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T Sakai

Publications and source records attributed to T Sakai.

At least 91 records · Page 5Linked to original sources
PubMed2001

Genetic characterization and geographic distribution of rabies virus isolates in Brazil: identification of two reservoirs, dogs and vampire bats.

We analyzed 50 rabies virus samples isolated in Brazil from 12 dogs, 11 cats, 5 vampire bats, 15 cattle, 2 horses, 1 pig, 1 sheep, and 3 humans to investigate the molecular epidemiology of rabies viruses. We sequenced 203 nucleotides on the nucleoprotein gene by direct sequencing of the PCR-amplified products. All the isolates belonged to the genotype 1 and homology of the 203 nucleotides was at least 83.7% among isolates. The main reservoirs were estimated based on the homology of nucleotide sequences. Brazilian rabies virus isolates were clustered into two reservoir groups: dogs and vampire bats. All the dog-related rabies virus isolates showed nucleotide homology greater than 99.0%. Vampire bat-related rabies virus isolates showed nucleotide homology greater than 96.6% and could be further divided into subgroups corresponding to areas where viruses were isolated. These data suggest that circulating rabies variants belong to at least two different genotype clusters in Brazil and that these two clusters are maintained independently among vampire bats and dogs.

Amino Acid Sequence↗
PubMed2001

Arabidopsis nph1 and npl1: blue light receptors that mediate both phototropism and chloroplast relocation.

UV-A/blue light acts to regulate a number of physiological processes in higher plants. These include light-driven chloroplast movement and phototropism. The NPH1 gene of Arabidopsis encodes an autophosphorylating protein kinase that functions as a photoreceptor for phototropism in response to low-intensity blue light. However, nph1 mutants have been reported to exhibit normal phototropic curvature under high-intensity blue light, indicating the presence of an additional phototropic receptor. A likely candidate is the nph1 homologue, npl1, which has recently been shown to mediate the avoidance response of chloroplasts to high-intensity blue light in Arabidopsis. Here we demonstrate that npl1, like nph1, noncovalently binds the chromophore flavin mononucleotide (FMN) within two specialized PAS domains, termed LOV domains. Furthermore, when expressed in insect cells, npl1, like nph1, undergoes light-dependent autophosphorylation, indicating that npl1 also functions as a light receptor kinase. Consistent with this conclusion, we show that a nph1 npl1 double mutant exhibits an impaired phototropic response under both low- and high-intensity blue light. Hence, npl1 functions as a second phototropic receptor under high fluence rate conditions and is, in part, functionally redundant to nph1. We also demonstrate that both chloroplast accumulation in response to low-intensity light and chloroplast avoidance movement in response to high-intensity light are lacking in the nph1 npl1 double mutant. Our findings therefore indicate that nph1 and npl1 show partially overlapping functions in two different responses, phototropism and chloroplast relocation, in a fluence rate-dependent manner.

Arabidopsis↗
PubMed2001

Impaired production of naive T lymphocytes in human T-cell leukemia virus type I-infected individuals: its implications in the immunodeficient state.

Opportunistic infections frequently occur in patients with adult T-cell leukemia (ATL) and human T-cell leukemia virus type I (HTLV-I) carriers. However, the underlying mechanisms of such infections remain unknown. To clarify the mechanism of immunodeficiency in those infected with HTLV-I, this study analyzed the T-cell subsets in HTLV-I carriers and patients with HTLV-I-associated myelopathy/tropical spastic paraparesis and ATL using 3-color fluorescence with CD62L and CD45RA coexpression either with CD4(+) or CD8(+) T cells. The number of naive T lymphocytes was markedly suppressed in patients with ATL, particularly in those with acute form, compared with uninfected control individuals. The number of naive T cells was low in HTLV-I-infected individuals under 50 years old compared with uninfected individuals, whereas the number of memory T lymphocytes was greater in HTLV-I-infected individuals. Although the increase of memory T lymphocytes correlated with HTLV-I provirus loads, no relationship was found between naive T-cell counts and provirus loads. T-cell receptor rearrangement excision circles (TRECs), which are generated by DNA recombination during early T lymphopoiesis, were quantified to evaluate thymic function in HTLV-I-infected individuals. TREC levels were lower in HTLV-I-infected individuals than in uninfected individuals. In HTLV-I carriers less than 70 years old, an increase of Epstein-Barr virus DNA in peripheral blood mononuclear cells was observed in 6 of 16 (38%) examined, whereas it was detectable in only 1 of 11 uninfected controls. These results suggested that the low number of naive T lymphocytes was due to suppressed production of T lymphocytes in the thymus, which might account for immunodeficiency observed in HTLV-I-infected individuals.

CD4-Positive T-Lymphocytes↗
PubMed2001

Promoter analysis for daily expression of Drosophila timeless gene.

Drosophila circadian clock gene timeless (tim) displays circadian oscillation in its mRNA level, and such oscillation is transcriptionally regulated. The promoter region up to -756 of tim is suggested to promote the circadian mRNA expression, however, the role of the sequence upstream of tim promoter region in the transcriptional regulation is still unrevealed. We novelly isolated and determined tim 5'-flanking sequence -2764 to -757, and found a putative cAMP-response element, six regions of the half site for PAR-basic leucine zipper transcription factors and six nonpalindromic E-boxes. Our in vivo reporter assay showed that 966 bp of tim promoter region, including a palindromic CACGTG E-box and a half site of PAR-basic leucine zipper transcription factors, was minimally required for its daily mRNA expression. While, the deletion of the 5'-flanking region -1970 to -967 caused a slight decrease in the reporter mRNA levels. These results indicate that the 5'-flanking sequence upstream of the promoter region have a role in the daily regulation of tim mRNA expression in Drosophila.

Animals↗
PubMed2001

Splenic cathepsin L is maturated from the proform by interferon-gamma after immunization with exogenous antigens.

The processing of foreign protein antigens into peptides requires the participation of various endo/lysosomal proteases in antigen-presenting cells (APCs). In this study, a proenzyme of cathepsin L, procathepsin L, was found to be present in the spleens of naive mice, as demonstrated by immunoblotting. Interestingly, the maturation of cathepsin L from procathepsin L was strongly induced when the host BALB/c mice were immunized with ovalbumin or soluble leishmanial antigen, despite the fact that mouse albumin, a kind of self-antigen, did not have such a potential. Furthermore, foreign antigens, but not self-antigens, could increase the activity of cathepsin L, probably being mediated by interferon-gamma, as demonstrated by in vivo and in vitro experiments. As cathepsin L matured, the efficiency of antigen processing was increased in APCs. These results suggest that endo/lysosomal cathepsin L plays an important role in the immune regulation via antigen processing even in peripheral lymphoid tissues as well as in the thymus.

Animals↗
PubMed2001

Missense mutation of the interleukin-12 receptor beta1 chain-encoding gene is associated with impaired immunity against Mycobacterium avium complex infection.

Interleukin-12 (IL-12) plays an important role in the production of interferon gamma (IFN-gamma) and is essential for protection against intracellular pathogens such as Mycobacterium and Salmonella. A 31-year-old man had disseminated Mycobacterium avium complex (MAC) infection. The production of IFN-gamma by peripheral blood mononuclear cells stimulated with phytohemagglutinin (PHA-PBMCs) was found severely impaired (40.7 pg/mL compared with 833 +/- 289 pg/mL for the patient's and healthy subjects' (n = 3) PHA- PBMCs, respectively), and the patient's PHA-PBMCs completely lacked surface IL-12 receptor beta1 (IL-12Rbeta1) chain. The IL-12Rbeta1 gene transcript in his PHA-PBMCs had an R213W substitution in each allele. Family history showed that both parents were heterozygotes in the R213W substitution. Transfection of a human embryonal kidney cell line 293 (HEKC293) with wild-type IL-12Rbeta1wt gene led to cell surface IL-12Rbeta1 expression; however, no expression was seen in HEKC293 transfected with the mutated IL-12Rbeta1R213W gene. The IL-12Rbeta1 gene transcript, but no IL-12Rbeta1 protein, was detected in PHA-PBMCs and T cells, suggesting a post-translational event(s), most likely a shortened turnover of the protein. The R213W substitution was not detected in the cells of 32 healthy persons or of 25 patients with tuberculosis or MAC infection. Six amino acid substitutions (Q214R, M365T, G378R, H438Y, A525T, and G594E) were identified, but the incidences of such substitutions were not significantly different between the groups. The Q214R substitution is reportedly linked to IL-12Rbeta1 deficiency; however, the study showed that 19 and 10 of 57 Japanese and 6 and 4 of 33 healthy white persons were heterozygous and homozygous for Arg-214, respectively, suggesting that the Q214R substitution represents a polymorphism and is not related to IL-12Rbeta1 deficiency but that the R213W substitution is responsible for IL-12Rbeta1 deficiency.

Adult↗
PubMed2001

Role of the cytoplasmic tyrosines of beta 1A integrins in transformation by v-src.

GD25 cells lacking beta 1 integrins or expressing beta 1A with mutations of conserved cytoplasmic tyrosines (Y783, Y795) to phenylalanine have poor directed migration to platelet-derived growth factor or lysophosphatidic acid when compared with GD25 cells expressing wild-type beta 1A. We studied the effects of v-src on these cells. Transformation with v-src caused tyrosine and serine phosphorylation of wild-type beta1 A but not of Y783/795F doubly mutated beta 1A. v-src-transformed cells had rounded and/or fusiform morphology and poor assembly of fibronectin matrix. Adhesion to fibronectin or laminin and coupling of focal contacts to actin-containing cytoskeleton were preserved in transformed Y783/795F cells but lost on transformation when beta 1A was wild type. Transformed Y783/795F cells also retained ability, albeit limited, to migrate across filters, whereas transformed cells with wild-type beta 1A were unable to transverse filters. Studies of single tyrosine mutants showed that the more important tyrosine for retaining ability to adhere, assemble focal contacts, and migrate is Y783. These results suggest that overactive phosphorylation of cytoplasmic residues of beta 1A, particularly Y783, accounts in part for the phenotype of v-src-transformed cells.

Actins↗
PubMed2001

Arabidopsis NPL1: a phototropin homolog controlling the chloroplast high-light avoidance response.

Chloroplasts relocate their positions in a cell in response to the intensity of incident light, moving to the side wall of the cell to avoid strong light, but gathering at the front face under weak light to maximize light interception. Here, Arabidopsis thaliana mutants defective in the avoidance response were isolated, and the mutated gene was identified as NPL1 (NPH-like 1), a homolog of NPH1 (nonphototropic hypocotyl 1), a blue light receptor used in phototropism. Hence, NPL1 is likely a blue light receptor regulating the avoidance response under strong light.

Alleles↗
PubMed2001

Local lattice distortion in high-Tc cuprates.

The dynamical local lattice distortion in the CuO2 plane of the cuprate high-temperature superconductors is investigated with the numerical diagonalization study for finite clusters of t-J model. It is found that the in-plane vibrations of the oxygens play an important role in the formation of some static orders like CDW and SDW through the polaronic self localization effect. The 1/8 problem is also discussed based on the present result.

Journal Article↗
PubMed2001

Risk factors for distant recurrence of hepatocellular carcinoma in the liver after complete coagulation by microwave or radiofrequency ablation.

BACKGROUND: In patients with hepatocellular carcinoma (HCC), recurrences in the distant liver often are observed after curative treatment. Microwave coagulation therapy (MCT) and radiofrequency ablation (RFA) have been developed as less invasive alternatives than surgical resection for small HCCs. In the current study, risk factors for distant recurrence of HCC were analyzed in patients in whom complete coagulation was achieved. METHODS: Ninety-two patients with HCCs < 3 cm in greatest dimension were treated by MCT or RFA percutaneously or laparoscopically. Eighty-four patients in whom complete coagulation was achieved without recurrence in the same subsegment as the primary nodule were included in this study. Distant recurrences were observed in 22 patients. Fifteen possible risk factors for a distant recurrence were analyzed. RESULTS: When comparing the patients with a recurrence of HCC nodules in the remnant liver to those without recurrence, the authors observed a statistically significant difference only in serum alpha-fetoprotein. The distant recurrence-free survival was analyzed by the Kaplan-Meier method. A statistically significant difference was observed in hepatitis C virus (HCV) infection as an etiopathic agent of underlying liver diseases (P < 0.005) and in the number of the primary HCC nodules (P < 0.05, log-rank test). A multivariate stepwise Cox hazard model revealed that HCV infection and the number of primary HCC nodules were statistically independent risk factors. CONCLUSIONS: Patients who had more than two HCC nodules accompanied by HCV infection had a high incidence of recurrence of HCC in the remnant liver, even when coagulation by microwave or ablation by radiofrequency was complete.

Aged↗
PubMed2001

Involvement of the Oct-1 regulatory element of the gadd45 promoter in the p53-independent response to ultraviolet irradiation.

The gadd45 gene, a growth arrest and DNA damage (gadd)-induced gene, is transcriptionally activated by UV irradiation through two distinct pathways. One requires the sequence-specific binding of the p53 tumor suppressor protein to a responsive element within the third intron of the gadd45 gene, and the other is p53-independent activation of the gadd45 promoter region, although the UV-response element that mediates this has yet to be defined. To investigate the sequences involved in induction of gadd45 by UV irradiation in a p53-independent pathway, we performed mutation analyses of the human gadd45 promoter fused to the luciferase reporter gene in cell lines in which p53 was inactivated. We found that the UV-responsive element was involved in the Oct-1 binding site at -99 bp relative to the transcription start site. Electrophoretic mobility shift assays showed that Oct-1, a transcription factor, bound this element on the gadd45 gene, although the intensity and mobility pattern of the retarded bands were not altered by UV irradiation. These results suggest that the Oct-1 regulatory element might be one of the essential elements involved in the activation of the gadd45 promoter by UV irradiation in a p53-independent pathway.

Cell Division↗
PubMed2001

Molecular cloning and functional expression of bottle-nosed dolphin (Tursiops truncatus) interleukin-1 receptor antagonist.

The bottle-nosed dolphin (Tursiops truncatus) interleukin-1 receptor antagonist IL-1ra cDNA was cloned from mitogen-stimulated peripheral blood mononuclear cells (PBMC) RNA utilizing the reverse transcription-polymerase chain reaction (RT-PCR). The sequence of this cDNA showed that dolphin IL-1ra clones contained open reading frames encoding 177 amino acids. Comparison of the deduced amino acids showed that dolphin IL-1ra sequence shared 87.6, 77.9, 77.4, 77.4, 76.4, and 75.8% similarity with the bovine, rabbit, equine, human, mouse, and rat IL-1ra sequences, respectively. Recombinant glutathione S-transferase (GST) dolphin IL-1ra produced in Escherichia coli (E. coli) was purified. This protein suppressed the cytostatic activity of dolphin IL-1beta on A375S2 cells, indicating that the dolphin IL-1ra cDNA obtained in the present study encodes biologically active dolphin IL-1ra.

Amino Acid Sequence↗
PubMed2001

Molecular cloning and identification of bottle-nosed dolphin p40(phox), p47(phox) and p67(phox).

The bottle-nosed dolphin NADPH oxidase cytosolic components, p40(phox), p47(phox) and p67(phox) cDNA's were cloned from mitogen stimulated peripheral white blood cell mRNA utilizing the reverse transcription-polymerase chain reaction. The sequences of these cDNAs showed that dolphin p40(phox), p47(phox) and p67(phox) clones contained open reading frames encoding predicted polypeptides of 339, 391 and 526 amino acids, respectively. Analysis of the p47(phox) and p67(phox) amino acid sequences showed two potential Src homology three domains and p40(phox) one. Comparison of the deduced amino acids showed that dolphin p40(phox) sequence shared 88.8% similarity with the human p40(phox), that dolphin p47(phox) sequence shared 87.7% similarity with the bovine p47(phox), and that dolphin p67(phox) shared 88.1% similarity with the bovine p67(phox). Western blot analysis using anti-human p40(phox), p47(phox) and p67(phox) antibodies demonstrated that dolphin neutrophil possesses p40(phox), p47(phox) and p67(phox) with similar molecular masses and structures, to each counterpart in human neutrophils, except for the p67(phox) COOH-terminus. These results suggest that dolphin NADPH oxidase cytosolic components have functional activities equivalent to those of human.

Amino Acid Sequence↗
PubMed2001

Highly sensitive immunohistological study for detection of estrogen receptor in human breast cancer.

BACKGROUND: Immunohistological methods for the detection of estrogen receptor in human breast cancer using formalin-fixed, paraffin-embedded tissue not only have the advantages of being cheaper and requiring less tissue but also have several theoretical advantages. For example, such methods enable direct histological visualization of the assessed tissue and thereby reduce sampling error. Traditional immunohistological methods (traditional LSAB), however, show false-negative reactions more often than the enzyme-binding immunoassay method (EIA). METHODS: After determining 25 estrogen receptor-positive cases and 50 estrogen receptor-negative cases by EIA, we analyzed these same cases using a traditional LSAB method and a new highly sensitive immunohistological method for assaying formalin-fixed, paraffin-embedded tissue. RESULTS: Using this new method, we detected 10 instances of positive estrogen receptors among the 50 EIA receptor-negative cases. The 25 EIA-positive cases were positive for estrogen receptor antigen except 1case using the new method. CONCLUSIONS: This new method not only has the advantage of use in paraffin-embedded tissue but also is more sensitive than the EIA. We recommend this new method as a choice to decrease false-negatives.

Adult↗
PubMed2001

Efficacy of early piston-type high-frequency oscillatory ventilation in infants with respiratory distress syndrome.

We investigated whether the combination of surfactant replacement therapy and early application of high-frequency oscillatory ventilation (HFOV) was more effective in patients with respiratory distress syndrome (RDS) than late application of HFOV and conventional mechanical ventilation (CMV). To determine this, we retrospectively reviewed the cases of 126 neonates with RDS who received surfactant replacement therapy within 4 hr after birth. Patients were grouped into those who received HFOV immediately after birth (HFOV group), those who initially were ventilated by CMV and subsequently received HFOV (CMV/HFOV group), and those who did not receive HFOV (CMV group). Changes in respiratory system compliance (Crs), arterial-alveolar oxygen gradient (a/ApO(2)), and mean airway pressure (MAP) were compared. Infants who received HFOV were less mature than those who received CMV. The a/ApO(2) measured immediately after birth before surfactant replacement therapy was significantly lower in the HFOV and CMV/HFOV group than in the CMV group. After 72 hr, the Crs in the HFOV group was higher than in any other group and was significantly higher than the CMV/HFOV group at 48 and 120 hr. These results suggest that initiating HFOV in combination with surfactant replacement therapy immediately after birth provides effective ventilatory support for infants with RDS.

Equipment Design↗
PubMed2001

Cockroach midgut peptides that regulate cell proliferation, differentiation, and death in vitro.

The number of insect midgut cells is maintained homeostatically in vivo and in vitro. However, during starvation, the midgut shrinks and the rate of cell replacement appears to be suppressed. When they undergo metamorphosis, the internal organs of insects are drastically remodeled by cell proliferation, differentiation, and apoptotic processes, and the net number of cells usually increases. An extract of 1650 midguts of Periplaneta americana was fractionated by high-performance liquid chromatography (HPLC) to obtain the peptides that regulate these processes. The HPLC fractions were tested for myotropic activity in the foregut and for effects on cell proliferation or loss in primary cultures of larval Heliothis virescens midgut cells and in a cell line derived from the last-instar larval fat body of Mamestra brassicae. Some fractions stimulated midgut stem cell proliferation and differentiation, while others caused loss of differentiated columnar and goblet cells. Other fractions stimulated cell proliferation in the larval fat body cells.

Animals↗
PubMed2001

Premedication modifies the quality of sedation with propofol during regional anesthesia.

PURPOSE: To determine the effects of diazepam or clonidine on the quality of sedation with propofol during regional anesthesia. METHODS: In a prospective randomised, controlled, double-blinded study, 60 patients undergoing elective gynecological surgery were studied. They were given premedication with 0.15-mg clonidine (Group-CL, n=20), 5-mg diazepam (Group-DZ, n = 20), or placebo (Group-P, n = 20) po. After spinal anesthesia was established, sedation was provided with propofol and controlled using a five-point sedation score at 3, "eyes closed but rousable to command", and 4, "eyes closed but rousable to mild physical stimulation". During sedation, blinded anesthesiologist recorded occurrence of complications. At two hours after end of sedation, patients were asked if they had intraoperative dream and memory. RESULTS: The loading dose, steady-state infusion rate, and overall mean infusion rate in Group-CL were 0.80 mg x kg(-1), 2.35 mg x kg(-1) x hr(-1) and 2.89 mg x kg(-1) x hr(-1), compared with 0.97 mg x kg(-1), 3.13 mg x kg(-1) x hr(-1) and 3.59 mg x kg(-1) x hr(-1) in Group-DZ, and 1.38 mg x kg(-1), 4.10 mg x kg(-1) x hr(-1) and 4.36 mg x kg(-1) x hr(-1) in Group-P, respectively. Indices of both Group-CL (P < 0.001) and Group-DZ (P < 0.05) were smaller than those of Group-P Moreover, clonidine reduced the incidence of uncontrolled movement (P < 0.01), while diazepam reduced the incidence of intraoperative memory and increased the incidence of dream (P < 0.05). Premedication did not affect the incidence of other complications. CONCLUSION: Both premedicants reduced propofol requirements and exerted beneficial effects on the incidence of some complications during sedation with propofol as an adjunct to regional anesthesia.

Adult↗