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At least 739 records · Page 41Linked to original sources

Falsely elevated ankle pressures in severe leg ischaemia: the pole test--an alternative approach.

Ankle-branchial pressure indices (ABPIs), measured by sphygmomanometer and Doppler probe, are an accepted index of chronic leg ischaemia. However, tibial artery sclerosis or calcification decreases compliance, producing falsely elevated cuff occlusion pressures. Arterial cannulation is invasive and impractical, but by elevating the foot and measuring the height at which the Doppler signal disappears, ankle systolic pressure in mmHg can be derived. Using an 8MHz Doppler apparatus and calibrated pole, ankle systolic pressures measured by sphygmomanometer and elevation were compared in 49 severely ischaemic legs (40 patients). ABPIs were derived by dividing ankle systolic pressure by brachial pressure. Median (interquartile range) ABPI assessed by sphygmomanometry was 0.46 (0.35-0.56). Median ABPI measured by leg elevation was significantly lower at 0.21 (0.14-0.30), p < 0.0001, Wilcoxon. In 20 patients undergoing in situ vein bypass grafting, direct transducer-derived pressure measurements were obtained. Median ABPI for this method was 0.15 (0.11-0.27). No significant difference was found when compared with ABPIs derived by elevation, median 0.2 (0.13-0.31), p = 0.324, however median ABPI measured by sphygmomanometry was significantly higher at 0.37 (0.27-0.6), p = 0.0008. Correlation of elevation with transducer-assessed pressure measurements (r = 0.88) was closer than with cuff-derived measurements (r = 0.69). Pressures derived by leg elevation provide a more accurate index of severe leg ischaemia than sphygmomanometry, although the technique is limited to assessing pressures of less than approximately 60 mmHg. Falsely elevated ABPIs may underestimate the extent of disease in patients assessed for vascular reconstruction.

Aged↗

The role of the yeast spindle pole body and the mammalian centrosome in regulating late mitotic events.

Centrosomes of vertebrate cells and spindle pole bodies (SPBs) of fungi were first recognized through their ability to organize microtubules. Recent studies suggest that centrosomes and SPBs also have a function in the regulation of cell cycle progression, in particular in controlling late mitotic events. Regulators of mitotic exit and cytokinesis are associated with the SPB of budding and fission yeast. Elucidation of the molecular roles played by these regulators is helping to clarify the function of the SPB in controlling progression though mitosis.

Animals↗

Development of the root pole and cell patterning in Arabidopsis roots.

The root forms at the basal end of an axis that is set up early in embryogenesis, and recent genetic analysis has indicated that auxin transport is required for the formation of the root pole. Drug studies show that auxin transport is also required for the maintenance of the tissue organisation in the seedling root. These studies support existing models for tissue patterning that involve canalised auxin flow. Molecular insights into the mechanism of cell-type specification and patterning has come from the epidermis where positionally controlled cell specification is regulated by a cascade of transcription factors.

Arabidopsis↗

Phosphorylation and spindle pole body localization of the Cdc15p mitotic regulatory protein kinase in budding yeast.

Cdc15p is an essential protein kinase and functions with a group of late mitotic proteins that includes Lte1p, Tem1p, Cdc14p and Dbf2p/Dbf20p to inactivate Cdc28p-Clb2p at the end of mitosis in budding yeast [1] [2]. Cdc14p is activated and released from the nucleolus at late anaphase/telophase to dephosphorylate important regulators of Cdc28p-Clb2p such as Hct1p/Cdh1p, Sic1p and Swi5p in a CDC15-dependent manner [3] [4] [5] [6] [7]. How Cdc15p itself is regulated is not known. Here, we report that both the phosphorylation and localization of Cdc15p are cell cycle regulated. The extent of phosphorylation of Cdc15p gradually increases during cell-cycle progression until some point during late anaphase/telophase when it is rapidly dephosphorylated. We provide evidence suggesting that Cdc14p is the phosphatase responsible for the dephosphorylation of Cdc15p. Using a Cdc15p fusion protein coupled at its carboxyl terminus to green fluorescent protein (GFP), we found that Cdc15p, like its homologue Cdc7p [8] in fission yeast, localizes to the spindle pole bodies (SPBs) during mitosis. At the end of telophase, a portion of Cdc15p is located at the mother-bud neck, suggesting a possible role for Cdc15p in cytokinesis.

Cell Cycle↗

Mitotic exit network controls the localization of Cdc14 to the spindle pole body in Saccharomyces cerevisiae.

Budding yeast Cdc14 phosphatase plays essential roles in mitotic exit. Cdc14 is sequestered in the nucleolus by its inhibitor Net1/Cfi1 and is only released from the nucleolus during anaphase to inactivate mitotic CDK. It is believed that the mitotic exit network (MEN) is required for the release of Cdc14 from the nucleolus because liberation of Cdc14 by net1/cfi1 mutations bypasses the essential role of the MEN. But how the MEN residing at the spindle pole body (SPB) controls the association of Cdc14 with Net1/Cfi1 in the nucleolus is not yet understood. We found that Cdc14-5GFP was released from the nucleolus in the MEN mutants (tem1, cdc15, dbf2, and nud1), but not in the cdc5 cells during early anaphase. The Cdc14 liberation from the nucleolus was inhibited by the Mad2 checkpoint and by the Bub2 checkpoint in a different manner when microtubule organization was disrupted. We observed Cdc14-5GFP at the SPB in addition to the nucleolus. The SPB localization of Cdc14 was significantly affected by the MEN mutations and the bub2 mutation. We conclude that Cdc14 is released from the nucleolus at the onset of anaphase in a CDC5-dependent manner and that MEN factors possibly regulate Cdc14 release from the SPB.

Cell Cycle Proteins↗

Rng2p, a protein required for cytokinesis in fission yeast, is a component of the actomyosin ring and the spindle pole body.

BACKGROUND: An actomyosin-based contractile ring plays a pivotal role in cytokinesis. Despite the identification of many components of the ring, the steps involved in its assembly are unknown. The fission yeast Schizosaccharomyces pombe is an attractive organism in which to study cytokinesis because its cell cycle has been well characterized; it divides by medial fission using an actomyosin ring; and a number of S. pombe mutants defective in actomyosin ring assembly have been isolated. Here, we have characterized one such mutant, rng2. RESULTS: Temperature-sensitive rng2 mutants accumulated F-actin cables in the medial region of the cell but failed to organize the cables into a ring. In rng2-null mutants, only a spot-like structure containing F-actin was detected. The rng2+ gene encodes a protein related to human IQGAP1, a protein that binds actin and calmodulin and is a potential effector for the Rho family of GTPases. Rng2p localized to the actomyosin ring and to the spindle pole body (SPB) of interphase and mitotic cells. Localization of Rng2p to the actomyosin ring but not the SPB required F-actin. Rng2p interacted with calmodulin, a component of the SPB and the actomyosin ring. The rng2 gene showed genetic interactions with three other actomyosin ring assembly mutants, cdc4, cdc12, and rng5. CONCLUSIONS: The S. pombe IQGAP-related protein Rng2p is a component of the actomyosin ring and the SPB and is required for actomyosin ring construction following assembly of F-actin at the division site.

Actomyosin↗

Differential effects of low-frequency rTMS at the occipital pole on visual-induced alpha desynchronization and visual-evoked potentials.

Visual-induced alpha desynchronization (VID) and visual-evoked potentials (VEPs) characterize occipital activation in response to visual stimulation but their exact relationship is unclear. Here, we tested the hypothesis that VID and VEPs reflect different aspects of cortical activation. For this purpose, we determined whether VID and VEPs are differentially modulated by low-frequency repetitive transcranial magnetic stimulation (rTMS) over the occipital pole. Scalp EEG responses to visual stimuli (flashed either to the left or to the right visual field) were recorded for 8 min in six healthy subjects (1) before, (2) immediately following, and (3) 20 min after left occipital rTMS (1 Hz, 10 min). The parameters aimed to reduce cortical excitability beyond the end of the TMS train. In addition, simple reaction times to visual stimulation were recorded (left or right hand in separate blocks). In all subjects, VID was significantly and prominently reduced by rTMS (P = 0.0001). In contrast, rTMS failed to modulate early VEP components (P1/N1). A moderate effect was found on a late VEP component close to manual response onset (P = 0.014) but this effect was in the opposite direction to the VID change. All changes were restricted to the targeted left occipital cortex. The effects were present only after right visual field stimulation when a right hand response was required, were associated with a behavioral effect, and had washed out 20 min after rTMS. We conclude that VID and early VEPs represent different aspects of cortical activation. The findings that rTMS did not change early VEPs and selectively affected VID and late VEPs in conditions where the visual input must be transferred intrahemispherically for visuomotor integration (right visual field/right hand) are suggestive of rTMS interference with higher-order visual functions beyond visual input. This is consistent with the idea that alpha desynchronization serves an integrative role through a corticocortical "gating function."

Adult↗

Evidence for a direct role of nascent basal bodies during spindle pole initiation in the green alga Spermatozopsis similis.

Basal body replication in the naked biflagellate green alga Spermatozopsis similis was analyzed using standard electron microscopy and immunogold localization of centrin, an ubiquitous centrosomal protein, and p210, a recently characterized basal apparatus component of S. similis. Fibrous disks representing probasal bodies appear at the proximal end of parental basal bodies at the end of interphase and development proceeds via a ring of nine singlet microtubules. Nascent basal bodies dock early to the plasma membrane but p210, usually present in basal body-membrane-linkers of S. similis, was already present on the cytosolic basal body precursors. In addition to the distal connecting fiber and the nuclear basal body connectors (NBBC) of the parental basal bodies, centrin was present on the fibrous probasal bodies, in a linker between probasal bodies and the basal apparatus, in the connecting fiber between nascent basal bodies and their corresponding parent, and, finally, a fiber linking the nascent basal bodies to the nucleus. This NBBC probably is present only in mitotic cells. During elongation a cartwheel of up to seven layers is formed, protruding from the proximal end of nascent basal bodies. Microtubules develop on the cartwheel indicating that it temporarily functions as a microtubule organizing center (MTOC). These microtubules and probably the cartwheels, touch the nuclear envelope at both sides of a nuclear projection. We propose that spindle assembly is initiated at these attachment sites. During metaphase, the spindle poles were close to thylakoid-free lobes of the chloroplast, and the basal bodies were not in the spindle axis. The role of nascent basal bodies during the initial steps of spindle assembly is discussed.

Algal Proteins↗

Potential inhibitors of S-adenosylmethionine-dependent methyltransferases. 5. Role of the asymmetric sulfonium pole in the enzymatic binding of S-adenosyl-L-methionine.

The configuration at the asymmetric sulfonium pole of S-adenosyl-L-methionine (SAM) necessary for optimal enzymatic binding and methyl donation has been elucidated in this study. For the transmethylations catalyzed by catechol O-methyltransferase, phenylethanolamine N-methyltransferase, histamine N-methyltransferase, and hydroxyindole O-methyltransferase, it was demonstrated that only the natural (-) enantiomer of SAM was active as a methyl donor. The corresponding (+)-SAM, which was prepared by enzymatic resolution of synthetic (+/-)-SAM, was shown to be inactive as a methyl donor in these enzymatic reactions. The (+)-SAM was found, however, to be a potent inhibitor of each of these enzyme-catalyzed transmethylations. These results suggest that the (+) enantiomer offers a nonproductive configuration for the methyl-transfer reaction itself; however, this configuration fails to hamper enzymatic binding. These results are discussed relative to the geometric requirements necessary for the methyl-transfer reaction and the requirements for enzymatic binding.

Animals↗

Can octupolar molecules be poled by an external electric field?

Octupolar molecules are generally believed to be of potential use in developing nonlinear optical materials owing to the fact that they do not easily form molecular aggregates. This is often put against the conjectured drawback that electric fields have no poling, or ordering, effect for this class of molecules because of the lack of a permanent ground state dipole moment. In this paper, we analyze this notion in some detail and present results from molecular dynamics computer simulations of an ensemble of a prototypical octupolar molecule, the 1,3,5-triamino-2,4,6-trinitrobenzene (TATB) molecule, dissolved in chloroform. It is found that TATB molecules indeed show rather significant dipole moments in solutions because of the dual action of the thermal motions of the atoms and the strong intermolecular interactions. Applied electric fields accordingly show significant effects on the orientations of the molecular dipole moments. We also find that TATB molecules can aggregate because of the strong hydrogen-bonding interactions between the molecules, though they lack a static permanent dipole moment. Thus, the simulation results for TATB molecules in solution present us with a totally different notion about the collective properties of octupolar molecules. Taking account of quantum chemistry results, we found that the collective molecular nonlinear optical (NLO) properties are enhanced after the onset of the electric field, showing significant anisotropic characteristics.

Journal Article↗

Theoretical evidence that more microtubules reach the cortex at the pole than at the equator during anaphase in sea urchin eggs.

Astral microtubules are rapidly elongated during anaphase and telophase in sea urchin eggs. The number of microtubules extending to the cell surface was calculated with a computer. For the calculations, microtubules were assumed to radiate from the astral center uniformly over angles. Although microtubules from two asters freely overlapped around the equator, the number per the unit area, i.e. the surface density, was larger in the polar region than in the equatorial region. The ratio of the theoretically calculated numbers in the two regions was close to the ratio obtained from the ultrastructural observations by Asnes and Schroeder in 1979. When counted in the longitudinal section including two astral centers, the microtubule number was a little larger in the equatorial region than in the polar region. However, the numbers do not represent the surface density because the two-dimensional section contains only a small portion of all the microtubules spreading in the three-dimensional space. The fluorescence image for tubulin, in most cases, provides the microtubule distribution in the longitudinal section. Therefore, from such an image, we cannot judge whether the surface density of astral microtubules is larger at the pole or at the equator.

Anaphase↗

The distal pole complex: a novel membrane domain distal to the immunological synapse.

While much interest has focused on the finding that T cell-antigen presenting cell (APC) interaction induces the recruitment of proteins to the immunological synapse (IS), we have recently discovered that APC binding induces the formation of a novel protein complex distal to the site of T-cell receptor ligation. This 'distal pole complex' (DPC) is important for appropriate T-cell activation, functioning either to remove proteins from the synapse or as a signaling complex in its own right. The first component of the DPC to be identified was CD43, a cell-surface mucin that has been proposed to function as a negative regulator of T-cell signaling. CD43 movement was found to depend on ezrin and moesin, members of the ERM family, which serve to link CD43 and other cargo molecules to the actin cytoskeleton. ERM proteins interact with several other important surface receptors and cytoplasmic signaling molecules, some of which we have identified as additional components of the DPC. Disruption of the DPC leaves early T-cell activation events intact but affects cytokine expression. Here, we review what is currently known about the formation and function of the DPC and speculate on how this novel protein complex serves to facilitate T-cell activation.

Animals↗

Visual learning suppressed by cooling the temporal pole.

Three monkeys were trained to remember colored photographs of objects over delays of 0, 15, 30, and 45 s. Then two pairs of cooling devices were implanted bilaterally over the anterior 9 mm of the temporal lobe. The devices consisted of 3 X 10 mm loops of stainless steel tubing into which cooled methanol could be pumped. One pair (anterior pair) covered the medial part of the temporal tip (area TG), starting at the rhinal sulcus and extending 3 mm laterally. The second pair (posterior pair) was placed 3 mm lateral to the anterior pair, covering the rest of TG and the anterior extreme of the inferotemporal gyri, anterior TE. Cooling either pair of probes produced a deficit at all delays, but the deficit was greater at the longest delays. There was no difference between cooling the anterior pair and cooling the posterior pair except that cooling the anterior pair greatly increased the disruption of recall that is produced by an interfering stimulus. When all four probes were cooled, which suppressed the function of the entire temporal tip, performance dropped to chance at all delays. While under this condition, the animals could not learn new visual discriminations but could perform previously learned visual discriminations. These results are consistent with the suggestion that the temporal pole is the store for the brief anterograde memory that is available to the medial temporal amnesics.

Animals↗

Persistent parainfluenza virus shedding during isolation at the South Pole.

Persistent parainfluenza virus shedding in healthy young adults occurred throughout the 8 1/2-month winter isolation period at Amundsen-Scott South Pole Station during 1978. Two episodes of respiratory illness were observed after 10 and 29 weeks of complete social isolation. Throat swabs collected both routinely, and during each outbreak of respiratory illness, were directly inoculated into cell cultures. Parainfluenza virus types 1 and 3 were recovered from both symptomatic and asymptomatic subjects throughout the winter. No other viruses were obtained by these efforts. The presence of parainfluenza virus in these subjects long after the accepted incubation period for viral upper respiratory illness, and when the introduction of new virus to this community was impossible, suggests its persistence in man.

Animals↗

Accretion rate of cosmic spherules measured at the South Pole.

Micrometeorites are terrestrially collected, extraterrestrial particles smaller than about 1 mm, which account for most of the mass being accreted to the Earth. Compared with meteorites, micrometeorites more completely represent the Earth-crossing meteoroid complex and should include fragments of asteroids, comets, Mars and our Moon, as well as pre-solar and interstellar grains. Previous measurements of the flux of micrometeoroids that survive to the Earth's surface have large uncertainties owing to the destruction of particles by weathering, inefficiencies in magnetic collection or separation techniques, low particle counts, poor age constraint or highly variable concentrating processes. Here we describe an attempt to circumvent these problems through the collection of thousands of well preserved and dated micrometeorites from the bottom of the South Pole water well, which supplies drinking water for the Scott-Amundsen station. Using this collection, we have determined precise estimates of the flux and mass distribution for 50-700-microm cosmic spherules (melted micrometeorites). Allowing for the expected abundance of unmelted micrometeorites in the samples, our results indicate that about 90% of the incoming mass of submillimetre particles evaporates during atmospheric entry. Our data indicate the loss of glass-rich and small stony spherules from deep-sea deposits, and they provide constraints for models describing the survival probability of micrometeoroids.

Antarctic Regions↗

Transcripts of one of two Drosophila cyclin genes become localized in pole cells during embryogenesis.

Cyclins, originally discovered in the eggs of marine invertebrates, are proteins which undergo dramatic cycles of synthesis followed by degradation at the metaphase-anaphase transition of cell division. That they participate in the G2-M transition is supported by the fact that when synthetic cyclin messenger RNAs from clam and sea urchin are microinjected into the G2-arrested oocytes of Xenopus, they induce maturation. The cyclin of fission yeast is the product of the cdc13 gene, which is known to interact with cdc2, a gene required for the entry into mitosis. We have cloned the genes that encode A-type and B-type cyclins from Drosophila melanogaster by virtue of their sequence similarity to oligonucleotides corresponding to conserved regions of the cyclin genes. We show that both genes encode abundant maternal mRNAs, but whereas the cyclin A mRNA is relatively uniformly distributed before cell formation, the cyclin B mRNA becomes localized to the developing pole cells. In larvae, cyclin A is expressed predominantly in brain and imaginal disks, whereas cyclin B transcripts are abundant in testes.

Amino Acid Sequence↗

Distinct nuclear and spindle pole body population of cyclin-cdc2 in fission yeast.

Cyclins, as subunits of the protein kinase encoded by the cdc2 gene are major controlling elements of the eukaryotic cell cycle. The fission yeast Schizosaccharomyces pombe has a B-type cyclin, which is a nuclear protein encoded by the cdc13 gene. Here we demonstrate the presence of two spatially distinct cdc13 cyclin populations in the nucleus of S. pombe, one of which is associated with the mitotic spindle poles. Both populations colocalize with the product of the cdc2 gene (p34cdc2). Treatment of cells with the antimicrotubule drug thiabendazole prevents cyclin degradation and blocks the tyrosine dephosphorylation and activation of cdc2. These results suggest a key regulatory role of the cdc2-cyclin complex in the initiation of mitotic spindle formation and also that mitotic microtubule function is required for cdc2 activation.

CDC2 Protein Kinase↗

Multi-step control of spindle pole body duplication by cyclin-dependent kinase.

Organelles called centrosomes in metazoans or spindle pole bodies (SPBs) in yeast direct the assembly of a bipolar spindle that is essential for faithful segregation of chromosomes during mitosis. Abnormal accumulation of multiple centrosomes leads to genome instability, and has been observed in both tumour cells and cells with targeted mutations in tumour-suppressor genes. The defects that lead to centrosome amplification are not understood. We have recapitulated the multiple-centrosome phenotype in budding yeast by disrupting the activity of specific cyclin-dependent kinase (CDK) complexes. Our observations are reminiscent of mechanisms that govern DNA replication, and show that specific cyclin/CDK activities function both to promote SPB duplication and to prevent SPB reduplication.

Cell Cycle↗