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Biomedical subjects

E Johannes

Publications and source records attributed to E Johannes.

At least 19 recordsLinked to original sources

Gravisensing: ionic responses, cytoskeleton and amyloplast behavior.

In Zea mays L., changes in orientation of stems are perceived by the pulvinal tissue, which responds to the stimulus by differential growth resulting in upward bending of the stem. Gravity is perceived in the bundle sheath cells, which contain amyloplasts that sediment to the new cell base when a change in the gravity vector occurs. The mechanism by which the mechanical signal is transduced into a physiological response is so far unknown for any gravity perceiving tissue. It is hypothesized that this involves interactions of amyloplasts with the plasma membrane and/or ER via cytoskeletal elements. To gain further insights into this process we monitored amyloplast movements in response to gravistimulation. In a pharmacological approach we investigated how the dynamics of plastid sedimentation are affected by actin and microtubule (MT) disrupting drugs. Dark grown caulonemal filaments of the moss Physcomitrella patens respond to gravity vector changes with a reorientation of tip growth away from the gravity vector. MT distributions in tip cells were monitored over time and MTs were seen to accumulate preferentially on the lower flank of the tip 30 min after a 90 degree turn. Using a self-referencing Ca2+ selective ion probe, we found that growing caulonemal filaments exhibit a Ca2+ influx at the apical dome, similar to that reported previously for other tip growing cells. However, in gravistimulated Physcomitrella filaments the region of Ca2+ influx is not confined to the apex, but extends about 60 micrometers along the upper side of the filament. Our results indicate an asymmetry in the Ca2+ flux pattern between the upper and side of the filament suggesting differential activation of Ca2+ permeable channels at the plasma membrane.

Actin Cytoskeleton↗

Cytoplasmic pH dynamics in maize pulvinal cells induced by gravity vector changes.

In maize (Zea mays) and other grasses, changes in orientation of stems are perceived by pulvinal tissue, which responds to the stimulus by differential growth resulting in upward bending of the stem. The amyloplast-containing bundle sheath cells are the sites of gravity perception, although the initial steps of gravity perception and transmission remain unclear. In columella cells of Arabidopsis roots, we previously found that cytoplasmic pH (pH(c)) is a mediator in early gravitropic signaling (A.C. Scott, N.S. Allen [1999] Plant Physiol 121: 1291-1298). The question arises whether pH(c) has a more general role in signaling gravity vector changes. Using confocal ratiometric imaging and the fluorescent pH indicator carboxy seminaphtorhodafluor acetoxymethyl ester acetate, we measured pH(c) in the cells composing the maize pulvinus. When stem slices were gravistimulated and imaged on a horizontally mounted confocal microscope, pH(c) changes were only apparent within the bundle sheath cells, and not in the parenchyma cells. After turning, cytoplasmic acidification was observed at the sides of the cells, whereas the cytoplasm at the base of the cells where plastids slowly accumulated became more basic. These changes were most apparent in cells exhibiting net amyloplast sedimentation. Parenchyma cells and isolated bundle sheath cells did not show any gravity-induced pH(c) changes although all cell types responded to external stimuli in the predicted way: Propionic acid and auxin treatments induced acidification, whereas raising the external pH caused alkalinization. The results suggest that pH(c) has an important role in the early signaling pathways of maize stem gravitropism.

Cytoplasm↗

Control of Cl- efflux in chara corallina by cytosolic pH, free ca2+, and phosphorylation indicates a role of plasma membrane anion channels in cytosolic pH regulation

Enhanced Cl- efflux during acidosis in plants is thought to play a role in cytosolic pH (pHc) homeostasis by short-circuiting the current produced by the electrogenic H+ pump, thereby facilitating enhanced H+ efflux from the cytosol. Using an intracellular perfusion technique, which enables experimental control of medium composition at the cytosolic surface of the plasma membrane of charophyte algae (Chara corallina), we show that lowered pHc activates Cl- efflux via two mechanisms. The first is a direct effect of pHc on Cl- efflux; the second mechanism comprises a pHc-induced increase in affinity for cytosolic free Ca2+ ([Ca2+]c), which also activates Cl- efflux. Cl- efflux was controlled by phosphorylation/dephosphorylation events, which override the responses to both pHc and [Ca2+]c. Whereas phosphorylation (perfusion with the catalytic subunit of protein kinase A in the presence of ATP) resulted in a complete inhibition of Cl- efflux, dephosphorylation (perfusion with alkaline phosphatase) arrested Cl- efflux at 60% of the maximal level in a manner that was both pHc and [Ca2+]c independent. These findings imply that plasma membrane anion channels play a central role in pHc regulation in plants, in addition to their established roles in turgor/volume regulation and signal transduction.

Journal Article↗

Kinetic analysis of Ca2+/K+ selectivity of an ion channel by single-binding-site models.

Current-voltage relationships of a cation channel in the tonoplast of Beta vulgaris, as recorded in solutions with different activities of Ca2+ and K+ (from Johannes & Sanders 1995, J. Membrane Biol. 146:211-224), have been reevaluated for Ca2+/K+ selectivity. Since conversion of reversal voltages to permeability ratios by constant field equations is expected to fail because different ions do not move independently through a channel, the data have been analyzed with kinetic channel models instead. Since recent structural information on K+ channels show one short and predominant constriction, selectivity models with only one binding site are assumed here to reflect this region kinetically. The rigid-pore model with a main binding site between two energy barriers (nine free parameters) had intrinsic problems to describe the observed current-saturation at large (negative) voltages. The alternative, dynamic-pore model uses a selectivity filter in which the binding site alternates its orientation (empty, or occupied by either Ca2+ or K+) between the cytoplasmic side and the luminal side within a fraction of the electrical distance and in a rate-limiting fashion. Fits with this model describe the data well. The fits yield about a 10% electrical distance of the selectivity filter, located about 5% more cytoplasmic than the electrical center. For K+ translocation, reorientation of the unoccupied binding site (with a preference of about 6:5 to face the lumenal side) is rate limiting. For Ca2+, the results show high affinity to the binding site and low translocation rates (<1% of the K+ translocation rate). With the fitted model Ca2+ entry through the open channel has been calculated for physiological conditions. The model predicts a unitary open channel current of about 100 fA which is insensitive to cytoplasmic Ca2+ concentrations (between 0.1 and 1 microM) and which shows little sensitivity to the voltage across the tonoplast.

Binding Sites↗

The voltage-gated Ca2+ release channel in the vacuolar membrane of sugar beet resides in two activity states.

The voltage-gated Ca2+ release channel in the vacuolar membrane of sugar beet tap roots resides in two states which show similar reversal potentials in biionic conditions but differ dramatically in their single channel current and open frequency. State I has a unitary Ca2+ conductance of 3.9 +/- 0.5 pS and channel openings are rare at vacuolar resting potentials. After spontaneous and reversible transition into State II the unitary Ca2+ conductance increases three-fold, accompanied by a concomitant rise in channel activity over the physiological voltage range. The 15- to 19-fold increase in total Ca2+ current after the State I to State II transition could play an essential role in channel activation during signal transduction.

Biological Transport↗

Lumenal calcium modulates unitary conductance and gating of a plant vacuolar calcium release channel.

The patch clamp technique has been used to investigate ion permeation and Ca(2+)-dependent gating of a voltage-sensitive Ca2+ release channel in the vacuolar membrane of sugar beet tap roots. Reversal potential measurements in bi-ionic conditions revealed a sequence for permeability ratios of Ca2+ approximately Sr2+ approximately Ba2+ > Mg2+ >> K+ which is inversely related to the size of the unitary conductances K+ >> Mg2+ approximately Ba2+ > Sr2+ approximately Ca2+, suggesting that ion movement is not independent. In the presence of Ca2+, the unitary K+ current is reduced in a concentration- and voltage-dependent manner by Ca2+ binding at a high affinity site (K0.5 = 0.29 mM at 0 mV) which is located 9% along the electric field of the membrane from the vacuolar side. Comparison of reversal potentials obtained under strictly bi-ionic conditions with those obtained in the presence of mixtures of the two ions indicates that the channel forms a multi-ion pore. Lumenal Ca2+ also has an effect on voltage-dependent channel gating. Stepwise increases of vacuolar Ca2+ from micromolar to millimolar concentrations resulted in a dramatic increase in channel openings over the physiological voltage range via a shift in threshold for channel activation to less negative membrane potentials. The steepness of the concentration dependence of channel activation by Ca2+ at -41 mV predicts that two Ca2+ ions need to bind to open the gate. The implications of the results for ion permeation and channel gating are discussed.

Calcium↗

Voltage-gated Ca2+ release channels in vacuolar membranes from beet storage roots and guard cells.

The ubiquity and wide-ranging roles of Ca(2+)-mediated stimulus-response coupling in plant cells is likely to demand flexibility in the duration and amplitude of the cytosolic Ca2+ signal. The presence of both ligand- and voltage-gated Ca2+ release channels at the vacuolar membrane suggests that this flexibility might be achieved through differential regulation of these discrete channel classes. The properties and distribution of voltage-gated vacuolar Ca2+ channels is described in this chapter. In beet storage roots, the channels display a single channel conductance of 12 pS (5-20 mM Ca2+ on the lumenal side), Ca2+:K+ selectivity = 15:1 and a unitary current which saturates as a function of (physiological) negative voltages at about 0.5 pA. Channel opening is enhanced by negative voltages and by intravacuolar (lumenal) Ca2+. Variation of lumenal Ca2+ concentration has enabled the binding sites both for ionic selectivity and for Ca2+ gating to be mapped with respect to their fractional electrical distance across the membrane. Voltage-gated Ca2+ channels with similar permeation properties also reside in the vacuolar membranes of Vicia faba and Commelina communis guard cells. Although these channels exhibit somewhat higher unitary conductances and voltage-saturating currents than the beet channel, they are similar in many other respects, including sensitivity to Gd3+. We conclude that the widespread distribution of an emerging class of voltage-gated Ca2+ channel in the vacuolar membrane of plants points towards a fundamental role of these channels in plant cell biology.

Calcium↗

Proton Gradient Across the Tonoplast of Riccia fluitans as a Result of the Joint Action of Two Electroenzymes.

Using pH-sensitive microelectrodes (in vitro) and acridine orange photometry (in vivo), the actions of the two tonoplast phosphatases, the tp-ATPase and the tp-PPase, were investigated with respect to how effectively they could generate a transtonoplast pH-gradient. Under standard conditions the vacuoles of the aquatic liverwort Riccia fluitans have an in vivo pH of 4.7 to 5.0. In isolated vacuoles a maximal vacuolar pH (pH(v)) of 4.74 +/- 0.1 is generated in the presence of 0.1 millimolar PP(i), but only 4.93 +/- 0.13 in the presence of 2.5 millimolar ATP. Both substrates added together approximate the value for PP(i). Cl(-)-stimulates the H(+)-transport driven by the tp-ATPase, but has no effect on the tp-PPase. The transport activity of the tp-ATPase approximates saturation kinetics (K((1/2)) approximately 0.5 millimolar), whereas transport by the tp-PPase yields an optimum around 0.1 millimolar PP(i). The transtonoplast pH-gradient is dissipated slowly by weak bases, from which a vacuolar buffer capacity of roughly 300 to 400 millimolar/pH(v) unit has been estimated. From the free energy (-11.42 kilojoules per mole) for the hydrolysis of PP(i) under the given experimental conditions, we conclude that the PPase-stoichiometry (transported H(+) per hydrolyzed substrate molecule) must be 1, and that in vivo this enzyme works as a H(+)-pump rather than as a pyrophosphate synthetase.

Journal Article↗

[Studies of voice stress in teachers with functional voice disorders caused by teaching activity].

30 female teachers of ten-years technical schools-15 with functional voice-disturbance an 15 with healthy voice-have been examined while teaching in respect of their strain. Duration of speech, intensity of their teachers voice an background noise, the frequency of heart rate and the arterial blood pressure were measured on the teaching day. Between both research groups no statistically significant differences were shown. There were no correlations between the durations of speech, intensity and vegetative parameters.

Adult↗

[Ventricular aneurysm and coronary heart disease. Pathophysiology, differential therapy, and postoperative haemodynamics (author's transl)].

In the course of coronary heart disease an aneurysmal dilatation of the left ventricle may occur. This may be an additional risk for the patient by changed haemodynamics. Depending on the extent of the aneurysm and the contractile potency of the remaining myocard the cardiac compensation may be sufficient. A reduction of the pump efficiency is not necessarily the consequence. In case there is an increasing cardiac insufficiency by means of a pathologic ventricular filling pressure pulse, the best therapy is digitalis in combination with a reduction of volume by sodium-selective diuretics. Under same haemodynamic conditions the treatment of angina pectoris consists of long acting nitrites in combination with a betablocking agent having some intrinsic activity. Special care for the choic of medicaments has to be taken in relation to the sufficiency of the remaining myocard, if an antiarrhythmic therapy is necessary. If there is no stabilisation of the haemodynamic parameters by conservative therapy, the left ventricular function is meliorated by surgical aneurysmectomy. The data demonstrate, that under resting condition a normalisation and under exercise condition at least a melioration of pulse pressure and circulation is achieved after resection of the aneurysma. A small but measurable decrease in cardiac output under exercise condition is the consequence of a persisting cardiodepressive effect due to the operation.

Adrenergic beta-Antagonists↗

[Course of coronary disease. Evaluation of prognosis and progression of coronary insufficiency with atrial pacing and ergometry].

Significant reduction of angina threshold (145 Imp./min to 134 Imp./min) and increase of ST-segment depression (0.13 to 0.17 mV) indicating progression of coronary artery disease was seen in 34 subjects studied by atrial pacing at intervals betion (0.22 mV to 0.12 mV) during exercise, which correlated significantly with decrease of heart rate (121 to 110 beats/min), is interpreted as consequence of diminished sympathetic activity and myocardial O(2)-demand. The change of hemodynamic parameters during controlled exercise does not allow evaluation concerning the progress of coronary artery disease, whereas cardiac stress test with atrial pacing is reproducible. There was no difference in relation to reduction of angina threshold between the group after combined longterm medication with nitrate and ss-blocking agent and the control group. Plasma lipid abnormalities were predictive of subsequent reduction of angina threshold. Severe 2 and 3 vessel obstruction was seen more frequently in patients exhibiting reduction of angina threshold. Level of uric acid, obesity, hypertension, age, combination of risk factors, the initially studied myocardial lactate production and angina threshold during exercise and atrial pacing had no predictive value concerning reduction of angina threshold.

Adult↗