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The rapid and facile generation of 4'-carbon radicals from oxime imidates of nucleosides via 1,5-hydrogen atom transfer induced by iminyl radicals was developed. The cyclization of 4'-carbon radicals with olefins, followed by the hydrolysis of imidate residues, provided various 2'-O,4'-C- and 3'-O,4'-C-bridged nucleosides. This operationally simple approach can be applied to the few-step syntheses of 6'S-methyl-2'-O,4'-C-ethylene-bridged 5-methyluridine (6'S-Me-ENA-T) and S-constrained ethyl-bridged 5-methyluridine (S-cEt-T).
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Hidrogênio , Nucleosídeos , Carbono , Etilenos/química , Alcenos , Oximas , ImidoésteresRESUMO
STUDY DESIGN: A retrospective multicenter observational study. OBJECTIVE: To investigate correction surgeries that were performed in relatively aged patients in terms of mechanical complications (MCs) and their predictive factors. SUMMARY OF BACKGROUND DATA: The risk factors associated with MCs have not yet been well examined, especially in aged populations. METHODS: We retrospectively reviewed 230 surgically treated ASD patients with an average age of 72.2 years. Twenty-eight patients with ASD caused by vertebral fractures were excluded. The minimum follow-up was 2 years. Postoperative MCs were defined as proximal junction kyphosis, distal junction kyphosis, pseudoarthrosis, rod breakage, and vertebral fractures. We divided all the ASD patients into two groups: patients with MC (the MC (+) group) and patients without MC (the MC (-) group). Radiographic parameters were evaluated before and immediately after surgery. The SRS-Schwab ASD classification and global alignment and proportion (GAP) score were also evaluated. RESULTS: Of the 202 patients, 91 (45.0%) had MCs. The age at surgery was significantly higher in the MC (+) group than in the MC (-) group. Regarding radiographic parameters, postoperative global tilt (GT), pre- and postoperative thoracolumbar kyphosis (TLK), and postoperative thoracic kyphosis were significantly higher in the MC (+) group than in the MC (-) group. Other parameters, such as the proposed ideal alignment target of PI-LL<10, did not significantly affect MC rates. The GAP score was high in both groups and not significantly related to a higher rate of MC. Forward stepwise logistic regression indicated that the age at surgery, postoperative GT, and preoperative TLK were significant risk factors for MCs. CONCLUSION: Older age, higher postoperative GT, and higher pre and postoperative TLK can be risk factors for MCs. The GAP score was high in both groups and not significantly related to a higher rate of MC. LEVEL OF EVIDENCE: 4.
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Fenômenos Biomecânicos/fisiologia , Procedimentos Neurocirúrgicos/efeitos adversos , Complicações Pós-Operatórias/diagnóstico por imagem , Complicações Pós-Operatórias/fisiopatologia , Doenças da Coluna Vertebral/diagnóstico por imagem , Doenças da Coluna Vertebral/cirurgia , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Cifose/diagnóstico por imagem , Cifose/cirurgia , Vértebras Lombares/diagnóstico por imagem , Vértebras Lombares/cirurgia , Masculino , Pessoa de Meia-Idade , Procedimentos Neurocirúrgicos/tendências , Complicações Pós-Operatórias/etiologia , Valor Preditivo dos Testes , Estudos Retrospectivos , Vértebras Torácicas/diagnóstico por imagem , Vértebras Torácicas/cirurgiaRESUMO
This study examined the effects of microtubule-targeting anticancer drugs (paclitaxel, cabazitaxel, and eribulin) on the expression of drug efflux transporter P-glycoprotein, which is encoded by MDR1. Paclitaxel and eribulin induced MDR1 promoter activity in a concentration-dependent manner, while cabazitaxel had little effect in human intestinal epithelial LS174T cells. Overexpression of the nuclear receptor pregnane X receptor (PXR) gene (NR1I2) enhanced paclitaxel- and eribulin-induced MDR1 activation, but expression of the nuclear receptor co-repressor silencing mediator for retinoid and thyroid receptors (SMRT) gene (NCOR2) repressed MDR1 activation. Eribulin increased the mRNA and protein expression of P-glycoprotein in LS174T cells. Cellular uptake of rhodamine 123 and calcein-acetoxymethyl ester (calcein-AM), P-glycoprotein substrates, decreased in paclitaxel- or eribulin-treated LS174T cells. Eribulin also increased MDR1 promoter activity in human breast cancer MCF7 cells. The results suggest that the microtubule-targeting anticancer drug eribulin can induce the drug efflux transporter P-glycoprotein via PXR in human intestinal and breast cancer cells and thus influence the efficacy of anticancer drugs.
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An Ir(III) polypyridyl complex-conjugated 14-mer oligonucleotide (IrIII-DNA) was synthesized and its hybridization properties with single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA) were evaluated by UV-melting experiments. The stabilities of the duplexes of IrIII-DNA with 14-, 20-, and 26-mer ssDNAs were higher than those of the unconjugated oligonucleotides. The triplex of IrIII-DNA with 14-mer dsDNA was also stabilized. However, the triplexes of IrIII-DNA with 20- and 26-mer dsDNAs, flanked by 3 and 6 base pairs at the both ends of 14-mer dsDNA target, were destabilized. This is presumably because of steric repulsion between the Ir(III) complex and the protruding 3- and 6-mer dsDNA moieties which are inflexible compared to ssDNA.
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Técnicas de Química Sintética , Irídio/química , Hibridização de Ácido Nucleico , Oligonucleotídeos/química , Sondas de DNA , Espectroscopia de Ressonância Magnética , Estrutura Molecular , Conformação de Ácido Nucleico , Oligonucleotídeos/síntese química , Oligonucleotídeos/efeitos da radiaçãoRESUMO
BACKGROUND: While several complications related to pressure-wire (PW) have been reported, mechanistic justification has not always been offered. Furthermore, interference between a PW and a protruding side-branch stent has not been previously reported. The purpose of this study was to evaluate interference between PW-pullback from a main-branch with a protruded ostial stent deployed in a side-branch. METHODS: In a polyurethane bifurcation vessel model, PW-pullback was performed in a main-branch following protruded ostial stenting in a side-branch. Tested PWs included PressureWire X, Comet, OptoWire, and Verrata. For each PW, pullback was performed through the same proximal cell of the protruded stent 20 times. Interference during PW-pullback was objectively analyzed with a fiberscope placed at the distal main-branch and classified into 3 grades according to the interaction with stent strut. RESULTS: There were significant differences in the rate of interference between the PWs. No-interference, interference without strut traction, and interference with strut traction (i.e. stent deformation) were observed as follows: 17/20, 3/20, and 0/20 in PressureWire X; 19/20, 1/20, and 0/20 in Comet; 8/20, 10/20, and 2/20 in OptoWire; and 13/20, 2/20, and 5/20 in Verrata, respectively (p for any differences: <0.001). Visually identifiable major stent deformation was observed once in OptoWire due to the deep concave sensor window and twice in Verrata due to the proximal gap between the sensor and coiled-wire. CONCLUSIONS: PW-pullback in the main-branch after side-branch ostial stenting should be carefully performed to avoid stent deformation. Consideration on the specific mechanical features of the PW is also essential.
Assuntos
Angioplastia Coronária com Balão/instrumentação , Cateterismo Cardíaco/instrumentação , Cateteres Cardíacos , Stents , Transdutores de Pressão , Angioplastia Coronária com Balão/efeitos adversos , Cateterismo Cardíaco/efeitos adversos , Tecnologia de Fibra Óptica , Humanos , Teste de Materiais , Modelos Cardiovasculares , Desenho de PróteseRESUMO
Previous reports showed that GuideLiner (GL) and Guidezilla (GZ) can accommodate bulky and multiple devices beyond the official profiles. However, feasibility of kissing balloon technique (KBT) through these devices is unknown. The tested devices included 7Fr-GL/GZ and respective three types of 2.5 mm semi-compliant (SC) and non-compliant (NC) balloons: conventional model (CM), tapered-tip model (TM) and latest model (LM). First, three experienced operators attempted to advance all 21 combinations of the 2 balloons through GL/GZ on the guidewires and assessed the crossability in 3 grades: easy, difficult and impossible. Second, the only balloon combinations graded as easy by all operators were tested in the polyurethane-made bifurcation model which required KBT following cross-over stenting. Within the total of 42 device combinations, only one balloon combination of double LM-NC balloons was classified as easy in both GL/GZ by consensus opinion of the operators. While two combinations of LM-SC and LM-SC/NC balloons were classified as difficult in both GL/GZ, all four combinations of LM-SC/NC and CM/TM-NC balloons were classified as difficult only in GL. Other 32 combinations were all classified as impossible. In the bifurcation model, the combination of double LM-NC balloons using GL achieved KBT while the same balloon combination with GZ failed. The feasibility of KBT using child-catheter is highly dependent on the device characteristics. The combination of latest small-profile NC balloons through GL could be clinically applicable.
Assuntos
Angioplastia Coronária com Balão/instrumentação , Angioplastia Coronária com Balão/métodos , Stents , Dispositivos de Acesso Vascular/classificação , Estudos de Viabilidade , HumanosRESUMO
New C,N,S-cyclometalated palladium(II) and platinum(II) complexes have been synthesized and their structural, electrochemical, and photochemical properties examined. The blue color of these complexes in solution changed to yellow under visible-light irradiation. By measurement of the absorption spectra for quantifying changes in color, isosbestic points for each complex clearly indicated the presence of only two species responsible for the change of color. X-ray analysis revealed that the visible-light-induced yellow species were S-oxygenated sulfinato complexes. Photosensitized generation of singlet oxygen (1O2) was confirmed by the direct detection of singlet oxygen luminescence at 1275 nm. The present cyclometalated palladium(II) and platinum(II) complexes are efficient photosensitizers of singlet oxygen, which rapidly reacts with coordinating sulfur atoms.
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INTRODUCTION: We describe 5 patients who underwent operative treatment for arachnoid web (AW) and discuss the postoperative clinical outcome in each case. AW is an extremely rare disease that causes cord compression and syringomyelia in the thoracic spine. To date, 14 cases only of AW have been reported, and the effect of surgical intervention on clinical and radiologic outcomes is unknown. METHODS: Five patients who underwent surgical treatment for AW were retrospectively reviewed. The clinical outcomes were evaluated using the thoracic Japanese Orthopaedic Association (T-JOA) score. Preoperative and postoperative images were reviewed. RESULTS: All the patients presented with spastic gait and numbness in the lower extremities. Two patients also presented with bladder-bowel dysfunction (BBD). AW, or the so-called "scalpel" sign, was seen dorsally in the thoracic spine on magnetic resonance imaging in all the patients. Syringomyelia adjacent to the web was observed in 4 patients. Fenestration and web resection without instrumentation was performed in all the cases. Overall, significant improvement was seen in locomotion and the total T-JOA score postoperatively. However, numbness in the lower extremities improved in 2 patients but was unchanged in 3 cases. BBD was ameliorated in 1 patient but remained unchanged in the other patient. CONCLUSIONS: Our experience suggests that surgical treatment, including the another patient and resection of the web, can correct the flow dynamics of cerebrospinal fluid and allow neurologic recovery, in particular locomotion, in patients with AW.
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BACKGROUND: Secondary prevention in patients with myocardial infarction (MI) is critically important to prevent ischaemic heart failure and reduce social burden. Pioglitazone improves vascular dysfunction and prevents coronary atherosclerosis, mainly via anti-inflammatory and antiatherogenic effects by enhancing adiponectin production in addition to antihyperglycemic effects, thus suggesting that pioglitazone attenuates cardiovascular events in patients with mild (HbA1c levelsâ¯<â¯6·5%) diabetes mellitus (DM). Therefore, we evaluated the effects of pioglitazone on cardiovascular events in patients with both previous MI and mild DM. METHODS: In this multicentre, prospective, randomised, open, blinded-endpoint trial, we randomly assigned 630 patients with mild DM with a history of MI to undergo either DM therapy with (pioglitazone group) or without (control group) pioglitazone. DM was diagnosed using the 75-g oral glucose tolerance test, and mild DM was defined if HbA1c level was < 6·5%. The primary endpoint was the composite of cardiovascular death and hospitalisation caused by acute MI, unstable angina, coronary revascularisation (including percutaneous coronary intervention and cardiac bypass surgery), and stroke. FINDINGS: HbA1C levels were 5·9 and 5·8% (pâ¯=â¯0·71) at baseline and 6·0 and 5·8% (pâ¯<â¯0·01) at 2â¯years for the control and pioglitazone groups, respectively.The primary endpoint was observed in 14·2% and 14·1% patients in the control and pioglitazone groups during two years (95% confidential interval (CI):0.662-1·526, pâ¯=â¯0·98), respectively; the incidence of MI and cerebral infarction was 0·3% and 2·2% (95%CI: 0·786-32·415, pâ¯=â¯0·09) and 1·0% and 0·3% (95%CI: 0·051-3·662, pâ¯=â¯0·44), respectively. Post-hoc analyses of the 7-year observation period showed that these trends were comparable (21·9% and 19·2% in the control and pioglitazone groups, 95%CI: 0.618-1·237, pâ¯=â¯0·45). INTERPRETATION: Pioglitazone could not reduce the occurrence of cardiovascular events in patients with mild DM and previous MI.
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INTRODUCTION: Pedicle subtraction osteotomy (PSO) is performed to correct sagittal plane deformity. This procedure is useful with revision cases in which the number of intact discs for correction is limited. METHODS: Forty-four patients (10 male and 34 female) with minimum follow-up of 2 years were reviewed; all had undergone PSO revision surgery for kyphosis following previous lumbar fusion surgery. The average age at operation was 72.8 years (range 42-85 years), and the average follow-up period was 4.1 years (2-9 years). The average fusion level was 7.5 (4-13 level), and the average previously fused level was 2.4 (1-7 level). RESULTS: The average operation time was 424 min, and average blood loss was 2880 g. The average JOA score of 14.0 before operation changed to 21.8 at 1-year follow-up and to 20.7 at final follow-up. The average recovery rate at final follow-up was 45.7%. Four patients underwent re-operations for proximal junctional kyphosis and 3 patients for rod fracture. The fusion rate was 88.6%, and 13 patients (29.5%) developed subsequent vertebral fracture. The average PI-LL (Pelvic incidence minus Lumbar lordosis) at pre-op of 52.9 degrees changed to 3.8 degrees at post-op, to 13.4 degrees at 1-year follow-up, and to 14.8 degrees at final follow-up. The average correction at the PSO site was 36.0 degrees at post-op, 36.7 degrees at 1-year follow-up, and 37.0 degrees at final follow-up. The average sagittal vertical axis at pre-op of 145.0 mm decreased to 51.2 mm at 1-year follow-up; however, it increased to 75.3 mm at final follow-up. CONCLUSION: PSO for correction of kyphosis following previous lumbar fusion surgery was an effective procedure without correction loss at the local osteotomy site; however, its surgical invasiveness and complication rate were high. Subsequent vertebral fracture, adjacent segment degeneration, and rod fracture contribute to deterioration of outcome that is evident at long-term follow-up.
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Stent placement for treating superficial femoral artery (SFA) lesions has been approved. The Zilver PTX stent, a drug-eluting stent (DES) for treating SFA lesions, has been available in Japan since 2012. However, the penetration rate of this DES has not yet been reported. This prospective multicenter registry study enrolled 314 patients (354 limbs) to be treated by stent placement in 2014 (UMIN000011551). The primary endpoint was the measurement of the penetration rate of the DES. The secondary endpoints were measuring the freedom from restenosis, freedom from target lesion revascularization (TLR), freedom from major adverse limb event (MALE), and the survival rate at 12 months postoperatively. Female patients comprised 28% participants. The mean age was 73.1 ± 9.2 years. A total of 56% patients had diabetes mellitus (DM), 36% patients were receiving hemodialysis, and 30% used cilostazol at baseline. The mean lesion length was 156 ± 101 mm, and the percentage of TASC II C/D lesions was 58%. Critical limb ischemia (CLI) was observed in 32% limbs. The penetration rates of the Zilver PTX stent were only 8%. The primary patency rate was similar between DES and bare-metal stents (BMS) at 12 months postoperatively (77 vs. 84%, p = 0.52). In this study, the rates of freedom from restenosis, freedom from TLR, freedom from MALE, and the survival rate at 12 months postoperatively were 83, 86, 85, and 89%, respectively. The penetration rate of a first-generation DES placement for treating SFA lesions is low in Japan. On the other hand, BMS is well utilized and its primary patency is acceptable.
Assuntos
Stents Farmacológicos , Procedimentos Endovasculares/instrumentação , Artéria Femoral/cirurgia , Doença Arterial Periférica/cirurgia , Grau de Desobstrução Vascular , Idoso , Intervalo Livre de Doença , Feminino , Artéria Femoral/diagnóstico por imagem , Artéria Femoral/fisiopatologia , Seguimentos , Humanos , Incidência , Japão/epidemiologia , Masculino , Doença Arterial Periférica/diagnóstico , Doença Arterial Periférica/epidemiologia , Estudos Prospectivos , Desenho de Prótese , Resultado do Tratamento , Ultrassonografia Doppler DuplaAssuntos
Angioplastia Coronária com Balão/efeitos adversos , Trombose Coronária/cirurgia , Anomalias dos Vasos Coronários/diagnóstico , Complicações Pós-Operatórias , Tomografia de Coerência Óptica/métodos , Ultrassonografia de Intervenção/métodos , Doenças Vasculares/congênito , Adulto , Angiografia Coronária , Trombose Coronária/diagnóstico , Anomalias dos Vasos Coronários/etiologia , Vasos Coronários/diagnóstico por imagem , Vasos Coronários/patologia , Feminino , Seguimentos , Humanos , Remissão Espontânea , Fatores de Tempo , Doenças Vasculares/diagnóstico , Doenças Vasculares/etiologiaRESUMO
Mediator release from mast cells is a critical step in allergic and inflammatory disease. However, the processes regulating the latter stages of granule release are yet to be fully understood. Rab27 small GTPases regulate release of secretory lysosomes in a variety of cells, including mast cell granules. In the present study, using murine bone marrow-derived mast cells (BMMC) from Rab27-deficient mutant mice, we found that, in contrast to Rab27b, Rab27a primarily plays an inhibitory role in regulating degranulation. Immunofluorescence analysis revealed that resting Rab27a-deficient (ashen) BMMCs display abnormal cortical F-actin distribution. Actin disassembly prior to IgE cross-linking increased wild-type BMMC secretion to ashen levels, suggesting that changes in the integrity of cortical F-actin underlie the ashen phenotype. Comparison of the secretory impairment of Rab27b knockout and Rab27a/b double knockout BMMCs highlighted a secondary positive role for Rab27a in enhancing degranulation. Rab27 is known to interact with actin via its effectors melanophilin (Mlph) and myosin Va (MyoVa) in other cell types. To better understand the differing roles of Rab27 proteins, we analysed the secretory phenotype of BMMCs derived from mice lacking Rab27 effector proteins. These experiments revealed that the phenotype of BMMCs deficient in Mlph (leaden) and BMMCs deficient in MyoVa (dilute) resembles the hyper-secretion of ashen BMMCs, while Munc13-4-deficient (jinx) BMMCs phenocopy the Rab27b knockout and double Rab27a/b knockout secretory impairment. We conclude that Rab27a and Rab27b regulate distinct steps in the BMMC degranulation pathway, with Rab27a/Mlph/MyoVa regulating cortical actin stability upstream of Rab27a/b/Munc13-4-dependent granule exocytosis.
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Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Mastócitos/metabolismo , Proteínas de Membrana/metabolismo , Cadeias Pesadas de Miosina/metabolismo , Miosina Tipo V/metabolismo , Proteínas rab de Ligação ao GTP/metabolismo , Actinas/metabolismo , Proteínas Adaptadoras de Transdução de Sinal/genética , Animais , Células da Medula Óssea/metabolismo , Células da Medula Óssea/fisiologia , Degranulação Celular/genética , Degranulação Celular/fisiologia , Células Cultivadas , Citoesqueleto/metabolismo , Exocitose/genética , Exocitose/fisiologia , Feminino , Immunoblotting , Masculino , Mastócitos/fisiologia , Proteínas de Membrana/genética , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Microscopia Confocal , Microscopia Eletrônica , Cadeias Pesadas de Miosina/genética , Miosina Tipo V/genética , Receptores de IgE/metabolismo , Proteínas rab de Ligação ao GTP/genética , Proteínas rab27 de Ligação ao GTPRESUMO
Cellulose synthesis in plants is believed to be carried out by the plasma membrane-associated rosette structure which can be observed by electron microscopy. Despite decade-long speculation, it had not been demonstrated whether the rosette is the site of catalytic activity of cellulose synthesis. To determine the relationship between this structure and cellulose synthesis, we successfully isolated detergent-insoluble rosettes from the plasma membrane of bean epicotyls. However, the purified rosettes did not possess cellulose synthesis activity in vitro. Conversely, detergent-soluble granular particles of approximately 9.5-10 nm diameter were also isolated and exhibited UDP-glucose binding activity and possessed beta-1,4-glucan (cellulose) synthesis activity in vitro. The particle, referred to as the catalytic unit of cellulose synthesis, was enriched with a 78 kDa polypeptide which was verified as sucrose synthase like by mass spectrometry and immunoblotting. The catalytic units were able to bind to the rosettes and retained the cellulose synthesis activity in the presence of UDP-glucose or sucrose plus UDP when supplemented with magnesium. The incorporation of the catalytic unit into the rosette structure was confirmed by immunogold labeling with anti-sucrose synthase antibodies under an electron microscope. Our results suggest that the plasma membrane-associated rosette anchors the catalytic unit of cellulose synthesis to form the functional cellulose synthesis machinery.
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Celulose/biossíntese , Fabaceae/enzimologia , Glucosiltransferases/metabolismo , Sequência de Aminoácidos , Membrana Celular/metabolismo , Glucanos/biossíntese , Microscopia Eletrônica de Transmissão , Dados de Sequência Molecular , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Sacarose/metabolismo , Uridina Difosfato Glucose/metabolismoRESUMO
In acentriolar higher plant cells, the surface of the nucleus acts as a microtubule-organizing center, substituting for the centrosome. However, the protein factors responsible for this microtubule organization are unknown. The nuclear surfaces of cultured tobacco BY-2 cells possess particles that generate microtubules. We attempted to isolate the proteins in these particles to determine their role in microtubule organization. When incubated with plant or mammalian tubulin, some, but not all, of the isolated nuclei generated abundant microtubules radially from their surfaces. The substance to induce the formation of radial microtubules was confirmed by SDS-PAGE to be a protein with apparent molecular mass of 38 kDa. Partial analysis of the amino acid sequences of the peptide fragments suggested it was a histone H1-related protein. Cloning and cDNA sequence analysis confirmed this and revealed that when the recombinant protein was incubated with tubulin, it could organize microtubules as well as the 38-kDa protein. Histone H1 and tubulin formed complexes immediately, even on ice, and then clusters of these structures were formed. These clusters generated radial microtubules. This microtubule-organizing property was confined to histone H1; all other core histones failed to act as organizers. On immunoblot analysis, rabbit antibodies raised against the 38-kDa protein cross-reacted with histone H1 proteins from tobacco BY-2 cells. These antibodies virtually abolished the ability of the nucleus to organize radial microtubules. Indirect immunofluorescence showed that the antigen was distributed at the nuclear plasm and particularly at nuclear periphery independently from DNA.
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Regulação da Expressão Gênica de Plantas , Histonas/metabolismo , Microtúbulos/metabolismo , Nicotiana/metabolismo , Animais , Anticorpos Monoclonais/química , Núcleo Celular/metabolismo , Células Cultivadas , Centrossomo/metabolismo , Clonagem Molecular , DNA/metabolismo , Eletroforese em Gel Bidimensional , Modelos Biológicos , Coelhos , Proteínas Recombinantes/metabolismoRESUMO
In higher plant cells, various microtubular arrays can be seen despite of their lack of structurally defined microtubule-organizing centers (MTOCs) like centrosomes in animal cells. Little is known about the molecular properties of the microtubule-organizing centers in higher plant cells. The nuclear surface contains one of these microtubule-organizing centers and generates microtubules radially toward the cell periphery (radial microtubules). Previously, we reported that histone H1 possessed the microtubule-organizing activity, and it was suggested that histone H1 localized on the nuclear surfaces in Tobacco BY-2 cells (Nakayama, T., Ishii, T., Hotta, T., and Mizuno, K. J. Biol. Chem. (submitted)). Here we show that histone H1 forms ring-shaped complexes with tubulin, and these complexes nucleated and elongated the radial microtubules continuously (processively) associating with their proximal ends where the incorporation of tubulin occurred. Furthermore, the polarity of radial microtubules was determined to be proximal end plus. Immunofluorescence microscopy of the isolated nuclei revealed that histone H1 localized on the nuclear surfaces, distinct from that in the chromatin. These results indicate that radial microtubules are organized by a novel MTOC that is totally different from MTOCs previously found in either plant or animal cells.
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Forma do Núcleo Celular/fisiologia , Histonas/fisiologia , Microtúbulos/fisiologia , Nicotiana/citologia , Nicotiana/fisiologia , Células Cultivadas , Células VegetaisRESUMO
The correlation between the insertion torque of a lumbar pedicle screw and the mechanical stability of the screw in the bone has been mentioned in in vitro studies. The purpose of this study was to confirm the factors affecting the insertion torque of such screws in vivo. Also, the contribution of insertion torque to the initial stability of the fusion area was to be analyzed in vivo. A series of 23 cases representing 50 lumbar vertebrae were included in this study, in which we examined bone mineral density using quantitative computed tomography (CT) prior to operation. Two screw shapes were utilized, with the insertion torque for each screw measured at two points in time. The correlation between insertion torque and mineral density was investigated. Screw positions were confirmed on postoperative CT scans, and the effect of the screw thread cutting into the cortex bone was investigated. Radiographic changes at three points during a period of 3 months were also measured, and we then evaluated the interrelations between these changes and insertion torque. Furthermore, the relation between insertion torque and instability at 3 months was investigated. Correlations of insertion torque and bone mineral density depended on screw shape. There was no correlation found with mineral density in the case of cylindrical screws. Insertion torque was not affected by the screw thread cutting into the cortex of bone. As for postoperative alignment changes, no definitive trends could be ascertained, and no interrelations with torque and alignment changes were observed. There is a possibility that insertion torque was related to early-stage stability, but no statistical relation could be determined.
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Parafusos Ósseos , Vértebras Lombares/diagnóstico por imagem , Vértebras Lombares/cirurgia , Idoso , Idoso de 80 Anos ou mais , Densidade Óssea , Desenho de Equipamento , Feminino , Humanos , Cuidados Intraoperatórios , Masculino , Pessoa de Meia-Idade , Cuidados Pós-Operatórios , Radiografia , Fatores de Tempo , TorqueRESUMO
Although seed plants have gamma-tubulin, a ubiquitous component of centrosomes associated with microtubule nucleation in algal and animal cells, they do not have discrete microtubule organizing centers (MTOCs) comparable to animal centrosomes, and the organization of microtubule arrays in plants has remained enigmatic. Spindle development in basal land plants has revealed a surprising variety of MTOCs that may represent milestones in the evolution of the typical diffuse acentrosomal plant spindle. We have isolated and characterized the gamma-tubulin gene from a liverwort, one of the extant basal land plants. Sequence similarity to the gamma-tubulin gene of higher plants suggests that the gamma-tubulin gene is highly conserved in land plants. The G9 antibody to fission yeast gamma-tubulin recognized a single band of 55 kD in immunoblots from bryophytes. Immunohistochemistry with the G9 antibody clearly documented the association of gamma-tubulin with various MTOC sites in basal land plants (e.g., discrete centrosomes with and without centrioles and the plastid surface in monoplastidic meiosis of bryophytes). Changes in the distribution of gamma-tubulin occur in a cell cycle-specific manner during monoplastidic meiosis in the liverwort Dumortiera hirsuta. gamma-Tubulin changes its localization from the plastid surface in prophase I to the spindle, from the spindle to phragmoplasts and the nuclear envelope in telophase I, and back to the plastid surfaces in prophase II. In vitro experiments show that gamma-tubulin is detectable on the surface of isolated plastids and nuclei of D. hirsuta, and microtubules can be repolymerized from the isolated plastids. gamma-Tubulin localization patterns on plastid and nuclear surfaces are not affected by the destruction of microtubules by oryzalin. We conclude that gamma-tubulin is a highly conserved protein associated with microtubule nucleation in basal land plants and that it has a cell cycle-dependent distribution essential for the orderly succession of microtubule arrays.
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Evolução Molecular , Centro Organizador dos Microtúbulos/metabolismo , Plantas/metabolismo , Sulfanilamidas , Tubulina (Proteína)/metabolismo , Sequência de Aminoácidos , Anticorpos/imunologia , Briófitas/genética , Briófitas/metabolismo , Núcleo Celular/metabolismo , Clonagem Molecular , Reações Cruzadas/imunologia , DNA de Plantas/química , DNA de Plantas/genética , Dinitrobenzenos/farmacologia , Hepatófitas/genética , Hepatófitas/metabolismo , Imuno-Histoquímica , Meiose/genética , Microscopia Imunoeletrônica , Centro Organizador dos Microtúbulos/efeitos dos fármacos , Dados de Sequência Molecular , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas/química , Plantas/genética , Plastídeos/metabolismo , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Tubulina (Proteína)/genética , Tubulina (Proteína)/imunologiaRESUMO
Xyloglucan is a key polymer in the walls of growing plant cells. Using split pea stem segments and stem segments from which the epidermis had been peeled off, we demonstrate that the integration of xyloglucan mediated by the action of wall-bound xyloglucan endotransglycosylase suppressed cell elongation, whereas that of its fragment oligosaccharide accelerated it. Whole xyloglucan was incorporated into the cell wall and induced the rearrangement of cortical microtubules from transverse to longitudinal; in contrast, the oligosaccharide solubilized xyloglucan from the cell wall and maintained the microtubules in a transverse orientation. This paper proposes that xyloglucan metabolism controls the elongation of plant cells.