Experimental forest soil warming: response of autotrophic and heterotrophic soil respiration to a short-term 10°C temperature rise

We warmed the top soil of a mature coniferous forest stand by means of heating cables on control and trenched plots within 24 h by 10°C at 1 cm soil depth (9°C at 5 cm depth) and measured the effect on the autotrophic (R_A) and heterotrophic (R_H) component of total soil CO₂ efflux (R_S). The short time frame of warming enabled us to exclude confounding fluctuations in soil moisture and carbon (C) flow from the canopy. The results of the field study were backed up by a lab soil incubation experiment. During the first 12 h of warming, R_A strongly responded to soil warming; The Q ₁₀ values were 5.61 and 6.29 for 1 and 5 cm soil depth temperature. The Q ₁₀ values for R_A were almost twice as high as the Q ₁₀ values of R_H (3.04 and 3.53). Q ₁₀ values above 5 are above reasonable plant physiological values for root respiration. We see interactions of roots, mycorrhizae and heterotrophic microbes, combined with fast substrate supply to the rhizosphere as an explanation for the high short-term temperature response of R_A. When calculated over the whole duration (24 h) of the field soil-warming experiment, temperature sensitivities of R_A and R_H were similar (no significant difference at P < 0.05); Q ₁₀ values were 3.16 and 3.96 for R_A and 2.94 and 3.35 for R_H calculated with soil temperatures at 1 and 5 cm soil depth, respectively. Laboratory incubation showed that different soil moisture contents of trenched and control plots affected rates of R_H, but did not affect the temperature sensitivity of R_H. We conclude that a single parameter is sufficient to describe the temperature sensitivity of R_S in soil C models which operate on larger temporal and spatial scales. The strong short-term response of R_A may be of relevance in soils suspected to experience increasingly strong diurnal temperature variations.     

Heritability of and correlations among genotype-by-environment stability statistics for grain yield in bread wheat

Several genotype-by-environment stability measures are in use, but little information exists about their inheritance or genetic inter-relationships. Among those measures in common use are the linear regression coefficient (b), deviations from regression (s_b), coefficient of determination (R²), coefficient of phenotypic variation (CPV) and, more recently, interaction principal components (IPCA) of the additive-main-effect-and-multiplicative-interaction (AMMI) model. Because of the factorial structure of the data, the diallel cross is well suited to study these parameters and their relationship to quantitative traits. For this study a complete diallel cross, derived by mating eight lines from a broad based bread wheat breeding population, was grown for several growing seasons at two Ugandan locations, one of which was prone to yellow rust. Stability parameters and grain yield were measured for each cross. CPV had the highest narrow-sense heritability (h²=0.522) followed by IPCA1 of the AMMI (h²=0.461). Lowest narrow-sense heritabilities were calculated for b and R² (h²=0.150 and 0.100 respectively). There were high additive genetic correlations (r_A) between grain yield and CPV (r_A=−0.933), grain yield and IPCA1 (r_A=0.707), and grain yield and IPCA2 (r_A=0.751). The genetic association between CPV and IPCA1 was also high and negative (r_A= −0.934). These results suggest that it may be possible to select simultaneously for high and stable grain yield in this broad-based bread wheat breeding pool by selecting outyielders that exhibit a low CPV. Received: 25 July 2000 / Accepted: 7 December 2000     

Autotrophic and heterotrophic respiration in needle fir and Quercus-dominated stands in a cool-temperate forest, central Korea

To investigate annual variation in soil respiration (R _S) and its components [autotrophic (R _A) and heterotrophic (R _H)] in relation to seasonal changes in soil temperature (ST) and soil water content (SWC) in an Abies holophylla stand (stand A) and a Quercus-dominated stand (stand Q), we set up trenched plots and measured R _S, ST and SWC for 2 years. The mean annual rate of R _S was 436 mg CO₂ m⁻² h⁻¹, ranging from 76 to 1,170 mg CO₂ m⁻² h⁻¹, in stand A and 376 mg CO₂ m⁻² h⁻¹, ranging from 82 to 1,133 mg CO₂ m⁻² h⁻¹, in stand Q. A significant relationship between R _S and its components and ST was observed over the 2 years in both stands, whereas a significant correlation between R _A and SWC was detected only in stand Q. On average over the 2 years, R _A accounted for approximately 34% (range 17–67%) and 31% (15–82%) of the variation in R _S in stands A and Q, respectively. Our results suggested that vegetation type did not significantly affect the annual mean contributions of R _A or R _H, but did affect the pattern of seasonal change in the contribution of R _A to R _S.     

Partitioning of Respiration in an Intensively Managed Grassland

Total (R_TOT) and heterotrophic (R_H) respiration were measured in an intensively managed perennial ryegrass (Lolium perenne L.) grassland. The overall aim of the study was to partition R_TOT into R_H and autotrophic respiration (R_A). This was achieved as follows: (1) analyse the effect of air temperature, soil moisture content and leaf area index on R_TOT and the influence of soil temperature and soil moisture content on R_H; (2) combine these effects into separate empirical models for R_TOT and R_H and; (3) use these models to determine temporal trends in R_TOT and R_H and to assess the relative contribution of R_H and R_A to R_TOT. CO₂ fluxes were measured using a vented and thermostatically controlled perspex chamber in conjunction with a portable infrared gas analyser. R_TOT was measured in plots with grass and R_H in plots with bare soil. R_TOT was related to air temperature and R_H to soil temperature using exponential relationships. Both R_TOT and R_H were related to soil moisture content using lognormal relationships. R_TOT was related to leaf area index using a linear relationship. These relationships were combined to produce statistical response functions that explained 87% and 84% of the variation in R_TOT and R_H, respectively. These relationships were combined with meteorological and leaf area index data to reconstruct daily and seasonal fluxes. R_TOT values in wintertime were ~4 g C m⁻² day⁻¹ increasing to ~10 g C m⁻² day⁻¹ in summertime when temperatures and leaf area index were higher and soils were drier. R_H has a similar seasonal trend to R_TOT but was consistently lower. Wintertime values were ~2 g C m⁻² day⁻¹ and increased to ~5 g C m⁻² day⁻¹ in summertime. Before day of year 143, and after day of year 259 R_H and R_A represented 62% and 38% of R_TOT, respectively. In the period between these days R_H and R_A both accounted for 50% of R_TOT. In total during 2004 R_TOT, R_H and R_A were 2.34, 1.31 and 1.03 kg C m⁻², respectively.     

Quantitative structure-activity relationship study on affinity profile of a series of 1,8-naphthyridine antagonists toward bovine adenosine receptors

The affinity profiles for the bovine adenosine receptors, A₁ and A_2A, of a series of 1,8-naphthyridine derivatives were quantitatively analyzed using physicochemical and structural parameters of the substituents, present at varying positions of the molecules. The derived significant correlation, for bovine A₁ receptor, suggested that a R₁ substituent having a higher van der Waals volume, a R₂ substituent being a hydrogen-bond donor and a R₃ substituent able to transmit a higher field effect are helpful in augmenting the pK_i of a compound. Similarly the study, pertaining to bovine A_2A receptor, revealed that a less bulky substituent at R₂ and a strong electron-withdrawing substituent at R₃ are desirable in improving the binding affinity of a compound while substituents at R₁ remain insignificant to any interaction.     

Transient changes in transpiration, and stem and soil CO2 efflux in longleaf pine (Pinus palustris Mill.) following fire-induced leaf area reduction

In 20-year-old longleaf pine, we examined short-term effects of reduced live leaf area (A _L) via canopy scorching on sap flow (Q; kg H₂O h⁻¹), transpiration per unit leaf area (E _L; mm day⁻¹), stem CO₂ efflux (R _stem; μmol m⁻² s⁻¹) and soil CO₂ efflux (R _soil; μmol m⁻² s⁻¹) over a 2-week period during early summer. R _stem and Q were measured at two positions (1.3-m or BH, and base of live crown—BLC), and R _soil was measured using 15 open-system chambers on each plot. E _L before and after treatment was estimated using Q measured at BLC with estimates of A _L before and after scorching. We expected Q to decrease in scorched trees compared with controls resulting from reduced A _L. We expected R _stem at BLC and BH and R _soil to decrease following scorching due to reduced leaf area, which would decrease carbon supply to the stem and roots. Scorching reduced A _L by 77%. Prior to scorching, Q at BH was similar between scorch and control trees. Following scorching, Q was not different between control and scorch trees; however, E _L increased immediately following scorching by 3.5-fold compared to control trees. Changes in E _L in scorched trees corresponded well with changes in VPD (D), whereas control trees appeared more decoupled over the 5-day period following treatment. By the end of the study, R _stem decreased to 15–25% in scorched trees at both stem positions compared to control trees. Last, we found that scorching resulted in a delayed and temporary increase in R _soil rather than a decrease. No change in Q and increased E _L following scorching indicates a substantial adjustment in stomatal conductance in scorched trees. Divergence in R _stem between scorch and control trees suggests a gradual decline in stem carbohydrates following scorching. The absence of a strong R _soil response is likely due to non-limiting supplies of root starch during early summer.     

Characteristics of leaf photosynthesis and simulated individual carbon budget in <Emphasis Type="Italic">Primula nutans</Emphasis> under contrasting light and temperature conditions

Primula nutans Georgi is widely distributed in hummock-and-hollow wetlands on the Qinghai-Tibetan Plateau. To assess the ecophysiology of this species in responding to microenvironments, we examined the photosynthetic characteristics and individual carbon gain of plants growing in different microsites from a hummock-and-hollow wetland on the Qinghai-Tibetan Plateau and under laboratory conditions. Plants from wetland hummock microsites showed significantly higher light-saturated photosynthetic CO₂ uptake (A _max) than those from microsites in hollows at a controlled temperature of 15°C in leaf chamber. Leaf dark respiration rate (R) was only significantly higher in plants from hummocks than hollows at the measuring temperature of 35°C. Optimum temperature for A _max was 15°C for all plants in the field despite different microsites. In plants growing under laboratory conditions differing in light and temperature, both A _max and R were significantly higher under higher growth light (photosynthetic photon flux density, PPFD: 800 or 400 μmol m⁻² s⁻¹) than low light of 90 μmol m⁻² s⁻¹. No statistically significant differences in A _max and R existed in plants differing in growing temperatures. Estimates derived from the photosynthetic parameters of field plants, and microsite environmental measures including PPFD, air temperature and soil temperature showed that the optimum mean daily temperature for net daily carbon gain was around 10°C and the net daily carbon gain was largely limited under lower daily total PPFD. These results suggest that the differences in A _max and R in P. nutans are strongly affected by growing light regimes but not by temperature regimes. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

毛竹林老竹水平和经营措施对新竹发育质量的影响

毛竹林是我国重要的森林资源类型,在森林固碳和林业应对气候变化中具有不可替代的重要作用。由于毛竹林的持续采伐与自我更新特性,在竹林经营过程中,新竹的发育数量和质量成为评价竹林固碳功能变化的决定性因子。利用两因素随机区组设计,排除地形因子等影响,选取施肥和采伐留养方式这两个因素,研究老竹水平和经营措施对2010年和2013年毛竹林新竹发育质量的影响。结果表明:无论2010年还是2013年,新竹平均胸径、株数和碳储量与3年生和5年生老竹的相关性均高于2年生和4年生老竹。新竹碳储量与3年生和5年生老竹碳储量呈线性相关,建立线性回归模型y=0.675x-2.2491,R~2=0.8561,而新竹碳储量与2年生和4年生老竹碳储量相关性较低,线性回归模型为y=-0.1109x+6.7287,R~2=0.0061。不同经营措施实施后,新老竹之间关系发生了很大的改变,新竹平均胸径与老竹的相关性大幅下降,新竹株数和碳储量与老竹几乎没有相关性,新竹碳储量与3年生和5年生老竹碳储量的线性回归模型为y=0.1036x+3.7539,R~2=0.0981,新竹碳储量与2年生和4年生老竹碳储量的线性回归模型为y=-0.0408x+5.9069,R~2=0.0151。不同经营措施的实施对新竹的平均胸径、株数和地上碳储量产生了很大的影响。处理A_1B_2(大量施肥中度采伐中密度留养)、A_2B_2(中等施肥中度采伐中密度留养)和A_3B_2(不施肥中度采伐中密度留养)新竹平均胸径、新竹株数和新竹碳储量都有所增加,新竹平均胸径增幅为:处理A_2B_2(8.78%)A_1B_2(2.43%)A_3B_2(2.06%),新竹株数增幅为:处理A_1B_2(81.0%)A3B2(35.4%)A2B2(15.2%),新竹地上碳储量增幅为:处理A_1B_2(90.8%)A_3B_2(35.7%)A_2B_2(49.7%),而其余处理基本都会减少,说明适度采伐留养最有利于提高毛竹林新竹的发育质量。仅仅从固碳最大化的角度出发,大量施肥中度采伐中密度留养最有利于新竹碳储量的增加,而从培养大径竹材的角度考虑,中等施肥中度采伐中密度留养能收到更好的效果。     

Estimates of subtropical forest biomass based on airborne LiDAR and Landsat 8 OLI data

 快速、定量、精确地估算区域森林生物量一直是森林生态功能评价以及碳储量研究的重要问题。该研究基于机载激光雷达(LiDAR)点云与Landsat 8 OLI多光谱数据, 借助江苏省常熟市虞山地区55块调查样地数据, 首先提取并分析了87个特征变量(53个OLI特征变量, 34个LiDAR特征变量)与森林地上、地下生物量的Pearson’s相关系数以进行变量优选, 然后利用多元逐步回归法建立森林生物量估算模型(OLI生物量估算模型和LiDAR生物量估算模型), 并与基于两种数据建立的综合生物量估算模型的结果进行比较, 讨论预测结果及其精确性。结果表明: 3种模型(OLI模型、LiDAR模型和综合模型)在所有样地无区分分析时, 地上和地下生物量的估算精度均达到0.4以上, 基于不同森林类型(针叶林、阔叶林、混交林)分析时地上和地下生物量的估算精度均有明显提高, 达到0.67及以上。利用分森林类型模型估算生物量, 综合生物量估算模型精度(地上生物量: R²为0.88; 地下生物量: R²为0.92)优于OLI生物量估算模型(地上生物量: R²为0.73; 地下生物量: R²为0.81)和LiDAR生物量估算模型(地上生物量: R²为0.86; 地下生物量: R²为0.83)。     

Growth and nitrate uptake properties of plants grown at different relative rates of nitrogen supply. II. Activity and affinity of the nitrate uptake system in Pisum and Lemna in relation to nitrogen availability and nitrogen demand

Abstract Net nitrate uptake rates were measured and the kinetics calculated in non-nodulated Pisum sativum L. cv. Marma and Lemna gibba L. adapted to constant relative rates of nitrate-N additions (R_A), ranging from 0.03 to 0.27 d^?1 for Pisum and from 0.05 to 0.40 d^?1 for Lemna, V_max of net nitrate uptake (measured in the range 10 to 100 mmol m^?3 nitrate, i.e. ‘system I’) increased with R_A in the growth limiting range but decreased when R_A exceeded the relative growth rate (RGR), K_m was not significantly related to changes in R_A. On the basis of previous ¹³N-flux experiments, it is concluded that the differences in V_max at growth limiting R_A are attributable to differences in influx rates. Linear relationships between V_max and tissue nitrogen concentrations were obtained in the growth limiting range for both species, and extrapolated intercepts relate well with the previously defined minimal nitrogen concentrations for plant growth (Oscarson, Ingemarsson & Larsson, 1989). Analysis of V_max for net nitrate uptake on intact plant basis in relation to nitrogen demand during stable, nitrogen limited, growth shows an increased overcapacity at lower R_A values in both species, which is largely explained by the increased relative root size at low R_A. A balancing nitrate concentration, defined as the steady state concentration needed to sustain the relative rate of increase in plant nitrogen (R_N), predicted by R_A, was calculated for both species. In the growth limiting range, this value ranges from 3.5 mmol m^?3 (R_A 0.03 d^?1) to 44 mmol m^?3 (R_A 0.21 d^?1) for Pisum and from 0.2 mmol m^?3 (R_A 0.05 d^?1) to 5.4 mmol m^?3 (R_A 0.03 d^?1) for Lemna. It is suggested that this value can be used as a unifying measure of the affinity for nitrate, integrating the performance of the nitrate uptake system with nitrate flux and long term growth and demand for nitrogen.     

Multiple sites of retardation of electron transfer in Photosystem II after hydrolysis of phosphatidylglycerol

Phosphatidylglycerol (PG), containing the unique fatty acid Δ3, trans-16:1-hexadecenoic acid, is a minor but ubiquitous lipid component of thylakoid membranes of chloroplasts and cyanobacteria. We investigated its role in electron transfers and structural organization of Photosystem II (PSII) by treating Arabidopsis thaliana thylakoids with phospholipase A₂ to decrease the PG content. Phospholipase A₂ treatment of thylakoids (a) inhibited electron transfer from the primary quinone acceptor Q_A to the secondary quinone acceptor Q_B, (b) retarded electron transfer from the manganese cluster to the redox-active tyrosine Z, (c) decreased the extent of flash-induced oxidation of tyrosine Z and dark-stable tyrosine D in parallel, and (d) inhibited PSII reaction centres such that electron flow to silicomolybdate in continuous light was inhibited. In addition, phospholipase A₂ treatment of thylakoids caused the partial dissociation of (a) PSII supercomplexes into PSII dimers that do not have the complete light-harvesting complex of PSII (LHCII); (b) PSII dimers into monomers; and (c) trimers of LHCII into monomers. Thus, removal of PG by phospholipase A₂ brings about profound structural changes in PSII, leading to inhibition/retardation of electron transfer on the donor side, in the reaction centre, and on the acceptor side. Our results broaden the simple view of the predominant effect being on the Q_B-binding site.     

Selection,structure and the heritability of behaviour

Characters which are closely linked to fitness often have low heritabilities (V_A/V_P). Low heritabilities could be because of low additive genetic variation (V_A), that had been depleted by directional selection. Alternatively, low heritabilities may be caused by large residual variation (V_R=V_P – V_A) compounded at a disproportionately higher rate than V_A across integrated characters. Both hypotheses assume that each component of quantitative variation has an independent effect on heritability. However, V_A and V_R may also covary, in which case differences in heritability cannot be fully explained by the independent effects of elimination‐selection or compounded residual variation. We compared the central tendency of published behavioural heritabilities (mean=0.31, median=0.23) with morphological and life history data collected by 26 ). Average behavioural heritability was not significantly different from average life history heritability, but both were smaller than average morphological heritability. We cross‐classified behavioural traits to test whether variation in heritability was related to selection (dominance, domestic/wild) or variance compounding (integration level). There was a significant three‐way interaction between indices of selection and variance compounding, related to the absence of either effect at the highest integration level. At lower integration levels, high dominance variance indicated effects of selection. It was also indicated by the low CV_A of domestic species. At the same time CV_R increased disproportionately faster than CV_A across integration levels, demonstrating variance compounding. However, neither CV_R nor CV_A had a predominant effect on heritability. The partial regression coefficients of CV_R and CV_A on heritability were similar and a path analysis indicated that their (positive) correlation was also necessary to explain variation in heritability. These results suggest that relationships between additive genetic and residual components of quantitative genetic variation can constrain their independent direct effects on behavioural heritability.     

An examination of structural interactions presumed to be of importance in the stabilization of phospholipase A2 dimers based upon comparative protein sequence analysis of a monomeric and dimeric enzyme from the venom ofAgkistrodon p. piscivorus

Phospholipases A₂ may exist in solution both as monomers and dimers, but enzymes that form strong dimers (K _D approximately 10^–9 M) have been found, thus far, only in venoms of the snake family Crotilidae. The complete amino acid sequences of a basic monomeric and an acidic dimeric phospholipase A₂ fromAgkistrodon piscivorus piscivorus (American cotton-mouth water moccasin) venom have been determined by protein sequencing methods as part of a search for aspects of structure contributing to formation of stable dimers. Both the monomeric and dimeric phospholipases A₂ are highly homologous to the dimeric phospholipases A₂ fromCrotalus atrox andCrotalus adamanteus venoms, and both have the seven residue carboxy-terminal extension characteristic of the crotalid and viperid enzymes. Thus, it is clear that the extension is not a prerequisite for dimerization. Studies to date have revealed two characteristic features of phosphilipases A₂ that exist in solution as strong dimers. One is the presence in the dimers of a Pro-Pro sequence at position 112 and 113 which just precedes the seven residue carboxy-terminal extension (residues 116–122). The other is a low isoelectric point; only the acidic phospholipases A₂ have been observed, thus far, to form stable dimers. These, alone or together, may be necessary, though not sufficient conditions for phospholipase A₂ dimer formation. Ideas regarding subunit interactions based upon crystallographic data are evaluated relative to the new sequence information on the monomeric and dimeric phospholipases A₂ fromA. p. piscivorus venom.     

Predicting tropical plant physiology from leaf and canopy spectroscopy

A broad regional understanding of tropical forest leaf photosynthesis has long been a goal for tropical forest ecologists, but it has remained elusive due to difficult canopy access and high species diversity. Here we develop an empirical model to predict sunlit, light-saturated, tropical leaf photosynthesis using leaf and simulated canopy spectra. To develop this model, we used partial least squares (PLS) analysis on three tropical forest datasets (159 species), two in Hawaii and one at the biosphere 2 laboratory (B2L). For each species, we measured light-saturated photosynthesis (A), light and CO₂ saturated photosynthesis (A _max), respiration (R), leaf transmittance and reflectance spectra (400–2,500 nm), leaf nitrogen, chlorophyll a and b, carotenoids, and leaf mass per area (LMA). The model best predicted A [r ²  = 0.74, root mean square error (RMSE) = 2.9 μmol m⁻² s⁻¹)] followed by R (r ²  = 0.48), and A _max (r ²  = 0.47). We combined leaf reflectance and transmittance with a canopy radiative transfer model to simulate top-of-canopy reflectance and found that canopy spectra are a better predictor of A (RMSE = 2.5 ± 0.07 μmol m⁻² s⁻¹) than are leaf spectra. The results indicate the potential for this technique to be used with high-fidelity imaging spectrometers to remotely sense tropical forest canopy photosynthesis.     

Inhibition by light of CO2 evolution from dark respiration: Comparison of two gas exchange methods

Two approaches to determine the fraction (μ) of mitochondrial respiration sustained during illumination by measuring CO₂ gas exchange are compared. In single leaves, the respiration rate in the light (`day respiration' rate R<sub>d</sub>) is determined as the ordinate of the intersection point of A–c<sub>i</sub> curves at various photon flux densities and compared with the CO<sub>2</sub> evolution rate in darkness (`night respiration' rate R_n). Alternatively, using leaves with varying values of CO₂ compensation concentration (Γ), intracellular resistance (r_i) and R_n, an average number for μ can be derived from the linear regression between Γ and the product r_iċR_n. Both methods also result in a number c* for that intercellular CO₂ concentration at which net CO₂ uptake rate is equal to –R_d. c* is an approximate value of the photocompensation point Γ* (Γ in the absence of mitochondrial respiration), which is related to the CO₂/O₂ specificity factor of Rubisco S_c/o. The presuppositions and limitations for application of both approaches are discussed. In leaves of Nicotiana tabacum, at 22 °C, single leaf measurements resulted in mean values of μ = 0.71 and c_* = 34 μmol mol⁻¹. At the photosynthetically active photon flux density of 960 μmol quanta m⁻² s⁻¹, nearly the same numbers were derived from the linear relationship between Γ and r_iċR_n. c* and R_d determined by single leaf measurements varied between 31 and 41 μmol mol⁻¹ and between 0.37 and 1.22 μmol m⁻² s⁻¹, respectively. A highly significant negative correlation between c* and R_d was found. From the regression equation we obtained estimates for Γ* (39 μmol mol⁻¹), S_c/o (96.5 mol mol⁻¹) and the mesophyll CO₂ transfer resistance (7.0 mol⁻¹ m² s). This revised version was published online in June 2006 with corrections to the Cover Date.     

An examination of structural interactions presumed to be of importance in the stabilization of phospholipase A2 dimers based upon comparative protein sequence analysis of a monomeric and dimeric enzyme from the venom ofAgkistrodon p. piscivorus

Phospholipases A₂ may exist in solution both as monomers and dimers, but enzymes that form strong dimers (K _D approximately 10^?9 M) have been found, thus far, only in venoms of the snake family Crotilidae. The complete amino acid sequences of a basic monomeric and an acidic dimeric phospholipase A₂ fromAgkistrodon piscivorus piscivorus (American cotton-mouth water moccasin) venom have been determined by protein sequencing methods as part of a search for aspects of structure contributing to formation of stable dimers. Both the monomeric and dimeric phospholipases A₂ are highly homologous to the dimeric phospholipases A₂ fromCrotalus atrox andCrotalus adamanteus venoms, and both have the seven residue carboxy-terminal extension characteristic of the crotalid and viperid enzymes. Thus, it is clear that the extension is not a prerequisite for dimerization. Studies to date have revealed two characteristic features of phosphilipases A₂ that exist in solution as strong dimers. One is the presence in the dimers of a Pro-Pro sequence at position 112 and 113 which just precedes the seven residue carboxy-terminal extension (residues 116–122). The other is a low isoelectric point; only the acidic phospholipases A₂ have been observed, thus far, to form stable dimers. These, alone or together, may be necessary, though not sufficient conditions for phospholipase A₂ dimer formation. Ideas regarding subunit interactions based upon crystallographic data are evaluated relative to the new sequence information on the monomeric and dimeric phospholipases A₂ fromA. p. piscivorus venom.     

Adenosine receptors regulate exosome production

Exosomes, small-sized extracellular vesicles, carry components of the purinergic pathway. The production by cells of exosomes carrying this pathway remains poorly understood. Here, we asked whether type 1, 2A, or 2B adenosine receptors (A₁Rs, A_2ARs, and A_2BRs, respectively) expressed by producer cells are involved in regulating exosome production. Preglomerular vascular smooth muscle cells (PGVSMCs) were isolated from wildtype, A₁R^?/?, A_2AR^?/?, and A_2BR^?/? rats, and exosome production was quantified under normal or metabolic stress conditions. Exosome production was also measured in various cancer cells treated with pharmacologic agonists/antagonists of A₁Rs, A_2ARs, and A_2BRs in the presence or absence of metabolic stress or cisplatin. Functional activity of exosomes was determined in Jurkat cell apoptosis assays. In PGVSMCs, A₁R and A_2AR constrained exosome production under normal conditions (p?=?0.0297; p?=?0.0409, respectively), and A₁R, A_2AR, and A_2BR constrained exosome production under metabolic stress conditions. Exosome production from cancer cells was reduced (p?=?0.0028) by the selective A_2AR agonist CGS 21680. These exosomes induced higher levels of Jurkat apoptosis than exosomes from untreated cells or cells treated with A₁R and A_2BR agonists (p?=?0.0474). The selective A_2AR antagonist SCH 442416 stimulated exosome production under metabolic stress or cisplatin treatment, whereas the selective A_2BR antagonist MRS 1754 reduced exosome production. Our findings indicate that A_2ARs suppress exosome release in all cell types examined, whereas effects of A₁Rs and A_2BRs are dependent on cell type and conditions. Pharmacologic targeting of cancer with A_2AR antagonists may inadvertently increase exosome production from tumor cells.

     

Adrenoceptors and signal transduction in neurons

The adrenergic system is an essential regulator of neuronal, endocrine, cardiovascular, vegetative, and metabolic functions. The endogenous catecholamines epinephrine and norepinephrine activate G-protein-coupled receptors to transmit their signal across the plasma membrane. These adrenoceptors can be divided into three different groups: the α₁-receptors (α_1A, α_1B, α_1D), α₂-receptors (α_2A, α_2B, α_2C), and β-receptors (β₁, β₂, β₃). This review summarizes recent findings in the field of adrenoceptor signaling in neurons and includes a discussion of receptor-associated proteins, receptor dimerization, subcellular trafficking, and fluorescence optical methods for studying the kinetics of adrenergic signaling. Spatio-temporal imaging may become an important future tool for identifying the physiological significance of these complex signaling mechanisms in vivo. Gene-targeted mouse models carrying deletions in α₂-adrenoceptor have provided detailed insights into specific neuronal functions of the three α₂-receptor subtypes.     

Recent improvements in the development of A<Subscript>2B</Subscript> adenosine receptor agonists

Adenosine is known to exert most of its physiological functions by acting as local modulator at four receptor subtypes named A₁, A_2A, A_2B and A₃ (ARs). Principally as a result of the difficulty in identifying potent and selective agonists, the A_2B AR is the least extensively characterised of the adenosine receptors family. Despite these limitations, growing understanding of the physiological meaning of this target indicates promising therapeutic perspectives for specific ligands. As A_2B AR signalling seems to be associated with pre/postconditioning cardioprotective and anti-inflammatory mechanisms, selective agonists may represent a new therapeutic group for patients suffering from coronary artery disease. Herein we present an overview of the recent advancements in identifying potent and selective A_2B AR agonists reported in scientific and patent literature. These compounds can be classified into adenosine-like and nonadenosine ligands. Nucleoside-based agonists are the result of modifying adenosine by substitution at the N ⁶-, C²-positions of the purine heterocycle and/or at the 5′-position of the ribose moiety or combinations of these substitutions. Compounds 1-deoxy-1-{6-[N′-(furan-2-carbonyl)-hydrazino]-9H-purin-9-yl}-N-ethyl-β-D-ribofuranuronamide (19, hA₁ K _i = 1050 nM, hA_2A K _i = 1550 nM, hA_2B EC₅₀ = 82 nM, hA₃ K _i > 5 μM) and its 2-chloro analogue 23 (hA₁ K _i = 3500 nM, hA_2A K _i = 4950 nM, hA_2B EC₅₀ = 210 nM, hA₃ K _i > 5 μM) were confirmed to be potent and selective full agonists in a cyclic adenosine monophosphate (cAMP) functional assay in Chinese hamster ovary (CHO) cells expressing hA_2B AR. Nonribose ligands are represented by conveniently substituted dicarbonitrilepyridines, among which 2-[6-amino-3,5-dicyano-4-[4-(cyclopropylmethoxy)phenyl]pyridin-2-ylsulfanyl]acetamide (BAY-60–6583, hA₁, hA_2A, hA₃ EC₅₀ > 10 μM; hA_2B EC₅₀ = 3 nM) is currently under preclinical-phase investigation for treating coronary artery disorders and atherosclerosis.