[Effect of ultraviolet irradiation on the vascular bed of the skin].
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Cell Adhesion Molecules↗
Biomedical subjects
M Heckmann
Publications and source records attributed to M Heckmann.
Distribution of desensitization time constants of mouse embryonic-like nicotinic and homomeric GLUR6 glutamate receptor channels.
The distribution of the desensitization time constant, tauD, of two different, molecular well defined ligand gated receptor channels was analyzed: the embryonic-like nicotinic receptor (nAChR) channel of cultured mouse myotubes and recombinant, homomeric GluR6 receptor channels transiently expressed in HEK293 cells. Experiments were performed using outside-out patches in combination with a system for fast application of agonists which allows solution exchange within 0.1 ms. In response to application of saturating concentrations of acetylcholine (ACh) or glutamate (Glu). the peak current was reached in a submillisecond range and decayed monexponentially in the presence of the agonist, due to desensitization. tauD varied from 10 ms to 100 ms with a mean value of 55.0 +/- 22.6 ms (n = 133) in response to pulses of 10(-4) M ACh for embryonic-like nAChR channels and from 2.6 ms to 8.9 ms with a mean value of 5.0 +/- 1.9 ms (n = 35) in response to pulses of 10(-2) M Glu for homomeric GluR6 receptor channels. The reason for the high variability of the time course of desensitization is at present unclear.
Animals↗
Desensitization and resensitization kinetics of glutamate receptor channels from Drosophila larval muscle.
Outside-out patches from wild-type Drosophila larval muscle were exposed to L-glutamate (glu) using a piezo-driven application system. Glu receptor channels opened and desensitized in response to rapid applications of 10 mM glu. Desensitization was fitted with an exponential function with a mean time constant of desensitization (tau d) of 15 ms in response to 10 mM glu. The tau d was concentration dependent and decreased to 6 ms (on average) with 0.7 mM glu and increased again to 12 ms (on average) in response to 0.5 mM glu. Desensitization in response to longer applications of glu was almost complete, but surprisingly, even a 1-ms pulse of 3 mM glu produced about 30% desensitization. In the presence of low glu concentrations, the response to a pulse was reduced and was about halved by preequilibration with 30 microM glu. Recovery from desensitization was not concentration dependent and was fitted with an exponential function with a mean time constant of 150 ms. During recovery the channels rarely opened. Kinetic schemes were fitted to these results, and a circular reaction scheme was found to fit the data best. An important feature of the scheme is desensitization from a lower ligated closed state. This allows substantial desensitization of synaptic receptor channels in response to quantal release of transmitter, in part without opening of the channels. Desensitization reduces the probability of the channels opening in response to a subsequent release for a period of time determined by the rate of recovery from desensitization and might serve as a form of molecular short-term memory.
Animals↗
Adhesion of leukocytes to dermal endothelial cells is induced after single-dose, but reduced after repeated doses of UVA.
Approximately 20-50% of ultraviolet A (UVA) irradiation delivered to the skin surface may reach the human dermal microvascular endothelial cells (HDMEC) that play a pivotal role in cellular inflammatory tissue; however, the pathophysiologic role of HDMEC in UVA-induced skin changes is largely unknown. Based on previous in vivo and in vitro studies revealing UVA-induced expression of endothelial adhesion molecules, we studied isolated HDMEC under various conditions in order to further delineate the impact of UVA on these cells. The expression of cell adhesion molecules was determined by flow cytometry and the resulting changes of stable adhesion of leukocytes to endothelial cells were quantitated for granulocytes, lymphocytes, and monocytes using a newly developed multicellular adhesion assay. Additionally, antibody blocking experiments were performed to delineate the role of individual cell adhesion molecules in UVA-induced leukocyte adherence. High-dose polychromatic UVA (25 J per cm2, maximal emission at 375 nm) induced intercellular adhesion molecule-1 and E-selectin with different kinetics but correlating the adhesion of leukocyte subsets. This effect subsided, however, in the course of 3-6 daily applied UVA doses. Moreover, pro-inflammatory cytokine challenge by tumor necrosis factor-alpha and interleukin-1-alpha resulted in significantly weaker induction of intercellular adhesion molecule-1 and E-selectin in repeatedly UVA-exposed HDMEC. Differential quantitation of peripheral blood derived granulocytes, lymphocytes, and monocytes revealed reduced adhesion particularly of lymphocytes followed by monocytes and granulocytes compared with leukocyte adhesion to nonirradiated but cytokine-stimulated HDMEC. It is concluded that UVA substantially influences endothelial cell adhesion molecules expression and thus directly interferes with leukocyte adhesion to endothelial cells. Divergent UVA-induced effects in this respect can be attributed to the mode of UV exposure as well as to the condition of endothelial cells prior to UVA exposure.
Cell Adhesion↗
Botulinum beyond wrinkles.
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Botulinum Toxins, Type A↗
Simultaneous analysis of peripheral blood granulocytes, lymphocytes, and monocytes adhering to human microvascular endothelial cells.
We describe a sensitive, reliable, and convenient procedure to assay cell adhesion based on flow cytometry which allows quantitative studies of all major peripheral blood leukocytes adhering to microvascular endothelial cells. Using this assay, we show that adhesion of mononuclear cells is significantly increased in the presence of granulocytes and that maximal quantitative adhesion of lymphocytes and monocytes requires 30-40 min compared to 5-10 min required for maximal granulocyte adhesion. Moreover, we demonstrate peripheral blood leukocytes from different healthy individuals displayed considerable variations in efficiency of adhesion to pooled microvascular endothelial cells. The here-described methodology may provide a useful tool to characterize the impact of defined mediators or pharmacological agents on stable cell-cell adhesion of peripheral blood leukocytes to microvascular endothelial cells. Moreover, it may hell to identify interindividual variations in cell adhesion efficiency relevant to cell-mediated immune reactions.
Cell Adhesion↗
[The vascular bed of the skin. Integration site of endogenous and exogenous stimuli].
Human skin is exposed to a great deal of environmental stimulation, which in predisposed individuals may cause specific inflammatory dermatosis. Typically, endothelial activation can be observed as the induced expression of cell adhesion molecules. This allows circulating immune cells to adhere to the endothelium and results in inflammatory, perivascular cell infiltrates in the affected tissue area. While tissue-derived cytokines have been discussed primarily as controlling factors of these inflammatory processes, recent evidence suggests direct endothelial activation by various exogenous impulses. Endothelial adhesion molecule expression has been found to be exclusively responsive to biochemical or biophysical stimulation, such as the concentration of metal ions, oxygen tension, UV radiation or mechanically operating shear stress without appreciable interaction of cytokines. These observations indicate that endothelial cells are capable of responding to exogenous stimulation, or rather transduce both exogenous and endogenous impulses, resulting in an integrative process. In view of this, individual deviations of endothelial cell functions need to be considered in pathophysiological concepts about diseases caused by external provocation, while their clinical relevance must be evaluated.
Cell Adhesion Molecules↗
Activation kinetics of glutamate receptor channels from wild-type Drosophila muscle.
Outside-out patches from wild-type Drosophila larval muscle were exposed briefly to L-Glutamate (Glu) using a piezo-driven application system. Glu in concentrations of 0.1 to 30 mM was applied and the responses to repeated applications of a given concentration were averaged. The peak current, î, and the current rise time, tr, from 0.1 î to 0.9 î were determined from the averages. Half-maximum activation of the channels was reached with approximately 2 mM Glu. î increased proportional to the power n = 3. 5 to n = 5.8 (average of four experiments, n = 4.4) for Glu concentrations between 0.3 and 0.5 mM. tr increased from approximately 0.2 ms at 10 mM Glu to a value of approximately 3.5 ms at 0.2 mM Glu. A linear reaction scheme with five binding steps preceding the channel-opening conformational change is proposed as the kinetic mechanism of channel activation and investigated in computer simulations. A set of rate constants assuming the same affinity for each binding site is found to describe the data better than one assuming positive cooperativity. The results are very similar to those for Glu-gated channels of crayfish and locust muscle, which is evidence for a common kinetic mechanism of these channels.
Animals↗
Kinetics of homomeric GluR6 glutamate receptor channels.
We studied the kinetics of the unedited version of rat GluR6 glutamate (glu) receptor channels, GluR6Q, in outside-out patches using a system for submillisecond solution exchange. Half-maximum activation of the channels was reached with approximately 0.5 microM glu. The maximum slope of the double-logarithmic plot of the peak current versus glu was approximately 1.3, indicating that at least two binding steps are necessary to open the channels. Currents in response to a pulse of 10 microM glu had a short rise time (10-90% of peak current) of approximately 220 microseconds at approximately 20 degrees C. The rise time increased with falling glu concentration, reaching approximately 6.0 ms with 10 microM glu. In the continued presence of glu, the channels desensitized, and this desensitization can be described with a single time constant of approximately 7.0 ms for a pulse of 10 microM glu. The steady-state current in response to a long pulse of 10 microM glu was below 1/280th of the peak current. The time constant of desensitization was found to be independent of concentration between 30.0 and 0.3 microM glu, but to be increased for lower concentrations. After a short pulse of 1 ms duration and 10 or 0.3 microM glu, currents decayed with a time constant of approximately 2.5 ms. Recovery from desensitization after a pulse took approximately 5 s, and the half-time of recovery was approximately 2.2 s. Continuous application of low concentrations of glutamate reduced the peak currents in response to a pulse of 10 microM glu markedly. Fifty percent response reduction was observed in the continuous presence of approximately 0.3 microM glu. Our results for homomeric GluR6 agree with a cyclical reaction scheme developed for completely desensitizing, glu-activated channels on crayfish muscles.
Animals↗
Transforming growth factor-beta controls cell-matrix interaction of microvascular dermal endothelial cells by downregulation of integrin expression.
Transforming growth factor-beta (TGF-beta) is a pleiotropic regulatory factor of tissue remodeling. Angiogenesis, a prerequisite of tissue repair and tissue expansion, is induced by TGF-beta in vivo, while proliferation and migration of cultured endothelial cells are inhibited by TGF-beta. Indirect mechanisms stimulating angiogenesis and modification of TGF-beta effects by cell-matrix interaction have been postulated to account for this paradigm. Because cellular behavior in tissue remodeling is decisively determined by cell-matrix interactions, which in turn is mediated via integrins, we investigated the effect of TGF-beta on matrix-dependent endothelial cell functions. Integrin expression of human dermal microvascular endothelial cells (HDMEC) was measured by Northern blot and fluorescence-activated cell sorter analysis after TGF-beta treatment and correlated to cell-matrix interactions, which were studied in a colorimetric cell attachment assay as well as the Boyden chamber chemotaxis assay. We found a cell-specific downregulation of integrin expression in HDMEC on the level of mRNA as well as on the cell surface. This effect correlated well with the reduction of integrin-dependent cell adhesion to several matrix proteins, in particular to fibronectin. Moreover, TGF-beta decreased fibronectin-induced chemotaxis of HDMEC. Thus, TGF-beta controls cell-matrix interaction of HDMEC by down-regulation of integrin expression. This effect of TGF-beta reflects direct and cell-specific control mechanisms on microvascular cells that may be critical for the coordinated process of angiogenesis requiring a balance of stimulatory and inhibitory factors.
Cell Adhesion↗
Cytological diagnosis of zosteriform skin metastases in undiagnosed breast carcinoma.
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Breast Neoplasms↗
Energy metabolism response to calcium activation in isolated rat hearts during development and regression of T3-induced hypertrophy.
The effect of calcium activation on energy production was investigated in isolated perfused hearts from rats treated with triiodothyronine (T3) during 15 days (0.2 mg/kg/day) and in hearts of rats allowed to recover after T3-treatment during 15 days. Changes in phosphorylated compound concentrations were followed in the isolated hearts perfused with a glucose-pyruvate medium by 31P-NMR spectroscopy, when the external calcium concentration was increased from 0.5-1, 1.5 and 2 mM. As expected, T3-treatment resulted in the hypertrophy of the heart (50% increase in HW/BW) that was nearly reversible 15 days after discontinuation of the treatment. When compared to controls, creatine, phosphocreatine (PCr) and glycogen contents were lower (58, 24 and 17% decrease respectively) in the hypertrophied hearts and higher (10, 14 and 18% respectively) after regression of hypertrophy. Intracellular pH, ATP, inorganic phosphate concentrations and the phosphorylation potential were not altered under T3-treatment and after regression of hypertrophy, while calculated free ADP concentration was lower in hypertrophied hearts (control: 40 +/- 2 microM, T3-treatment: 21 +/- 1 microM, regression: 37 +/- 1 microM). Increasing the calcium concentration induced a similar increase in left ventricular developed pressure in the three groups of hearts, with inorganic phosphate concentration increasing with cardiac work. The PCr concentration slightly decreased while the ATP concentration did not change. In spite of different initial PCr concentrations, the evolutions of PCr and Pi concentrations for each stepwise increase in external calcium were similar in the three groups. It is concluded that, in spite of the well-known decrease in efficiency induced by the drug, the mechanisms of PCr (ATP) production remain able to respond to an acute moderate increase in energy demand provoked by a physiological stimulus. This adaptation also persists after the treatment when the energy metabolism balance is apparently improved.
Adenosine Diphosphate↗
Recordings of glutamate-gated ion channels in outside-out patches from Drosophila larval muscle.
Outside-out patches of membrane were excised from muscle fibers 6 and 7 of third-instar wild-type Drosophila larvae. Channels were observed to open in response to short pulses of L-glutamate. At a holding potential of -60 mV, the channels opened to one main conductance level of about 120 pS. The current-voltage plot for the channels was linear and reversed around 0 mV holding potential. The channels were also activated by quisqualate but not by aspartate, N-methyl-D-aspartate (NMDA), kainate, glycine, gamma-aminobutyric acid (GABA) and acetylcholine. At high glutamate concentrations (3 or 10 mM), channel activation reached a peak within 0.3 ms. The channels opened in 'bursts' flickering between open and closed states. The channels opened only for a few milliseconds after switching on the glutamate and channel activity declined after the initial surge to zero with time constants between 5 and 20 ms. During applications of low glutamate concentrations (0.2-0.5 mM) to the same patch the channels opened much less frequently and during most applications no openings were observed. The openings observed were short and 'bursts' of openings were rare. Two exponential components were identified in the open-time distribution obtained with pulsed applications of glutamate (0.5-10 mM) with time constants of about 0.1 and 2.0 ms. The kinetics of the channels seem to be similar to the kinetics of certain glutamate gated channels found on muscle of crayfish and locust.
Animals↗
[The differential diagnosis of nodular tumors of the hand].
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Diagnosis, Differential↗
Ultraviolet-A radiation induces adhesion molecule expression on human dermal microvascular endothelial cells.
Ultraviolet radiation is capable of inducing numerous skin reactions. Considerable amounts of UVA radiation penetrate the epidermis and reach the microvascular endothelium of the papillary dermis. In order to investigate putative direct effects of UV radiation on endothelial cells, we studied adhesion molecule expression by immunostaining procedures and FACS analysis, following irradiation of normal human skin and cultured human dermal endothelial cells. Enhanced immunostaining for ICAM-1 and E-selectin was detected in biopsies taken after in vivo UVA and UVB irradiation, compared with non-irradiated control skin. On cultured human dermal endothelial cells, however, ICAM-1 and E-selectin were inducible by UVA but not UVB. The induction was dose-dependent, peaking at 20 J/cm2 for both adhesion molecules, and time-dependent, peaking after 6 and 24 h for E-selectin and ICAM-1, respectively. Expression of VCAM-1 and PECAM/EndoCAM/CD31 was unaffected by any UV-radiation modality. The functional integrity of irradiated cells was monitored by an exclusion assay of the fluorescent dye 7-AAD, and by staining for the cytoskeletal proteins actin and vimentin. Our results demonstrate that dermal microvascular endothelial cells are a critical and direct target of UVA, and suggest they may play a pivotal role in UV-induced inflammatory skin conditions.
Cell Adhesion↗
Function and energy metabolism of isolated hearts obtained from hyperthyroid spontaneously hypertensive rats (SHR). A 31P-nuclear magnetic resonance study.
It was the aim of this study to evaluate the effects of hyperthyroidism on heart function and cardiac energy metabolism of spontaneously hypertensive (SHR) rats. Hyperthyroidism was induced by daily injections of T3 (0.2 mg/kg s.c.) for 14 days. The hearts were then isolated and perfused in the Langendorff mode. ATP, phosphocreatine (PCr), and inorganic phosphate (Pi) were measured continuously by means of 31P-nuclear magnetic resonance (NMR) spectroscopy. Work load was altered by varying stepwise the Ca++ concentration in the perfusion fluid from 0.5 to 1.0, 1.5, and 2.0 mM, respectively. At every elevation of the Ca++ concentration, the increase in left ventricular developed pressure (LVDP) was higher in the hyperthyroid SHR than in the untreated SHR hearts. The ATP and PCr concentrations were lower in the hyperthyroid SHR compared to the untreated SHR hearts throughout the perfusion period. PCr decreased at every Ca++ elevation in both the untreated and hyperthyroid SHR hearts. The PCr/ATP ratio was not altered at any Ca++ concentration neither in the untreated SHR nor in the hyperthyroid SHR hearts. The Ca(++)-induced stepwise elevation in LVDP was higher at any given PCr/Pi ratio in the hyperthyroid SHR than in the untreated SHR hearts. Thus, the Ca(++)-inducible contractile reserve was greater in the hyperthyroid SHR heart.
Adenosine Triphosphate↗
Biphasic effects of interleukin-1 alpha on dermal fibroblasts: enhancement of chemotactic responsiveness at low concentrations and of mRNA expression for collagenase at high concentrations.
Fibrotic reactions in the skin are frequently preceded by infiltration of inflammatory cells and subsequent migration of fibroblastic cells. Interleukin-1 is secreted by inflammatory cells and can regulate proliferation and protein synthesis of fibroblasts. Its role in fibroblast chemotaxis has not been elucidated in any detail. Using the well-established Boyden chamber assay for measurement of chemotaxis in vitro, we studied a wide range of recombinant human interleukin-1 alpha concentrations to assess intrinsic chemotactic activity of interleukin-1 alpha and to determine the capacity of this mediator to modify the chemotactic response of fibroblasts to other chemoattractants. This was compared with the interleukin-1 alpha dose required for enhancement of mRNA expression for collagenase. Although interleukin-1 alpha was not chemoattractive for fibroblasts, it specifically augmented migration toward fibroblast-conditioned medium and toward platelet-derived growth factor but not toward epidermal growth factor, fibronectin, or transforming-growth factor-beta. Interleukin-1 alpha did not measurably alter the expression of mRNA for the platelet-derived growth factor receptor or its platelet-derived growth factor-binding characteristics. Doses required to enhance fibroblast chemotaxis were distinctly lower than those required for stimulation of collagenase mRNA expression.
Cell Division↗
Alteration of cardiac energy state during development of hypertension in rats of the Lyon strain: a 31P-NMR study on the isolated rat heart.
The effect of different chronic blood pressure levels on cardiac energy metabolism was studied by 31P-NMR spectroscopy in perfused hearts from the Lyon strains of hypertensive (LH), normotensive (LN) and hypotensive (LL) rats at the ages of 12 and 21 weeks. The in vivo assessment of haemodynamic parameters measured at 21 weeks in anaesthetized rats with an ultraminiature catheter pressure transducer confirmed that left ventricular systolic pressure and mean aortic pressure were significantly greater (+25%) in LH rats than in LN and LL rats. In the LL animals, left ventricular systolic pressure was slightly reduced (-10%) and cardiac contractility (estimated by LV dP/dtmax) showed a 24% decreased compared to normotensive animals. The energy state of the cardiomyocytes was characterized at different work levels of isolated rat hearts, by determining the concentration of the free phosphorylated compounds at each work level. Changes in workload were induced by varying the calcium concentration in the perfusion fluid. Increasing extracellular calcium concentration resulted in a similar increase in left ventricular developed pressure (LVDP) in all groups studied. Intracellular pH was not influenced by either the age of the animals or the level of cardiac work, in the three groups of animals. ATP content of the LN and LL rats remained constant during the whole perfusion period while the 12 week-old LH rats showed a decreased ATP content with increasing cardiac work. In the older LH rats, ATP content was decreased at the highest work level (corresponding to 2 mM calcium). In response to the increase in work, phosphocreatine (PCr) content diminished and inorganic phosphate (Pi) content increased in both LN, LH and LL animals. PCr degradation and Pi accumulation were higher in the LH rats and less in LL rats compared to the LN. These changes were more important in the younger than in the older hypertensive animals. The relationship between LVDP and [Pi]/[PCr] indicates that oxidative metabolism is maximally activated in the young hypertensive rats and suggests that this maximal activation represents an adaptive phase to the increase in blood pressure. Since the difference between the metabolic pattern of the 21 week-old LH rats and age-matched LN rats was less pronounced, it is likely that a compensatory stage has been reached at that age.
Animals↗