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T Volk

Publications and source records attributed to T Volk.

At least 55 records · Page 3Linked to original sources

Intracellular Ca2+ dependence of nitric oxide mediated enhancement of interleukin-8 secretion in human endothelial cells.

Nitric oxide (NO.) can induce transient [Ca2+] changes in endothelial cells not different from receptor mediated signalling. Whether this Ca2+ signal may provide a link with IL-8 secretion induced by NO. donors was investigated in human endothelial cells. Sodium nitroprusside (SNP) and S-nitroso-N-acetyl-DL-penicillamine (SNAP) dose dependently increased IL-8 production in this cell type. Additive IL-8 secretion was found with TNFalpha. Buffering intracellular Ca2+ with MAPT/AM suppressed NO. induced [Ca2+]i changes and reduced subsequent IL-8 secretion. The additive effect of both NO. donors on TNFalpha induced IL-8 secretion was completely blocked in the presence of MAPT/AM. SKF 96365, which has been shown to block receptor mediated Ca2+ entry, and TMB-8, which blocks intracellular Ca2+ release, both inhibited IL-8 secretion, particularly when TNFalpha was used as a costimulator, indicating that [Ca2+]i changes are important components of IL-8 induction by NO..

Calcium↗

Nitric oxide induces transient Ca2+ changes in endothelial cells independent of cGMP.

Ca2+ changes induced by nitric oxide (NO.) were investigated in cultured human endothelial cells. Sodium nitroprusside (SNP) (1-100 mumol/L) and S-Nitroso-N-acetylpenicillamine (SNAP) (100 mumol/L) were used as NO. donors. The cytoplasmatic Ca2+ concentration was calculated using ratiometric FURA2 fluorescence measurements. Both NO. donors caused transient oscillatory Ca2+ changes, which were not detectable in the presence of oxyhemoglobin (50 mumol/L). Digital ratio imaging revealed initiation sites within cells where Ca2+ increases started spreading, which indicates that nonuniformly distributed targets might be involved in these reactions. Calcium was released from intracellular stores as indicated by experiments performed in Ca(2+)-free buffer. L-type Ca(2+)-channel blocker diltiazem (100 mumol/L) was not able to block these responses. NO.-induced Ca2+ release from intracellular stores caused capacitative Ca2+ entry. Both thapsigargin (1 mumol/L) and cyclopiazonic acid (10 mumol/L) inhibited the SNP response completely, whereas neither ryanodine (up to 100 mumol/L) nor dantrolene (100 mumol/L) was able to inhibit Ca2+ changes induced by SNP, indicating that primarily inositol 1,4,5-triphosphate (IP3)-dependent stores are released upon stimulation with NO.. A small inhibitory effect of ATP- and SNP-induced peak [Ca2+]i increase was measured in the presence of both caffeine (20 mmol/L) and procaine (1 mmol/L). Evidence is presented that cGMP is not involved in NO.-induced Ca2+ signals, as neither inhibitors of guanylate cyclase (methylene blue and LY 83583) nor cell permeant analogues of cGMP altered or simulated [Ca2+] changes. An inhibitor of cGMP-dependent protein kinase was also ineffective. We therefore propose that endothelial cells have specific targets proximal or at IP3 receptors to induce Ca2+ changes in endothelial cells stimulated with NO..

Adenosine Triphosphate↗

Reliability audit of a regional cardiac surgery registry.

OBJECTIVE: The British Columbia Provincial Cardiac Registry collects demographic and clinical data on all patients who undergo cardiac surgery procedures in the province. The purpose of this study was to compare the reliability of data contained in Registry with data contained in hospital charts. METHODS: Registry and hospital charts were compared for 480 cases. Thirty cases were randomly selected for the province's 16 cardiac surgeons. For each case, 10 distinct fields were selected for analysis and classified as consistent, inconsistent, or rejected (data unavailable in one or other source). RESULTS: The overall rate of consistency between charts and the Registry was 86.4%, with an inconsistency rate of 9.9% and a rejection rate of 3.7%. Consistency rates varied significantly across the 10 fields and among the 16 surgeons. Pairwise comparisons of rates between fields indicated that specific field types were problematic and should be targeted for improvement. In addition, pairwise comparisons of rates between surgeons indicated that further education on Registry use is required. CONCLUSIONS: Recommendations for database design and management include provision of standard definitions for all fields; education of users; extension of the number of mandatory fields; revision of check-off box fields to yes/no/unsure fields; and collection of data close to the time that it is generated.

British Columbia↗

Transient Ca2+ changes in endothelial cells induced by low doses of reactive oxygen species: role of hydrogen peroxide.

Cultured human and rat endothelial cells were used to study cellular toxicity and Ca2+ signalling upon exposure to reactive oxygen species. Superoxide and hydrogen peroxide (O2.-/H2O2) were produced by the hypoxanthine/xanthine oxidase system (HX/XO) and caused intracellular Ca2+ concentration ([Ca2+]i) to rise steadily when activities above 2 mU/ml were used. These Ca2+ increases were also measured when the glucose/glucose oxidase (G/GO) system above 5 mU/ml was used to produce hydrogen peroxide (H2O2). Gross morphological changes appeared to parallel elevated [Ca2+]i levels preceding cell death. However, when HX/XO or G/GO were used at non toxic doses rapid and transient changes in [Ca2+]i were measured. These treatments did not alter subsequent receptor mediated Ca2+ signalling induced by ATP (10 microM) or histamine (100 microM). Superoxide dismutase (50 U/ml), which dismutates O2.- into H2O2 also had no influence, whereas catalase (50 U/ml), which removes H2O2, completely diminished transient [Ca2+]i responses. H2O2 added directly was able to induce similar Ca2+ transients when concentrations of at least 500 microM were used. Buffering trace amounts of iron (o-phenanthroline; 200 microM) in order to inhibit .OH radical formation was not effective to alter Ca2+ changes. Experiments performed in Ca(2+)-free buffer showed a similar rise in [Ca2+]i and readdition of Ca2+ to the extracellular medium indicated the activation of store operated Ca2+ entry. Blocking Ca(2+)-ATPases of the endoplasmatic reticulum with thapsigargin (1 microM) inhibited ROS induced transient increases and cells preincubated with pertussis toxin (200 nM) showed unchanged Ca2+ transients after exposure to both enzyme systems. Phospholipase C inhibitor U73122 (2 microM) effectively reduced hydrogen peroxide induced emptying of intracellular stores. Taken together, we demonstrate that enzymatically produced non-toxic H2O2 rather than O2.- or .OH causes calcium signalling from thapsigargin sensitive stores, and activates store operated Ca2+ entry at least partially by activating phospholipase C. These changes clearly differ from pathological 'oxidative stress' associated with a progressive increase in [Ca2+]i.

Animals↗

Reciprocal signaling between Drosophila epidermal muscle attachment cells and their corresponding muscles.

Directed intercellular interactions between distinct cell types underlie the basis for organogenesis during embryonic development. This paper focuses on the establishment of the final somatic muscle pattern in Drosophila, and on the possible cross-talk between the myotubes and the epidermal muscle attachment cells, occurring while both cell types undergo distinct developmental programs. Our findings suggest that the stripe gene is necessary and sufficient to initiate the developmental program of epidermal muscle attachment cells. In stripe mutant embryos, these cells do not differentiate correctly. Ectopic expression of Stripe in various epidermal cells transforms these cells into muscle-attachment cells expressing an array of epidermal muscle attachment cell-specific markers. Moreover, these ectopic epidermal muscle attachment cells are capable of attracting somatic myotubes from a limited distance, providing that the myotube has not yet been attached to or been influenced by a closer wild-type attachment cell. Analysis of the relationships between muscle binding and differentiation of the epidermal muscle attachment cell was performed in mutant embryos in which loss of muscles, or ectopic muscles were induced. This analysis indicated that, although the initial expression of epidermal muscle-attachment cell-specific genes including stripe and groovin is muscle independent, their continuous expression is maintained only in epidermal muscle attachment cells that are connected to muscles. These results suggest that the binding of a somatic muscle to an epidermal muscle attachment cell triggers a signal affecting gene expression in the attachment cell. Taken together, our results suggest the presence of a reciprocal signaling mechanism between the approaching muscles and the epidermal muscle attachment cells. First the epidermal muscle attachment cells signal the myotubes and induce myotube attraction and adhesion to their target cells. Following this binding, the muscle cells send a reciprocal signal to the epidermal muscle attachment cells inducing their terminal differentiation into tendon-like cells.

Animals↗

A modified CROPGRO model for simulating soybean growth in controlled environments.

The CROPGRO crop growth model is adapted in order to analyze experimental data from a soybean (cv. Hoyt) experiment conducted at elevated CO2 levels (1200 micromol mol-1) at Kennedy Space Center, FL. The following adaptations to original CROPGRO produced model agreement with gas-exchange data: the input of square-wave temperature and photosynthetically active radiation (PAR) profiles; the input of the appropriate hydroponic substrate PAR albedo; modified biomass partitioning and developmental parameters; an increased leaf area expansion rate through the fifth vegetative node; a decreased specific leaf area after the fifth vegetative node; and an increased incident diffuse PAR fraction over typical field values. The model demonstrated here suggests that with continued development, modified CROPGRO will be a useful tool in the analysis and eventual optimization of legume production in bioregenerative life support systems.

Biomass↗

Phasic temperature control appraised with the Ceres-Wheat model.

Phasic control refers to the specification of a series of different environmental conditions during a crop's life cycle, with the goal of optimizing some aspect of productivity. Because of the enormous number of possible scenarios, phasic control is an ideal situation for modeling to provide guidance prior to experiments. Here we use the Ceres-Wheat model, modified for hydroponic growth chambers, to examine temperature effects. We first establish a baseline by running the model at constant temperatures from 10 degrees C to 30 degrees C. Grain yield per day peaks at 15 degrees C at a value that is 25% higher than the yield at the commonly used 23 degrees C. We then show results for phasic control limited to a single shift in temperature and, finally, we examine scenarios that allow each of the five phases of the life cycle to have a different temperature. Results indicate that grain yield might be increased by 15-20% over the best yield at constant temperature, primarily from a boosted harvest index, which has the additional advantage of less waste biomass. Such gains, if achievable, would help optimize food production for life support systems. Experimental work should first verify the relationship between yield and temperature, and then move to selected scenarios of phasic control, based on model predictions.

Biomass↗

Diffuse light and wheat radiation-use efficiency in a controlled environment.

Radiation-use efficiency (dry matter produced per unit absorbed radiation) of a spring wheat (Triticum aestivum L., cv. Veery-10) was 40% higher in controlled growth chamber experiments than under optimal field conditions. Simulations with CERES-Wheat, a field model modified to account for growth chamber conditions, suggest that the observed increase in radiation-use efficiency was due to the large fraction of diffuse light in the experimental chamber. Under optimal conditions in the field, the highest crop growth rates occur when the daily photosynthetic photon flux (PPF) is at its highest levels (50-60 mol m-2 d-1). However, these high growth rates do not appear to be associated with the highest radiation-use efficiency. High PPF levels in the field occur on clear days when the fraction of direct radiation is high and the diffuse fraction is low. In controlled environments with reflective walls, high PPF levels with a large fraction of diffuse radiation can be obtained. Diffuse radiation penetrates to the lower leaves of a canopy better than direct radiation, with the result that the upper leaves are less light saturated and the lower leaves receive more light, increasing radiation-use efficiency, and thus growth rates. The data and model simulations presented here suggest that when diffuse light is a high fraction of the total PPF crop productivity can exceed the highest values attainable in the field under optimal conditions.

Computer Simulation↗

Heartless, a Drosophila FGF receptor homolog, is essential for cell migration and establishment of several mesodermal lineages.

A Drosophila FGF receptor homolog (DFGF-R2/DFRI) termed Heartless (Htl) is expressed in the embryonic mesoderm. The phenotypes of null mutant embryos demonstrated that Htl is a central player that is required for the development of several mesodermal lineages. No abnormalities in the primary specification of the mesoderm were observed. The first defects were seen as irregular migration and spreading of the mesoderm over the ectoderm. Subsequently, cell fates were not induced in several lineages including the visceral mesoderm, heart, and the dorsal somatic muscles. The defects in the induction of cell fates are likely to result from failure of the mesoderm to spread over the ectoderm and receive patterning signals. The defective spreading could be circumvented in htl mutant embryos by providing an ectopic Dpp patterning signal, leading to the formation of heart and dorsal muscle cells. Htl appears to be required also subsequently during the migration and morphogenesis of the different lineages. Expression of a dominant-negative htl construct after the initial induction of cell fates gave rise to aberrant migration and organization of the visceral mesoderm, heart, and somatic muscles. Thus, a common role for Htl in cell migration and tissue organization may account for the pleiotropic defects of the htl mutation.

Animals↗

Epidermal egr-like zinc finger protein of Drosophila participates in myotube guidance.

We have cloned and molecularly characterized the Drosophila gene stripe (sr) required for muscle-pattern formation in the embryo. Through differential splicing, sr encodes two nuclear protein variants which contain a zinc finger DNA-binding domain in common with the early growth response (egr) family of vertebrate transcription factors. The sr transcripts and their protein products are exclusively expressed in the epidermal muscle attachment cells and their ectodermal precursors, but not in muscles or muscle precursors. The results suggest that sr activity induces a subset of ectodermal cells to develop into muscle attachment sites and to provide spatial cues necessary to orient myotubes along the basal surface of the epidermis to their targeted attachment cells.

Amino Acid Sequence↗

Assessing long-term impacts of increased crop productivity on atmospheric CO2.

A full assessment of the impacts of land clearance and crop production on atmospheric CO2 requires a systems approach. By considering long-term soil carbon changes and fossil fuel energy inputs, we show that increased crop productivity will alleviate CO2 release to the atmosphere primarily by preventing additional land cultivation. Each hectare of cropland undergoing a simulated threefold crop productivity increase here prevents a net release on the order of 150-200 Mg C to the atmosphere over 100 years by avoiding additional land cultivation which would otherwise be required. This effective carbon sink would slowly diminish with time due to fossil fuel energy input requirements. However, future self-containment of the energy needs of high-yield crop production may displace on the order of 1.0 Pg C per year of fossil fuel carbon, in addition to the carbon sink attributable to avoided land cultivation. By avoiding land cultivation, high yield crop systems also preserve natural ecosystems.

Agriculture↗

Considerations in miniaturizing simplified agro-ecosystems for advanced life support.

Miniaturizing the Earth's biogeochemical cycles to support human life during future space missions is the goal of the NASA research and engineering program in advanced life support. Mission requirements to reduce mass, volume, and power have focused efforts on (1) a maximally simplified agro-ecosystem of humans, food crops, and microbes; and, (2) a design for optimized productivity of food crops with high light levels over long days, with hydroponics, with elevated carbon dioxide and other controlled environmental factors, as well as with genetic selection for desirable crop properties. Mathematical modeling contributes to the goals by establishing trade-offs, by analyzing the growth and development of experimental crops, and by pointing to the possibilities of directed phasic control using modified field crop models to increase the harvest index.

Agriculture↗

A Drosophila dystrophin-related protein, MSP-300, is required for embryonic muscle morphogenesis.

Proteins from the spectrin superfamily contribute to cell polarity and shape during the morphogenetic that accompany embryogenesis. Drosophila MSP-300, a member of the spectrin superfamily, is expressed in somatic, visceral and heart embryonic muscles. Cloning and sequence analysis of various spliced forms of MSP-300 reveals functional and structural similarities between MSP-300 and vertebrate Dystrophin, the product of the Duchenne Muscular Dystrophy gene. The identification of a strain mutant for the MSP-300 gene is described. Analysis of the somatic muscle phenotype in MSP-300 mutant embryos suggests that the protein contributes to the integrity of the somatic and visceral muscle during periods of significant morphogenetic change. Functional synergism between MSP-300 and laminin is demonstrated by the analysis of the phenotype of embryos mutant for both genes. The enhancement of aberrant muscle phenotype in the double mutants suggests a link between MSP-300 and laminin function in mediating proper extension of the myotube towards the epidermal muscle attachment site. In addition, both genes function to establish gut integrity. In view of the functional and structural similarities between MSP-300 and Dystrophin, it is postulated that Dystrophin is not only required for proper muscle function in adult life but also contributes to muscle morphogenesis during the development of the vertebrate embryo.

Amino Acid Sequence↗

[Changes in coagulation physiology and rheology after preoperative normovolemic hemodilution].

AIM: In a prospective randomised controlled trial the effect of preoperative normovolaemic haemodilution on coagulation, plasma viscosity and plasma protein levels was examined. METHOD: 50 patients undergoing gastrectomies were investigated (haemodilution group, n = 30; control group, n = 20). In the haemodilution group a haematocrit of 30% was aimed at. Blood was replaced by normovolaemic infusion of 6% hydroxyethyl starch 200/0.5. MAIN RESULTS: Haematocrit, colloid osmotic pressure, total serum protein, serum albumin and platelet count were significantly decreased intra- and postoperatively in the haemodilution group compared with control group (p < 0.01). All of these showed no differences between the two groups on the 7th postoperative day. Global coagulation parameters showed dilutional influences without significant differences between the two groups. Measurements of rheological parameters showed a statistically significant decrease in plasma viscosity in the haemodilution group compared with control group (p < 0.01). Haemodilution led to a marked reduction in the use of homologous blood (1 unit/haemodilution group; 10 units/ control group). The average volume of 6% hydroxyethyl starch 200/0.5 administered per patient was 15.2 ml/kgKM/d (7.6-22.2 ml/kgKM/d) in the haemodilution group and 12.7 ml/ kgKM/d (8.4-17.7 ml/kgKM/d) in the control group. CONCLUSION: Haemodilution induced decreases in plasma coagulation, platelet count and plasma proteins did not cause any functional impairement and may just reflect dilution of these parameters. It seems that infusion of 6% hydroxyethyl starch 200/0.5 in an amount of 10-20 ml/kgKM/d does not result in a relevant decrease in coagulation parameters.

Adult↗

Hypertonicity activates nonselective cation channels in mouse cortical collecting duct cells.

We investigated the effect of cell shrinkage on whole-cell currents of M-1 mouse cortical collecting duct cells. Addition of 100 mM sucrose to an isotonic NaCl bath solution induced cell shrinkage and increased whole-cell currents within 5-10 min by approximately 12-fold. The effect was reversible upon return to isotonic solution and could also be elicited by adding 100 mM urea or 50 mM NaCl. Replacement of bath Na+ by K+, Cs+, Li+, or Rb+ did not significantly affect the stimulated inward current, but replacement by N-methyl-D-glucamine reduced it by 88.1 +/- 1.3% (n = 34); this demonstrates that hypertonicity activates a nonselective alkali cation conductance. The activation was independent of extra- and intracellular Ca2+, but 1 or 10 mM ATP in the pipette suppressed it in a concentration-dependent manner, indicating that intracellular ATP levels may modulate the degree of channel activation. Flufenamic acid (0.1 mM) and gadolinium (0.1 mM) inhibited the stimulated current by 68.7 +/- 5.9% (n = 9) and 32.4 +/- 11.7% (n = 6), respectively, whereas 0.1 mM amiloride had no significant effect. During the early phase of hypertonic stimulation single-channel transitions could be detected in whole-cell current recordings, and a gradual activation of 30 and more individual channels with a single-channel conductance of 26.7 +/- 0.4 pS (n = 29) could be resolved. Thus, we identified the nonselective cation channel underlying the shrinkage-induced whole-cell conductance that may play a role in volume regulation.

Adenine Nucleotides↗

Endothelial damage induced by nitric oxide: synergism with reactive oxygen species.

Interactions of reactive oxygen and nitrogen species to mediate endothelial cell damage were studied in vitro. S-Nitroso-N-acetyl-DL-penicillamine (SNAP), 3-morpholinosydnonimine-N-ethylcarbamide (SIN-1) and sodiumnitroprusside (SNP) were used as NO.-donating agents. The toxicity of SIN-1 (5 mM), which produces both O2-. and NO., was reduced when catalase was added to remove H2O2 whereas superoxide dismutase had a marginal protective influence. Low doses of H2O2 producing enzymes added to low doses of SNAP (1 mM) or SNP (5 mM) substantially increased toxicity. Such damage was absent when catalase was present, but was still seen in the presence of superoxide dismutase. Non toxic doses of KCN (1 mM), antimycin A (1 microM), and rotenone (0.5 microM) in order to increase endogeneously produced reactive oxygen species increased toxic effects by 20-30% (p < 0.05). In our experiments we provide evidence that extracellularly produced H2O2 rather than O2-. enhances toxicity of NO. against endothelial cells. Likewise, endogeneous production of reactive oxygen species may increase toxicity of NO..

Analysis of Variance↗

Laminin is required for heart, somatic muscles, and gut development in the Drosophila embryo.

Laminin, an extracellular matrix glycoprotein, has been implicated in a wide array of biological activities. Its specific roles during development, however, remain to be elucidated. In this report we describe the specific contribution of laminin to histogenesis and organogenesis during Drosophila embryogenesis. In particular, the function of laminin during mesoderm development and morphogenesis was dissected by analyzing embryos deficient in laminin A chain alone or in both laminin A and B2 chains. We show that laminin function is not required for the early phases of mesoderm patterning and morphogenesis in embryos at the extended germ band phase (stage 11). However, laminin function is required for the proper morphogenesis of the heart, somatic mesoderm, and the gut during later stages of embryonic development. In laminin A-deficient embryos, the heart is "broken" as a result of dissociation of the pericardial cells. The somatic myotubes are defective and do not extend properly, and the ventral oblique muscles never reach their proper attachment sites. Finally, during midgut formation, the endoderm fails to undergo the initial columnar polarization in the absence of functional laminin. These studies indicate that the primary function of laminin is to provide structural support for the various cell types, thereby enabling their proper organization into the various organs.

Animals↗