Angiopoietin-1 treatment reduces inflammation but does not prevent ventilator-induced lung injury

Background

Loss of integrity of the epithelial and endothelial barriers is thought to be a prominent feature of ventilator-induced lung injury (VILI). Based on its function in vascular integrity, we hypothesize that the angiopoietin (Ang)-Tie2 system plays a role in the development of VILI. The present study was designed to examine the effects of mechanical ventilation on the Ang-Tie2 system in lung tissue. Moreover, we evaluated whether treatment with Ang-1, a Tie2 receptor agonist, protects against inflammation, vascular leakage and impaired gas exchange induced by mechanical ventilation.

Methods

Mice were anesthetized, tracheotomized and mechanically ventilated for 5 hours with either an inspiratory pressure of 10 cmH₂O (‘low’ tidal volume ∼7.5 ml/kg; LV_T) or 18 cmH₂O (‘high’ tidal volume ∼15 ml/kg; HV_T). At initiation of HV_T-ventilation, recombinant human Ang-1 was intravenously administered (1 or 4 µg per animal). Non-ventilated mice served as controls.

Results

HV_T-ventilation influenced the Ang-Tie2 system in lungs of healthy mice since Ang-1, Ang-2 and Tie2 mRNA were decreased. Treatment with Ang-1 increased Akt-phosphorylation indicating Tie2 signaling. Ang-1 treatment reduced infiltration of granulocytes and expression of keratinocyte-derived chemokine (KC), macrophage inflammatory protein (MIP)-2, monocyte chemotactic protein (MCP)-1 and interleukin (IL)-1β caused by HV_T-ventilation. Importantly, Ang-1 treatment did not prevent vascular leakage and impaired gas exchange in HV_T-ventilated mice despite inhibition of inflammation, vascular endothelial growth factor (VEGF) and Ang-2 expression.

Conclusions

Ang-1 treatment downregulates pulmonary inflammation, VEGF and Ang-2 expression but does not protect against vascular leakage and impaired gas exchange induced by HV_T-ventilation.     

Collection of Gravitropic Effectors from Mucilage of Electrotropically-stimulated Roots of Zea mays L.

We placed agar blocks adjacent to tips of electrotropicallystimulated primary roots of Zea mays. Blocks placed adjacentto the anode-side of the roots for 3 h induced significant curvaturewhen subsequently placed asymmetrically on tips of vertically-orientedroots. Curvature was always toward the side of the root ontowhich the agar block was placed. Agar blocks not contactingroots and blocks placed adjacent to the cathode-side of electrotropicallystimulated roots did not induce significant curvature when placedasymmetrically on tips of vertically-oriented roots. Atomicabsorption spectrophotometry indicated that blocks adjacentto the anode-side of electrotropically-stimulated roots containedsignificantly more calcium than (1) blocks not contacting roots,and (2) blocks contacting the cathode-side of roots. These resultsdemonstrate the presence of a gradient of endogenous Ca in mucilageof electrotropically-stimulated roots (i.e. roots undergoinggravitropic-like curvature). Zea mays, corn, mucilage, root gravitropism, electrotropism     

Gravity perception in decapped roots of Zea mays

Time-lapse photography and light microscopy were used to determine whether or not sedimentation of the newly developed amyloplasts in the apex of Zea mays L. roots occurred at the time when geotropic responsiveness reappears following removal of the cap. All decapped roots exhibiting a geotropic response had some amyloplast sedimentation in the apical cortical cells. Exposing decapped roots to a centrifugal acceleration of 25 g for 4 h showed that amyloplasts of a similar size and development were not displaced within the cytoplasm when this treatment began 12 h after decapping, whereas displacement did occur when the treatment began 24 h after decapping. This finding indicates the occurrence of a change in the physical characteristics of the cytoplasm between 12 h and 24 h after removing of the cap, which allows amyloplast movement and thus restores gravity perception.     

Entry and distribution of aluminum in Zea mays

Summary The electron microprobe X-ray analyzer (microprobe) has been used to determine the mode of entry of aluminum (Al) and its distribution and localization in the corn plant. Microprobe analysis is a non-destructive method allowing for multiple element analysis in the same tissues, cells or cell organelles.Al was found to be precipitated on the surface of the epidermal cells of the root with no penetration into the cortex as long as the root surface remained intact. The root cap was freely permeable and contained the highest concentration of Al. The epidermal layer behind the root cap prevented movement into the cortex and conductive tissue.The penetration of a lateral root through the endodermis, cortex and epidermis provided a channel of entry for Al into the cortex and conducting tissues of both the lateral and main root. Essentially no Al was found in the transition zone and only small quantities were present in the above-ground plant parts.The localization of phosphorus was exactly the same as that of Al. This suggested that there was a precipitation of P by Al. A similar analysis for calcium and phosphorus on control plants did not reveal such a precipitation.The method of sample preparation was critical in retaining and localizing the elements in question and is discussed in that light.Michigan Agricultural Experiment Station Journal Article No. 4269.     

玉米根冠粘胶和铝的结合及有机酸累积

玉米根系分泌较多的植物粘胶 ,用 0 .0 1%中性红溶液使粘胶染成红色。 30 μmol/L的AlCl3 处理根系 1h后 ,不除去粘胶的根尖的铝含量大于除去粘胶的根尖的铝含量。粘胶的铝含量随着处理的铝浓度的增加而增加。用高效液相色谱仪能使粘胶 铝复合物分离成均含糖和铝的两种组分。 5 μmol/L的AlCl3 处理根系 2 4h后 ,粘胶中积累了铝诱导分泌的有机酸。粘胶中含有酸性磷酸酶 ,它的活性随着处理的铝浓度的增加而降低。这些结果证明了植物粘胶能够和铝结合、粘胶液滴能够积累有机酸及磷酸酶的假说     

A Gradient of Endogenous Calcium Forms in Mucilage of Graviresponding Roots of Zea mays

Agar blocks that contacted the upper sides of tips of horizontally-orientedroots of Zea mays contain significantly less calcium (Ca) thanblocks that contacted the lower sides of such roots. This gravity-inducedgradient of Ca forms prior to the onset of gravicurvature, anddoes not form across tips of vertically-oriented roots or rootsof agravitropic mutants. These results indicate that (1) Cacan be collected from mucilage of graviresponding roots, (2)gravity induces a downward movement of endogenous Ca in mucilageoverlying the root tip, (3) this gravity-induced gradient ofCa does not form across tips of agravitropic roots, and (4)formation of a Ca gradient is not a consequence of gravicurvature.These results are consistent with gravity-induced movement ofCa being a trigger for subsequent redistribution of growth effectors(e.g. auxin) that induce differential growth and gravicurvature. Atomic absorption, calcium, corn, gravitropism (root), Zea mays     

Isolation and characterization of Azotobacter chroococcum from the roots of Zea mays

Abstract Bacteria showing rapid growth on a nitrogenfree medium and acetylene-reducing activity were isolated from maize roots collected from agricultural soils in Spain. The isolates were Gram-negative motile rods and were identified as Azotobacter chroococcum . Acetylene-reducing activity and microbial counts were determined on root segments from 7- and 30-day-old plants. Rates obtained were in the range of 0.0053–0.848 nmol C₂H₂· g⁻¹· h⁻¹. Root populations were 1.4–6.0 × 10⁴ micro-organisms · g⁻¹. These results showed that there was an association between A. chroococcum strains and roots of maize planted in some Spanish soils.     

Auxin biosynthesis and metabolism in isolated roots of Zea mays

The synthesis and metabolism of indole-3-acetic acid (IAA) was investigated in isolated roots of corn, Zea mays L. Roots were cultured aseptically in media supplemented with either ¹⁴C-tryptophan or ¹⁴C-IAA. Exogenously supplied IAA is rapidly and completely metabolized by root tissues. The main site in the root for the synthesis of IAA is in the apex. Removal of either the root cap or the quiescent center, or the root cap and the quiescent center from the apex has no effect on the IAA-synthesizing ability of the apex. Subdividing the terminal 2.1 cm of the root into various segments and culturing them separately stimulates IAA synthesis in these isolated root tissues. Roots in culture maintain relatively constant IAA levels, reflecting the precise controls of the level of this hormone.     

Localization and identification of auxin in roots of Zea mays

Summary Roots of 3.5-day-old seedlings of Zea mays cv. Giant White Horsetooth contain an extractable auxin which has chromatographic properties and reactions to chromogenic sprays identical with those of indole-3-acetic acid (IAA). By separating stele from cortex (and root tips) before extraction it was shown that the auxin is localized predominantly in the stele, with little being found in the cortex. Whole roots, isolated cortices and isolated steles accumulate and metabolize exogenously applied IAA-1-¹⁴C. The stelar tissue is distinguished from whole roots and cortical tissue in having a different pattern of IAA metabolism.     

Chlorophyll deficiency delays but does not prevent melanogenesis in barley seed melanoplasts

Protoplasma - Plant melanin is a dark polymerized polyphenolic substance that can by synthesized in seed tissues. Unlike well-defined enzymatic browning reaction leading to melanin synthesis in...     

Mitotic activity and cell elongation in geostimulated roots of Zea mays

Mitotic activity was investigated in the primary meristem of horizontally oriented excised root tips of Zea mays during the first six hours of their georeaction. The only statistically significant change that could be detected in the meristem was a decrease of the length of its upper half. No significant difference in mitotic activity was found between the upper and lower halves of roots kept continuously horizontal for 6 h. Cell proliferation thus seems relatively insensitive to changes in the redistribution of endogenous growth regulators that are believed to occur within the meristem during the onset of geotropism. In the zone of bending proximal to the meristem cell length was significantly greater in the upper half than in either the lower half or in the equivalent position in vertical control roots. Thus, cell elongation seems to be promoted in the upper half of the horizontal root. Thus, The differences in cell length were not accompanied by any change in the proportion of nuclei synthesising DNA in these elongating, non-meristematic cells.     

Hydroxylation reactions in roots of maize (Zea mays L.)

The authors succeeded in demonstrating accumulation of tyrosine and p-coumaric acid in three day-old roots of maize (Zea mays L.) fed with L-phenylalanine and cinnamic acid. Phenylpyruvic acid applied under the same conditions gave rise to phenylalanine, indicating the presence of the corresponding transferase activity. Even simultaneous application of inhibitors of transaminase activity—hydroxylamine and isonicotinic acid hydrazide—did not result in the formation of p-hydroxyphenylpyruvic acid.     

Hydroxylation reactions in roots of maize (Zea mays L.)

The authors succeeded in demonstrating accumulation of tyrosine and p-coumaric acid in three day-old roots of maize (Zea mays L.) fed with L-phenylalanine and einnamie acid. Phenylpyruvic acid applied under the same conditions gave rise to phenylalanine, indicating the presence of the corresponding transferase activity. Even simultaneous application of inhibitors of transaminase activity — hydroxylamine and isonicotinic acid hydrazide — did not result in the formation of p-hydroxyphenylpyruvic acid.     

Morphometric analysis of epidermal differentiation in primary roots of Zea mays

Epidermal differentiation in primary roots of Zea mays was divided into six cell types based on cellular shape and cytoplasmic appearance. These six cell types are: 1) apical protoderm, located at the tip of the root pole and characterized by periclinally flattened cells; 2) cuboidal protoderm, located approximately 230 microns from the root pole and characterized by cuboidal cells; 3) tabular epidermis, located approximately 450 microns from the root pole and characterized by anticlinally flattened cells; 4) cuboidal epidermis, located approximately 900 microns from the root pole and characterized by cuboidal cells having numerous small vacuoles; 5) vacuolate cuboidal epidermis, located approximately 1,500 microns from the root pole and characterized by cuboidal cells containing several large vacuoles; and 6) columnar epidermis, located approximately 2,200 microns from the root pole (i.e., at the beginning of the zone of elongation) and characterized by elongated cells. We also used stereology to quantify the cellular changes associated with epidermal differentiation. The quiescent center and the apical protoderm have significantly different ultrastructures. The relative volume of dictyosomes increases dramatically during the early stages of epidermal differentiation. This increase correlates inversely with the amount of coverage provided by the root cap and mucilage.     

Inhibition of gravitropism in primary roots of Zea mays by chloramphenicol

Primary roots of Zea mays seedlings germinated and grown in 0.1 mM chloramphenicol (CMP) were significantly less graviresponsive than primary roots of seedlings germinated and grown in distilled water. Elongation rates of roots treated with CMP were significantly greater than those grown in distilled water. Caps of control and CMP-treated roots possessed extensive columella tissues comprised of cells containing numerous sedimented amyloplasts. These results indicate that the reduced graviresponsiveness of CMP-treated roots is not due to reduced rates of elongation, the absence of the presumed gravireceptors (i.e., amyloplasts in columella cells), or reduced amounts of columella tissue. These results are consistent with CMP altering the production and/or transport of effectors that mediate gravitropism.