Biomedical subjects

Akira Ito

Publications and source records attributed to Akira Ito.

At least 73 records · Page 4Linked to original sources

PubMed2005

Heat immunotherapy using magnetic nanoparticles and dendritic cells for T-lymphoma.

Dendritic cells (DCs) are potent antigen-presenting cells that play important roles in regulating immune responses in cancer. Immunotherapy using these immunocytes has become an accepted therapeutic modality. We previously reported that hyperthermia using magnetic nanoparticles induces antitumor immunity, which could be activated by adjuvant including cytokines. In the present study, we investigated the therapeutic effects of hyperthermia combined with DC immunotherapy on mouse EL4 T-lymphoma. Magnetite cationic liposomes (MCLs) have a positive surface charge and generate heat in an alternating magnetic field (AMF) due to hysteresis loss. MCLs were injected into an EL4 nodule in C57BL/6 mice, which were subjected to AMF for 30 min. The temperature at the surface of the tumor reached 45 degrees C and was maintained by controlling the magnetic field intensity. Hyperthermia treatment was repeated twice with 24 h intervals. After hyperthermia, immature DCs were directly injected into the EL4 nodule. As a result, complete regression of tumors in 75% (6/8) of the mice was observed, while the percentage of complete regression of tumors was 12.5% (1/8) in the case of mice treated by hyperthermia alone. This novel cancer therapy, which we have termed "heat immunotherapy", may be applicable to patients with advanced malignancies.

Animals↗

PubMed2005

Screening of cytokines to enhance vaccine effects of heat shock protein 70-rich tumor cell lysate.

Heat shock proteins (HSPs) have been recognized as significant participants in immune reactions. We have previously reported that heat-treated cells expressing HSP70 can mediate potent antitumor immune responses. As successful immunotherapy is dependent on the host immune system, the present study evaluated whether systemic administration of immunocyte stimulatory and growth promoting cytokines could enhance heat-treated cell lysate vaccine (HCLV) immunization to further promote the antitumor immunity. After heating mouse melanoma B16 cells (43 degrees C, 30 min) to elicit increased HSP70 expression, cells were lysed by freeze thawing to prepare HCLV. In approaches using a poorly immunogenic melanoma B16, the effects of various cytokines (IL-1beta, -2, -4, -6 and -12, IFN-beta and -gamma, GM-CSF and TNF-alpha) were assessed in combination with HCLV. Syngenic C57BL/6 mice were immunized subcutaneously with HCLV twice, on days -14 and -7, while cytokines were injected intraperitoneally on day -7. Subcutaneous B16 cell challenge was performed on day 0. IL-12 significantly enhanced the efficacy of HCLV, compared to non-heated cell lysate vaccine (CLV) and non-vaccination. Systemic administration of recombinant IL-12 augmented the efficacy of HCLV, inducing protective immunity against tumor challenge and enhancing cytotoxicity assessed in primed splenocytes against B16 cells in treated mice. These results suggest that IL-12 represents an important modulator of antitumor immune responses induced by HCLV, and may facilitate further efforts to develop novel cancer immunotherapies based on HSP70-mediated vaccination.

Animals↗

PubMed2005

Increase of antibody titer against Leucocytozoon caulleryi by oral administration of recombinant R7 antigen.

In this study, we examined oral administration with recombinant R7 (rR7) antigen expressed in Escherichia coli using chicken leucocytozoonosis subunit vaccine (LV)-vaccinated and unvaccinated chickens. Only LV-vaccinated chickens showed re-induction of anti-second-generation schizont (2GS) antibody. Also, LV-vaccinated chickens whose anti-2GS antibody titer was middle-level showed increases of the antibody titer compared to vaccinated-control chickens (P>0.01, >0.05) after oral administration with rR7 antigen.

Administration, Oral↗

PubMed2005

Echinococcosis in Tibetan populations, western Sichuan Province, China.

We screened 3,199 people from Shiqu County, Sichuan Province, China, for abdominal echinococcosis (hydatid disease) by portable ultrasound combined with specific serodiagnostic tests. Both cystic echinococcosis (CE) (Echinococcus granulosus infection) and alveolar echinococcosis (AE) (E. multilocularis) were co-endemic in this area at the highest village prevalence values recorded anywhere in the world: 12.9% were infected with one or the other form (6.8% CE and 6.2% AE). Prevalences of both CE and AE were significantly higher in female than male patients and increased with the age of the person screened. Pastoral herdsmen were at highest risk for infection (prevalence 19.0%). Prevalence of CE varied in 5 townships from 0% to 12.1%, whereas AE prevalence ranged from 0% to 14.3%. Risk factors associated with both infections included the number of owned dogs, frequency of contact with dogs, and sources of drinking water.

Adolescent↗

PubMed2005

Prevalence of toxocariasis in northeastern Brazil based on serology using recombinant Toxocara canis antigen.

To evaluate the prevalence of toxocariasis in children in Jaboatão dos Guararapes, Pernambuco in northeastern Brazil, 215 serum samples were examined by an enzyme-linked immunosorbent assay (ELISA) using a recombinant Toxocara canis antigen. In the ELISA, 26 (12.1%) of 215 subjects were positive. In a dot-blot assay using 53 of 215 serum samples, the diagnostic results correlated with those obtained by the ELISA. Moreover, it has been confirmed that the recombinant T. canis antigen was highly specific for toxocariasis by ELISA using serum samples positive for antibody to Ascaris lumbricoides. Considering the specificity of the recombinant antigen to toxocariasis, the ELISA or dot-blot assay using the recombinant T. canis antigen is recommended in tropical and sub-tropical regions where various parasitic infections are commonly endemic.

Animals↗

PubMed2005

Short report: Echinococcus multilocularis confirmed on Kunashiri Island, 15 kilometers from the eastern part of Hokkaido, Japan.

There are two foci of alveolar echinococcosis (AE) caused by Echinococcus multilocularis in Japan. The first focus is on Rebun Island where AE patients were found from 1937, and the second is in eastern Hokkaido where patients have been found since the 1960s. The origin of the second focus is unknown. To further investigate AE in eastern Hokkaido, wild rodents (Muridae) were captured and examined for infection on Kunashiri Island, which is located 15 km off the northeastern coast of Hokkaido. Metacestodes of E. multilocularis were isolated from two of 31 voles, all of which were identified to be Clethrionomys rufocanus. Mitochondrial DNA sequencing data of recovered cestodes showed total identity with the cestode reported from Hokkaido. These results suggest that E. multilocularis may have been introduced to Hokkaido from Kunashiri Island during or after 1965.

Animals↗

PubMed2005

Experimental evaluation of conversion factors, N(D,w)/N(X), for Roos-type and Advanced-Markus-type plane-parallel ionization chambers.

A Japanese code of practice for clinical dosimetry, titled "Standard Dosimetry of Absorbed Dose in External Beam Radiotherapy" was published by the Japan Society of Medical Physics (JSMP) in 2002. It mostly followed IAEA Technical Reports Series No. 398, which was based on N(D,w), i.e., the calibration factor in terms of absorbed dose to water for a dosimeter. The Japanese primary standard dosimetry laboratory, however, has not supplied N(D,w) but N(X), as the calibration factor in terms of exposure. The unique feature of the Japanese code was provision of a data table of calculated conversion factors, N(D,w) / N(X) values, for many types of ionization chambers, excluding new plane-parallel ionization chambers. This paper describes the experimental evaluation of the conversion factors for the new plane-parallel ionization chambers, such as the Roos-type and Advanced Markus chambers. The obtained N(D,w) / N(X) values for PTW 34001, Wellhöfer PPC 40 and PTW 34045 were 37.96 +/- 0.19, 37.85 +/- 0.36 and 37.90 +/- 0.26 (Gy/C kg(-1)), respectively. They agreed with estimations based on Monte Carlo calculations.

Calibration↗

PubMed2005

Taeniasis and cysticercosis in Asia and the Pacific: present state of knowledge and perspectives.

Several topics on taeniasis and cysticercosis in Asia and the Pacific are overviewed. In Asia and the Pacific, three human taeniid species have been recognized: Taenia solium, Taenia saginata and Taenia asiatica. The first topic is on evolution of T. solium. Mitochondrial DNA polymorphisms of T. solium worldwide are discussed with emphasis of two specific genotypes: American-African and Asian. The second topic is recent major advances in sero- and molecular-diagnosis of T. solium cysticercosis in humans, pigs and dogs. The third is the present situation of T. solium taeniasis/cysticercosis in Papua (Irian Jaya), Indonesia. The forth is the present situation of T. solium cysticercosis and T. saginata taeniasis in Bali, Indonesia. The fifth is the present situation of T. asiatica taeniasis in Asia and the Pacific and in North Sumatra, Indonesia. The sixth is on the debate of the exact definition of T. asiatica. Because T. asiatica can not be differentiated from T. saginata morphologically, it is time to re-evaluate T. saginata in Asia and the Pacific. New and broad-based surveys across this region are necessary from epidemiological and public health perspectives, based on evidence.

Animals↗

PubMed2005

Molecular identification of Taenia solium cysticercus genotype in the histopathological specimens.

PCR-based molecular diagnosis was done for identification of causative agents found in paraffin-embedded specimens that were resected from two suspected neurocysticercosis patients. DNA samples were extracted from tissues or sections and cytochrome c oxidase subunit 1 gene and cytochrome b gene were amplified for the detection of taeniid DNA. Two different genes were successfully amplified in both specimens, but the sizes of amplified products seemed to depend on the quality of DNA. Based on the nucleotide sequences of the PCR-amplified genes, the causative agents from two cases were identified as T. solium Asian genotype. When infection with T. solium is not confirmed by histopathological examination, molecular diagnosis will be more useful for definitive diagnosis.

Child↗

PubMed2005

Fundamental study of expression-library immunization against Taenia taeniaeformis in rats.

Expression-library immunization has been proposed as an effective means to screen a large number of genes of the pathogen as candidate protective molecules. In this study, we examined the efficacy of expression-library immunization using a T. taeniaeformis rat model system. Total RNAs were isolated from the last 15 segments of adult T. taeniaeformis and poly A RNA was purified. cDNA library was produced using SuperScript Plasmid System, which contains a mammalian expression vector, pCMV*SPORT6. From about 3,500 clones examined, more than 800 clones were found to contain DNA fragments. About 200 clones were sequenced and the homology search was carried out. The blast search revealed that 29% of the expression genes were mitochondrial genes (rRNA; 17%, protein; 12%). Nuclear rRNA genes (10%), nuclear protein (9%) and genes from Escherichia coli were also detected. Forty-two percent of sequences did not show a significant similarity to any genes deposited in the public database. Rats were immunized with expression-library and injected orally with 1,000 T. taeniaeformis eggs. However the protective effect of expression-library vaccine was not confirmed.

Animals↗

PubMed2004

Characterization of the Alzheimer's disease-associated CLAC protein and identification of an amyloid beta-peptide-binding site.

Amyloid beta-peptide (Abeta) deposition into amyloid plaques is one of the invariant neuropathological features of Alzheimer's disease. Other proteins co-deposit with Abeta in plaques, and one recently identified amyloid-associated protein is the collagen-like Alzheimer amyloid plaque component CLAC. It is not known how CLAC deposition affects Abeta plaque genesis and the progress of the disease. Here, we studied the in vitro properties of CLAC purified from a mammalian expression system. CLAC displays features characteristic of a collagen protein, e.g. it forms a partly protease-resistant triple-helical structure, exhibits an intermediate affinity for heparin, and is glycosylated. Purified CLAC was also used to investigate the interaction between CLAC and Abeta. Using a solid-phase binding assay, we show that CLAC bound with a similar affinity to aggregates formed by Abeta-(1-40) and Abeta-(1-42) and that the interaction was impaired by increasing salt concentrations. An 8-residue-long sequence located in non-collagenous domain 2 of CLAC was found to be crucial for the interaction with Abeta. These findings may be useful for future therapeutic interventions aimed at finding compounds that modulate the binding of CLAC to Abeta deposits.

Alzheimer Disease↗

PubMed2004

Nicastrin, presenilin, APH-1, and PEN-2 form active gamma-secretase complexes in mitochondria.

Mitochondria are central in the regulation of cell death. Apart from providing the cell with ATP, mitochondria also harbor several death factors that are released upon apoptotic stimuli. Alterations in mitochondrial functions, increased oxidative stress, and neurons dying by apoptosis have been detected in Alzheimer's disease patients. These findings suggest that mitochondria may trigger the abnormal onset of neuronal cell death in Alzheimer's disease. We previously reported that presenilin 1 (PS1), which is often mutated in familial forms of Alzheimer's disease, is located in mitochondria and hypothesized that presenilin mutations may sensitize cells to apoptotic stimuli at the mitochondrial level. Presenilin forms an active gamma-secretase complex together with Nicastrin (NCT), APH-1, and PEN-2, which among other substrates cleaves the beta-amyloid precursor protein (beta-APP) generating the amyloid beta-peptide and the beta-APP intracellular domain. Here we have identified dual targeting sequences (for endoplasmic reticulum and mitochondria) in NCT and showed expression of NCT in mitochondria by immunoelectron microscopy. We also showed that NCT together with APH-1, PEN-2, and PS1 form a high molecular weight complex located in mitochondria. gamma-secretase activity in isolated mitochondria was demonstrated using C83 (alpha-secretase-cleaved C-terminal 83-residue beta-APP fragment from BD8 cells lacking presenilin and thus gamma-secretase activity) or recombinant C100-Flag (C-terminal 100-residue beta-APP fragment) as substrates. Both systems generated an APP intracellular domain, and the activity was inhibited by the gamma-secretase inhibitors l-685,458 or Compound E. This novel localization of NCT, PS1, APH-1, and PEN-2 expands the role and importance of gamma-secretase activity to mitochondria.

Adenosine Triphosphate↗

PubMed2004

Magnetite nanoparticle-loaded anti-HER2 immunoliposomes for combination of antibody therapy with hyperthermia.

Anti-HER2 antibody can induce antitumor responses, and can be used in delivering drugs to HER2-overexpressing cancer. Previously, we produced hyperthermia using magnetite nanoparticles that generate heat in an alternating magnetic field. In the present study, we constructed anti-HER2 immunoliposomes containing magnetite nanoparticles, which act as tumor-targeting vehicles, combining anti-HER2 antibody therapy with hyperthermia. The magnetite nanoparticle-loaded anti-HER2 immunoliposomes exerted HER2-mediated antiproliferative effects on SKBr3 breast cancer cells in vitro. Moreover, 60% of magnetite nanoparticles were incorporated into SKBr3, and the cells were then heated at 42.5 degrees C under an alternating magnetic field, resulting in strong cytotoxic effects. These results suggest that this novel therapeutic tool is applicable to treatment of HER2-overexpressing cancer.

Antibodies, Monoclonal↗

PubMed2004

Prevention of UVB-induced photoinflammation and photoaging by a polymethoxy flavonoid, nobiletin, in human keratinocytes in vivo and in vitro.

Exposure to ultraviolet B (UVB) irradiation induces acute skin inflammation such as erythema (sunburn) and edema, and prostaglandin (PG)E2 in the epidermis plays an important role as its prominent mediator. In the present study, we investigated the effect of nobiletin (5,6,7,8,3',4'-hexamethoxy flavone) from Citrus depressa, on the production of PGE2 in UVB-irradiated human keratinocytes. When keratinocytes were irradiated with 60mJ of UVB/cm2, the production and gene expression of cyclooxygenase (COX)-2, but not COX-1, were augmented along with an increase in PGE2 levels. The augmented COX-2 production was transcriptionally suppressed by nobiletin. In addition, neither the release of [14C]arachidonic acid from membrane phospholipids nor the gene expression of cytosolic phospholipase A2 (cPLA2) was altered in UVB-irradiated human keratinocytes. However, nobiletin was found to inhibit the release of [14C]arachidonic acid by decreasing the Ca2+ -dependent activity of cPLA2. Furthermore, topical treatment of nobiletin on the skin of the back prevented the UVB-induced increase of transepidermal water loss and hyperplasia of the epidermis in hairless mice. Therefore, these results suggest that nobiletin inhibits the UVB-induced production of PGE2 not only by suppressing the expression of COX-2 but also by decreasing the activity of cPLA2 in human keratinocytes. Furthermore, nobiletin may be useful as a novel sunscreen reagent to be applied for protection against photoinflammation and photoaging.

Animals↗

PubMed2004

Functional expression and characterization of Echinococcus granulosus thioredoxin peroxidase suggests a role in protection against oxidative damage.

A full-length cDNA sequence coding for Echinococcus granulosus thioredoxin peroxidase (EgTPx) was isolated from a sheep strain protoscolex cDNA library by immunoscreening using a pool of sera from mice infected with oncospheres. EgTPx expressed as a fusion protein with glutathione S-transferase (GST) exhibited significant thiol-dependent peroxidase activity that protected plasmid DNA from damage by metal-catalyzed oxidation (MCO) in vitro. Furthermore, the suggested antioxidant role for EgTPx was reinforced in an in vivo assay, whereby its expression in BL21 bacterial cells markedly increased the tolerance and survival of the cells to high concentrations of H2O2 compared with controls. Immunolocalization studies revealed that EgTPx was specifically expressed in all tissues of the protoscolex and brood capsules. Higher intensity of labelling was detected in many, but not all, calcareous corpuscle cells in protoscoleces. The purified recombinant EgTPx protein was used to screen sera from heavily infected mice and patients with confirmed hydatid infection. Only a portion of the sera reacted positively with the EgTPx-GST fusion protein in Western blots, suggesting that EgTPx may form antibody-antigen complexes or that responses to the EgTPx antigen may be immunologically regulated. Recombinant EgTPx may prove useful for the screening of specific inhibitors that could serve as new drugs for treatment of hydatid disease. Moreover, given that TPx from different parasitic phyla were phylogenetically distant from host TPx molecules, the development of antiparasite TPx inhibitors that do not react with host TPx might be feasible.

Animals↗

PubMed2004

The Tibetan hare Lepus oiostolus: a novel intermediate host for Echinococcus multilocularis.

Multicystic hydatids have been found in the livers of hares ( Lepus oiostolus) examined from the Qinghai-Tibet plateau of China. In this study, the causative species was definitively identified as Echinococcus multilocularis by mitochondrial DNA sequencing. This is the first confirmation of larval E. multilocularis from hares. The hydatids contained protoscolices, suggesting that the hare may contribute to the transmission of E. multilocularis on the Tibetan plateau.

Animals↗

PubMed2004

Functional magnetic particles for medical application.

Magnetic particles for medical applications have been developed by many researchers. Since magnetic particles have unique magnetic features not present in other materials, they can be applied to special medical techniques. Separation, immunoassay, magnetic resonance imaging (MRI), drug delivery, and hyperthermia are enhanced by the use of magnetic particles. Magnetite cationic liposomes (MCLs), one of the groups of cationic magnetic particles, can be used as carriers to introduce DNA into cells since their positively charged surface associates with the negatively charged DNA. MCLs can also be used as heat mediators for cancer therapy. Magnetic particles conjugated with tumor-specific antibodies have enabled tumor-specific contrast enhancement in MRI. In addition, antibody-conjugated magnetic particles were shown to target renal cell carcinoma cells, and are applicable to the hyperthermic treatment of carcinomas. It was also found that the hyperthermic treatment using magnetic particles induced an antitumor immunity. Thus, the use of magnetic particles with their unique features will further improve medical techniques.

Animals↗

PubMed2004

Giant adrenal myelolipoma: report of a case.

Adrenal myelolipomas are rare benign tumors composed of mature adipose tissue and hematopoietic elements that resemble bone marrow. They are usually asymptomatic, and most cases are incidentally found at radiological examination or autopsy. Symptoms such as abdominal pain and increasing girth occur only when the tumor grows large. We report the case of a giant adrenal myelolipoma in a 51-year old man who presented with a huge abdominal mass and abdominal pain. The resected tumor weighed 6,000 g and could represent the largest such tumor ever documented in the literature. We discuss the diagnosis and treatment of this unusual tumor.

Adrenal Gland Neoplasms↗