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Biomedical subjects

T Ishida

Publications and source records attributed to T Ishida.

At least 55 records · Page 3Linked to original sources

Expression of thyroid transcription factor-1 in 16 human lung cancer cell lines.

It has been suggested that thyroid transcription factor-1 (TTF-1) is frequently expressed in human lung cancer, especially in adenocarcinoma and small cell lung cancer, and the TTF-1 expression is closely related with the expression of surfactant protein. We hypothesized that TTF-1 is expressed in human lung cancer cell lines and its expression might be related to the expression of surfactant protein. To test this, expressions of TTF-1 and surfactant protein A (SP-A) were immunohistochemically evaluated in 16 human lung cancer cell lines. In addition, expressions of mRNAs for TTF-1 and SP-A were analyzed by reverse transcriptase-polymerase chain reaction (RT-PCR) and sequencing. As a result, nuclear staining of TTF-1 was observed in two of six adenocarcinoma cell lines, none of seven small cell lung cancer cell lines, and none of three squamous lung cancer cell lines. Among the 16 cell lines, six cell lines (PC3, LC2/Ad, A549, RERF-LC-OK, HI1017, and PC9) expressed significant amounts of mRNA for TTF-1. In contrast, cytoplasmic staining of TTF-1 was observed in five of six adenocarcinoma cell lines, in six of seven small cell lung cancer cell lines, and in all three squamous cell lung cancer cell lines. One of the two adenocarcinoma cell lines those showed positive nuclear staining and cytoplasmic SP-A staining released a significant amount of SP-A in culture supernatant. Our present study demonstrates that the frequency of TTF-1 expression in the nucleus was very low in human lung cancer cell lines; however, their cytoplasmic positivities should be further investigated.

Adenocarcinoma↗

KL-6, a human MUC1 mucin, is expressed early in premature lung.

KL-6, one of the MUC1 antigens, is a mucin-like high-molecular-weight glycoprotein, which is strongly expressed on type II pneumocytes. Serum levels of KL-6 have been shown to correlate with activity of interstitial pneumonia (IP). During embryonic development, MUC1 expression coincides with the onset of epithelial sheet and glandular formation. To investigate the potential role of KL-6 in lung morphogenesis, we examined KL-6 expression by immunohistochemistry on autopsied lung tissue specimens of 35 neonates and infants with gestational age from 23 to 40 weeks. Hyaline membranes (HMs) were detected in 13 of 35 cases. Simultaneously, antibody against surfactant protein A (SP-A) was employed in the study which is a distinct marker for type II pneumocytes. In all cases studied with gestational age above 23 weeks, staining for KL-6 was strongly positive in alveolar epithelial cells and in HMs found in 13 cases, whereas immunoreaction for PE10 varied depending on gestational age and duration of postnatal survival. Our findings suggest that KL-6 is expressed earlier in premature lung and may act as an important factor contributing to morphogenesis and function of developing lung in early gestation.

Antigens↗

The SDF1-G801A polymorphism is not associated with SDF1 gene expression in Epstein-Barr virus-transformed lymphoblastoid cells.

The effects of the SDF1-3'A on AIDS progression have been attributed to the altered amount of stromal cell-derived factor 1 (SDF-1). However, the contribution of the SDF1-G801A polymorphism to SDF-1 expression is still unclear. In contrast to fresh peripheral blood mononuclear cells (PBMCs), Epstein-Barr virus (EBV)-transformed lymphoblastoid cell lines (LCLs) express the SDF-1 mRNA. Using EBV-transformed LCLs from 42 individuals with different genotypes, we investigated the SDF-1 mRNA levels and methylation status in the SDF1 gene. Both in PBMCs and in EBV-transformed LCLs, CpG dinucleotides in the 5' region of the SDF1 gene were unmethylated. As for the 3' untranslated region (3'UTR), by contrast, CpG dinucleotides were methylated in PBMCs, whereas site-specific demethylation around the polymorphic site was detected in EBV-transformed LCLs. The levels of the demethylation were correlated with the SDF-1 mRNA levels. However, the genotype for the SDF1-G801A polymorphism did not significantly alter the SDF-1 mRNA levels. The allele preferences in transcription and methylation were also absent in the heterozygous cells. In conclusion, this study suggested a contribution of site-specific demethylation in the 3'UTR to the SDF1 gene expression, but did not show any evidence for the contribution of the SDF1-G801A polymorphism to the amount of the SDF-1 mRNA.

Alleles↗

Significance of early biopsy in pediatric kidney transplantation.

Long-term renal allograft survival is limited mainly by the progressive process termed chronic allograft nephropathy (CAN) or chronic rejection. A pathological feature of CAN is characterized by progressive interstitial fibrosis. Transforming growth factor (TGF)-beta(1) plays an important role in fibrogenesis. We investigated whether the degree of TGF-beta(1) expression in early biopsy specimens routinely obtained from stable allografts at 100 days could predict fibrosis and graft dysfunction in the late phase by immunohistochemistry. Patients were children with a graft from related donors. We immunohistochemically determined intracellular and extracellular expression of TGF-beta(1) in the graft at 100 days using LC antibody (LC) for intracellular TGF-beta(1) and CC antibody (CC) for extracellular TGF-beta(1). We used the change in creatinine clearance between 100 days and 3 years after transplantation (Delta Ccr) as an index of long-term graft function. Image analysis was used to calculate the relative area involved by interstitial fibrosis in trichrome-stained sections of graft biopsy specimens at 100 days and 3 years, designating the change as Delta FI. Delta Ccr was - 4.2 +/- 9.4 mL/min in subjects with minimal early immunoreactivity for CC and - 20.5 +/- 5.9 mL/min in subjects with strong reactivity (p < 0.05). Delta Ccr was - 14.5 +/- 18.6 mL/min in subjects with minimal early immunoreactivity for LC and - 11.7 +/- 12.8 mL/min in those with strong reactivity. Delta FI in subjects with minimal CC reactivity (1.28 +/- 4.11 %) tended to be lower than in subjects with strong reactivity (8.45 +/- 15.47 %). Neither fibrosis at 100 days nor Delta FI differed between subjects with minimal and strong LC reactivity. Thus, extracellular TGF-beta(1) expression in grafts at 100 days after transplantation has an influence on long-term graft function and tends to be associated with increased graft fibrosis at 3 years.

Adolescent↗

Gene arrangement at the Rhesus blood group locus of chimpanzees detected by fiber-FISH.

The Rhesus (Rh) blood group system in humans is encoded by two genes with high sequence homology. These two genes, namely, RHCE and RHD, have been implied to be duplicated during evolution. However, the genomic organization of Rh genes in chimpanzees and other nonhuman primates has not been precisely studied. We analyzed the arrangement of the Rh genes of chimpanzees (Pan troglodytes) by two-color fluorescence in situ hybridization on chromatin DNA fibers (fiber-FISH) using two genomic DNA probes that respectively contain introns 3 and 7 of human RH genes. Among the five chimpanzees studied, three were found to be homozygous for the two-Rh-gene type, in an arrangement of Rh (5'-->3') - Rh (3'<--5'). Although a similar gene arrangement can be detected in the RH gene locus of typical Rh-positive humans, the distance between the two genes in chimpanzees was about 50 kb longer than that in humans. The remaining two chimpanzees were homozygous for a four-Rh-gene type, in an arrangement of Rh (5'-->3') - Rh (3'<--5') - Rh (3'<--5') - Rh (3'<--5') within a region spanning about 300 kb. This four-Rh-gene type has not been detected in humans. Further analysis of other great apes showed different gene arrangements: a bonobo was homozygous for the three-Rh-gene type; a gorilla was heterozygous for the one-Rh- and two-Rh-gene types; an orangutan was homozygous for the one-Rh-gene type. Our findings on the intra- and interspecific genomic variations in the Rh gene locus in Hominoids would shed further light on reconstructing the genomic pathways of Rh gene duplication during evolution.

Animals↗

[Usefulness of low tube operating potential (TOP) for improved detection of minimally contrast-enhanced nodules CT].

PURPOSE: We recently observed that hepatic nodules which were only minimally enhanced on dynamic CT were much better visualized using a low TOP in spite of some increase of the background noise. The purpose of this study is to evaluate the potential superiority of low TOP in detecting minimally contrast-enhanced hepatic nodules on CT. METHODS AND MATERIALS: A phantom was made of anger and contrast medium. Four anger columns were made four different concentrations of contrast media (0.8%, 1.0% 1.5% 1.8%). The phantom was scanned using different TOP (80, 100, 120, 140 kVp). Detectability of the columns were also evaluated contrast to noise ratio (CNR) and visually by radiologists in a doubleblind fashion. Weighted computed tomography dose index (CTDIw) was compared with different TOP. In the clinical study, seven patients were studied. The diagnostic usefulness of a TOP of 80 kVp was compared with that of 120 kVp in same patient by Dynamic CT which has better detectability of minimally contrast enhancing nodules. RESULT: With using lower TOP, CNR was increased. On visual evaluation by radiologists, images taken at 80 kVp achieved the highest detectability of the columns. CTDIw associated with the 80, 100, 120, 140 kVp at one scan were 5.78 mGy, 6.64 mGy, 8.13 mGy, 11.02 mGy, respectively. In the clinical study a lesion could be detected clearly with a low TOP of 80 kVp though the existence of the lesion could not be detected at 120 kVp. The existence of the HCC was proven with DSA, CTA and CTAP. CONCLUSION: It was demonstrated in the phantom study that if up-to-date equipment is used, CT operated at a TOP of 80 kVp has potentially better detectability of minimally contrast-enhanced nodules than 120 kVp.

Contrast Media↗

[Development of computerized method for automated classification of the body parts in digital radiographs].

PURPOSE: In picture archiving and communication systems (PACS), the information on the body parts included in radiographs is often not or incorrectly recorded in an image header. In order to apply the computer-aided diagnosis (CAD) system in the PACS environment, the body parts in radiographs need to be recognized correctly by computer. The purpose of this study is to develop a computerized method for correctly classifying the body parts in digital radiographs based on a template matching technique. METHODS/MATERIALS: The image database used in this study was 1032 digital radiographs (14 x 17 inches) obtained with a computed radiography, and included 505 chest of postetroanterior view, 39 chest of lateral view, 241 abdomen, 108 pelvis, 10 upper limbs, 125 lower limbs, and 4 thoracic spine. In this method, test images were classified into four body parts, i.e., (1) chest, (2) abdomen, (3) pelvis, and (4) upper/lower limbs and thoracic spine. This computerized method was tested with 852 images, since 180 images were employed for creation of 98 templates, which represented the average radiographs for various body parts. Our approach was to examine the similarity of a given test image with templates by use of the cross-correlation values as the similarity measures. The body part of the test image was identified as the body part in the template yielding the maximum correlation value. Our method consisted of the following five steps. First, test images were classified into one of three groups; i.e. 1) chest and abdomen, 2) pelvis, and 3) upper/lower limbs and thoracic spine by using the templates obtained from images with the average size and position. Second, the remaining uncertain images were classified by using additional templates in various directions. Third, the chest and abdomen group was separated into two subgroups; i.e.chest and abdomen. Fourth, in order to classify some uncertain images, templates were shifted horizontally and vertically. Fifth, outer pixels of templates were eliminated to avoid the misclassification due to x-ray collimation. RESULTS: Our preliminary results indicated that the body parts for 850 cases (99.8%) were correctly classified with our method. CONCLUSIONS: This method would be useful for automated identification of the body parts in radiographs when various CAD systems would be implemented in the PACS environment.

Diagnosis, Computer-Assisted↗

Nonspecific interstitial pneumonia as pulmonary involvement of primary Sjögren's syndrome.

The pathologic patterns of lung involvement in nine patients with Sjögren's syndrome (SjS) are evaluated. The patients consisted of three males and six females, with a median age of 59 years. The SjS was diagnosed according to the criteria of the First International Seminar on SjS. In all patients, high-resolution computed radiographic scanning (HRCT) of the lungs was performed, and apparent honeycomb or microhoneycomb formation was observed in six patients. Pathologically, six patients were diagnosed with usual interstitial pneumonia (UIP), and three were diagnosed with nonspecific interstitial pneumonia/fibrosis (NSIP) (group II). There were no apparent honeycomb formations on HRCT in patients diagnosed with NSIP. In conclusion, NSIP is also a possible histologic classification of interstitial pneumonia associated with SjS.

Aged↗

Normal nephrogenesis occurs in the early stage of bilateral multicystic dysplastic kidneys.

We present a case report about bilateral multicystic dysplastic kidneys (MCDKs). Pregnancy was terminated at 21 weeks' gestation and the fetus was examined by autopsy. The kidneys were replaced by many cysts and there was no parenchyma. However, on histological examination, there was normal nephrogenesis amid the cystic/dysplastic tissue. This suggests that normal nephrogenesis occurs before the secondary changes leading to MCDKs.

Abortion, Induced↗

Effect of hot water extract from Agaricus blazei Murill on antibody-producing cells in mice.

We investigated the immunopotentiating activities of boiled water-soluble extracts from desiccated Agaricus blazei Murill (ABM). Effect of ABM extract on antibody production was investigated by method of hemolytic plaque-forming cells (PFC) against sheep red blood cells (SRBC) antigen. ABM extracts significantly (p<0.01) increased the number of PFC in spleen with intraperitoneal administration at doses of 25 mg/kg as compared with control group. The populations of Mac-1- or CD25-positive cells significantly (p<0.01, p<0.001) increased, but in CD19-positive cells, there were no differences in ABM-treated mice as compared with control mice. The expressions of IL-6 and IL-1beta mRNA were augmented by ABM extract in both peritoneal macrophages and spleen cells. These results suggested that ABM extract might be an effective stimulator for T cell and macrophage to IL-1beta and IL-6 release, resulting in augmentation of antibody production against SRBC antigen.

Agaricus↗

Effect of asymmetric modification on the conformation of ascidiacyclamide analogs.

Ascidiacyclamide (ASC), cyclo(-Ile1-Oxz2-d-Val3-Thz4-)2 (Oxz=oxazoline and Thz=thiazole) has a C2-symmetric sequence, and the relationships between its conformation and symmetry have been studied. In a previous study, we performed asymmetric modifications in which an Ile residue was replaced by Gly, Leu or Phe to disturb the symmetry [Doi et al. (1999) Biopolymers49, 459-469]. In this study, the modifications were extended. The Ile1 residue was replaced by Gly, Ala, aminoisobutyric acid (Aib), Val, Leu, Phe or d-Ile, and the d-Val3 residue was replaced by Val. The structures of these analogs were analyzed by X-ray diffraction, 1H NMR and CD techniques. X-Ray diffraction analyses revealed that the [Ala1], [Aib1] and [Phe1]ASC analogs are folded, whereas [Val1]ASC has a square form. These structures are the first examples of folded structures for ASC analogs in the crystal state and are similar to the previously reported structures of [Gly1] and [Phe1]ASC in solution. The resonances of amide NH and Thz CH protons linearly shift with temperature changes; in particular, those of [Aib1], [d-Ile1] and [Val3]ASCs exhibited a large temperature dependence. DMSO titration caused nonlinear shifts of proton resonances for all analogs and largely affected [d-Ile1] and [Val3]ASCs. A similar tendency was observed upon the addition of acetone to peptide solutions. Regarding peptide concentration changes, amide NH and Thz CH protons of [Gly1]ASC showed a relatively large dependence. CD spectra of these analogs indicated approximately two patterns in MeCN solution, which were related to the crystal structures. However, all spectra showed a similar positive Cotton effect in TFE solution, except that of [Val3]ASC. In the cytotoxicity test using P388 cells, [Val1]ASC exhibited the strongest activity, whereas the epimers of ASC ([d-Ile1] and [Val3]ASCs), showed fairly moderate activities.

Amino Acid Substitution↗

Detection of antivimentin antibody in sera of patients with idiopathic pulmonary fibrosis and non-specific interstitial pneumonia.

It has been suggested that the humoral immune system plays a role in the pathogenesis of non-specific interstitial pneumonia (NSIP). Although some circulating autoantibodies to cytoskeletal protein(s) have been suggested, the antimyofibroblast antibody has not been investigated in patients with idiopathic pulmonary fibrosis (IPF) and NSIP. The purpose of this study is to evaluate the existence of antimyofibroblast antibody in the sera of patients with IPF and NSIP. The MRC5 cell line was used as a model of myofibroblast. The anti-MRC5 cell antibody was characterized in a patient with NSIP using Western blotting. Since we found that one of the anti-MRC5 antibodies was an antivimentin antibody, we established an enzyme-linked immunosorbent assay (ELISA) to measure the levels of antivimentin antibody in the sera of patients with IPF (n = 12) and NSIP (n = 23). Initially, two anti-MRC5 cell antibodies were detected in the sera of patients with NSIP, one of which was characterized as the antivimentin antibody by Western blotting. The other was characterized as an antivimentin fragment antibody. We established an ELISA to measure the antivimentin antibody and found significantly higher levels in patients with IPF and NSIP than in normal volunteers. One of the anti-MRC5 cell antibodies in the serum of a patient with NSIP was against vimentin. The serum levels of antivimentin antibody were increased in patients with IPF and NSIP compared with that of normal volunteers. These results suggest that the antivimentin antibody may be involved in the process of lung injury in IPF and NSIP.

Autoantibodies↗

Relationship between the flow of bolus and occlusal condition during mastication--computer simulation based on the measurement of characteristics of the bolus.

The purpose of the present study was to clarify the relationship between the flow of a bolus and occlusal condition during mastication. First, the characteristics of a bolus under mastication was measured in subjects having different occlusal conditions. Secondly, the flow of a bolus between the upper and lower first molars under mastication was simulated using finite element non-linear dynamic analysis. Measurement of the elasticity of the bolus clarified the phenomenon of its communition. The measurement of the viscosity of the bolus clarified the phenomenon of its mixing with saliva. In addition, a relationship between the elasticity and the viscosity of the bolus at the point of just before swallowing was investigated. The flow of the bolus under mastication was revealed to vary according to the occlusal condition. These results suggest a close relationship between the occlusal condition, the flow of the bolus and its characteristics.

Adult↗

Analysis of gene expression during maturation of immature dendritic cells derived from peripheral blood monocytes.

Dendritic cells (DCs) are the most important antigen-presenting cells. Many recent studies have compared the function of immature DCs (iDCs) and mature DCs (mDCs), but there have been few reports of the molecular changes that occur in DCs during maturation. Here, we report on differential gene expression in iDCs generated from peripheral blood monocytes compared with mDCs. Gene expression was evaluated using the differential display method after activation of iDCs with a low concentration of lipopolysaccharide (LPS) to induce maturation. Proteasome subunit alpha type 3 (PSMA3), transcription factor EC (TFEC) isoform and BTK region clone 2f10-rpi were transiently upregulated. Tryptophanyl-tRNA synthetase and CD63 antigen were upregulated for at least 24 h. Neuronal apoptosis inhibitory protein (NAIP) and transforming growth factor-beta-induced 68 kDa protein were downregulated. This is the first report of NAIP expression in human DCs. By comparing the expression of NAIP with that of other members of the inhibitor of apoptosis protein (IAP) family and the Bcl-2 family, only NAIP was found to be strongly expressed in iDCs before stimulation by LPS. PSMA3 was also induced in the DCs stimulated with immune complex. These findings might contribute to our understanding of DC maturation and the effectiveness of DC-based vaccines.

Apoptosis↗

Nosocomial contamination by Mycobacterium gordonae in hospital water supply and super-oxidized water.

We experienced contamination by Mycobacterium gordonae of the hospital water of our surgical ward. The contamination was discovered following detection of the organism in operative lung samples, washed with super-oxidized water. Repeated examination of water demonstrated contamination by M. gordonae occurred only in the surgical ward, related to the apparatus for making super-oxidized water. No patients were infected by M. gordonae. After changing the water supply equipment and cleaning the water tubes, M. gordonae in the water disappeared.

Cross Infection↗

Characteristics of mastication in the anodontic mouse.

Teeth and periodontal mechanoreceptors play important roles in regulating jaw movements during mastication. However, little is known concerning how jaw movements develop without tooth eruption. To answer this question, we studied masticatory behavior in the osteopetrotic mouse, where tooth eruption does not occur and periodontal mechanoreceptors are missing. A masticatory sequence of the osteopetrotic mouse was divided into two stages: incision and chewing. Incision is characterized by small amplitude and rapid (7 Hz) open-close jaw movements, while slow (5 Hz) and large amplitude open-close jaw movements characterize chewing. The frequency and properties of jaw movements were comparable with those in the normal mouse, though the osteopetrotic mouse had a higher cycle number during incision than did the normal mouse. These results indicate that conversion from sucking to mastication occurs in the anodontic mouse, and the central pattern generator producing the masticatory rhythm develops almost normally without tooth eruption.

Animals↗

The multiple endocrine neoplasia type 1 gene product, menin, inhibits the human prolactin promoter activity.

Menin is a protein encoded by the gene mutated in multiple endocrine neoplasia type 1 (MEN1) characterized by multiple endocrine tumors of the parathyroid glands, pancreatic islets and the anterior pituitary, especially prolactinoma. In this study, we examined the effects of menin on human prolactin (hPRL) expression. In rat pituitary GH3 cells stably expressing menin, both PRL gene expression/secretion and thymidine incorporation into DNA were inhibited as compared with mock-transfected cells. The transcriptional activity of PRL promoter in GH3 cells co-transfected with menin was significantly decreased. A deletion mutation (569 delC), which we identified in a Japanese MEN1 family, was introduced into menin. When GH3 cells were transfected with a mutant menin expression vector, inhibition of hPRL promoter activity was partially reversed. These observations suggest that menin inhibits hPRL promoter activity and cell proliferation, raising the possibility that menin might play an important role in the tumorigenesis of prolactinoma.

Animals↗