Biomedical subjects

Y Saijo

Publications and source records attributed to Y Saijo.

At least 55 records · Page 3Linked to original sources

PubMed1998

[Gamma-knife radiosurgery for metastatic brain tumors from primary lung cancer].

Forty patients with metastatic brain tumors from primary lung cancer underwent radiosurgery (gamma-knife). We retrospectively compared their prior treatment history number of metastatic foci, and performance status, to evaluate the effects of, and indications for, gamma-knife therapy. After both the primary and the metastatic tumors were controlled, performance status could be used as an index in the choice of gamma-knife therapy. Our results demonstrate that repeated gamma-knife radiosurgeries prolonged survival time. Gamma-knife radiosurgery improves quality of life and prognosis of patients with metastatic brain tumors.

Adult↗

PubMed1997

cDNA cloning and prokaryotic expression of maize calcium-dependent protein kinases.

Using degenerate oligonucleotide primers corresponding to conserved regions of the calcium-dependent protein kinase (CDPK) family, we carried out a polymerase chain reaction and obtained four distinct partial-length cDNAs from a maize leaf library. We then used these clones as probes for conventional screening and isolated 19 longer clones from another cDNA library of maize seedlings. These clones were classified into four groups based on their DNA cross-hybridization. Two full-length cDNAs, designated as ZmCDPK9 and ZmCDPK7, were sequenced and characterized. The predicted protein of each clone was a typical CDPK with eleven canonical subdomains of protein kinases, and four EF-hand calcium-binding motifs in its N-terminal and C-terminal halves, respectively. The catalytic and regulatory domains were linked by a well-conserved junction domain. The N-terminus of the protein also contained a consensus sequence for an N-myristoylation signal. Northern blot analysis showed that the transcription level of each gene was higher in roots and etiolated leaves than in green leaves. To confirm the calcium dependency of the maize enzymes, the entire coding region of ZmCDPK9 was subcloned into an expression vector so that it was in frame with the vector-encoded peptide tags. A cell-free extract of Escherichia coli transformed with the recombinant plasmid exhibited calcium-dependent phosphorylation activity, using casein as a substrate.

Amino Acid Sequence↗

PubMed1997

Characterization of renal angiomyolipoma by scanning acoustic microscopy.

A scanning acoustic microscope system was used to differentiate renal angiomyolipoma from renal cell carcinoma. The ultrasonic frequency used ranged from 100 to 200 MHz, and the attenuation constant and sound speed were measured on a two-dimensional distribution. The sound speed was significantly lower for lipoma cells than for vessels, smooth muscle fibres, clear cell renal cancer or granular cell renal cancer. The attenuation constant was significantly lower for lipoma cells than for vessels or clear cells. Both acoustic parameters for smooth muscle fibres were significantly lower than for vessels. The heterogeneity of the microacoustic field in renal angiomyolipoma is closely related to the high intensity echo observed on clinical echography. Renal angiomyolipoma and renal cell carcinoma can thus be distinguished by acoustic examination.

Angiomyolipoma↗

PubMed1997

Production, crystallization, and preliminary X-ray analysis of rabbit skeletal muscle troponin complex consisting of troponin C and fragment (1-47) of troponin I.

Troponin is a ternary protein complex consisting of subunits TnC. TnI, and TnT, and plays a key role in calcium regulation of the skeletal and cardiac muscle contraction. In the present study, a partial complex (CI47) was prepared from Escherichia coli-expressed rabbit skeletal muscle TnC and fragment 1-47 of TnI, which is obtained by chemical cleavage of an E. coli-expressed mutant of rabbit skeletal muscle TnI. Within the ternary troponin complex, CI47 is thought to form a core that is resistant to proteolytic digestion, and the interaction within CI47 likely maintains the integrity of the troponin complex. Complex CI47 was crystallized in the presence of sodium citrate. The addition of trehalose improved the diffraction pattern of the crystals substantially. The crystal lattice belongs to the space group P3(1)(2)21, with unit cell dimensions a = b = 48.2 A, c = 162 A. The asymmetric unit presumably contains one CI47 complex. Soaking with p-chloromercuribenzenesulfonate (PCMBS) resulted in loss of isomorphism, but enhanced the quality of the crystals. The crystals diffracted up to 2.3 A resolution, with completeness of 91% and R(merge) = 6.4%. The crystals of PCMBS-derivative should be suitable for X-ray studies using the multiple-wavelength anomalous diffraction technique. This is the first step for elucidating the structure of the full troponin complex.

Animals↗

PubMed1997

Ultrasonic tissue characterization of infarcted myocardium by scanning acoustic microscopy.

The purpose of this study was to ultrasonically characterize infarcted human myocardial tissue at the microscopic level by scanning acoustic microscopy. Infarcted myocardial specimens from ten cases with acute myocardial infarction were studied. Specimens were formalin fixed, paraffin embedded and sectioned to 10-micron thickness. A specially developed scanning acoustic microscope system, operating in the 100- to 200-MHz ultrasound frequency range, was used for the measurements. The values of the attenuation constant were 0.94 +/- 0.04 dB/mm/MHz in normal myocardium, 0.71 +/- 0.12 dB/mm/MHz in degenerated myocardium, 0.88 +/- 0.47 dB/mm/MHz in granulation tissue and 1.75 +/- 0.11 dB/mm/MHz in fibrosis. The values of sound speed were 1620.2 +/- 8.2 m/s in normal myocardium, 1572.4 +/- 10.6 m/s in degenerated myocardium, 1590.2 +/- 32.5 m/s in granulation tissue and 1690.3 +/- 9.1 m/s in fibrosis. The ultrasonic properties of the diseased myocardium at the microscopic level will provide important information for ultrasonic tissue characterization at the macroscopic level.

Acoustics↗

PubMed1997

Acoustic properties of dialysed kidney by scanning acoustic microscopy.

BACKGROUND: A correlation between acquired renal cysts in the dialysed kidney and renal cancer has long been debated, but no changes in the physical properties of kidneys at the microscopic level have been reported. The purpose of the present study was to classify the physical properties of the kidneys of patients undergoing haemodialysis at several stages of pathology by use of the scanning acoustic microscope. METHODS: Sixteen surgically excised kidneys of dialysis patients were investigated. Tissues were fixed in 10% formalin, frozen in acetone, and cut 10 microns thick on a cryostat. We used a scanning acoustic microscope operated in the frequency range of 100-200 MHz. Attenuation constant and sound speed were measured on a two-dimensional distribution. RESULTS: The attenuation constant for inflammatory granulation tissue was significantly higher than that for hyaline degeneration tissue (P < 0.001). Sound speed was high for granulation tissue, but tended to diminish gradually for hyaline degeneration. Sound speed increased again with progression to cystic degeneration (P < 0.001), but the attenuation constant remained low. When a cystic kidney contained a malignant lesion, the previously low attenuation constant rose at that site (P < 0.001), and the previously high sound speed was diminished (P < 0.001). CONCLUSION: Our data suggest that the physical properties of dialysed kidneys at different stages of pathology can be classified by their acoustic properties. Simultaneous evaluation of attenuation constant and sound speed is considered applicable to determining whether tissues contain malignant elements.

Acoustics↗

PubMed1997

Contiguous four-guanosine sequence in c-myc antisense phosphorothioate oligonucleotides inhibits cell growth on human lung cancer cells: possible involvement of cell adhesion inhibition.

A contiguous four-guanosine (4G) sequence in c-myc antisense phosphorothioate oligonucleotides caused an antiproliferative effect in smooth muscle cells. To investigate the antiproliferative effect of c-myc antisense oligonucleotides on human lung cancer cell lines, we synthesized oligonucleotides of various lengths and sequences, focusing on the contiguous four-guanosine (4G) sequence. While a c-myc antisense oligonucleotide (20AS1 (4G)) targeted to the translation initiation codon of c-myc mRNA inhibited cell growth of A549 cells by 69% at 10 microM, a scrambled oligonucleotide (20SCR1 (4G)) containing the contiguous four-guanosine (4G) sequence also inhibited cell growth by 72% at the same dose. Although treatment with either 20AS1 (4G) or 20SCR1 (4G) inhibited cell adhesion by 70% at 10 microM, expression of c-myc protein was significantly suppressed only by 20AS1 (4G) (62%), and was only weakly inhibited by 20SCR1 (4G) (32%). Furthermore, a small cell lung carcinoma cell line, Lu65, which can grow in suspension form, was highly resistant to 20AS1 (4G) treatment (IC50>20 microM). These results suggest that the cell growth inhibition by c-myc antisense oligonucleotides containing the contiguous four-guanosine (4G) sequence was possibly correlated with inhibition of cell adhesion, but not with inhibition of c-myc protein expression, via a sequence-specific non-antisense mechanism.

Base Sequence↗

PubMed1997

[Pulmonary thromboembolism associated with antiphospholipid syndrome in scleroderma].

A 51-year-old woman was referred to our hospital with dyspnea. Chest roentgenogram on admission showed dilation of the pulmonary arteries and hyperlucency in the lung fields. An ultrasonic cardiographic examination showed that the right atrium and ventricle were dilated. Pulmonary thromboembolism due to left popliteal vein thrombosis was diagnosed by perfusion scintigram of the lung, which showed multiple wedge-shaped defects, and by digital subtraction angiogram, which showed a filing defect in the left popliteal vein. Antiphospholipid syndrome was diagnosed after IgG anticardiolipin antibody was defected. Scleroderma was subsequently diagnosed because the patient exhibited Raynaud's phenomenon and proximal scleroderma. Although closely associated with lupus erythematosus and other lupus variants, antiphospholipid syndrome has not been recognized as a common complication of scleroderma. This is the first report of a patient with pulmonary thromboembolism associated with antiphospholipid syndrome and scleroderma.

Antibodies, Anticardiolipin↗

PubMed1997

Expression of nucleolar protein p120 in human lung cancer: difference in histological types as a marker for proliferation.

The function of proliferation-associated nucleolar protein p120 is unclear. A recent report that a yeast protein, NOP2, 67% homologous to human p120, is up-regulated during the onset of growth and influences the morphology of the nucleolus supports the notion that this protein could serve as a marker for proliferation in neoplastic cells. Lung cancer is characteristic in that different histological types show different biological features. We attempted to evaluate the levels of p120 expression in resected human lung cancer tissues of different histological types and the relation of p120 expression and cell proliferation using human lung cancer cell lines. When 37 frozen specimens of human lung cancer and normal lung were stained with a p120 monoclonal antibody, the nucleoli of cancer cells were positively stained, whereas a few macrophages in normal lung revealed only weak staining. The labeling index of p120 in squamous cell carcinoma (67.7 +/- 12.4%) was significantly higher than that in adenocarcinoma (35.3 +/- 12.6%) or in large cell carcinoma (30.1 +/- 17.3%; P < 0.01). In six human lung cancer cell lines and one normal lung fibroblast cell line cultured in vitro, there was a significant correlation between S-phase fraction and p120 mRNA (r = 0.851, P < 0.02)/p120 protein (r = 0.869, P < 0.01) or between doubling time and p120 protein (r = -0.928, P < 0.01). In the context of the reports that indicate higher [3H]thymidine incorporation and shorter doubling time in the squamous cell carcinoma, these results indicate that p120 can be a marker for proliferation in human lung cancer cells in vivo and in vitro, and that it has an important function in the cell cycle of tumor proliferation.

Adenocarcinoma↗

PubMed1996

Influence of tissue preparation on the high-frequency acoustic properties of normal kidney tissue.

The influence of various tissue preparations on the acoustic properties of normal kidney tissue at high frequencies was investigated. Eight surgically excised normal kidney tissue specimens were classified into three groups: (i) fresh, frozen section, (ii) formalin-fixed, frozen section and (iii) formalin-fixed, paraffin section. Scanning acoustic microscopy operating in the frequency range of 100-200 MHz was used to display the two-dimensional distribution of attenuation constant and sound speed. Our results indicate that (i) there is no significant variation in both acoustic parameters between the three tissue groups, (ii) fixation by 10% formalin produces no significant change in the acoustic parameters, (iii) in fat-free tissue regions, the acoustic parameters are independent of preparation method and (iv) frozen sections must be used to assess the acoustic parameters in fat-rich tissues.

Histocytological Preparation Techniques↗

PubMed1996

cDNA cloning and tissue-specific expression of a novel basic helix-loop-helix/PAS factor (Arnt2) with close sequence similarity to the aryl hydrocarbon receptor nuclear translocator (Arnt).

We isolated mouse cDNA clones (Arnt2) that are highly similar to but distinct from the aryl hydrocarbon receptor (AhR) nuclear translocator (Arnt). The composite cDNA covered a 2,443-bp sequence consisting of a putative 2,136-bp open reading frame encoding a polypeptide of 712 amino acids. The predicted Arnt2 polypeptide carries a characteristic basic helix-loop-helix (bHLH)/PAS motif in its N-terminal region with close similarity (81% identity) to that of mouse Arnt and has an overall sequence identity of 57% with Arnt. Biochemical properties and interaction of Arnt2 with other bHLH/PAS proteins were investigated by coimmunoprecipitation assays, gel mobility shift assays, and the yeast two-hybrid system. Arnt2 interacted with AhR and mouse Sim as efficiently as Arnt, and the Arnt2-AhR complex recognized and bound specifically the xenobiotic responsive element (XRE) sequence. Expression of Arnt2 successfully rescued XRE-driven reporter gene activity in the Arnt-defective c4 mutant of Hepa-1 cells. RNA blot analysis revealed that expression of Arnt2 mRNA was restricted to the brains and kidneys of adult mice, while Arnt mRNA was expressed ubiquitously. In addition, whole-mount in situ hybridization of 9.5-day mouse embryos showed that Arnt2 mRNA was expressed in the dorsal neural tube and branchial arch 1, while Arnt transcripts were detected broadly in various tissues of mesodermal and endodermal origins. These results suggest that Arnt2 may play different roles from Arnt both in adult mice and in developing embryos. Finally, sequence comparison of the currently known bHLH/PAS proteins indicates a division into two phylogenetic groups: the Arnt group, containing Arnt, Arnt2, and Per, and the AhR group, consisting of AhR, Sim, and Hif-1alpha.

Amino Acid Sequence↗

PubMed1996

Ultrasonic tissue characterization of renal cell carcinoma tissue.

The purpose of this study was to characterize renal cell carcinoma tissue by the measurement of microacoustic properties. A scanning acoustic microscope system operating in the frequency range of 100-200 MHz was employed and the attenuation constant and sound speed were measured on the two-dimensional distribution. The values of attenuation constant and sound speed were lower in both kinds of cancer cells than those in normal kidney, although a significant difference was not found between the clear cell and granular cell. Also, both acoustic parameters of cancer cells were significantly lower than those in hemorrhage and fibrosis. These data suggest that the elasticity of renal cell carcinoma tissue may be lower than that of normal kidney. Moreover, the high intensity echo in clinical echography may be related to the heterogeneity of the microacoustic field in the carcinoma tissue.

Acoustics↗

PubMed1996

Human T-cell lymphotropic virus type I (HTLV-I) carrier with clinical manifestations characteristic of diffuse panbronchiolitis.

A 45-year-old male was referred due to prolonged productive cough. Despite the fact that bronchoalveolar lavage fluid suggested lymphocytic and neutrophilic alveolitis, the histologic diagnosis of his biopsied lung was diffuse panbronchiolitis-like lesion with infiltration of lymphocytes and plasma cells into respiratory bronchioles but few foamy cells. Serologic examination revealed that he was a carrier of human T-cell lymphotropic virus type I (HTLV-I). Together with uveitis and ulcerative colitis in his past history, the persistent respiratory symptoms of this patient can be attributed to non-neoplastic inflammation due to the chronic HTLV-I infection.

Bronchiolitis↗

PubMed1995

[Acoustic microscope and its medical applications].

There are two kinds of the acoustic microscope, the one is the Scanning Laser Acoustic Microscope, SLAM and the other the Mechanical Scanned Acoustic Microscope, SAM. In the acoustic microscope, high frequency ultrasound of 100 MHZ to 500 MHZ was used and the resolving power was in the range of 5 to 10 microns. The propagating speed and attenuation of ultrasound from the C-mode image which was obtained by mechanically scan with laser or ultrasound. From the results of the experiments by using the SAM to analyze acoustic characteristics of the tissue specimens which were taken from the patients with cardiac sarcoidosis, HCM, DCM, gastric and kidney cancer, clinical significances and usefulness of the evaluation for physical characteristics of the tissue were discussed in this paper.

Cardiomyopathies↗

PubMed1995

[Ultrasonic tissue characterization of diseased myocardium by scanning acoustic microscopy].

Ultrasonic tissue characterization of the myocardium has become very important because the requirement for evaluating local cardiac function has increased. In the present study, the acoustic properties of the tissue elements in myocardial infarction were measured by a specially developed scanning acoustic microscope (SAM) system, and the echo intensity of the infarcted myocardium was assessed. Ten samples of infarcted myocardium obtained at autopsy were formalin-fixed, paraffin-embedded and cut into 10 microns thick slices. The SAM system was operated in the frequency range of 100 to 200 MHz. The values of the attenuation constant and sound speed were measured in five tissue elements, normal myocardium, degenerated myocardium, fibrosis, granulation, and hemorrhage by comparing the optical microscopic image and acoustic images. The density of each tissue element was measured by the CuSO4 method, and the specific acoustic impedance was calculated by multiplying the sound speed and the density of each tissue element. The specific acoustic impedance was 1.75 x 10(6) Ns/m3 in normal myocardium, 1.69 x 10(6) Ns/m3 in degenerated myocardium, and 1.85 x 10(6) Ns/m3 in fibrosis. The intensity of the reflected ultrasound at the interface between two different tissue elements was calculated by the difference of specific acoustic impedance. Using the sensitivity changing method in clinical echocardiography, the intensity of myocardial scar was -15 dB compared to the pericardial echo. The corrected ideal ultrasonic reflection between degenerated myocardium and fibrosis was calculated as -15.4 dB. The results suggest that measurement of acoustic properties is very important to provide basic data for ultrasonic characterization of myocardium tissue.

Acoustics↗

PubMed1994

Identification of the nuclear and nucleolar localization signals of the protein p120. Interaction with translocation protein B23.

The human p120 nucleolar protein is a cell cycle-related protein that peaks during the S phase and has been shown to be associated with a beaded fibrillar structure. To study domains responsible for the nucleolar localization of protein p120, initially deletion mutants were made that defined sequences containing the localization signals; then, fusion genes that were composed of segments of the p120 molecule joined to the N-terminal end of the Escherichia coli beta-galactosidase were constructed. In the absence of the localization signals the beta-galactosidase remained in the cytoplasm. When the identified nuclear localization signal containing the amino acid sequence 99-110 (NAPRGKKRPAPG) was fused to the beta-galactosidase, the protein localized to the nucleus. When only the identified nucleolar localization signal containing the amino acid sequence 40-57 (SKRLSSRARKRAAKRRLG) was fused to the beta-galactosidase, the fusion protein remained in the cytoplasm. When both the nuclear and nucleolar localization signals were fused to the beta-galactosidase it localized predominantly to the nucleolus. Nucleolar protein B23, a putative "shuttle protein," bound to amino acid sequence 24-56 of protein p120. Deletion analysis showed that amino acids 187-215 of protein B23 bound to protein p120. The results suggest that protein B23 may be part of the mechanism of protein targeting to the nucleolus.

3T3 Cells↗

PubMed1994

Pharmacokinetics, tissue distribution, and stability of antisense oligodeoxynucleotide phosphorothioate ISIS 3466 in mice.

Phosphorothioate oligonucleotides have a potential as therapeutic agents. The pharmacokinetics, tissue distribution, stability, and cellular uptake by LOX ascites tumor of p120 antisense phosphorothioate oligonucleotide, ISIS 3466, were studied in vivo. The oligonucleotide, which was quickly cleared from the circulation in the normal mice after IV injection, was readily absorbed into the systemic circulation from the peritoneum. The oligonucleotide was found in most tissues 48 h after IP administration. The highest concentrations were in kidney and liver, but the brain had a very low concentration. The phosphorothioate oligonucleotide was intact even after 48 h. When the oligonucleotide was complexed with cationic lipid DOTMA, the DOTMA did not affect the oligonucleotide uptake or tissue distribution in normal mice. However, DOTMA significantly increased the oligonucleotide cellular uptake (4-10 times) in LOX ascites tumors in an IP/IP model. These results indicate that the phosphorothioate oligonucleotide is stable, has favourable kinetics for use as an therapeutic agent, and that DOTMA could be useful in local delivery of the oligonucleotide in vivo.

Animals↗

PubMed1993

Nucleolar and nuclear aberrations in human lox tumor cells following treatment with p120 antisense oligonucleotide ISIS-3466.

Previous reports from this laboratory have shown marked cytocidal effects of the ISIS-3466 antisense phosphorothioate oligodeoxynucleotide to the human nucleolar protein p120 on human cancer cell lines in vitro and inhibition of tumor growth in vivo in an i.p/i.p. LOX cell model (L. Perlaky et al. Anti-Cancer Drug Design 8:3-14, 1993). In this study, light and fluorescence microscopy showed that the number of LOX cells in mitosis decreased by 50% after incubation for 4 h in 0.2-0.4 microM antisense oligonucleotide; a 70% reduction in cell number was found from 8-72 h post-treatment. In addition, marked unravelling of nucleolar structures and chromatin fragmentation was found after a 4-h incubation. The nucleolar unravelling occurred in varying degrees ranging from partial unfolding to almost complete separation of the strands of nucleolar residues. Twenty four hours post-treatment, immunofluorescence staining with the anti-p120 monoclonal antibody showed reduced nucleolar protein p120 and translocation of the p120 protein from the nucleoli to the nucleoplasm. Analysis of the mechanisms of the nucleolar unravelling and inhibition of mitosis will provide further understanding of the cytocidal effects of the ISIS-3466 antisense oligonucleotide.

Antigens, Neoplasm↗