atherothrombotic infarction, and in 9 patients (25.7%) of 35 with cardioembolic infarction. However, symptomatic intracerebral hematoma was recognized in 4 patients (11.4%) with cardioembolic infarction. Indication for acute surgical or endovascular treatment for brain infarction was very limited because of the time factor from the onset to admission. It is suggested that neurosurgeons might enlighten citizens about the necessity for acute surgical or endovascular therapy for stroke.

Angioplasty, Balloon, Coronary↗

[Malignant peripheral nerve sheath tumor with divergent cartilage differentiation from the acoustic nerve: case report].

Malignant peripheral nerve sheath tumors(MPNSTs) of the acoustic nerve are very rare. Only seven cases of MPNST arising from the acoustic nerve have been reported. The authors present a case of MPNST with divergent cartilage and melanotic differentiation of the acoustic nerve. The patient was a 69 year old man admitted to our neurosurgical service in January 1999 complaining of left facial nerve palsy and hearing difficulty of his left ear. The initial CT showed a tumor at the left cerebellopontine angle region. On MRI the tumor was depicted as low intensity on T 1-weighted image and high intensity on T 2-weighted image, the mass was heterogeneously enhanced after administration of Gd-DTPA. The partial removal of the tumor was performed in January 1999. He was discharged February 1999. But he was admitted again because of progressive cerebellar ataxia. MRI showed the rapid regrowth of the residual tumor. In March 1999, complete removal of the tumor was performed. Histopathological analysis revealed a malignant spindle cell neoplasm with divergent cartilage and melanotic differentiation. We review the relevant literature concerning MPNST of the acoustic nerve and discuss the clinical features of malignant eighth cranial nerve tumor.

Aged↗

[Common carotid artery dissection propagated from acute aortic dissection: a case successfully treated by PTA].

We reported a rare case, which was successfully treated by PTA, of right common carotid artery dissection propagated from acute aortic dissection (AAD) type A. A 45-year-old male with a past history of hypertension and an artificial graft replacement of the abdominal aorta due to AAD type B, 7 years ago, was brought into our hospital by ambulance 30 minutes after an attack of fainting and left hemiparesis. On admission, the patient complained not of chest pain or left hemiparesis, but nausea. At that time his consciousness level was JCS 1. During examinations, he had the same attack twice and his consciousness level deteriorated to JCS 2. Brain MRI showed no abnormality, but cervical MRA did not visualize the right carotid artery and thoracic CT depicted acute aortic dissection including branches of the aorta. Emergent angiography disclosed that the dissecting 99% stenosis of the right common carotid artery had developed from AAD type A with poor collateral blood flow. PTA was carried out 8 times and reduced the residual stenosis to about 50% with shortened circulation time. The patient's consciousness disturbance improved. After the replacement of the whole aortic arch in an artificial graft, the residual stenosis disappeared. The patient recovered without neurological deficit but right frontal silent embolic infarction caused by the artificial graft replacement was detected. AAD is a catastrophic illness and sometimes accompanied by devastating ischemic cerebral disease (ICD) because of propagation of dissecting to extracranial vessels. This is the first report that shows the efficacy of PTA for treatment of ICD associated with AAD.

Acute Disease↗

Candida albicans: adherence, signaling and virulence.

The focus of this symposium was to present new information on the morphogenesis of Candida albicans, particularly how it relates to signal transduction pathways and other genes involved in the regulation of morphogenesis. In addition, we discuss the role of adherence and colonization of the oral cavity by the organism and discuss the role of mannan as an adhesin that recognizes the human red blood cell. C. albicans utilizes at least two signal pathways to regulate its conversion from a yeast form to filamentous growth (hyphae). One of these two pathways is similar to the Saccharomyces cerevisiae pseudohyphal/mating pathway, which utilizes the regulatory protein, Cphlp. The other pathway is not totally defined but requires a second regulatory protein, referred to as Efg1p. Other signal pathways may exist, which include a two-component histidine kinase and response regulator proteins. The latter pathway(s) may include proteins such as Chk1p, Ssk1p, Shi1p and Cos1p/Nik1p. Mutations in strains, which specifically target these proteins, result in morphogenesis defects and avirulence or attenuation of strains. A growth regulatory gene has also been recently defined whose expression is associated with growth cessation and which appears to be a necessary prerequisite in conversion of the organism to a filamentous growth form. Starvation of yeast cells induces exponentially grown cells (and usually non-germinative) to germinate. This phenomenon is also observed in cells that are transiently treated with metabolic inhibitors. During each of these treatments (starvation, metabolic inhibition), expression of a growth regulatory gene (CGRI) increases. Adherence of C. albicans to host cells and tissues is complex; several proteins, which appear to have host recognition functions, have been defined. In the oral cavity, C. albicans selectively adheres to salivary proteins, which are absorbed to many oral surfaces. This mechanism enables the cells to colonize surfaces of the oral cavity. An understanding of these interactions may lead to strategies to prevent oral disease. Mannan from C. albicans may provide a host recognition function for C. albicans. Recent experiments indicate that mannan binds to human red blood cells and causes hemolysis. Binding of mannan to the band 3 protein of human red blood cells has been established. This activity may be associated with the ability of the organism to utilize hemoglobin (and iron).

Candida albicans↗

Interleukin 4 is a crucial cytokine in controlling Trypanosoma brucei gambiense infection in mice.

The role of interleukin 4 (IL-4) was studied in relation to host defense during Trypanosoma brucei gambiense IL3253 (IL3253) infection in mice. BALB/c/A-+/+ (BALB/c), BALB/c/A-nu/nu (nude) and C.B-17/Icr-scid/scid (SCID) mice were infected intraperitoneally with 5 x 10(3) bloodstream forms (BSFs) of the trypanosome. The BALB/c mice showed high resistance to IL3253 infection with sporadic parasitemia. The nude mice were also able to control IL3253 infection and experienced low, but persistent parasitemia. However, the SCID mice, which have no functional T- and B-cells, showed high susceptibility to IL3253 infection with more than 1 x 10(8) BSFs/ml. Serum IL-4 levels in the infected BALB/c mice were increased on days 12-18 post-infection (PI). In BALB/c mice depleted of CD4+ T-cells by monoclonal antibody (mAb) treatment, parasitemia was persistent, ranging from 1 x 10(4) to 1 x 10(6) BSFs/ml and was significantly higher than that of the other groups. IL-4 was not detected in the serum of CD4+ T-cells-depleted mice. On the other hand, anti-IL-4-treated IL3253-infected BALB/c mice relapsed significantly longer than the control mice (p < 0.01). These findings suggest that the CD4+ T-cells may control the levels of parasitemia in IL3253 infection through the IL-4 pathway.

Animals↗

Tandem organization of medaka fish soluble guanylyl cyclase alpha1 and beta1 subunit genes. Implications for coordinated transcription of two subunit genes.

We determined the complete nucleotide sequences of the alpha1 subunit gene (OlGCS-alpha1) and the beta1 subunit gene (OlGCS-beta1) of medaka fish soluble guanylyl cyclase. In the genome, OlGCS-alpha1 and OlGCS-beta1 are organized in tandem. The two genes are only 986 base pairs apart and span approximately 34 kilobase pairs in the order of OlGCS-alpha1 and OlGCS-beta1. The nucleotide sequence of a large part of the 5'-upstream region of OlGCS-alpha1 is complimentarily conserved in that of OlGCS-beta1. To analyze the promoter activity of each gene, a fusion gene construct in which the 5'-upstream region was fused with the green fluorescent protein gene was injected into medaka fish 2-cell embryos. When the fusion gene containing the OlGCS-alpha1 upstream region was injected, green fluorescent protein fluorescence was detected in the embryonic brain. The 5'-upstream region of OlGCS-beta1 alone was insufficient for the reporter gene expression in the embryos. When the OlGCS-alpha1 upstream region was located upstream of the OlGCS-beta1-green fluorescence protein fusion gene, the reporter gene was expressed in the brain and trunk region of the embryos. These results suggest that the 5'-upstream region of OlGCS-alpha1 can affect the expression of OlGCS-beta1. It is therefore possible that the expression of OlGCS-alpha1 and OlGCS-beta1 is coordinated.

Animals↗