Biomedical subjects

K Sandau

Publications and source records attributed to K Sandau.

16 recordsLinked to original sources

PubMed1998

Nitrosative and oxidative stress induced heme oxygenase-1 accumulation in rat mesangial cells.

The formation of nitric oxide (NO.) and superoxide (O2-) promotes rat mesangial cell death. Apoptotic death is characterized by DNA fragmentation, caspase-3 activation and concomitant poly(ADPribose) polymerase cleavage, as well as accumulation of the tumor suppressor protein p53. In close association with apoptotic parameters we noticed upregulation of heme oxygenase by the NO donor S-nitrosoglutathione (GSNO) and the redox cycler 2,3-dimethoxy-1,4-naphthoquinone (DMNQ) in a time- and concentration-dependent manner. In response to the NO. donor, heme oxygenase-1 expression was more easily obtained than initiation of apoptosis. Radical (NO./O2-) cogeneration abrogated DNA fragmentation, suppressed caspase activation and lowered p53 accumulation, thereby promoting cell survival of mesangial cells. In contrast, heme oxygenase-1 expression remained elevated under conditions of GSNO/DMNQ coadministration. Conclusively, heme oxygenase-1 is a stress marker for both nitrosative and oxidative stress. Accumulation of heme oxygenase-1 is found under conditions of both, apoptotic cell death and cell survival, thereby questioning a specific cytoprotective role of heme oxygenase-1 under conditions of NO. and/or O2- formation in rat mesangial cells.

Animals↗

PubMed1998

Automated evaluation of angiogenic effects mediated by VEGF and PlGF homo- and heterodimers.

The effects of growth factors on the blood vessel pattern of chick chorioallantoic membrane (CAM) were assessed with a fast and automated method (extended counting method, XCM; Sandau, 1996) that measures complexity, without assumptions about a fractal structure. XCM is a reliable measure of complexity not only in theory but also in practice: (1) it is robust with respect to thresholding; (2) it shows reduced variance due to pattern translation and rotation; (3) its properties come close to requirements of fractal geometry. It hence is superior to established fractal methods for distinguishing effects induced by various isoforms of vascular endothelial growth factor (VEGF121 and VEGF165), placenta growth factor (PlGF) isoforms, and control treatment. We here show that VEGF homo- and heterodimers and VEGF121/PlGF1 heterodimers increase vascular complexity, whereas PlGF1 and PlGF2 are not effective. PlGF1 and VEGF121 did not mutually influence each other when applied in adjacent fields on the same CAM. Since blood vessels in the CAM originate via nonfractal growth processes, their growth should be analyzed accordingly.

Amnion↗

PubMed1998

Apoptotic cell death and nitric oxide: activating and antagonistic transducing pathways.

Nitric oxide (NO) is a unique diffusible molecular messenger that occupies central roles in mammalian pathophysiology. Overproduction of NO is important for nonspecific "host" defense, helping to kill tumors and intracellular pathogens. Cytotoxicity as a result of long-lasting NO generation is now established to initiate apoptosis. Apoptotic cell death defines morphological alterations and distinctive biochemical events that lead to cell demise. NO-mediated apoptosis comprises upregulation of the tumor suppressor protein p53, activation of proteases known as caspases, chromatin condensation, DNA laddering, and is associated with alterations in the expression of apoptotic associated proteins that belong to the Bc1-2 family. An active role of NO was established by blocking adverse effects by NO-synthase inhibitors. Overexpression of the classical antiapoptotic protein Bc1-2 rescued cells from apoptosis by attenuating signaling downstream of p53 and upstream of caspase activation. Accumulating evidence suggests that transducing mechanisms can intersect and therefore a cell response to a given stimulus may alter significantly. As a result, transducing pathways of NO are not only adapted to cytotoxicity but also refer to cell protection. Protection from NO-elicited apoptosis may result as a consequence of a diffusion controlled NO/O2- (superoxide) interaction. The NO/O2- interaction redirects the apoptotic initiating activity of radicals (NO or O2-) towards protection as long as reduced glutathione compensates the resultant oxidative stress. Further, NO-mediated protective principles are understood on the basis of gene transcription of protective proteins such as heat shock proteins, hemeoxygenase-1, or cyclooxygenase-2 that attenuate cell injury in a cell specific way. The crosstalk between destructive and protective principles as a result of NO formation will determine the role of NO in cell injury. The balance between pro- and anti-apoptotic signaling mechanisms, their activation or deactivation as a result of NO formation, will allow cells to cope with NO or to exit into apoptosis.

Animals↗

PubMed1997

The balance between nitric oxide and superoxide determines apoptotic and necrotic death of rat mesangial cells.

Nitric oxide (NO.) and superoxide (O2-) are inflammatory mediators. Their formation seems to be associated with apoptotic and/or necrotic cell death in diseases such as mesangioproliferative glomerulonephritis in which the early phase of mesangiolysis is linked to significant NO. production. Notably, mesangial cells (MC) not only generate NO. but also O2- after cytokine stimulation. Here we investigated the interrelation between NO. and O2- in MC death by generating both radicals with the use of NO donors (S-nitrosoglutathione, spermine-NO) and O2(-)-generating systems (2,3-dimethoxy-1,4-naphtoquinone, hypoxanthine/xanthine oxidase). Exogenously supplied NO. or O2- in a concentration-dependent manner induced apoptosis and/or necrosis. Apoptosis is characterized by chromatin condensation and DNA fragmentation in contrast to necrotic cytoplasmatic membrane rupture. Noteworthy, coincubation of NO. and O2- was cross-protective. Maximum protection required the existence of a balanced NO./O2- ratio. Analysis in cytokine-stimulated MC suggests endogenous radical formation, which may participate in modulating apoptosis. Manipulation of the endogenous NO./O2- ratio by exogenous, sublethal S-nitrosoglutathione in addition to cytokines produced death, which was antagonized by inducible nitric oxide synthase (iNOS) inhibition. Moreover, pyrrolidine dithiocarbamate supplementation, which down-regulates iNOS expression and blocks superoxide dismutase activity, initiates apoptosis. Our results imply the participation of reactive nitrogen and oxygen species in determining life and death of MC.

Animals↗

PubMed1997

Superoxide formation and macrophage resistance to nitric oxide-mediated apoptosis.

RAW 264.7 macrophages, when challenged with a combination of lipopolysaccharide (10 microg/ml) and interferon-gamma (100 units/ml), respond with endogenous NO. formation, which ultimately results in apoptotic cell death. Apoptosis is detected morphologically by chromatin condensation. Concomitantly we noticed the accumulation of the tumor suppressor protein p53. NO.-derived apoptosis was blocked by the NO.-synthase inhibitor NG-monomethyl-L-arginine. Repetitive treatment of RAW 264.7 macrophages with lipopolysaccharide/interferon-gamma, followed by subculturing viable cells, allowed us to select resistant macrophages which we called RES. RES cells still produced comparable amounts of nitrite/nitrate in response to agonist treatment but showed no apoptotic markers, i.e. chromatin condensation or p53 accumulation. However, RES macrophages undergo apoptosis in the presence of exogenously supplied NO., released from the NO-donors S-nitrosoglutathione or spermine-NO. Assessment of cytochrome c reduction established that RES cells released twice the amount of superoxide compared to RAW 264.7 macrophages under both resting and stimulated conditions. We linked increased superoxide production to cellular macrophage resistance by demonstrating decreased apoptosis after simultaneous application of S-nitrosoglutathione or spermine-NO and the redox cycler 2,3-dimethoxy-1,4-naphthoquinone. Our results suggest that macrophage resistance toward NO.-mediated apoptosis is, at least in part, due to increased superoxide formation. Therefore, the balance between reactive nitrogen and reactive oxygen species regulates RAW 264.7 macrophage apoptosis.

Animals↗

PubMed1997

On the bifurcation of blood vessels--Wilhelm Roux's doctoral thesis (Jena 1878)--a seminal work for biophysical modelling in developmental biology.

Wilhelm Roux's doctoral thesis described the relationship between the angle and diameter of bifurcating blood vessels. We have re-read this work in the light of biophysics and developmental biology and found two remarkable aspects hidden among a multitude of observations, rules and exceptions to these rules. First, the author identified the major determinants involved in vascular development; genetics, cybernetics, and mechanics; moreover, he knew that he could not deal with the genetic and regulatory aspects, and could hardly treat the mechanical part adequately. Second, he was deeply convinced that the laws of physics determine the design of organisms, and that a necessity for optimality was inherent in development. We combined the analysis of diameter relationships with the requirement for optimality in a stochastic biophysical model, and concluded that a constant wall-stress condition could define a minimum wall-tissue optimum during arterial development. Hence, almost 120 years after Wilhelm Roux's pioneering work, our model indicates one possible way in which physical laws have determined the evolution of regulatory and structural properties in vessel wall development.

Animals↗

PubMed1997

Nitric oxide and superoxide induced p53 and Bax accumulation during mesangial cell apoptosis.

During proliferative glomerulonephritis, the early phase of mesangiolysis is linked to increased nitric oxide (NO) production. NO. as well as superoxide (O2-) are inflammatory mediators that are generated by mesangial cells (MC) after cytokine stimulation. Added individually, both radicals induce MC apoptosis. However, the co-existence of a defined NO./O2- ratio is cross-protective. Apoptosis is characterized by specific features such as chromatin condensation, DNA strand breaks, and the occurrence of apoptotic regulating proteins. The tumor suppressor p53 and Bax (Bcl-2 associated protein x) are considered to be classical death promotors, which accumulate after toxic insults. To study p53 and Bax protein accumulation in NO. and/or O2(-)-induced apoptosis, we used the NO-donor S-nitrosoglutathione (GSNO) and the redox cycler 2,3-dimethoxy-1,4-naphtoquione (DMNQ). Both agonists initiated DNA fragmentation in a concentration dependent manner associated with transient p53 and Bax up-regulation. Co-generation of NO./O2- resulted not only in reduced DNA fragmentation, but also in decreased Bax accumulation. Comparable to the NO./O2- co-generation, cytokines failed to induce apoptosis. In contrast, cytokines in combination with pyrrolidine dithiocarbamate, which blocks endogenous superoxide dismutase, allowed p53 and Bax accumulation as well as DNA fragmentation. Our results demonstrate p53 and Bax as early components in NO. and O2(-)-induced rat MC apoptosis and point to the NO./O2- interaction as a naturally occurring cell defense mechanism.

Animals↗

PubMed1997

Measuring fractal dimension and complexity--an alternative approach with an application.

Fractal dimension has often been applied as a parameter of complexity, related to, for example, surface roughness, or for classifying textures or line patterns. Fractal dimension can be estimated statistically, if the pattern is known to be self-similar. However, the fractal dimension of more general patterns cannot be estimated, even though the concept may be retained to characterize complexity. We here show that the usual statistical methods, e.g. the box counting method, are not appropriate to measure complexity. A recently developed approach, the extended counting method, whose properties are closer to what fractal dimension means, is considered here in more detail. The methods are applied to geometric and to blood vessel patterns. The weak assumptions about the structure, and the lower variance of the estimate, suggest that the extended counting method has beneficial properties for comparing complexity of naturally occurring patterns.

Allantois↗

PubMed1996

Nitric oxide donors induce apoptosis in glomerular mesangial cells, epithelial cells and endothelial cells.

Renal mesangial cells exposed to inflammatory cytokines produce high concentrations of nitric oxide (NO) which may exert cytotoxic actions. We report here that glomerular mesangial cells, endothelial cells and epithelial cells in culture are themselves targets for NO and undergo apoptotic cell death upon exposure to high concentrations of NO. NO generated from different NO-releasing compounds as well as NO-saturated solution induce apoptosis in all three cell types as demonstrated by internucleosomal DNA fragmentation, an enrichment of cytosolic DNA/histone complexes, an increasing number of cellular 3'-OH-fragmented DNA ends and typical nuclear chromatin condensation. Induction of apoptosis was found to be dependent on protein synthesis and is preceded by expression of the tumour suppressor gene product p53 in mesangial cells. Induction of inducible NO synthase in mesangial cells by interleukin-1 beta leads to excessive formation of NO by the cells as measured by nitrite production. However, there was no evidence for apoptotic changes in mesangial cells triggered by endogenously produced NO. Co-cultures of glomerular endothelial or epithelial cells with interleukin-1 beta-activated mesangial cells expressing inducible NO synthase do not show apoptotic alterations in endothelial or epithelial cells. Moreover, preincubation of mesangial cells with interleukin-1 beta protects the cells from apoptosis induced by subsequent addition of exogenous NO thus suggesting that interleukin-1 beta not only triggers the expression of inducible NO synthase and massive NO formation but simultaneously stimulates a protecting principle in the cells. In summary, these results suggest that exogenous NO can induce apoptosis in all three types of intrinsic glomerular cells. However, whether endogenously produced NO can fulfil this function critically depends on a balance between a yet to be defined protective mechanism and inducible NO synthase expression in mesangial cells in response to interleukin-1 beta and eventually other inflammatory cytokines.

Animals↗

PubMed1996

Apoptosis is triggered by the cyclic AMP signalling pathway in renal mesangial cells.

Glomerular mesangial cells are regarded as specialized smooth muscle cells located within the renal glomeruli and fulfilling important functions in glomerular physiology and pathophysiology. Here, we report that activation of the cyclic AMP signalling pathway by dibutyryl cyclic AMP, forskolin, or the beta 2-adrenergic receptor agonist salbutamol results in induction of apoptosis in mesangial cells. Activation of the apoptotic programme results in DNA fragmentation which is visible for most forms of apoptosis and is paralleled by enrichment of cytosolic DNA/histone complexes, an increasing number of cellular 3'-OH-fragmented DNA ends and typical nuclear chromatin condensation. Induction of apoptosis was found to be dependent on translation and independent of nitric oxide synthase activity.

Adrenergic beta-Agonists↗

PubMed1996

The dual role of S-nitrosoglutathione (GSNO) during thymocyte apoptosis.

Nitric oxide (NO) is known to regulate redox-sensitive signalling pathways in physiology and pathophysiology. Depending on its concentration, the NO-releasing compound S-nitrosoglutathione (GSNO) causes negative and positive regulation of thymocyte apoptosis. At levels below 0.6 mM, GSNO produces deoxyribonucleic acid (DNA) laddering, which is inhibited by activation of protein kinase C (PKC), cycloheximide treatment, and calcium chelation. Higher concentrations of the NO donor (1-2 mM) suppress thymocyte apoptosis initiated by the classical agonist dexamethasone. Inhibition of apoptosis by NO is analogous to the action of the thiol-blocking compound N-ethylmaleimide (NEM) and the glutathione-S-transferase substrate 1-chloro-2,4-dinitrobenzene (CDNB). Inhibition of apoptosis results from thiol modification of critical proteins in response to NO treatment. Depending on the concentration, GSNO can be involved either in toxic or in protective signalling in thymocyte biology.

Animals↗

PubMed1995

The role of nitric oxide in cell injury.

Nitric oxide (NO) is a pathophysiological mediator with unique signal transducing properties. Signaling mechanisms are categorized as cGMP-dependent or cGMP-independent. Multiple interactions of NO with oxygen, superoxide, and transition metals determine the biological activity. Cyclic GMP-independent responses of NO account for the antimicrobial, the cytostatic, and the cytotoxic capacity of NO. Cytotoxicity is not only directed to harmful cells but also affects the NO-producing cell in a self-destructing loop. For macrophages and pancreatic beta-cells (RINm5F), we established NO-mediated apoptotic cell death. Endogenously generated or exogenously applied NO causes DNA cleavage after endonuclease activation. NO-mediated accumulation of the tumor suppressor p53 precedes apoptotic cell death.

Animals↗

PubMed1994

Modelling of vascular growth processes: a stochastic biophysical approach to embryonic angiogenesis.

In a computer simulation, growth of a capillary network is driven by a stochastic process on a planar hexagonal grid. Starting at a point source, the probabilities for the formation of new capillary elements depend on local biophysical knowledge. This knowledge is mainly derived from the flow theorem of Hagen-Poiseuille and the diameter exponent delta. The hexagonal grid is visualized as being supported by a cylinder or a sphere. An arterial tree results from the adaptive diameter augmentation, and is considered to have limited fractal properties. The dimension of its border, and the time course of growth and of blood pressure are compared with biological data from the chorioallantoic membrane (CAM) of incubated chicken eggs. The model is discussed in view of mechanosensitivity and cell-matrix interactions of endothelial cells, and CAM haemodynamics.

Allantois↗

PubMed1992

An approach based on two-dimensional graph theory for structural cluster detection and its histopathological application.

An approach based on graph theory is described for detecting clusters of cells in tissue specimens (two-dimensional space). With a set of discrete basic elements (cell nuclei) having several measurable features (area, surface, main and minor axis of best-fitting ellipses) a graph is defined as having attributes associated with edges. Different minimum spanning trees (MSTs) can be constructed using different weight functions on the attributes (attributed MST). Analysis of the MST and of an attributed MST by use of a decomposition function allows detection of image areas with similar local properties. These clusters, which are then clusters of the tree, describe, for example, partial growth in different directions in a case of a human fibrosarcoma assuming that tumour cell nuclei are homogeneous with respect to their configuration and size. The model allows the separation of clusters of tumour cells growing in different directions and the approximation of the different growth angles. This decomposition also allows us to create new (higher) orders of structure (cluster tree).

Algorithms↗

PubMed1992

Measuring the surface area of a cell by the method of the spatial grid with a CSLM--a demonstration.

The spatial grid is a method for estimating the surface area of particles. A stack of perfectly registered sections is the essential prerequisite for its use. The confocal scanning light microscope provides such a stack by optical sectioning. The spatial grid method is briefly described and applied to an osteocyte lacuna in dry mineralized human mandible. This type of cell was chosen because of its very complex shape. The variance of the area estimate is studied and compared with the results of a simulation.

Humans↗

PubMed1991

Analysis of soft tissue tumors by an attributed minimum spanning tree.

Histologic slides of 22 soft tissue tumors (9 malignant fibrous histiocytoma, 8 fibrosarcoma, 2 rhabdomyosarcoma, 2 osteosarcoma, 1 Askin tumor) were Feulgen stained. Using an automated image analyzing system (Cambridge 570) at low magnification (25x), the tumor cell nuclei were segmented. The geometrical center of the nuclei was considered the vertex. A basic graph was constructed according to the neighborhood condition of O'Callaghan. Neighboring tumor cell nuclei were visualized by connecting edges. Several features of tumor cell nuclei were measured, including area, surface, major and minor axis of best fitting ellipsis and extinction (DNA content). Nuclear features are attributed to the vertices. The differences, or "distances," between features of connected vertices are attributed to the corresponding edges, which are dependent on the attributes. Thus, different minimum spanning trees (MST) result. Each MST can be decomposed into clusters using a suitable decomposition function on the edges, which rejects an edge if its attributes differ from the mean of the attributed values of surrounding edges more than a neighbor dependent bound (lower limit). Taking into account the length and other attributes of edges (e.g., differences in orientation of the major axis), clusters of different nuclear orientation can be detected. A cluster tree can be constructed by defining the geometric center of a cluster as a new vertex, and by computing the neighborhood of the cluster vertices. The result is an attributed MST containing characteristic structural properties of the image (in cases of sarcomatous tumors, local orientation of tumor cell nuclei and local DNA abnormalities).

Cell Nucleus↗