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Biomedical subjects

Masaki Takeuchi

Publications and source records attributed to Masaki Takeuchi.

At least 19 recordsLinked to original sources

Tailoring elution of tetraalkylammonium ions. Ideal electrostatic selectivity elution order on a polymeric ion exchanger.

Although ion exchange is often depicted as a process driven by electrostatic forces, ionic solvation or hydrophobic forces contribute greatly to ion exchange selectivity and is often the dominant factor. On a variety of commercial anion exchange columns, monovalent ClO4- elutes after doubly charged SO42- and even triply charged PO43-. For identically charged alkali metal ions, electrostatic charge densities based on crystal radii would suggest Li+ to be the most strongly retained on a cation exchanger. In practice, it is typically the least strongly held cation on most cation exchangers, because of its very high hydration energy and with most eluents its capacity factor approaches zero. Even when the ion is very poorly solvated, as with tetraalkylammonium (NR4+) cations, there has never been a report on a polymeric ion exchanger of an ideal electrostatic selectivity order where NR4+ cations elute in their increasing charge density order: R = n-butyl first, followed by n-propyl, ethyl, and last, methyl. We show that this selectivity order is easily achieved on recently described methracrylate-based monolithic capillary cation exchange columns (Ueki, Y.; Umemura, T.; Li, J. X.; Odake, T; Tsunoda, K. Anal. Chem. 2004, 76, 7007-7012) with minor amounts of hydroorganic modifiers. Indeed, under such conditions, Li+ (and other alkali cations) elutes after NMe4+.

Journal Article↗

Methods for imaging and analyses of intracellular organelles using fluorescent and luminescent proteins.

One of the important features of eukaryotic cells is the presence of intracellular compartments, organelles. Each organelle plays an essential role in the organelle-specific function; each cooperates with others for maintaining the vital activity of living cells. To analyze the organelle-specific functions, researchers have used many fluorescent and luminescent reporter proteins extensively. The reporter proteins have provided unprecedented insights into the movement of proteins and their interactions in the organelles. In this review, we summarize recently validated methods for visualizing protein localization, dynamics and protein-protein interactions, and for identifying a series of proteins localized in organelles using newly developed reporter proteins.

Animals↗

Antigen-receptor genes of the agnathan lamprey are assembled by a process involving copy choice.

Jawless vertebrates have acquired immunity but do not have immunoglobulin-type antigen receptors. Variable lymphocyte receptors (VLRs) have been identified in lamprey that consist of multiple leucine-rich repeat (LRR) modules. An active VLR gene is generated by the assembly of a series of variable gene segments, including many that encode LRRs. Stepwise assembly of the gene segments seems to occur by replacement of the intervening DNA between the 5' and 3' constant-region genes. Here we report that lamprey (Lethenteron japonicum) assemble their VLR genes by a process involving 'copy choice'. Regions of short homology seemed to prime copying of donor LRR-encoding sequences into the recipient gene. Those LRR-encoding germline sequences were abundant and shared extensive sequence homologies. Such genomic organization permits initiation of copying anywhere in an LRR-encoding module for the generation of various hybrid LRRs. Thus, a vast repertoire of recombinant VLR genes could be generated not only by copying of various LRR segments in diverse combinations but also by the use of multiple sites in an LRR gene segment for priming.

Alleles↗

Live imaging of yeast Golgi cisternal maturation.

There is a debate over how protein trafficking is performed through the Golgi apparatus. In the secretory pathway, secretory proteins that are synthesized in the endoplasmic reticulum enter the early compartment of the Golgi apparatus called cis cisternae, undergo various modifications and processing, and then leave for the plasma membrane from the late (trans) cisternae. The cargo proteins must traverse the Golgi apparatus in the cis-to-trans direction. Two typical models propose either vesicular transport or cisternal progression and maturation for this process. The vesicular transport model predicts that Golgi cisternae are distinct stable compartments connected by vesicular traffic, whereas the cisternal maturation model predicts that cisternae are transient structures that form de novo, mature from cis to trans, and then dissipate. Technical progress in live-cell imaging has long been awaited to address this problem. Here we show, by the use of high-speed three-dimensional confocal microscopy, that yeast Golgi cisternae do change the distribution of resident membrane proteins from the cis nature to the trans over time, as proposed by the maturation model, in a very dynamic way.

COP-Coated Vesicles↗

Protein aggregates are transported to vacuoles by a macroautophagic mechanism in nutrient-starved plant cells.

When a fusion protein of cytochrome b5 (Cyt b5) and the red fluorescent protein (RFP) are expressed in tobacco BY-2 cells, the expressed protein forms intracellular aggregates that emit red fluorescence. When such cells are grown to the stationary phase or incubated in nutrient limited medium, RFP fluorescence can be detected in the vacuolar lumen. We investigated this transport mechanism using a limited-nitrogen model. E-64 and 3-methyladenine, which inhibit autophagic processes, blocked the transport of the RFP signal to the vacuole. We next traced the autophagic process in tobacco cells using YFP fused with the tobacco Atg8 homologue (YFP-NtAtg8) and analyzed the contribution of autophagy to the vacuolar transport of the aggregates. Under limited-nitrogen conditions, the aggregates were degraded in preference to other organelles, and the autophagosomes colocalized with the aggregates at a higher frequency than with mitochondria. This is the first demonstration that selective macroautophagic degradation can occur in plant cells.

Adenine↗

Novel application method of artificial dermis: one-step grafting procedure of artificial dermis and skin, rat experimental study.

BACKGROUND: Currently, to treat skin defects with artificial dermis (AD), two surgical procedures where the artificial dermis grafting and another secondary skin grafting are required. The purpose of this study was to achieve simultaneous grafting of the artificial dermis and the split-skin. To enhance the wound angiogenesis, cultured endothelial cells, fibroblasts and PDWHF (platelet derived wound healing factor) were employed. METHODS: The experiment consists of following two parts: (1) Investigation to obtain faster angiogenesis into the bilayer artificial dermis: full-thickness wounds created on the back of the rats were treated with the artificial dermis (Terudermis, with silicone sheet, TERUMO Co., Japan). Prior to the artificial dermis grafting, following four groups were established; control group (AD alone, n=6), PDWHF group (AD treated with PDWHF, n=6), cultured cells group (AD treated with cultured endothelial cells and fibroblasts, n=6), combination group (AD treated with PDWHF and cultured cells, n=6). (2) Trial of one-stage grafting of the AD and the skin: simultaneous grafting of the artificial dermis and skin was performed using the same rat model. Before making skin defects, split thickness skin were harvested. Then the skin grafting was carried out immediately after the AD grafting. To allow grafting of the skin onto the artificial dermis, the AD without silicone sheet (Terudermis without silicone sheet, TERUMO Co., Japan) were used. Two groups, control group (AD alone, n=3) and treatment group (AD with PDWF and cultures, n=3) were established. RESULTS: (1) When the artificial dermis were treated with PDWHF, cultured endothelial cells and fibroblasts, vascular invasion into the artificial dermis was observed 5 days after the surgery. (2) In the treatment group, the skin grafted immediately after the artificial dermis grafting was completely taken. CONCLUSIONS: The present study revealed that treatment with PDWHF, combined with cultured endothelial cells and fibroblasts, accelerated wound angiogenesis. By this method, one-step grafting procedure of the artificial dermis and the skin is possible.

Animals↗

Versatile gas/particle ion chromatograph.

A new, compact gas/particle ion chromatograph has been developed for measuring ionic constituents in PM2.5 (particulate matter of aerodynamic diameter < or = 2.5 microm) and water-soluble ionogenic gases. The instrument has separate sampling channels for gases and particles. In one, a membrane denuder collects soluble gases for preconcentration and analysis. In the other, a cyclone removes larger particles, a membrane denuder removes soluble gases, and a continuously wetted hydrophilic filter collects particles. A single, multiport, syringe pump handles liquid transport, and one conductivity detector measures anions and ammonium for both channels. Electrodialytically generated gradient hydroxide eluent permits 20 min chromatographic runs. Gas/particle samples are each collected for 40 min, butthe sampling intervals are staggered by 20 min. Liquid samples from the gas denuder and particle collector are aspirated and preconcentrated on sequential cation and anion concentrators and transferred respectively to an ammonia transfer device and an anion separation column. The flow configuration results in an ammonium peak before anion peaks in the chromatogram. The system measures ammonia, organic acids (such as acetic, formic, and oxalic acids), HCl, HONO, SO2, HNO3, and the corresponding ions in the aerosol phase. Low ng/m3 to sub-ng/m3 limits of detection (LODs) are attained for most common gases and particulate constituents, the LODs for gaseous SO2 to NH3 range, for example, from sub parts per trillion by volume (sub-pptv) to approximately 5 pptv.

Aerosols↗

Comparison study of a traditional pulsed dye laser versus a long-pulsed dye laser in the treatment of early childhood hemangiomas.

BACKGROUND AND OBJECTIVE: The role of pulsed dye laser (PDL) in the treatment of childhood hemangiomas is still controversial because of the inherent characteristics of hemangiomas as well as the side effects of the PDL. Recently, the long pulsed dye laser (LPDL) with cryogen spray cooling (CSC) has been found relatively more effective and safer than the PDL in the treatment of port-wine stains and telangiectasia. This study was designed to compare the efficacy and complication rate of PDL versus LPDL for treating childhood hemangiomas. STUDY DESIGN/MATERIALS AND METHODS: We did a prospective, randomized, controlled trial in which we enrolled 52 Asian infants, aged 1-3 months, with early hemangiomas. These infants were assigned to PDL treatment (n = 26) or LPDL treatment (n = 26) and followed to age 1 year. A PDL with a wavelength of 585 nm and spot size of 7 mm and a LPDL with a wavelength of 595 nm and spot size of 7 mm were used. Each patient in the PDL group was treated with energy fluence between 6 and 7 J/cm(2) and a pulse duration of 0.45 milliseconds without epidermal cooling. Each patient in the LPDL group was treated with energy fluence between 9 and 15 J/cm(2) and a pulse duration of 10-20 milliseconds, utilizing CSC to protect the epidermis. Each group was treated at 4-week intervals until the lesion cleared. When each patient reached an age of 1 year, outcome measures such as clearance rate, time period of maximum proliferation, and complications were assessed. RESULTS: The number of children whose lesions showed complete clearance or minimal residual signs at 1 year of age was 14 (54%) in the PDL group and 17 (65%) in the LPDL group (P = 0.397). Compared with the LPDL, PDL treated children had more hypopigmentation (3, 12% vs. 8, 31%; P = 0.001), more hyperpigmentation (2, 8% vs. 4, 15%; P = 0.005), and more textural changes (1, 4% vs. 6, 23%; P = 0.001). The average time period of maximum proliferation in the LPDL group was significantly shorter than that of the PDL group (106 days vs. 177 days; P = 0.01). CONCLUSION: Early treatment of childhood hemangiomas with the LPDL is safer and more effective than the PDL.

Asian People↗

Treatment of facial angiofibroma of tuberous sclerosis using cultured epithelial autografts.

Treatment of facial angiofibroma of tuberous sclerosis is problematic, because the skin lesions involve entire dermis. Five patients aged from 14 to 33 (mean: 23.6) years old with angiofibroma of tuberous sclerosis were treated with cultured epithelial autografts between 1995 and 2004. The entire area of the facial lesions was excised using a razor to remove large nodules, and then the remaining lesions were further abraded to a rather deep layer of the dermis to smooth the skin and remove small nodules. Then a cultured autologous epithelium was grafted onto the wound. In all patients, epithelization was complete within 10 (mean: 9) days after the surgery. All patients were followed up for more than 6 months and showed neither depigmentation due to scar formation nor hypertrophic scars. In some patients, some pebbly regrowth had occurred at 5 years postoperatively, but the appearance was quite acceptable.

Adolescent↗

Evaluation of fluence and pulse-duration on purpuric threshold using an extended pulse pulsed-dye laser in the treatment of port wine stains.

Laser therapy of port-wine stains (PWS) using the extended pulse pulsed-dye laser (EPPDL) is accepted as the optimal approach because the thermal relaxation time for the vessels in PWS is actually 1-10 msec. The purpose of this study is to elucidate the purpuric threshold using the EPPDL for treatment of PWS. One hundred and seventy-seven Japanese patients with PWS were recruited for this study. All the patients were dark-skinned with skin phototype III (n = 103) and IV (n = 74). PWS were treated with the EPPDL with a pulse duration ranging 1.5-10 msec, fluence ranging 9-15 J/cm(2), and a spot size of 7 mm. Cryogen spray cooling (CSC) was fixed to 30 msec of delay and 30 msec of spray duration. Patients returned to our clinic within 1 week after their initial laser therapy and the treatment sites were examined for the evidence of purpura formation. Of the 177 patients, 108 developed purpura. The lowest fluences that caused purpura and were seen in more than 50% of patients were 10 J/cm(2) with a pulse duration of 1.5 msec, 12 J/cm(2) with a pulse duration of 3 msec, 13 J/cm(2) with a pulse duration of 6 msec, and 13 J/cm(2) with a pulse duration of 10 msec. The fluence and pulse duration thresholds were 12.5 J/cm(2) and 1.65 msec, respectively. Because purpura is one of the treatment endpoints when using a pulsed-dye laser for PWS, higher fluences are necessary when using a long pulse duration.

Adolescent↗

Transcription elongation factor S-II is required for definitive hematopoiesis.

Transcription elongation factor S-II/TFIIS promotes readthrough of transcriptional blocks by stimulating nascent RNA cleavage activity of RNA polymerase II in vitro. The biologic significance of S-II function in higher eukaryotes, however, remains unclear. To determine its role in mammalian development, we generated S-II-deficient mice through targeted gene disruption. Homozygous null mutants died at midgestation with marked pallor, suggesting severe anemia. S-II(-/-) embryos had a decreased number of definitive erythrocytes in the peripheral blood and disturbed erythroblast differentiation in fetal liver. There was a dramatic increase in apoptotic cells in S-II(-/-) fetal liver, which was consistent with a reduction in Bcl-x(L) gene expression. The presence of phenotypically defined hematopoietic stem cells and in vitro colony-forming hematopoietic progenitors in S-II(-/-) fetal liver indicates that S-II is dispensable for the generation and differentiation of hematopoietic stem cells. S-II-deficient fetal liver cells, however, exhibited a loss of long-term repopulating potential when transplanted into lethally irradiated adult mice, indicating that S-II deficiency causes an intrinsic defect in the self-renewal of hematopoietic stem cells. Thus, S-II has critical and nonredundant roles in definitive hematopoiesis.

Animals↗

Impact of nicorandil to prevent reperfusion injury in patients with acute myocardial infarction: Sigmart Multicenter Angioplasty Revascularization Trial (SMART).

BACKGROUND: Nicorandil in conjunction with percutaneous coronary intervention (PCI) has been reported to reduce reperfusion injury events and improve cardiac function in patients with acute myocardial infarction (AMI). This multicenter study was undertaken to determine the effectiveness and optimal administration of nicorandil in AMI patients. METHODS AND RESULTS: Ninety-two patients with first AMI were randomly assigned to 1 of 3 groups: intracoronary administration of nicorandil (Group A), combined intravenous and intracoronary administration of nicorandil (Group B), and no nicorandil administration (Group C). The primary endpoint was a composite of the incidence of reperfusion-induced arrhythmia, chest pain, and no-reflow/slow-reflow. The secondary endpoint was the combined rate of improvement in the Thrombolysis in Myocardial Infarction frame count (cTFC) and ST resolution (STR). A significant difference was observed in the primary endpoint for Group B as compared with Group C (p<0.05). In the meantime, a significant improvement was shown in the secondary endpoint for Group B compared with Group C (p=0.04 and 0.006 for cTFC and STR, respectively). CONCLUSIONS: Combined intravenous and intracoronary administration of nicorandil reduces reperfusion injury during PCI and improves the cTFC and STR in AMI, and appears to be preferable to intracoronary administration alone.

Aged↗

Oncostatin m maintains the hematopoietic microenvironment and retains hematopoietic progenitors in the bone marrow.

Bone marrow (BM) functions as the primary hematopoietic tissue throughout adult life by providing a microenvironment for the proliferation, differentiation, and retention of hematopoietic stem cells and progenitors. We describe novel roles for oncostatin M (OSM) in the BM hematopoietic microenvironment. Hematopoietic progenitor activity in OSM-deficient mice was reduced in BM but elevated in the spleen and peripheral blood. The level of circulating granulocyte colony-stimulating factor (G-CSF) was increased, whereas that of stromal cell-derived factor 1 (SDF-1) was decreased in OSM-deficient mice. Moreover, the ability of OSM-deficient BM stromal cells to support hematopoiesis in vitro was significantly reduced. These results indicate that OSM plays a unique role in hematopoiesis by maintaining the proper microenvironment for BM hematopoiesis; it also retains hematopoietic progenitors in BM by regulating G-CSF and SDF-1 levels.

Animals↗

Continuous collection of soluble atmospheric particles with a wetted hydrophilic filter.

Approximately one-third of the area (14-mm diameter of a 25-mm diameter) of a 5-microm uniform pore size polycarbonate filter is continuously wetted by a 0.25 mL/min water mist. The water forms a continuous thin film on the filter and percolates through it. The flowing water substantially reduces the effective pore size of the filter. At the operational air sampling flow rate of 1.5 standard liters per minute, such a particle collector (PC) efficiently captures particles down to very small size. As determined by fluorescein-tagged NaCl aerosol generated by a vibrating orifice aerosol generator, the capture efficiency was 97.7+% for particle aerodynamic diameters ranging from 0.28 to 3.88 microm. Further, 55.3 and 80.3% of 25- and 100-nm (NH4)2SO4 particles generated by size classification with a differential mobility analyzer were respectively collected by the device. The PC is integrally coupled with a liquid collection reservoir. The liquid effluent from the wetted filter collector, bearing the soluble components of the aerosol, can be continuously collected or periodically withdrawn. The latter strategy permits the use of a robust syringe pump for the purpose. Coupled with a PM2.5 cyclone inlet and a membrane-based parallel plate denuder at the front end and an ion chromatograph at the back end, the PC readily operated for at least 4-week periods without filter replacement or any other maintenance.

Journal Article↗

Prospective evaluation of a screening protocol to exclude deep vein thrombosis on the basis of a combination of quantitative D-dimer testing and pretest clinical probability score.

BACKGROUND: Clinical signs and symptoms such as swelling, pain, and redness are unreliable markers of deep vein thrombosis (DVT). Because of this venous duplex scanning (VDS) has been heavily used in DVT detection. The purpose of this study was to determine if a combination of D-dimer testing and pretest clinical score could reduce the use of VDS in symptomatic patients with suspected DVT. STUDY DESIGN: One hundred seventy-four consecutive patients with suspected DVT were prospectively evaluated using pretest clinical probability (PCP) score and D-dimer testing before VDS. After calculating clinical probability scores developed by Wells and associates, patients were divided into low risk ( or=3 points) PCP. RESULTS: One hundred fifty-eight patients were enrolled. The prevalence of DVT in this study was 37%. Thirty-eight patients (24%) were classified as low risk, 64 (41%) as moderate risk, and 56 (35%) as high risk PCP. DVT was identified in only one patient (2.6%) with low risk PCP. In contrast, DVT was found in 22 (34%) with moderate risk, and 35 (63%) with high risk PCP. In the high and moderate risk PCP groups, positive scan patients had a markedly higher value of D-dimer assay than negative scan patients (p=0.0001 and p=0.0057, respectively). In the low risk PCP patients, D-dimer testing provided 100% sensitivity, 46% specificity, 4.8% positive predictive value, and 100% negative predictive value in the diagnosis of DVT. Similarly, in the moderate risk PCP, the D-dimer testing showed 100% sensitivity, 45% specificity, 49% positive predictive value, and 100% negative predictive value. In the high risk group, D-dimer testing achieved 100% sensitivity, 57% specificity, 80% positive predictive value, and 100% negative predictive value in the diagnosis of DVT. These results suggested that 36 of 158 patients who had a non-high PCP (low and moderate PCP) and a normal D-dimer concentration were considered to have no additional investigation, so VDS could have been reduced by 23% (36/158). CONCLUSIONS: A combination of D-dimer testing and clinical probability score may be effective in avoiding unnecessary VDS in suspected symptomatic DVT in the low and moderate PCP patients. The need for VDS could be reduced by 23% despite a relatively high prevalence of DVT.

Adult↗

Total face reconstruction with one expanded free flap.

Deformities of a totally burned face present a profound challenge to the reconstructive plastic surgeon. Skin grafting has been used traditionally for resurfacing with limited success, especially when the burns were so severe the deeper structures were destroyed. Total face reconstruction, using bilateral extended scapular free flap, has been reported previously for severe deformities following an extensive facial burn. Although this method obtained better aesthetic and functional results than skin grafting, the donor-site morbidity was relatively high, with a large scar that extended across the entire back. In addition, the nose needed to be reconstructed separately with a forehead flap or free radial forearm flap. The authors experienced a case in which a totally burned face was reconstructed successfully with a single free-expanded flap. A 54-year-old man sustained a severe facial burn with gasoline that involved the face, anterior neck, anterior chest, and bilateral upper extremities. Sequential debridement and skin grafting were required to close the burn wound. A tissue expander was inserted in his left back before the facial reconstruction. Six months after insertion of the tissue expander, the left dorsal skin was transferred to the face as one large flap, size 28x27 cm, with three sets of vascular anastomoses. The flap totally survived with abundant tissue at the central area to reconstruct the nose. With five complementary procedures, including a costal cartridge graft, the shape of the nose was restored, and acceptable functional and aesthetic results were obtained. This method did not require a separate tissue transfer for nasal reconstruction. To our knowledge, this is the first case of successful reconstruction with one flap for total face reconstruction that included the nose.

Burns↗

Membrane-based parallel plate denuder for the collection and removal of soluble atmospheric gases.

A continuously wetted cellulose acetate membrane-based parallel plate diffusion denuder is described. This is the first membrane-based denuder that has a small enough internal liquid holdup volume to permit reasonably rapid response time (10 --> 90% rise time of approximately 1.2 min for a transient event at a liquid flow rate of 500 microL/min) while permitting quantitative removal of common soluble atmospheric trace gases at flow rates up to 1.7 L/min. The latter attribute permits the use of the device as the first element in a particle sampling and analysis system for the quantitative removal of potentially interfering soluble trace gases. Particle losses in the denuder range from 0.9 to 2.9% over an aerodynamic diameter range of 0.38-3.48 microm, averaging 1.8%. However, only approximately 0.5% of the particles actually appears in the denuder effluent liquid. The relatively compact (300 mm H x 57 mm W x 26 mm D) wet denuder should be attractive in a number of applications. We show excellent agreement for HONO measurements with a conventional larger parallel plate wetted denuder in field measurements.

Journal Article↗