A spatially dynamic cohort of regulatory genes in the endomesodermal gene network of the sea urchin embryo

A gene regulatory network subcircuit comprising the otx, wnt8, and blimp1 genes accounts for a moving torus of gene expression that sweeps concentrically across the vegetal domain of the sea urchin embryo. Here we confirm by mutation the inputs into the blimp1cis-regulatory module predicted by network analysis. Its essential design feature is that it includes both activation and autorepression sites. The wnt8 gene is functionally linked into the subcircuit in that cells receiving this ligand generate a β-catenin/Tcf input required for blimp1 expression, while the wnt8 gene in turn requires a Blimp1 input. Their torus-like spatial expression patterns and gene regulatory analysis indicate that the genes even-skipped and hox11/13b are also entrained by this subcircuit. We verify the cis-regulatory inputs of even-skipped predicted by network analysis. These include activation by β-catenin/Tcf and Blimp1, repression within the torus by Hox11/13b, and repression outside the torus by Tcf in the absence of Wnt8 signal input. Thus even-skipped and hox11/13b, along with blimp1 and wnt8, are members of a cohort of torus genes with similar regulatory inputs and similar, though slightly out-of-phase, expression patterns, which reflect differences in cis-regulatory design.     

Incidence of lameness and abrasions in piglets in identical farrowing pens with four different types of floor

Background  

Lameness in piglets is a major animal welfare issue. Floor abrasiveness is a common cause of superficial injury in piglets in farrowing pens. The abrasion achieved may act as a gate for infections, which in turn may induce development of infectious arthritis. In this study, the influence of improvements of the floor quality and of increased ratios of straw in identical farrowing pens was measured.     

Composite organization of the cobalamin binding and cubilin recognition sites of intrinsic factor

Intrinsic factor (IF(50)) is a cobalamin (Cbl)-transporting protein of 50 kDa, which can be cleaved into two fragments: the 30 kDa N-terminal peptide IF(30) and the 20 kDa C-terminal glycopeptide IF(20). Experiments on binding of Cbl to IF(30), IF(20), and IF(50) revealed comparable association rate constants (k(+)(Cbl) = 4 x 10(6), 14 x 10(6), and 26 x 10(6) M(-1) s(-1), respectively), but the equilibrium dissociation constants were essentially different (K(Cbl) = 200 microM, 0.2 microM, and 相似文献   

Studies on lytic activities of Chondrococcus coralloides (Myxobacteriales)

Summary When cells of Chondrococcus coralloides were grown on different bacteria and yeast as the only substrates, a pronounced lysis zone in the agar was observed after 3 days of incubation. Gelatine was liquified by the growing organism after 12 h. Enzyme preparations from the culture medium of Chondrococcus lysed only cells from Micrococcus lysodeikticus, at reasonable rates, and they were very active against casein and N--benzoyl-l-arginine-ethylester · HCl. The lytic activity in the culture medium decreased rapidly after the culture reached the stationary growth phase. A fifty-fold purification of the bacteriolytic activity was achieved by step-wise acetone precipitation. Chromatography of this preparation on Sephadex G-100 revealed the presence of four fractions, one proteolytic, two bacteriolytic and one fraction with both bacteriolytic and proteolytic activity. The bacteriolytic activity had a pH-optimum of pH 8, a stability optimum at pH 8 and was more stable in the alkaline region than in the acid. EDTA and Mg⁺⁺ inhibited the lysis. The proteolytic activity against N--benzoyl-l-arginine-ethylester · HCL had a pH-optimum of pH 7. The K _mof this reaction was 6.5×10^-4.

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Zusammenfassung Bei der Anzucht von Chondrococcus coralloides auf Agar, der diverse Bakterien und Hefen als das einzige Nährsubstrat enthielt, wurde nach 3 Tagen Kulturdauer ein deutlicher Lysishof sichtbar; Gelatine wurde nach 12 Std verflüssigt. Enzympräparationen aus dem Kulturmedium von Chondrococcus lysierten nur Micrococcus lysodeikticus-Zellen mit gut meßbarer Geschwindigkeit, sie waren sehr aktiv gegenüber Casein und N--Benzoyl-l-argininäthylester · HCL. Die lytische Aktivität im Kulturmedium sank sehr stark nach dem Erreichen der stationären Wachstumsphase ab. Eine 50fache Reinigung der bakteriologischen Aktivität konnte durch stufenweise erfolgende Acetonfällung erreicht werden. Eine Chromatographie dieser Präparation an Sephadex G-100 zeigte das Vorliegen von vier Fraktionen, einer proteolytischen, zweier bakteriolytischer und einer Fraktion mit bakteriolytischer und proteolytischer Aktivität. Die bakteriolytische Aktivität hatte ein pH-Optimum von pH 8, ein Stabilitätsoptimum von pH 8 und war im alkalischen Bereich stabiler als im sauren. EDTA und Mg⁺⁺ inhibierten die Lysis. Die proteolytische Aktivität gegenüber N--Benzoyl-l-argininäthylester · HCL wies ein pH-Optimum von pH 7 auf. Für diese Reaktion wurde eine K _mvon 6,5 · 10^-4 gefunden.

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Part of a Doctoral Thesis at the Fakultät für Bio- und Geowissenschaften der Universität Karlsruhe.     

Methanol and other VOC fluxes from a Danish beech forest during late springtime

In-canopy mixing ratio gradients and above-canopy fluxes of several volatile organic compounds (VOCs) were measured using a commercial proton transfer reaction mass spectrometer (PTR-MS) in a European beech (Fagus sylvatica) forest in Denmark. Fluxes of methanol were bidirectional: Emission occurred during both day and night with highest fluxes (0.2 mg C m⁻² h⁻¹) during a warm period; deposition occurred dominantly at daytime. Confirming previous branch-level measurements on beech, the forest’s monoterpene emissions (0–0.5 mg C m⁻² h⁻¹), and in-canopy mixing ratios showed a diurnal cycle consistent with light-dependent emissions; a result contrasting temperature-only driven emissions of most conifer species. Also emitted was acetone, but only at ambient temperatures exceeding 20°C. Slow deposition dominated at lower temperatures. Our in-canopy gradient measurements contrast with earlier results from tropical and pine forest ecosystems in that they did not show this beech ecosystem to be a strong sink for oxygenated VOCs (OVOCs). Instead, their gradients were flat and only small deposition velocities (<0.2 cm s⁻¹) were observed to the onsite soil. However, as methanol soil uptake was consistent and possibly related to soil moisture, more measurements are needed to evaluate its soil sink strength. In turn, as canopy scale fluxes are net fluxes with stomatal emissions from photosynthesizing leaves potentially affecting non-stomatal oxygenated VOC uptake, only independent, controlled laboratory experiments may be successful in separating gross fluxes.     

Angiogenic synergism,vascular stability and improvement of hind-limb ischemia by a combination of PDGF-BB and FGF-2

The establishment of functional and stable vascular networks is essential for angiogenic therapy. Here we report that a combination of two angiogenic factors, platelet-derived growth factor (PDGF)-BB and fibroblast growth factor (FGF)-2, synergistically induces vascular networks, which remain stable for more than a year even after depletion of angiogenic factors. In both rat and rabbit ischemic hind limb models, PDGF-BB and FGF-2 together markedly stimulated collateral arteriogenesis after ligation of the femoral artery, with a significant increase in vascularization and improvement in paw blood flow. A possible mechanism of angiogenic synergism between PDGF-BB and FGF-2 involves upregulation of the expression of PDGF receptor (PDGFR)-alpha and PDGFR-beta by FGF-2 in newly formed blood vessels. Our data show that a specific combination of angiogenic factors establishes functional and stable vascular networks, and provides guidance for the ongoing clinical trials of angiogenic factors for the treatment of ischemic diseases.     

Comparative analysis of cobalamin binding kinetics and ligand protection for intrinsic factor, transcobalamin, and haptocorrin.

Changes in the absorbance spectrum of aquo-cobalamin (Cbl x OH(2)) revealed that its binding to transcobalamin (TC) is followed by slow conformational reorganization of the protein-ligand complex (Fedosov, S. N., Fedosova, N. U., Nex?, E., and Petersen, T. E. (2000) J. Biol. Chem. 275, 11791-11798). Two phases were also observed for TC when interacting with a Cbl-analogue cobinamide (Cbi), but not with other cobalamins. The slow phase had no relation to the ligand recognition, since both Cbl and Cbi bound rapidly and in one step to intrinsic factor (IF) and haptocorrin (HC), namely the proteins with different Cbl specificity. Spectral transformations observed for TC in the slow phase were similar to those upon histidine complexation with Cbl x OH(2) and Cbi. In contrast to a closed structure of TC x Cbl x OH(2), the analogous IF and HC complexes revealed accessibility of Cbl's upper face to the external reagents. The binders decreased sensitivity of adenosyl-Cbl (Cbl x Ado) to light in the range: free ligand, IF x, HC x, TC x Cbl x Ado. The spectrum of TC x Cbl small middle dotAdo differed from those of IF and HC and mimicked Cbl x Ado participating in catalysis. The above data suggest presence of a histidine-containing cap shielding the Cbl-binding site in TC. The cap coordinates to certain corrinoids and, possibly, produces an incapsulated Ado-radical when Cbl small middle dotAdo is bound.     

Closed-Cell Stent-Assisted Coiling of Intracranial Aneurysms: Evaluation of Changes in Vascular Geometry Using Digital Subtraction Angiography

BackgroundStent-assisted coil embolization (SACE) plays an important role in the treatment of intracranial aneurysms. The purpose of this study was to investigate geometrical changes caused by closed-cell design stents in bifurcation and sidewall aneurysms.Methods31 patients with 34 aneurysms underwent SACE with closed-cell design stents. Inflow angle α, determined by aneurysm neck and afferent vessel, and angle between afferent and efferent vessel close to (δ₁), respectively, more remote from the aneurysm neck (δ₂) were graphically determined in 2D angiography projections.ResultsStent assisted coiling resulted in a significant increase of all three angles from a mean value (±SEM) of α = 119° (±6.5°) pretreatment to 130° (±6.6°) posttreatment (P ≤ .001), δ₁ = 129° (±6.4°) to 139° (±6.1°), (P ≤ .001) and δ₂ = 115° (±8.4°) to 126° (±7.5°), (P ≤ .01). Angular change of δ₁ in AcomA aneurysms was significant greater compared to sidewall aneurysms (26°±4.9° versus 8°± 2.3°, P ≤ .05). The initial angle of δ₁ and δ₂ revealed a significantly inverse relationship to the angle increase (δ₁: r = -0.41, P ≤ .05 and δ₂: r = -0.47, P ≤ .01). Moreover, angle δ₁ was significantly higher in unruptured compared to ruptured aneurysms (135°±7.1° versus 103°±10.8°, P ≤ .05).ConclusionStent deployment modulates the geometry of the aneurysm-vessel complex, which may lead to favorable hemodynamic changes more similar to unruptured than to ruptured aneurysms. Our findings also suggest that the more acute-angled aneurysm-vessel anatomy, the larger the angular change. Further studies are needed to investigate whether these changes improve the clinical outcome.