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Biomedical subjects

M L Evans

Publications and source records attributed to M L Evans.

At least 91 records · Page 5Linked to original sources

Induction of curvature in maize roots by calcium or by thigmostimulation: role of the postmitotic isodiametric growth zone.

We examined the response of primary roots of maize (Zea mays L. cv Merit) to unilateral application of calcium with particular attention to the site of application, the dependence on growth rate, and possible contributions of thigmotropic stimulation during application. Unilateral application of agar to the root cap induced negative curvature whether or not the agar contained calcium. This apparent thigmotropic response was enhanced by including calcium in the agar. Curvature away from objects applied unilaterally to the extreme root tip occurred both in intact and detipped roots. When agar containing calcium chloride was applied to one side of the postmitotic isodiametric growth zone ( a region between the apical meristem and the elongation zone), the root curved toward the side of application. This response could not be induced by plain agar. We conclude that curvature away from calcium applied to the root tip results from a thigmotropic response to stimulation during application. In contrast, curvature toward the calcium applied to the postmitotic isodiametric growth zone results from direct calcium-induced inhibition of growth.

Agar↗

Ca2+ efflux mechanisms following depolarization evoked calcium transients in cultured rat sensory neurones.

1. We have used a combination of microfluorimetry and patch-clamp techniques to investigate cytoplasmic Ca2+ ([Ca2+]i) buffering in response to physiological Ca2+ loads in neurones cultured from the dorsal root ganglia of the rat. 2. In cells loaded with Indo-1 AM and using high resistance microelectrodes to initiate and record action potentials, single action potentials were associated with a measurable rise in [Ca2+]i. Short trains of action potentials evoked [Ca2+]i transients with monoexponential recovery rates with time constants of around 5 s. 3. Similar Ca2+ buffering properties were seen in cells perfused with patch-clamp pipettes in the whole-cell recording mode suggesting that the slow (seconds) Ca2+ buffering properties were not seriously perturbed by the recording technique. 4. In cells held under voltage clamp, reversal of the Na(+)-Ca2+ exchanger driving force had a small but significant effect on the rate of Ca2+ removal. 5. Increasing extracellular pH or adding vanadate (200 microM) to the internal solution dramatically slowed the rate of recovery. Addition of calmidazolium to the pipette solution also produced a significant but much less dramatic slowing of Ca2+ efflux. 6. The results demonstrate that the activity of a plasmalemmal Ca(2+)-ATPase is important for the removal of somatic Ca2+ loads of a similar amplitude to those generated by the firing of a few action potentials.

Action Potentials↗

Vascular response to fractionated irradiation in the rat lung.

The effects of fractionated hemithorax irradiation on normal lung tissue were examined by measuring changes in the vascular permeability surface area product (PS) and relative lung blood flow in Sprague-Dawley rats. The rats received five daily fractions per week of either 3.0 or 4.0 Gy for 4 weeks to the left lung. Between 3 and 5 weeks after the start of irradiation, the average PS was approximately 50% above normal for the group of rats that received 3.0 Gy/day and 200-300% above normal in the group of rats that received 4.0 Gy/day. Treatment with cyproheptadine, indomethacin, or theophylline had no effect, but treatment with dexamethasone significantly reduced PS to near normal levels. Left-to-right blood flow ratios in the group of rats that received 3.0 Gy/day decreased to 66% of normal levels by 4 weeks. In the group of rats that received 4.0 Gy/day, blood flow decreased to 46% of normal levels by 4 weeks. Treatment with dexamethasone maintained normal blood flow until the drug dose was reduced. These results agree with earlier studies using single-dose irradiation and indicate that the methods used to measure PS and blood flow are sensitive at low doses.

Animals↗

Abnormalities of chromosome 16q in myeloid malignancy: 14 new cases and a review of the literature.

Fourteen patients with abnormalities of chromosome 16q, 13 with acute myelogenous leukaemia (AML), and one with refractory anaemia with excess of blasts (RAEB), are described. Seven patients had inv(16)(p13q22), two had del(16)(q22), and five had other abnormalities of 16q. Six of the seven patients with inv(16) had AML M4Eo and, following treatment with adriamycin, cytosine arabinoside, and 6-thioguanine, all achieved complete remission (CR). Neither patient with del(16)(q22) had typical M4Eo morphology at diagnosis; CR was achieved in one and one had resistant leukaemia. Patients with other abnormalities of 16q had blasts of diverse morphology and, although morphologically abnormal eosinophils were seen in three patients, this was not as marked as in the patients with inv(16). CR was achieved in two of the four patients with other abnormalities of 16q but duration of remission was short in both cases. These results suggest that most patients with del(16)(q22) and other abnormalities of 16q22 do not have typical AML M4Eo. Such patients tend to have a worse prognosis, and are more likely to have complex karyotypes typical of secondary leukaemia.

Adult↗

Vascular response to radiation injury in the rat lung.

Changes in relative left-to-right lung blood flow ratios were followed as an index of vascular radiation injury in left-hemithorax-irradiated Sprague-Dawley rats. Single doses of 11 to 21 Gy gamma radiation resulted in a dose-dependent decrease in relative blood flow to the irradiated lung from 3 to 5 weeks after exposure during the development of pneumonitis. Blood flow returned to near normal by 5 weeks after lower doses (11-13.5 Gy). After a single dose of 15 Gy the left-to-right blood flow ratio recovered to 75% of normal at 12 weeks and leveled off. Following 18 Gy irradiation a second period of reduced flow began 16 weeks after exposure. After 21 Gy irradiation flow to the irradiated side remained low for 1 year after exposure. Rats that received a single dose of 18 Gy to the left hemithorax were also treated with one or two of the following drugs: captopril, cyproheptadine, dexamethasone, diethylcarbamazine, penicillamine, or theophylline. Dexamethasone was most effective at preventing the decrease in blood flow to the irradiated lung when treatment was continued through the pneumonitis period and dose was not tapered until 8 weeks after radiation exposure. All other drugs and drug combinations were, for the most part, virtually ineffective after the pneumonitis period. There was a relatively poor correlation with earlier vascular permeability surface area product studies. This suggests that endothelial damage, as well as damage to other cell types, contributes to the development of post-irradiation fibrosis in the lung.

Animals↗

Pb2+ blocks calcium currents of cultured dorsal root ganglion cells.

The divalent cation lead (Pb2+) blocks sustained and transient voltage sensitive calcium channel currents of cultured rat dorsal root ganglion cells. The IC50 for inhibition of the total peak current evoked by a step depolarization from -80 to 0 mV was 0.6 microM, compared to an IC50 of 2.2 microM for Cd2+. The current activated by a depolarization from -40 to 0 mV was inhibited by 50% by 1.0 microM Pb2+. Low threshold currents activated by a step from -100 to -30 mV were blocked by Pb2+ at higher concentrations (IC50 = 6 microM). The block progressed in the absence of channel activation and showed little voltage dependence. Peak sodium current was reduced by 6.6% at 1 microM Pb2+ while at 20 microM the peak current was reduced by 40% with marked slowing of the time course of activation. The potassium rectifier current was reduced by 4.1% at 1 microM Pb2+. Thus, Pb2+ selectively blocks calcium currents at concentrations in the range of those causing toxicity in man.

Animals↗

Correlation of 125I-LSD autoradiographic labeling with serotonin voltage clamp responses in Aplysia neurons.

Autoradiographic receptor binding studies using 125I-LSD (2-[125I]lysergic acid diethyamide) revealed intense labelling on the soma of a symmetrically located pair of cells in the abdominal ganglion of Aplysia californica. This binding was blocked by micromolar concentrations of serotonin and lower concentrations of the serotonergic antagonists, cyproheptadine and mianserin (Kadan and Hartig, 1988). Electrophysiological investigation of responses to serotonin of neurons in the left upper quadrant, where one of the labeled neurons is located, revealed a range of serotonin responses. Cells L3 and L6 have a K+ conductance increase in response to serotonin that is not blocked by cyproheptadine or mianserin. Cells L2 and L4 have a biphasic response to serotonin: a Na+ conductance increase, which can be blocked by cyproheptadine and mianserin, followed by a voltage dependent Ca2+ conductance which is blocked by Co2+ but not the serotonergic antagonists. Cell L1, and its symmetrical pair, R1, have in addition to the Na+ and Ca2+ responses observed in L2 and L4, a Cl- conductance increase blocked by LSD, cyproheptadine and mianserin. LSD had little effect on the other responses. We conclude that the symmetrically located cells L1 and R1 have a Cl- channel linked to a cyproheptadine- and mianserin-sensitive serotonin receptor that is selectively labelled by 125I-LSD. This receptor has many properties in common with the mammalian serotonin 1C receptor.

Acetylcholine↗

Myeloablative therapy with autologous bone marrow transplantation as consolidation of remission in patients with follicular lymphoma.

A study has been in progress since June 1985 to evaluate the use of myeloablative therapy (cyclophosphamide [60 mg/kg x 2] and total body irradiation [200 cGy x 6]) followed by reinfusion of autologous bone marrow in patients in second or subsequent remission of B-cell non-Hodgkin's lymphoma. The marrow mononuclear cell fraction is being treated in vitro with three cycles of the monoclonal antibody anti-CD20 (anti-B1, Coulter Immunology) and baby rabbit complement (Pel-Freez). Thirty-eight patients with follicular lymphoma (age range 29-61 years, median 43) have been treated to date. At the time of treatment, 28 patients were in second remission, 7 were in third, and 3 were in more than third remission. Twenty-three patients were in complete remission, 15 had residual disease (7 had lymph nodes less than 2 cm diameter, 4 had less than 10% bone marrow infiltration, 1 had involvement of lymph nodes and bone marrow, and 3 had involvement at other sites). Of the 38 study patients, 32 are alive; 6 have died, 4 in remission. Two of the deaths were treatment related: 1 resulted from cerebral haemorrhage at 29 days; 1 resulted from systemic fungal infection at three months). One patient died from secondary acute myelogenous leukaemia at four years, and another from an unrelated cause. Two patients died following relapse. The median time to engraftment was 28 days (range 15-45 days) for neutrophils greater than 0.5 x 10(9)/L and 28 days (range 15-46 days) for platelets greater than 20 x 10(9)/L.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Computer-based video digitizer analysis of surface extension in maize roots: kinetics of growth rate changes during gravitropism.

We used a video digitizer system to measure surface extension and curvature in gravistimulated primary roots of maize (Zea mays L.). Downward curvature began about 25 +/- 7 min after gravistimulation and resulted from a combination of enhanced growth along the upper surface and reduced growth along the lower surface relative to growth in vertically oriented controls. The roots curved at a rate of 1.4 +/- 0.5 degrees min-1 but the pattern of curvature varied somewhat. In about 35% of the samples the roots curved steadily downward and the rate of curvature slowed as the root neared 90 degrees. A final angle of about 90 degrees was reached 110 +/- 35 min after the start of gravistimulation. In about 65% of the samples there was a period of backward curvature (partial reversal of curvature) during the response. In some cases (about 15% of those showing a period of reverse bending) this period of backward curvature occurred before the root reached 90 degrees. Following transient backward curvature, downward curvature resumed and the root approached a final angle of about 90 degrees. In about 65% of the roots showing a period of reverse curvature, the roots curved steadily past the vertical, reaching maximum curvature about 205 +/- 65 min after gravistimulation. The direction of curvature then reversed back toward the vertical. After one or two oscillations about the vertical the roots obtained a vertical orientation and the distribution of growth within the root tip became the same as that prior to gravistimulation. The period of transient backward curvature coincided with and was evidently caused by enhancement of growth along the concave and inhibition of growth along the convex side of the curve, a pattern opposite to that prevailing in the earlier stages of downward curvature. There were periods during the gravitropic response when the normally unimodal growth-rate distribution within the elongation zone became bimodal with two peaks of rapid elongation separated by a region of reduced elongation rate. This occurred at different times on the convex and concave sides of the graviresponding root. During the period of steady downward curvature the elongation zone along the convex side extended farther toward the tip than in the vertical control. During the period of reduced rate of curvature, the zone of elongation extended farther toward the tip along the concave side of the root. The data show that the gravitropic response pattern varies with time and involves changes in localized elongation rates as well as changes in the length and position of the elongation zone. Models of root gravitropic curvature based on simple unimodal inhibition of growth along the lower side cannot account for these complex growth patterns.

Computer Systems↗

Cytoplasmic calcium levels in protoplasts from the cap and elongation zone of maize roots.

Calcium has been implicated as a key component in the signal transduction process of root gravitropism. We measured cytoplasmic free calcium in protoplasts isolated from the elongation zone and cap of primary roots of light-grown, vertically oriented seedlings of Zea mays L. Protoplasts were loaded with the penta-potassium salts of fura-2 and indo-1 by incubation in acidic solutions of these calcium indicators. Loading increased with decreasing pH but the pH dependence was stronger for indo-1 than for fura-2. In the case of fura-2, loading was enhanced only at the lowest pH (4.5) tested. Dyes loaded in this manner were distributed predominantly in the cytoplasm as indicated by fluorescence patterns. As an alternative method of loading, protoplasts were incubated with the acetoxymethylesters of fura-2 and indo-1. Protoplasts loaded by this method exhibited fluorescence both in the cytoplasm and in association with various organelles. Cytoplasmic calcium levels measured using spectrofluorometry, were found to be 160 +/- 40 nM and 257 +/- 27 nM, respectively, in populations of protoplasts from the root cap and elongation zone. Cytoplasmic free calcium did not increase upon addition of calcium to the incubation medium, indicating that the passive permeability to calcium was low.

Calcium↗

A simple, dual tracer method for the measurement of transvascular flux of albumin into the lung.

A simple, first-order model of albumin kinetics in the rat lung is presented and validated with a more sophisticated model. The simple model assumes that intravascular concentration of tracer albumin is constant over 30 min after injection and transvascular flux of tracer albumin is unidirectional and proportional to the permeability-surface area product (PS). 125I-albumin is injected initially and 131I-albumin at 20 min. At 30 min the rat is sacrificed and plasma and tissue samples are obtained for gamma counting. Simultaneous equations are set up for the two tracers and solved for PS and plasma volume. The accuracy of this approach is examined with data generated from a more complete model. This model uses the concepts of hydraulic conductivity, solvent drag, reflection coefficients, hydrostatic and osmotic pressures, exclusion volumes, and lymph flow, as well as PS. Based on known PS and clearance rates from the complex model, the simple model estimates tracer albumin leakage rate with less than 5% error over the range of PS encountered in rat studies.

Albumins↗

Gravitropism: interaction of sensitivity modulation and effector redistribution.

Our increasing capabilities for quantitative hormone analysis and automated high resolution growth studies have allowed a reassessment of the classical Cholodny-Went hypothesis of gravitropism. According to this hypothesis, gravity induces redistribution of auxin toward the lower side of the organ and this causes the growth asymmetry that leads to reorientation. Arguments against the Cholodny-Went hypothesis that were based primarily on concerns over the timing and magnitude of the development of hormone asymmetry are countered by recent evidence that such asymmetry develops early and is sufficiently large to account for curvature. Thus, it appears that the Cholodny-Went hypothesis is fundamentally valid. However, recent comparative studies of the kinetics of curvature and the timing of the development of hormone asymmetry indicate that this hypothesis alone cannot account for the intricacies of the gravitropic response. It appears that time-dependent gravity-induced changes in hormone sensitivity as well as changes in sensitivity of the gravity receptor play important roles in the response.

Biological Transport↗

Red light-regulated growth. I. Changes in the abundance of indoleacetic acid and a 22-kilodalton auxin-binding protein in the maize mesocotyl.

We examined the changes in the levels of indoleacetic acid (IAA), IAA esters, and a 22-kilodalton subunit auxin-binding protein (ABP1) in apical mesocotyl tissue of maize (Zea mays L.) during continuous red light (R) irradiation. These changes were compared with the kinetics of R-induced growth inhibition in the same tissue. Upon the onset of continuous irradiation, growth decreased in a continuous manner following a brief lag period. The decrease in growth continued for 5 hours, then remained constant at 25% of the dark rate. The abundance of ABP1 and the level of free IAA both decreased in the mesocotyl. Only the kinetics of the decrease in IAA within the apical mesocotyl correlated with the initial change in growth, although growth continued to decrease even after IAA content reached its final level, 50% of the dark control. This decrease in IAA within the mesocotyl probably occurs primarily by a change in its transport within the shoot since auxin applied as a pulse move basipetally in R-irradiated tissue at the same rate but with half the area as dark control tissue. In situ localization of auxin in etiolated maize shoots revealed that R-irradiated shoots contained less auxin in the epidermis than the dark controls. Irradiated mesocotyl grew 50% less than the dark controls even when incubated in an optimal level of auxin. However, irradiated and dark tissue contained essentially the same amount of radioactivity after incubation in [14C]IAA indicating that the light treatment does not affect the uptake into the tissue through the cut end, although it is possible that a small subset of cells within the mesocotyl is affected. These observations support the hypothesis that R causes a decrease in the level of auxin in epidermal cells of the mesocotyl, consequently constraining the growth of the entire mesocotyl.

Biological Transport↗

Lead and zinc block a voltage-activated calcium channel of Aplysia neurons.

1. The effects of Pb2+ and Zn2+ on the peak of the voltage-activated calcium current of Aplysia neurons were examined. Calcium currents were reversibly blocked by Pb2+ at concentrations that did not significantly affect potassium and sodium currents and by Zn2+ at concentrations associated with a delay and reduction of peak sodium and potassium currents. 2. The block by both was concentration dependent, and percentage blockade was reduced in elevated Ca2+. The threshold Pb2+ concentration for blockade in 20 mM Ca artificial sea water (ASW) was approximately 1 microM, whereas for Zn2+ it was 2 mM. The Hill coefficient for Pb2+ action was near 1.0 under all conditions, whereas for Zn2+ it was 1.4-1.6. 3. With addition of Pb2+, the voltage at which peak calcium current was generated shifted to hyperpolarized voltages, an effect similar to that caused by reduction of Ca2+ concentration in the absence of Pb2+. Zn2+ shifted the voltage at which peak current was generated in a depolarizing direction. 4. Pb2+ did not significantly change inactivation but shifted the voltage dependence of activation to hyperpolarized voltages in a dose-dependent manner. Zn2+ shifted both activation and inactivation in a depolarizing direction in a dose-dependent fashion. 5. The blockade of calcium currents by Pb2+ but not Zn2+ was highly voltage dependent and increased with depolarization. 6. Our results suggest that Pb2+ is a specific, potent, competitive, and reversible blocker of calcium currents. These observations are consistent with a competition by Pb2+ with Ca2+ at a binding site within the calcium channel. In contrast, the blockade of calcium currents by Zn2+ is probably through actions at fixed charge sites external to the channel.

Animals↗

The pH profile for acid-induced elongation of coleoptile and epicotyl sections is consistent with the acid-growth theory.

The acid-growth theory predicts that a solution with a pH identical to that of the apoplast of auxin-treated tissues (4.5.-5.0) should induce elongation at a rate comparable to that of auxin. Different pH profiles for elongation have been obtained, however, depending on the type of pretreatment between harvest of the sections and the start of the pH-incubations. To determine the acid sensitivity under in vivo conditions, oat (Avena sativa L.) coleoptile, maize (Zea mays L.) coleoptile and pea (Pisum sativum L.) epicotyl sections were abraded so that exogenous buffers could penetrate the free space, and placed in buffered solutions of pH 3.5-6.5 without any preincubation. The extension, without auxin, was measured over the first 3 h. Experiments conducted in three laboratories produced similar results. For all three species, sections placed in buffer without pretreatment elongated at least threefold faster at pH 5.0 than at 6.0 or 6.5, and the rate elongation at pH 5.0 was comparable to that induced by auxin. Pretreatment of abraded sections with pH-6.5 buffer or distilled water adjusted to pH 6.5 or above gave similar results. We conclude that the pH present in the apoplast of auxin-treated coleoptile and stems is sufficiently low to account for the initial growth response to auxin.

Avena↗