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Biomedical subjects

T Tani

Publications and source records attributed to T Tani.

At least 37 records · Page 2Linked to original sources

Nuclear transfer of mouse follicular epithelial cells pretreated with spermine, protamine, or putrescine.

The in vitro and in vivo developmental potential of nuclear transferred embryos receiving follicular epithelial cells pretreated with spermine (5 and 20 mM), protamine (0.25 and 25 mg/ml), or putrescine (1 and 100 microg/ml) at room and reduced temperatures was examined in the mouse. The pretreated donor cells were first fused with enucleated oocytes, and then nuclei from reconstituted eggs at the two-cell stage were fused with the enucleated fertilized two-cell embryos. The proportion of reconstituted embryos that developed into blastocysts was not significantly different among groups. After transfer to recipients, implantation rates were not different between groups and fetuses were obtained in protamine- and spermine-treated groups as well as in control groups. These results demonstrate that pretreatment of nuclear donor cells with spermine, protamine, or putrescine does not enhance the developmental potential in vitro or in vivo in the mouse. J. Exp. Zool. 289:208-212, 2001.

Animals↗

Mouse cloned from embryonic stem (ES) cells synchronized in metaphase with nocodazole.

Full-term development occurred when nuclei from mouse embryonic stem (ES) cells, synchronized in metaphase with nocodazole, were fused with enucleated oocytes or nuclei of reconstituted eggs and again fused with the enucleated blastomeres of fertilized two-cell embryos using inactivated Sendai virus. Two surviving male mice were derived from undifferentiated ES cell nuclei, one from single nuclear transfer and another from serial nuclear transfer. Both were noticeably small and died within 24 hr of birth for unknown reasons. These findings demonstrate that nuclear transfer of ES cells using the fusion method produces young, as does the piezoelectric-actuated nuclear transfer. J. Exp. Zool. 289:139-145, 2001.

Animals↗

Lipopolysaccharide: neutralization by polymyxin B shuts down the signaling pathway of nuclear factor kappaB in peripheral blood mononuclear cells, even during activation.

BACKGROUND: There have been many studies on anti-lipopolysaccharide (LPS) agents and LPS-neutralizing agents; however, there have been no reports on the changes in clinical status and mediators that occur when these agents are used. Polymyxin (PMX) (treatment using a column containing polymyxin B-immobilized fiber) removed circulating endotoxin, and reduced various cytokines within 120 min, even in patients with high levels of plasma cytokines. Our purpose was examine the mechanisms of PMX treatment by which plasma cytokines are reduced by endotoxin neutralization with polymyxin B, even during therapy for sepsis and/or endotoxin shock. METHODS: We studied the interaction between nuclear factor kappaB (NF-kappaB) binding activity and tumor necrosis factor alpha (TNF-alpha) secretion in an experimental system using LPS-stimulated human peripheral blood mononuclear cells (PBMCs), after neutralization of LPS with polymyxin B. PBMCs were incubated with LPS in vitro, and TNF-alpha secretion and NF-kappaB activation were assessed. We then studied the changes in NF-kappaB activation and TNF-alpha secretion when both polymyxin B and LPS were added simultaneously and when polymyxin B was added after 30 or 120 min of incubation with LPS. RESULTS: Immediate inhibition of NF-kappaB binding activity and suppression of TNF-alpha secretion were observed after LPS neutralization with polymyxin B regardless of whether PBMCs were already producing TNF-alpha. CONCLUSIONS: These findings may indicate one of the mechanisms operating in the clinical changes that occur after circulating endotoxin removal, and are likely to have therapeutic value, even for patients with high proinflammatory cytokine levels.

Anti-Bacterial Agents↗

Correlation between plasma endotoxin, plasma cytokines, and plasminogen activator inhibitor-1 activities in septic patients.

The objective of this study was to evaluate the relation between the clinical and plasma parameters and the changes in plasma endotoxin activity with 2 hours of endotoxin-adsorbing therapy using polymyxin B (PMX). A total of 88 consecutive patients were admitted for PMX treatment of severe sepsis or septic organ failure. Standard supportive care was continued without alteration during PMX treatment. Endotoxin, tumor necrosis factor-alpha (TNFalpha), interleukin-6 (IL-6), IL-10, and plasminogen activator inhibitor-1 (PAI-1) activities and clinical parameters were measured before, immediately after, and the day after PMX treatment. The mean APACHE II and III scores were 24.2 +/- 1.0 and 85.8 +/- 3.0, respectively. The 2-week survival rate was 51.1%. In survivors, TNFalpha, IL-6, IL-10, and PAI-1 activities were significantly decreased during the 2-hour PMX treatment, the following day, or both times. There was no significant change in the parameters, except for TNFalpha, after PMX in nonsurvivors. In the subgroup whose plasma endotoxin decreased more than 30%, IL-6, TNFalpha, and PAI-1 significantly decreased after 2 hours of PMX or the following day (or both), but all four parameters in nonsurvivors showed no significant change. Hence PMX adsorbed plasma endotoxins and contributed to reductions in plasma proinflammatory cytokine levels and to improved clinical parameters during the 2-hour treatment. Changes in these parameters correlated with changes in plasma endotoxin activity in survivors whose plasma endotoxin levels were adequately reduced.

APACHE↗

Sequential changes of orthodromic sensory nerve action potentials induced by experimental compression of the median nerve at the wrist.

OBJECTIVE: We studied sequential waveform changes associated with a progressive conduction block to elucidate the relative vulnerability of slow versus fast conducting fibers to a focal compression. METHODS: In 12 healthy men, orthodromic compound sensory nerve action potentials (SNAPs) of the median nerve were recorded unipolarly at 4 sites over the forearm during a 30 min period of constant, localized compression of the nerve at the wrist. RESULTS: Initial findings at the compression site consisted of nearly immediate reduction in size of the negative component accompanied by progressive enlargement of the initial-positive component. Recording at 2 cm proximal to the compression and at the elbow showed no change in onset latency initially, indicating at least partial preservation of the fast conducting fibers. Amplitude also remained unchanged for about 20 min, presumably because loss of negative and positive peaks compensated each other until conduction block began to involve a greater number of the fast conducting fibers. CONCLUSIONS: The analysis of waveform changes and their time course suggests that a focal compression initially affects the slow conducting small diameter fibers. Partial conduction block gives rise to complex waveform changes depending on recording sites. A reduction in one polarity of constituent nerve fiber action potentials may enhance the other polarity of the SNAP.

Action Potentials↗

Mutation in the prp12+ gene encoding a homolog of SAP130/SF3b130 causes differential inhibition of pre-mRNA splicing and arrest of cell-cycle progression in Schizosaccharomyces pombe.

prp12-1 is one of the mutants defective in pre-mRNA splicing at a nonpermissive temperature in Schizosaccharomyces pombe. We found that the prp12+ gene encodes a protein highly homologous with a human splicing factor, SAP130/SF3b130, a subunit of a U2 snRNP-associated complex SF3b. Prp12p was shown to interact genetically with Prp10p that is a homolog of SAP155/SF3b155, another subunit in SF3b, suggesting that Prp12p is a functional homolog of human SAP130/SF3b130. Prp12p tagged with GFP is uniformly localized in the nuclear DNA region. In addition to pre-mRNA splicing defects, the prp12-1 mutant produced elongated cells, a typical phenotype of cell division cycle (cdc) mutants, suggesting a possible link between pre-mRNA splicing and cell-cycle progression. We examined kinetics of splicing defects in prp12-1 and several other prp mutants using northern blot hybridization and found that, among all the tested pre-mRNAs, only Tflld pre-mRNA with low splicing efficiency showed detectable splicing defects at the nonpermissive temperature in prp12-1. In addition, we found that other prp mutants with the cdc phenotype also showed differential splicing defects in tested pre-mRNAs at the nonpermissive temperature. On the other hand, prp mutants that do not exhibit the cdc phenotype showed a rapid and complete block of pre-mRNA splicing in all the tested pre-mRNAs at the nonpermissive temperature, indicating that prp mutants with weak splicing defects have a tendency to exhibit the cdc phenotype. These results suggest that the cdc phenotype in prp12-1 is caused by a selective reduction of spliced transcripts encoding a protein (or proteins) required for G2/M transition.

Amino Acid Sequence↗

Integrin laminin receptors and breast carcinoma progression.

This review explores the mechanistic basis of breast carcinoma progression by focusing on the contribution of integrins. Integrins are essential for progression not only for their ability to mediate physical interactions with extracellular matrices but also for their ability to regulate signaling pathways that control actin dynamics and cell movement, as well as for growth and survival. Our comments center on the alpha6 integrins (alpha6beta1 and alpha6beta4), which are receptors for the laminin family of basement membrane components. Numerous studies have implicated these integrins in breast cancer progression and have provided a rationale for studying the mechanistic basis of their contribution to aggressive disease. Recent work by our group and others on mechanisms of breast carcinoma invasion and survival that are influenced by the alpha6 integrins are discussed.

Antigens, Neoplasm↗

Femtosecond near-field scanning optical microscopy study of molecular thin films.

A near-field scanning optical microscope has been combined with a two-colour time-resolved pump-probe measurement system. It has a noise-equivalent transmittance change of 5.0 x 10-5 for a probe pulse with an intensity of 30 nW. The system has been used for evaluating molecular thin films that have a domain structure, particularly for observing a gate action of the single domains. The results include key features to understand an origin of the domains and suggest that the film composition is uniform over a distance of several micrometres.

Journal Article↗

Sub-wavelength region spectroscopy and local morphology of individual mesoscopic quantum systems.

Results of reflection microscopy and local reflection spectroscopy of J-aggregates of two pseudoisocyanine dyes in a thin film polymer matrix are reported. The individual J-aggregates assemble into fibre-like shapes of large structural heterogeneity. Reflectance spectra obtained at different samples, different locations on one sample and even at different positions of the same aggregate fibre reveal a wide distribution of optical properties. The shapes and absolute reflectivities of the spectra are suggested to originate from varying strengths of exciton-photon interaction and from the effect of finite thickness of the aggregate fibres. Polarization dependence measurements of local reflectivities provide orientations of the exciton transition dipole moments at 572 nm and 540 nm with respect to the orientation of the aggregate fibres. Furthermore, modified synthesis of capped semiconductor nanocrystals of CdSe is presented. The method yields high quality quantum dots suitable for microscopic imaging and spectroscopy.

Journal Article↗

Off-line leukapheresis using leukofiltration for active ulcerative colitis: a case report.

The method of leukocytapheresis for ulcerative colitis (UC) by using extracorporeal circulation (on-line system) has been reported. To perform leukocytapheresis, we have applied leukocyte elimination filters for blood transfusion to leukocytapheresis without using extracorporeal circulation (off-line leukocytapheresis system). Four hundred milliliters of peripheral blood was collected and reinfused through a leukocyte elimination filter. This procedure was repeated 5 times, and up to 2,000 ml of peripheral blood was treated. This method has been applied once a week for 5 weeks. We applied the off-line leukapheresis system to a 31-year-old male ulcerative colitis patient. As a result, the frequency of defecation and the dose of medicine were effectively decreased, and endoscopic finding was also improved. Because of the absence of complications observed with the on-line system, the off-line leukocytapheresis system that we have applied to the clinical patient is simple, safe, and useful.

Adult↗

Poor outcome of 44 cemented total hip arthroplasties with alumina ceramic heads: clinical evaluation and retrieval analysis after 10-16 years.

We evaluated the long-term results of cemented total hip arthroplasty with alumina ceramic heads in 39 patients (44 hips) from 1981 to 1985. The study comprised 8 men (8 hips) and 31 women (36 hips). Their mean age was 54 (37-76) years and mean weight 52 (34-93) kg. The preoperative diagnoses were osteoarthrosis in 34 patients (38 hips), rheumatoid arthritis in 3 (4 hips) and idiopathic osteonecrosis of the femoral head in 2 (2 hips). The mean duration of follow-up was 13 (10-16) years. At this time, 6 cases had been revised because of aseptic loosening (5 hips showed loosening of both components and 1 acetabular loosening alone). Kaplan-Meier survivorship analysis with radiographic loosening as end-point showed 10-year survival rates of 75% for the acetabular component and 86% for the femoral. The mean linear wear rate of polyethylene was 0.10 mm/year radiographically--i.e., about the same as reported for many metal to polyethylene bearings. 5 alumina ceramic heads were retrieved and showed excellent surface roughness and roundness without scratches. However, debris stuck to the gaps was detected. The debris may act as third-bodies and affect polyethylene wear.

Adult↗

Analysis of plasticizers in cap-sealing resins for bottled foods.

A survey of plasticizers in cap-sealing resins for bottled foods has been undertaken. During 1997-1999 di-(2-ethylhexyl)phthalate (DEHP) was found in seven out of 21 samples on the Japanese domestic market and in 10 out of 61 imported samples as well as a further two samples which contained di-(2-ethylhexyl)adipate (DEHA). In the period 1993-1999, of the other plasticizers diacetyl lauroyl glycerol (DALG) was only detected in domestic samples whereas diisodecyl phthalate (DIDP) and diisononyl phthalate (DINP) were only in imported samples. It was observed overall that DEHP and DEHA were restricted to use in cap-sealing resins for bottled foods. Whilst phthalates, DEHA or DALG were detected in samples in 1993 and 1995, the investigation in 1997-1999 showed fewer samples in which these plasticizers were found.

Chromatography, Gas↗

Fission yeast Prp4p kinase regulates pre-mRNA splicing by phosphorylating a non-SR-splicing factor.

We provide evidence that Prp4p kinase activity is required for pre-mRNA splicing in vivo and show that loss of activity impairs G1-S and G2-M progression in the cell cycle. Prp4p interacts genetically with the non-SR (serine/arginine) splicing factors Prp1p and Prp5p. Bacterially produced Prp1p is phosphorylated by Prp4p in vitro. Prp4p and Prp1p also interact in the yeast two-hybrid system. In vivo labelling studies using a strain with a mutant allele of the prp4 gene in the genetic background indicate a change in phosphorylation of the Prp1p protein. These results are consistent with the notion that Prp4p kinase is involved in the control of the formation of active spliceosomes, targeting non-SR splicing factors.

Alleles↗

Fission yeast Mog1p homologue, which interacts with the small GTPase Ran, is required for mitosis-to-interphase transition and poly(A)(+) RNA metabolism.

We have cloned and characterized the Schizosaccharomyces pombe gene mog1(+), which encodes a protein with homology to the Saccharomyces cerevisiae Mog1p participating in the Ran-GTPase system. The S. pombe Mog1p is predominantly localized in the nucleus. In contrast to the S. cerevisiae MOG1 gene, the S. pombe mog1(+) gene is essential for cell viability. mog1(+) is required for the mitosis-to-interphase transition, as the mog1-1 mutant arrests at restrictive temperatures as septated, binucleated cells with highly condensed chromosomes and an aberrant nuclear envelope. FACS analysis showed that these cells do not undergo a subsequent round of DNA replication. Surprisingly, also unlike the Delta mog1 mutation in S. cerevisiae, the mog1-1 mutation causes nucleolar accumulation of poly(A)(+) RNA at the restrictive temperature in S. pombe, but the signals do not overlap with the fibrillarin-rich region of the nucleolus. Thus, we found that mog1(+) is required for the mitosis-to-interphase transition and a class of RNA metabolism. In our attempt to identify suppressors of mog1-1, we isolated the spi1(+) gene, which encodes the fission yeast homologue of Ran. We found that overexpression of Spi1p rescues the S. pombe Delta mog1 cells from death. On the basis of these results, we conclude that mog1(+) is involved in the Ran-GTPase system.

Active Transport, Cell Nucleus↗

Direct exposure of chromosomes to nonactivated ovum cytoplasm is effective for bovine somatic cell nucleus reprogramming.

We examined the in vitro developmental potential of nonactivated and activated enucleated ova receiving cumulus cells at various stages of the cell cycle. Eleven to 29% of activated ova receiving donor cells stopped developing at the 8-cell stage but 21% to 50% of nonactivated ova receiving donor cells at either the G(0), G(1), G(2), or M phase, or cycling cells developed into blastocysts. One normal calf was born after transferring five blastocysts that had developed from ova receiving donor cells at the M phase. The present study demonstrated that direct exposure of donor chromosomes to nonactivated ovum cytoplasm is effective for somatic cell nucleus reprogramming, and activated ovum cytoplasm does not reprogram the nucleus.

Animals↗

The role of bacterial translocation on neutrophil activation during hemorrhagic shock in rats.

Some biological responses to hemorrhage have been reported to be associated with bacterial translocation (BT). While the relationship between peripheral blood neutrophils and BT in the late phase of hemorrhagic shock or burn injury has been reported, this relationship in the early phase has not been fully elucidated. We investigate the role of BT in neutrophil activation and priming during hemorrhagic shock. The experimental rats were divided into three groups: a group with normal intestinal flora (NF group), an antibiotic-decontaminated group (AD group), and a sham shock group with normal intestinal flora (sham group). Hemorrhagic shock was induced in the NF and AD groups (MAP 30 mm Hg for 30-90 min). The rats were sacrificed at 30, 60, or 90 min following the shock induction. Cultures were taken from the liver, spleen, mesenteric lymph nodes (MLNs), and systemic blood to assess the occurrence of BT. Hydrogen peroxide generation and CD11b/c expression were assayed by flow cytometry to evaluate peripheral blood neutrophil activation and priming, respectively. In the NF group, significant BT to the MLNs and spleen was noted from 30 min after the shock induction, and significant hydrogen peroxide generation was also noted from 30 min. The expression of CD11b/c on neutrophils was significantly up-regulated at 90 min after the shock induction. Furthermore, BT, as also the aforementioned parameters of neutrophil function, was significantly suppressed in the AD group. We, therefore, concluded that neutrophil activation and priming during hemorrhagic shock might be closely related to BT, and that infectious factors possibly influence the host responses starting from the early phase of damage, even in noninfectious stress-inducing conditions.

Animals↗