Emission of volatile sulfur compounds from spruce trees

Spruce (Picea Abies L.) trees from the same clone were supplied with different, but low, amounts of plant available sulfate in the soil (9.7-18.1 milligrams per 100 grams of soil). Branches attached to the trees were enclosed in a dynamic gas exchange cuvette and analyzed for the emission of volatile sulfur compounds. Independent of the sulfate supply in the soil, H₂S was the predominant reduced sulfur compound continuously emitted from the branches with high rates during the day and low rates in the night. In the light, as well as in the dark, the rates of H₂S emission increased exponentially with increasing water vapor flux from the needles. Approximately 1 nanomole of H₂S was found to be emitted per mole of water. When stomata were closed completely, only minute emission of H₂S was observed. Apparently, H₂S emission from the needles is highly dependent on stromatal aperture, and permeation through the cuticle is negligible. In several experiments, small amounts of dimethylsulfide and carbonylsulfide were also detected in a portion of the samples. However, SO₂ was the only sulfur compound consistently emitted from branches of spruce trees in addition to H₂S. Emission of SO₂ mainly proceeded via an outburst starting before the beginning of the light period. The total amount of SO₂ emitted from the needles during this outburst was correlated with the plant available sulfate in the soil. The diurnal changes in sulfur metabolism that may result in an outburst of SO₂ are discussed.     

Anabolic effect of human parathyroid hormone fragment on trabecular bone in involutional osteoporosis: a multicentre trial.

After baseline studies, 21 patients with osteoporosis were treated with human parathyroid hormone fragment (PTH 1-34) given as once-daily subcutaneous injections for 6-24 months. The dose used did not cause hypercalcaemia even in the first few hours after injection. Calcium and phosphate balances improved in some patients, but there was no significant improvement in the group values. There were, however, substantial increases in iliac trabecular bone volume: the mean increase, confirmed by repeat blind measurements, was 70% above mean baseline volume. The new bone was histologically normal. Those patients who had the largest increases in 47Ca-kinetic and histomorphometric indices of new bone formation showed the greatest increases in trabecular bone volume, suggesting that treatment with human parathyroid hormone fragment caused a dissociation between formation and resorption rates that was confined to trabecular bone. Since vertebrae are four-fifths composed of trabecular bone, this hormone fragment may prove useful in treating patients with the crush fracture syndrome.     

Uptake of jasmonic acid and related compounds by mesophyll protoplasts of the barley leaf

Jasmonic acid (JA) permeates the plasma membrane of mesophyll cells by diffusion as the lipophilic undissociated JAH molecule probably without the participation of a saturable uptake component. The mesophyll plasma membrane is nearly impermeable to the JA anion. The permeability coefficients of JA and several JA derivatives (its methyl ester (JAMe), 7-iso-cucurbic acid (7-iso-CA), 6-epi-7-iso-cucurbic acid (6-epi-7-iso-CA), and both stereoisomers of the JA leucine conjugate ((+)-JA-Leu and (-)-JA-Leu)) were determined and used in a simplified mathematical model to predict stressdependent JA redistribution between cytosol and apoplast in comparison with ABA. The redistribution of JA takes place similar to ABA; however, its velocity is much higher because of the high JA membrane permeability. When the permeability coefficients for the mesophyll plasma membrane are plotted double-logarithmically against the ratio of the distribution coefficient and the molecular ratio to the power of 1.5 (K_DM_r ^–1.5), two straight lines result for two different classes of compounds. The permeability coefficients of JA conjugates are lower than that of the free acid by approximately one order of magnitude, but they are still significantly higher than that of ABA.     

Compartmental distribution and redistribution of abscisic acid in intact leaves

Using a computer model written for whole leaves (Slovik et al. 1992, Planta 187, 14–25) we present in this paper calculations of abscisic acid (ABA) redistribution among different leaf tissues and their compartments in relation to stomatal regulation under drought stress. The model calculations are based on experimental data and biophysical laws. They yield the following results and postulates: (i) Under stress, compartmental pH-shifts come about as a consequence of the inhibition of the pH component of proton-motive forces at the plasmalemma. There is a decrease of net proton fluxes by about 8.6 nmol · s^–1 · m^–2. (ii) Using stress-induced pH-shifts we demonstrate how stress intensities can be quantified on a molecular basis. (iii) As the weak acid ABA is the only phytohormone which behaves in vivo and in vitro ideally according to the Henderson-Hasselbalch equation, pH-shifts induce a complicated redistribution amongst compartments in the model leaf. (iv) The final accumulation of ABA in guard-cell walls is intensive: up to 16.1-fold compared with only up to 3.4-fold in the guard-cell cytosol. We propose that the binding site of the guard-cell ABA receptor faces the apoplasm. (v) A twoto three-fold ABA accumulation in guard-cell walls is sufficient to induce closure of stomata. (vi) The minimum time lag until stomata start to close is 1–5 min; it depends on the stress intensity and on the guard-cell sensitivity to ABA: the more moderate the stress is, the later stomata start to close or they do not close at all. (vii) In the short term, there is almost no influence of the velocity of pH-shifts on the velocity of the ABA redistribution, (viii) Six hours after the termination of stress there is still an ABA concentration 1.4-fold the initial level in the guard-cell cytosol (delay of ABA relaxation, aftereffect), (ix) The observed induction of net ABA synthesis after onset of stress may be explained by a decrease in cytosolic ABA degradation. About 1 h after onset of stress the model leaf would start to synthesise ABA (and its conjugates) automatically, (x) This ABA net synthesis serves to inform roots via an increased ABA concentration in the phloem sap. The stress-induced ABA redistribution is per se not sufficient to feed the ploem sap with ABA. (xi) The primary target membrane of stress is the plasmalemma, not thylakoids. (xii) The effective stress sensor, which induces the proposed signal chain finally leading to stomatal closure, is located in epidermal cells. Mesophyll cells are not capable of creating a significant ABA signal to guard cells if the epidermal plasmalemma conductance to undissociated molecular species of ABA (HABA) is indeed higher than the plasmalemma conductance of the mesophyll (plasmodesmata open), (xiii) All model conclusions which can be compared with independent experimental data quantitatively fit to them. We conclude that the basic experimental data of the model are consistent. A stress-induced ABA redistribution in the leaf lamina elicits stomatal closure.Abbreviations ABA abscisic acid - CON vacuolar ABA conjugates We are grateful to Prof. U. Heber (Lehrstuhl Botanik I, University of Würzburg, FRG) for stimulating discussions. This work has been performed within the research program of the Sonderforschungsbereich 251 (TP 3 and 4) of the University of Würzburg. It has been also supported by the Fonds der Chemischen Industrie.     

Activation and inhibition of mast cells degranulation affect their morphometric parameters

Activation of mast cells, the key cells of allergic inflammation, causes typical morphological changes associated with an increase in volume, that is a function of area and perimeter. The purpose of this study was to evaluate the effect of mast cell activation to degranulate, carried out by the secretagogue Compound 48/80, and of inhibition of this activation carried out by Nedocromil sodium, a mast cell stabilizing drug, on mast cell area, perimeter and shape factor by a computerized image analyzer. Mast cells were isolated and purified by peritoneal lavage of rats (purity >98%) and co-cultured with mouse 3T3 fibroblasts to which they adhere. Cultures were incubated for 10 min at 37 degrees C with culture medium alone (Enriched Medium) or Enriched Medium containing either Nedocromil (10(-4) M) or Compound 48/80 (0.3 microg/ml) or Compound 48/80 and Nedocromil (0.3 microg/ml and 10(-4) M respectively). Supernatants were then assessed for histamine release, as a marker of mast cell activation and the cell monolayers were fixed and stained with an alcoholic-acidic toluidine blue solution and examined with a computerized image analyzer connected with a light microscope. Mast cells incubated in Enriched Medium or Nedocromil possessed similar morphometric parameters. Mast cells activated with Compound 48/80 (70% histamine release) had a significant increase in area and perimeter and a decrease in shape factor in comparison to mast cells in Enriched Medium alone. Simultaneous incubation of mast cells with Compound 48/80 and Nedocromil significantly inhibited their histamine release (36% histamine release) and the increase in area and perimeter, but did not affect significantly their shape factor, in comparison with mast cells incubated with Compound 48/80 alone. These data clearly show that there is a relationship between mast cell activation, consequent histamine release and changes in cell area, perimeter and shape factor and that Nedocromil not only inhibits mast cell histamine release but also the activation induced morphometric changes in mast cells.     

Closed-loop deep brain stimulation is superior in ameliorating parkinsonism

Continuous high-frequency deep brain stimulation (DBS) is a widely used therapy for advanced Parkinson's disease (PD) management. However, the mechanisms underlying DBS effects remain enigmatic and are the subject of an ongoing debate. Here, we present and test a closed-loop stimulation strategy for PD in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) primate model of PD. Application of pallidal closed-loop stimulation leads to dissociation between changes in basal ganglia (BG) discharge rates and patterns, providing insights into PD pathophysiology. Furthermore, cortico-pallidal closed-loop stimulation has a significantly greater effect on akinesia and on cortical and pallidal discharge patterns than standard open-loop DBS and matched control stimulation paradigms. Thus, closed-loop DBS paradigms, by modulating pathological oscillatory activity rather than the discharge rate of the BG-cortical networks, may afford more effective management of advanced PD. Such strategies have the potential to be effective in additional brain disorders in which a pathological neuronal discharge pattern can be recognized.     

Canopy throughfall of Picea abies (L.) Karst. as depending on trace gas concentrations

At six sites in central Germany consequences of SO₂, NO_X and O₃ deposition and of acid precipitation on canopy throughfall of sulphate, nitrate, ammonium, organic acids and of metal cations from Norway spruce crowns were investigated in the field. Measured canopy throughfall rates (mmol ion kg^-1 needle dw a^-1 are separated in (i) background ion throughfall rates in clean air and (ii) trace gas-(or acid interception)-dependent throughfall rates at ambient trace gas concentrations. Based on synchronously measured pollution, precipitation and canopy throughfall data, statistical response functions are given, which allow the separate estimation of annual rates of sulphur and nitrogen deposition into spruce canopies if only annual means of SO₂ or NO₂ concentrations in air are known. The specific SO₂ deposition rate of (0.841±0.214) mmol S kg^-1 needle dw a^-1 (nPa SO₂ Pa^-1)^-1 is 2.3 times higher than the specific stomatal SO₂ uptake. The NO₂-dependent nitrogen deposition of (2.464±0.707) mmol N kg^-1 needle dw a^-1 (nPa NO₂ Pa^-1)^-1 is 2.2 times higher than the specific stomatal NO_X (NO₂+NO) uptake. These ratios (2.32.2) are explained by the percentage of annual hours with open needle stomata. The shape of observed epicuticular SO₂ and NO_X deposition curves and of stomatal SO₂ and NO_X uptake curves are congruent. As for stomatal NO_X uptake, there is an apparent compensation point at (5 to 8) nPa NO₂ Pa^-1. There is significant SO₂-dependent canopy throughfall of Ca>K>Al>Mg>Fe in this order of relative importance. NO_X deposition in spruce canopies reduces K⁺ throughfall and it weakly promotes throughfall of Mn²⁺ and Zn²⁺. There was no significant codeposition of sulphate and ammonium and no ion exchange of intercepted H₃O⁺ with nutrient cations at the measured ambient pH values of the precipitation water. In the presence of O₃, throughfall of Mn²⁺ is reduced and throughfall of K⁺, Ca²⁺ and Al³⁺ is enhanced. In the cooperative presence of SO₂, NO₂ and O₃ pollution in the field there is a 1.3-fold increase of the annual K⁺ demand and a 1.5-fold Mg²⁺ demand of spruce canopies relative to the situation in clean air. This trace gas-dependent additional cation demand of spruce canopies corresponds to a needle loss percentage of (23 to 33)% if the additional K⁺ and Mg²⁺ throughfall could not be recycled in spruce ecosystems. Observed canopy thinning ranges from (13 to 26)% at the investigated six spruce stands.Abbreviations A_spec Specific needle surface area per kg needle dry matter (m²kg^-1 needle dw) - A_tot Total needle surface of spruce stands (ha ha^-1) - [gas]_a Ambient trace gas concentration (gas=SO₂; NO₂ or O₃) in air (nPa Pa^-1=ppb) - GP Number of days per annual growth period d a^-1) - IC_H30 ⁺ Acid interception rate (Eq H₃O⁺ kg^-1 needle dw a^-1) - k_o Trace gas-independent ion throughfall rate constant (mmol kg^-1 needle dw a^-1) - k_gas SO₂-,NO₂-or O₃-dependent ion throughfall rate per unit of trace gas pollution (mmol kg^-1 needle dw a^-1 (nPa Pa^-1)^-1) - k_H30 Specific H₃O⁺/Me⁺ exchange ratio (mol mol^-1) - L_o Background throughfall rate at [gas]_a=0 (mmol kg^-1 needle dw a^-1) - L_ion Canopy throughfall rate of ions (mmol kg^-1 needle dw a^-1) - L'_ion Trace gas dependent ion throughfall (mEq kg^-1 needle dw a^-1 (nPa Pa^-1)^-1) - LAI Leaf area index of the canopy (m² projected needle surface m^-2 ground) - Me⁺ Equivalents of metal cations (Eq) - N Stock of needles of spuce stands in the field (kg needle dw ha^-1) - P_% Percentage of needle loss relative to a healthy reference (%) - r Pearson correlation coefficient (no dimension) - R COO^--Sum of all organic anion equivalents Cat⁺ - An^- (Eq kg^-1 needle dw a^-1) - An^- Sum of all measured inorganic anion equivalents (Eq kg^-1 needle dw a^-1) - Cat⁺ Sum of all measured inorganic cation equivalents (Eq kg^-1 needle dw a^-1)     

Generation of iPSCs as a Pooled Culture Using Magnetic Activated Cell Sorting of Newly Reprogrammed Cells

Although significant advancement has been made in the induced pluripotent stem cell (iPSC) field, current methods for iPSC derivation are labor intensive and costly. These methods involve manual selection, expansion, and characterization of multiple clones for each reprogrammed cell sample and therefore significantly hampers the feasibility of studies where a large number of iPSCs need to be derived. To develop higher throughput iPSC reprogramming methods, we generated iPSCs as a pooled culture using rigorous cell surface pluripotent marker selection with TRA-1-60 or SSEA4 antibodies followed by Magnetic Activated Cell Sorting (MACS). We observed that pool-selected cells are similar or identical to clonally derived iPSC lines from the same donor by all criteria examined, including stable expression of endogenous pluripotency genes, normal karyotype, loss of exogenous reprogramming factors, and in vitro spontaneous and lineage directed differentiation potential. This strategy can be generalized for iPSC generation using both integrating and non-integrating reprogramming methods. Our studies provide an attractive alternative to clonal derivation of iPSCs using rigorously selected cell pools and is amenable to automation.     

An improved device for posterior rhinomanometry to measure nasal resistance

Rhinomanometry is a method for measuring nasal resistance for the purpose of providing an objective evaluation of nasal patency. Posterior rhinomanometry is accomplished without the use of a mask, thus allowing the patient to breathe naturally. Here, we report on the improvements we have made to the existing posterior rhinomanometry system. In this system, nasal airflow is measured indirectly by measuring the pressure differential across a small mesh window in the body plethysmograph. We have calibrated this measurement and developed software that automatically provides the correct values for all airflow rates. Also, we have developed software that automatically corrects for the phase shift caused by the plethysmograph structure. These refinements should provide more accurate values for nasal resistance.     

Emission of hydrogen sulfide by twigs of coniferes — acomparison of Norway spruce (Picea abies (L.) Karst.), Scotch pine (Pinus sylvestris L.) and Blue spruce (Picea pungens Engelm.)

The emission of reduced volatile sulfur compounds from twigs of Norway spruce (Picea abies (L.) Karst.) was measured in the field by cryosampling and gaschromatographic analysis. Trees were growing in the Erzgebirge (E-Germany) at Oberbärenburg and at the Kahleberg and at a third stand in NW-Bavaria (S-Germany). Emission rates were also measured for Scotch pine (Pinus sylvestris L.) and Blue spruce (Picea pungens Engelm.) at the Kahleberg. Twigs still attached to the trees were enclosed in a flow-through gas exchange cuvette. H₂S was detected as the predominant reduced sulfur compound emitted from the twigs. The mean H₂S emission rate from twigs of Norway spruce varied between 0.04 pmol kg^-1 dw s^-1 at Würzburg and 6.21 pmol kg^-1 dw s^-1 at the Kahleberg. Comparing different species at the Kahleberg, the mean H₂S emission rate was almost the same from twigs of Norway spruce (6.2 pmol kg^-1 dw s^-1) and Blue Spruce trees (5.9 pmol kg^-1 dw s^-1) but it was approximately 18 times higher for Scotch pine (110 pmol kg^-1 dw s^-1). The percentage of SO₂-exclusion via H₂S-emission of the tree species investigated at the Kahleberg is calculated on the basis of data on SO₂ fluxes. It is very small for Norway spruce and Blue spruce. However, for Scotch pine, H₂S emission contributes about 10% to the detoxification of SO₂.