Effectiveness in the field of Bt rice lines against target pests under various cultural regimes

China has a long history of rice cultivation, incorporating several cultural practices known to influence damage by insect pests. Transgenic Bt rice expresses lepidopteran‐specific insecticidal proteins that primarily target lepidopteran insect pests. However, the effectiveness of Bt rice against target insect pests under different cultural regimes has not been evaluated. In this study, the effectiveness of Bt rice lines against rice leaffolder, Cnaphalocrocis medinalis (Guenée) (Lepidoptera: Pyralidae), and striped stem borer, Chilo suppressalis Walker (Lepidoptera: Crambidae), was evaluated under various transplanting densities, crop establishment methods, and planting times. The results showed that Bt rice lines (T2A‐1 and T1C‐19, containing Cry2A and Cry1C, respectively) could prevent damage by these target pests under a range of cultural practices. Injury by C. medinalis or C. suppressalis on rice did not differ with the rice lines under various transplanting densities. Direct‐seeded non‐Bt rice MH63 suffered heavier injury by C. medinalis and C. suppressalis than it did with transplanting, whereas injury to the two Bt rice lines did not differ with planting methods. Planting time significantly affected injury by C. medinalis or C. suppressalis on non‐Bt rice, whereas injury to Bt rice lines did not differ with planting time. These results suggest that transplanting density, planting method, and planting time did not significantly affect the resistance of two Bt rice lines, due to their high insecticidal activity against target insects.     

Effects of Obesity on Substrate Utilization during Exercise

Objective: The capacity for lipid and carbohydrate (CHO) oxidation during exercise is important for energy partitioning and storage. This study examined the effects of obesity on lipid and CHO oxidation during exercise. Research Methods and Procedures: Seven obese and seven lean [body mass index (BMI), 33 ± 0.8 and 23.7 ± 1.2 kg/m², respectively] sedentary, middle‐aged men matched for aerobic capacity performed 60 minutes of cycle exercise at similar relative (50% Vo _2max) and absolute exercise intensities. Results: Obese men derived a greater proportion of their energy from fatty‐acid oxidation than lean men (43 ± 5% 31 ± 2%; p = 0.02). Plasma fatty‐acid oxidation determined from recovery of infused [0.15 μmol/kg fat‐free mass (FFM) per minute] [1‐¹³C]‐palmitate in breath CO₂ was similar for obese and lean men (8.4 ± 1.1 and 29 ± 15 μmol/kg FFM per minute). Nonplasma fatty‐acid oxidation, presumably, from intramuscular sources, was 50% higher in obese men than in lean men (10.0 ± 0.6 versus 6.6 ± 0.8 μmol/kg FFM per minute; p < 0.05). Systemic glucose disposal was similar in lean and obese groups (33 ± 8 and 29 ± 15 μmol/kg FFM per minute). However, the estimated rate of glycogen‐oxidation was 50% lower in obese than in lean men (61 ± 12 versus 90 ± 6 μmol/kg FFM per minute; p < 0.05). Discussion: During moderate exercise, obese sedentary men have increased rates of fatty‐acid oxidation from nonplasma sources and reduced rates of CHO oxidation, particularly muscle glycogen, compared with lean sedentary men.     

控制大田和自然大田稻纵卷叶螟危害水稻生理生态参数估算模型

为了准确监测和客观评估稻纵卷叶螟对水稻生长发育和产量形成的危害,利用ASD Field Spec3地物波谱仪和SPAD-502叶绿素仪分别采集控制大田试验（2015年和2019年）和自然大田试验（2020年）在各生育期（拔节期、孕穗期、灌浆期、成熟期）水稻的冠层高光谱数据和SPAD值,调查采集样点的虫量和水稻卷叶率,对比分析两种试验中稻纵卷叶螟的虫害发生特征、水稻冠层光谱特征和水稻生理生态参数特征,建立基于高光谱参数的水稻受稻纵卷叶螟危害的生理生态参数估算模型。结果表明,（1）两种试验的水稻SPAD值和冠层的红边至近红外波段的反射率均随着稻纵卷叶螟虫害程度的加重而降低,而可见光波段的反射率则相反;（2）自然大田试验的SPAD值和红光至近红外波段的冠层反射率在水稻生长发育前期要显著低于控制大田试验,而到了后期则反而要略高于控制大田试验;（3）综合分析筛选出自然大田试验和控制大田试验中的多个虫害特征参数和植被指数分别构建出了SPAD的单因子和多因子估算模型,各模型均达到了较好的估算效果,在单因子模型中EVI的二项式函数模拟效果最好,而多因子线性回归估测模型的模拟效果优于所有的单因子模型;（4）通过2021年对这些模型的应用检验发现：这些模型中基于虫量、卷叶率、OSAVI、EVI和DVI的单因子估算模型的SPAD估算值与实测值拟合度很高,其R_v²均超过了0.8,达到了比较理想的估算效果,这为稻纵卷叶螟危害下的水稻SPAD值估测提供了一种精度较高且可行的估算方法。     

An investigation of position 3 in arginine vasopressin with aliphatic,aromatic, conformationally‐restricted,polar and charged amino acids

We report the solid‐phase synthesis and some pharmacological properties of 23 new analogs of arginine vasopressin (AVP) which have the Phe³ residue replaced by a broad variety of amino acids. Peptides 1–9 have at position 3: (1) the mixed aromatic/aliphatic amino acid thienylalanine (Thi) and the aliphatic amino acids; (2) cyclohexylalanine (Cha); (3) norleucine (Nle); (4) Leu; (5) norvaline (Nva); (6) Val; (7) alpha‐aminobutyric acid (Abu); (8) Ala; (9) Gly. Peptides 10–23 have at position 3: the aromatic amino acids, (10) homophenylalanine (Hphe); (11) Tyr; (12) Trp; (13) 2‐naphthylalanine (2‐Nal); the conformationally‐restricted amino acids (14) Pro; (15) 2‐aminotetraline‐2‐carboxylic acid (Atc); the polar amino acids (16) Ser; (17) Thr; (18) Gln; and the charged amino acids (19) Asp; (20) Glu; (21) Arg; (22) Lys; (23) Orn. All 23 new peptides were evaluated for agonistic and, where appropriate, antagonistic activities in in vivo antidiuretic (V₂‐receptor) and vasopressor (V_1a‐receptor) assays and in in vitro (no Mg²⁺) oxytocic assays. The corresponding potencies (units/mg) in these assays for AVP are: 323±16; 369±6 and 13.9±0.5. Peptides 1–9 exhibit the following potencies (units/mg) in these three assays: (1) 379±14; 360±9; 36.2±1.9; (2) 294±21; 73.4±2.7; 0.33±0.02; (3) 249±28; 84.6±4.3; 4.72±0.16; (4) 229±19; 21.4±0.6; 2.1±0.2; (5) 134±5; 31.2±0.9; 28.4±0.2; (6) 114±9; 45.3±2.3; 11.3±1.6; (7) 86.7±2.5; 4.29±0.13; 0.45±0.03; (8) 15.5±1.5; 0.16±0.01; ∼0.02; (9) 3.76±0.03; <0.02; in vitro oxytocic agonism was not detected. These data show that the aliphatic amino acids Cha, Nle, Leu, Nva and Val are well‐tolerated at position 3 in AVP with retention of surprisingly high levels of antidiuretic activity. Peptides 2–9 exhibit significant gains in both antidiuretic/vasopressor (A/P) and antidiuretic/oxytocic (A/O) selectivities relative to AVP. [Thi³]AVP appears to be a more potent antidiuretic and oxytocic agonist than AVP and is equipotent with AVP as a vasopressor agonist. The antidiuretic potencies of peptides 10–23 exhibit drastic losses relative to AVP. They range from a low of 0.018±0.001 units/mg for the Lys³ analog (peptide 22) to a high of 24.6±4.6 units/mg for the Hphe³ analog (peptide 10). Their vasopressor potencies are also drastically reduced. These range from a low of <0.002 units/mg for peptide 22 to a high of 8.99±0.44 units/mg for the Atc³ analog (peptide 15). Peptides 10–23 exhibit negligible or undetectable in vitro oxytocic agonism. The findings on peptides 10–23 show that position 3 in AVP is highly intolerant of changes with aromatic, conformationally‐restricted, polar and charged amino acids. Furthermore, these findings are in striking contrast to our recent discovery that position 3 in the potent V₂/V_1a/OT antagonist d(CH₂)₅d ‐Tyr(Et)²VAVP tolerates a broad latitude of structural change at position 3 with many of the same amino acids, to give excellent retention of antagonistic potencies. The data on peptides 1–4 offer promising clues to the design of more potent and selective AVP V₂ agonists. Copyright © 1999 European Peptide Society and John Wiley & Sons, Ltd.     

Biological Evaluation of Secondary Metabolites from the Root of Machilus obovatifolia

Bioassay‐guided fractionation of the root of Machilus obovatifolia led to the isolation of four new lignans, epihenricine B ( 1 ), threo‐(7′R,8′R) and threo‐(7′S,8′S)‐methylmachilusol D ( 2 and 3 ), and isofragransol A ( 4 ), along with 23 known compounds. The compounds were obtained as isomeric mixtures (i.e., 2 / 3 and 4 / 20 , resp.). The structures were elucidated by spectral analyses. Among the isolates, 1 , licarin A ( 12 ), guaiacin ( 14 ), (±)‐syringaresinol ( 21 ), and (?)‐epicatechin ( 23 ) showed ABTS (=2,2′‐azinobis(3‐ethylbenzothiazoline‐6‐sulfonic acid) cation radical‐scavenging activity, with SC₅₀ values of 11.7±0.5, 12.3±1.1, 11.0±0.1, 10.6±0.3, and 9.5±0.2 μM in 20 min, respectively. In addition, kachirachirol B ( 17 ) showed cytotoxicity against the NCI‐H460 cell line with an IC₅₀ value of 3.1 μg/ml.     

Development of leaffolder resistant transgenic rice expressing <Emphasis Type="Italic">cry2AX1</Emphasis> gene driven by green tissue-specific <Emphasis Type="Italic">rbcS</Emphasis> promoter

The insecticidal cry genes of Bacillus thuringiensis (Bt) have been successfully used for development of insect resistant transgenic rice plants. In this study, a novel cry2AX1 gene consisting a sequence of cry2Aa and cry2Ac gene driven by rice rbcS promoter was introduced into a rice cultivar, ASD16. Among 27 putative rice transformants, 20 plants were found to be positive for cry2AX1 gene. The expression of Cry2AX1 protein in transgenic rice plants ranged from 5.95 to 122.40 ng/g of fresh leaf tissue. Stable integration of the transgene was confirmed in putative transformants of rice by Southern blot hybridization analysis. Insect bioassay on T₀ transgenic rice plants against rice leaffolder (Cnaphalocrosis medinalis) recorded larval mortality up to 83.33 %. Stable inheritance and expression of cry2AX1 gene in T₁ progenies was demonstrated using Southern and ELISA. The detached leaf bit bioassay with selected T₁ plants showed 83.33–90.00 % mortality against C. medinalis. The whole plant bioassay for T₁ plants with rice leaffolder showed significant level of resistance even at a lower level of Cry2AX1 expression varying from 131 to 158 ng/g fresh leaf tissue during tillering stage.     

Scavenging of hydroxyl radical by catecholamines

The direct effects of the four catecholamines (CATs), adrenaline (A), noradrenaline (NA), dopamine (D) and isoproterenol (I), on free radicals were investigated using the free radical 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH^?) and hydroxyl radial (HO^?). The CATs examined were found to inhibit the ESR signal intensity of DPPH^? in a dose‐dependent manner over the range 0.1–2.5 mmol/L in the following order: NA > A > I > D, with IC₅₀ = 0.30 ± 0.03 for noradrenaline and IC₅₀ = 0.86 ± 0.02 for dopamine. Hydroxyl radicals were produced using a Fenton reaction in the presence of the spin trap 5,5‐dimethyl‐1‐pyrroline N‐oxide (DMPO), and ESR technique was applied to detect the CATs reactivity toward the radicals. The reaction rates constant (k_r) of CATs with HO^? were found to be in the order of 10⁹ L/mol/s, and the k_r value for noradrenaline was the highest (k_r = 8.4 × 10⁹ L/mol/s). The CATs examined exhibited also a strong decrease in the light emission (62–73% at 1 mmol/L concentration and 79–89% at 2 mmol/L concentration) from a Fenton‐like reaction. These reactions may be relevant to the biological action of these important polyphenolic compounds. Copyright © 2012 John Wiley & Sons, Ltd.     

Crystallographic characterization of the α,γ C12 helix in hybrid peptide sequences

The solid‐state conformations of two αγ hybrid peptides Boc‐[Aib‐γ⁴(R)Ile]₄‐OMe 1 and Boc‐[Aib‐γ⁴(R)Ile]₅‐OMe 2 are described. Peptides 1 and 2 adopt C₁₂‐helical conformations in crystals. The structure of octapeptide 1 is stabilized by six intramolecular 4 → 1 hydrogen bonds, forming 12 atom C₁₂ motifs. The structure of peptide 2 reveals the formation of eight successive C₁₂ hydrogen‐bonded turns. Average backbone dihedral angles for αγ C₁₂ helices are peptide 1 , Aib; φ (°) = ?57.2 ± 0.8, ψ (°) = ?44.5 ± 4.7; γ⁴(R)Ile; φ (°) = ?127.3 ± 7.3, θ₁ (°) = 58.5 ± 12.1, θ₂ (°) = 67.6 ± 10.1, ψ (°) = ?126.2 ± 16.1; peptide 2 , Aib; φ (°) = ?58.8 ± 5.1, ψ (°) = ?40.3 ± 5.5; ψ⁴(R)Ile; φ (°) = ?123.9 ± 2.7, θ₁ (°) = 53.3 θ 4.9, θ ₂ (°) = 61.2 ± 1.6, ψ (°) = ?121.8 ± 5.1. The tendency of γ⁴‐substituted residues to adopt gauche–gauche conformations about the C^α–C^β and C^β–C^γ bonds facilitates helical folding. The αγ C₁₂ helix is a backbone expanded analog of α peptide 3₁₀ helix. The hydrogen bond parameters for α peptide 3₁₀ and α‐helices are compared with those for αγ hybrid C₁₂ helix. Copyright © 2016 European Peptide Society and John Wiley & Sons.     

In vivo quantification of neuro‐glial metabolism and glial glutamate concentration using 1H‐[13C] MRS at 14.1T

Astrocytes have recently become a major center of interest in neurochemistry with the discoveries on their major role in brain energy metabolism. An interesting way to probe this glial contribution is given by in vivo ¹³C NMR spectroscopy coupled with the infusion labeled glial‐specific substrate, such as acetate. In this study, we infused alpha‐chloralose anesthetized rats with [2‐¹³C]acetate and followed the dynamics of the fractional enrichment (FE) in the positions C4 and C3 of glutamate and glutamine with high sensitivity, using ¹H‐[¹³C] magnetic resonance spectroscopy (MRS) at 14.1T. Applying a two‐compartment mathematical model to the measured time courses yielded a glial tricarboxylic acid (TCA) cycle rate (V_g) of 0.27 ± 0.02 μmol/g/min and a glutamatergic neurotransmission rate (V_NT) of 0.15 ± 0.01 μmol/g/min. Glial oxidative ATP metabolism thus accounts for 38% of total oxidative metabolism measured by NMR. Pyruvate carboxylase (V_PC) was 0.09 ± 0.01 μmol/g/min, corresponding to 37% of the glial glutamine synthesis rate. The glial and neuronal transmitochondrial fluxes (V_x^g and V_xⁿ) were of the same order of magnitude as the respective TCA cycle fluxes. In addition, we estimated a glial glutamate pool size of 0.6 ± 0.1 μmol/g. The effect of spectral data quality on the fluxes estimates was analyzed by Monte Carlo simulations.

     

Cholecystokinin and tryptic activity in the gut and body of developing Atlantic halibut larvae: evidence for participation in the regulation of protein digestion

At 7 days after first feeding (DAFF), the peptide hormone cholecystokinin (CCK) content (fmol individual^?1) and the tryptic activity [μmol arginine‐methyl‐coumarinyl‐7‐amide (MCA) min^?1 individual^?1] per individual gut of Atlantic halibut Hippoglossus hippoglossus larvae were low: 0·2 ± 0·1 and 0·14 ± 0·10, respectively. Thereafter, both parameters increased with the increase in gut mass and reached 19·67 ± 5·58 and 2·71 ± 0·64 at 26 DAFF, respectively. Due to the small sample size, the dry mass (M_G, mg) of the individual gut could not be determined accurately at 7 DAFF. At 13 DAFF M_G represented 5·5% of whole body dry mass (M_w, mg) while at 26 DAFF it had increased to 23%. The mass specific tryptic activity [μmol MCA min^?1 per mg dry mass (M)] in the gut increased from 2·74 ± 1 ± 98 at 13 DAFF to 5·00 ± 0·78 at 26 DAFF. There was more individual variation in the mass specific CCK content (fmol M^?1) but no significant differences were found, although the data indicated an increase (from 23·38 ± 11·26 at 13 DAFF to 36·27 ± 8·96 fmol M^?1 at 26 DAFF). At 7 DAFF the CCK content of the gut represented c. 2% of the whole body CCK content while it increased to c. 62% of the whole body CCK content at 26 DAFF. This demonstrates that it is necessary to separate neural and gastrointestinal sources of CCK in order to determine its alimentary role in fish larvae. Trypsin activity was only found in the gut compartment. In larvae aged 45 DAFF dietary proteins delivery into the gut by tube‐feeding appeared to stimulate post‐prandial secretion of CCK from the gut as well as stimulate pancreatic trypsin secretion, suggesting that both factors contribute to protein digestion.     

High‐performance liquid chromatographic enantioseparation of N‐aryl‐thioureidoalkylphosphonates and thiourylenedi(alkylphosphonates) on polysaccharide‐based chiral stationary phases

The first successful enantioseparation of representative O,O‐diphenyl‐N‐arylthioureidoalkylphosphonates, (±)‐Ptc‐Val^P(OPh)₂ & (±)‐Ptc‐Leu^P(OPh)₂ and thiourylenedi(isobutyl phosphonate), Tcm[Val^P(OPh)₂]₂ on analytical and semipreparative scale was achieved by high‐performance liquid chromatography using polysaccharide‐based chiral stationary phases (CPs). Atc‐AA^P(OPh)₂ was obtained using modified tricomponent condensations of the corresponding aldehydes, N‐arylthiourea and triphenyl phosphite whereas Tcm[Val^P(OPh)₂]₂ by the condensations of aldehydes, thiourea, and triphenyl phosphite. The prepared, racemic (±)‐Atc‐AA^P(OPh)₂ [(±)‐Ptc‐Val^P(OPh)₂, (±)‐Ptc‐Leu^P(OPh)₂, (±)‐Ptc‐Pgly^P(OPh)₂ and (±)‐Ntc‐Pgly^P(OPh)₂] and racemic (±)‐Tcm[AA^P(OPh)₂]₂ [(±)‐Tcm[Nva^P(OPh)₂]₂ & (±)‐Tcm[Val^P(OPh)₂]₂] were adequately characterized and used for chromatographic separations on high‐performance liquid chromatography–chiral stationary phases. The best results were obtained for (±)‐Ptc‐Val^P(OPh)₂, (±)‐Ptc‐Leu^P(OPh)₂ and (±)‐Tcm[Val^P(OPh)₂]₂.     

Acute Effects of Dietary Glycemic Index on Antioxidant Capacity in a Nutrient‐controlled Feeding Study

Oxidative stress, caused by an imbalance between antioxidant capacity and reactive oxygen species, may be an early event in a metabolic cascade elicited by a high glycemic index (GI) diet, ultimately increasing the risk for cardiovascular disease and diabetes. We conducted a feeding study to evaluate the acute effects of low‐GI compared with high‐GI diets on oxidative stress and cardiovascular disease risk factors. The crossover study comprised two 10‐day in‐patient admissions to a clinical research center. For the admissions, 12 overweight or obese (BMI: 27–45 kg/m²) male subjects aged 18–35 years consumed low‐GI or high‐GI diets controlled for potentially confounding nutrients. On day 7, after an overnight fast and then during a 5‐h postprandial period, we assessed total antioxidant capacity (total and perchloric acid (PCA) protein‐precipitated plasma oxygen radical absorbance capacity (ORAC) assay) and oxidative stress status (urinary F_2α‐isoprostanes (F₂IP)). On day 10, we measured cardiovascular disease risk factors. Under fasting conditions, total antioxidant capacity was significantly higher during the low‐GI vs. high‐GI diet based on total ORAC (11,736 ± 668 vs. 10,381 ± 612 µmol Trolox equivalents/l, P = 0.002) and PCA‐ORAC (1,276 ± 96 vs. 1,210 ± 96 µmol Trolox equivalents/l, P = 0.02). Area under the postprandial response curve also differed significantly between the two diets for total ORAC and PCA‐ORAC. No diet effects were observed for the other variables. Enhancement in plasma total antioxidant capacity occurs within 1 week on a low‐GI diet, before changes in other risk factors, raising the possibility that this phenomenon may mediate, at least in part, the previously reported effects of GI on health.     

Field studies of ammonia excretion in Aphanius iberus (Pisces; Cyprinodontidae): body size and habitat effects

The effects of body size and habitat variability on ammonia excretion rates (R_AMs) of Aphanius iberus were analyzed in situ for the first time. At hourly intervals during a 5‐h field experiment, ammonia excretion was measured in 75 mature specimens from three sampling sites (small creek, marine salt‐mine, and salt‐marsh) established in a gradient of water salinity (0–5; 35–40; 65–70‰). Our results showed a specific size dependence pattern of R_AMs in the reproduction period, which might reflect an effect of the reproductive effort. In addition, the results point to a significant decrease in mean R_AM values of each population from freshwater aquatic systems (3.81 ± 0.58 μmol g⁻¹ h⁻¹ in fish of 2.8 ± 0.3 mm total length, TL) to salt aquatic systems with significantly higher alkalinity (2.52 ± 0.35 μmol g⁻¹ h⁻¹ in fish of 3.1 ± 0.5 mm TL in marine salt‐mine; 1.98 ± 0.55 μmol g⁻¹ h⁻¹ in fish of 3.1 ± 0.4 mm TL in salt‐marsh). Due to the size‐dependent pattern, R_AM in different habitats cannot be compared directly; ancova , followed by residual compared analysis (regression‐related techniques), is seen as a valid method for this purpose. This work presents the first field data on ammonia excretion in the Aphanius genus and the flexible physiologic response characteristic of Cyprinodontids has been demonstrated.     

Workshop 7: 2

Glutamine, the preferred precursor for neurotransmitter glutamate, is likely to be the principal substrate for the neuronal System A transporter SAT1 in vivo. By measuring currents associated with SAT1 expression in Xenopus oocytes, we found that SAT1 mediates transport of small, neutral, aliphatic amino acids including glutamine, alanine and the System A‐specific analogue 2‐(methylamino) isobutyrate, each with K_0.5 of 0.3–0.5 mm . Amino acid transport is driven by the Na⁺ electrochemical gradient. Kinetic data indicates that Na⁺/cotransport comprises the ordered binding first of Na⁺ (a voltage‐dependent step), then alanine, then simultaneous translocation. Li⁺ (but not H⁺) can substitute for Na⁺ but results in reduced V_max. In the absence of amino acid, SAT1 mediates a cation leak with selectivity Na⁺, Li⁺, H⁺, K⁺. The temperature‐dependence of the leak current (E_a = 17 ± 3 kcal/mol) is consistent with carrier‐mediated Na⁺ uniport activity (cf 13 ± 2 kcal/mol for Na⁺/alanine cotransport) but the leak does not saturate at physiological [Na⁺], suggesting channel activity. Despite a Na⁺ Hill coefficient of 1, we obtained Na⁺/amino acid coupling coefficients greater than 1 from simultaneous measurement of charge and [³H]alanine or [³H]glutamine uptake. Interpretation of these data is model‐dependent and consistent with either (1) an all‐carrier model in which Na⁺/amino acid cotransport is thermodynamically coupled 2 : 1, cotransport is preferred over Na⁺ uniport, and in which there is little cooperativity between Na⁺ binding events, or (2) 1 : 1 coupling in parallel with an always‐on Na⁺ channel activity. In either scenario, the presence of SAT1 at the plasma membrane and resultant Na⁺ fluxes will place a significant energy burden on the cell.     

Seasonal variability of the parameters of the Ball–Berry model of stomatal conductance in maize (Zea mays L.) and sunflower (Helianthus annuus L.) under well‐watered and water‐stressed conditions

The Ball–Berry (BB) model of stomatal conductance (g_s) is frequently coupled with a model of assimilation to estimate water and carbon exchanges in plant canopies. The empirical slope (m) and ‘residual’ g_s (g₀) parameters of the BB model influence transpiration estimates, but the time‐intensive nature of measurement limits species‐specific data on seasonal and stress responses. We measured m and g₀ seasonally and under different water availability for maize and sunflower. The statistical method used to estimate parameters impacted values nominally when inter‐plant variability was low, but had substantial impact with larger inter‐plant variability. Values for maize (m = 4.53 ± 0.65; g₀ = 0.017 ± 0.016 mol m^?2 s^?1) were 40% higher than other published values. In maize, we found no seasonal changes in m or g₀, supporting the use of constant seasonal values, but water stress reduced both parameters. In sunflower, inter‐plant variability of m and g₀ was large (m = 8.84 ± 3.77; g₀ = 0.354 ± 0.226 mol m^?2 s^?1), presenting a challenge to clear interpretation of seasonal and water stress responses – m values were stable seasonally, even as g₀ values trended downward, and m values trended downward with water stress while g₀ values declined substantially.     

The onset of photosynthetic acclimation to elevated CO2 partial pressure in field-grown Pinus radiata D. Don. after 4 years

The effects of CO₂ enrichment on photosynthesis and ribulose‐1,5‐bisphosphate carboxylase/oxygenase (rubisco) were studied in current year and 1‐year‐old needles of the same branch of field‐grown Pinus radiata D. Don trees. All measurements were made in the fourth year of growth in large, open‐top chambers continuously maintained at ambient (36 Pa) or elevated (65 Pa) CO₂ partial pressures. Photosynthetic rates of the 1‐year‐old needles made at the growth CO₂ partial pressure averaged 10·5 ± 0·5 μmol m^?2 s^?1 in the 36 Pa grown trees and 11·8 ± 0·4 μmol m^?2 s^?1 in the 65 Pa grown trees, and were not significantly different from each other. The photosynthetic capacity of 1‐year‐old needles was reduced by 25% from 23·0 ± 1·8 μmol m^?2 s^?1 in the 36 Pa CO₂ grown trees to 17·3 ± 0·7 μmol m^?2 s^?1 in the 65 Pa grown trees. Growth in elevated CO₂ also resulted in a 25% reduction in V_cmax (maximum carboxylation rate), a 23% reduction in J_max (RuBP regeneration capacity mediated by maximum electron transport rate) and a 30% reduction in Rubisco activity and content. Total non‐structural carbohydrates (TNC) as a fraction of total dry mass increased from 12·8 ± 0·4% in 1‐year‐old needles from the 36 Pa grown trees to 14·2 ± 0·7% in 1‐year‐old needles from the 65 Pa grown trees and leaf nitrogen content decreased from 1·30 ± 0·02 to 1·09 ± 0·10 g m^?2. The current‐year needles were not of sufficient size for gas exchange measurements, but none of the biochemical parameters measured (Rubisco, leaf chlorophyll, TNC and N), were effected by growth in elevated CO₂. These results demonstrate that photosynthetic acclimation, which was not found in the first 2 years of this experiment, can develop over time in field‐grown trees and may be regulated by source‐sink balance, sugar feedback mechanisms and nitrogen allocation.     

Effects of salinity,light intensity and sediment on growth,pigments, agar production and reproduction in Gracilaria tenuistipitata from Songkhla Lagoon in Thailand

The effects of salinity, light intensity and sediment on Gracilaria tenuistipitata C.F. Chang & B.M. Xia on growth, pigments, agar production, and net photosynthesis rate were examined in the laboratory under varying conditions of salinity (0, 25 and 33 psu), light intensity (150, 400, 700 and 1000 µmol photons m^?2 s^?1) and sediment (0, 0.67 and 2.28 mg L^?1). These conditions simulated field conditions, to gain some understanding of the best conditions for cultivation of G. tenuistipitata. The highest growth rate was at 25 psu, 700 µmol photons m^?2 s^?1 with no sediments, that provided a 6.7% increase in weight gain. The highest agar production (24.8 ± 3.0 %DW) was at 25 psu, 150–400 µmol photons m^?2 s^?1 and no sediment. The highest pigment contents were phycoerythrin (0.8 ± 0.5 mg g^?1FW) and phycocyanin (0.34 ± 0.05 mg g^?1 FW) produced in low light conditions, at 150 µmol photons m^?2 s^?1. The highest photosynthesis rate was 161.3 ± 32.7 mg O₂ g^?1 DW h^?1 in 25 psu, 400 µmol photons m^?2 s^?1 without sediment in the short period of cultivation, (3 days) and 60.3 ± 6.7 mg O₂ g^?1 DW h^?1 in 25 psu, 700 µmol photons m^?2 s^?1 without sediment in the long period of cultivation (20 days). The results indicated that salinity was the most crucial factor affecting G. tenuistipitata growth and production. This would help to promote the cultivation of Gracilaria cultivation back into the lagoon using these now determined baseline conditions. Extrapolation of the results from the laboratory study to field conditions indicated that it was possible to obtain two crops of Gracilaria a year in the lagoon, with good yields of agar, from mid‐January to the end of April (dry season), and from mid‐July to the end of September (first rainy season) when provided sediment was restricted.     

Effects and mechanisms of ghrelin on cardiac microvascular endothelial cells in rats

Ghrelin is thought to directly exert a protective effect on the cardiovascular system, specifically by promoting vascular endothelial cell function. Our study demonstrates the ability of ghrelin to promote rat CMEC (cardiac microvascular endothelial cell) proliferation, migration and NO (nitric oxide) secretion. CMECs were isolated from left ventricle of adult male Sprague—Dawley rat by enzyme digestion and maintained in endothelial cell medium. Dil‐ac‐LDL (1,1′‐dioctadecyl‐3,3,3′,3′‐ tetramethylindocarbocyanine‐labelled acetylated low‐density lipoprotein) intake assays were used to identify CMECs. Cells were split into five groups and treated with varying concentrations of ghrelin as follows: one control non‐treated group; three ghrelin dosage groups (1×10^?9, 1×10^?8, 1×10^?7 mol/l) and one ghrelin+PI3K inhibitor group (1×10^?7 mol/l ghrelin+20 μmol/l LY294002). After 24 h treatment, cell proliferation capability was measured by MTT [3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyl‐2H‐tetrazolium bromide] assay and Western blot for PCNA (proliferating cell nuclear antigen) protein expression. Migration of CMECs was detected by transwell assays, and NO secretion of CMECs was measured via nitrate reduction. Protein expression of AKT and phosphorylated AKT in CMECs was measured by Western blot after exposure to various concentrations of ghrelin and the PI3K inhibitor LY294002. Our results indicate that ghrelin significantly enhanced cell growth at concentrations of 10^?8 mol/l (0.271±0.041 compared with 0.199±0.021, P=0.03) and 10^?7 mol/l (0.296±0.039 compared with 0.199±0.021, P<0.01). However, addition of the PI3K/AKT inhibitor LY294002 inhibited the ghrelin‐mediated enhancement in cell proliferation (0.227±0.042 compared with 0.199±0.021, P=0.15). At a concentration between 10^?8 and 10^?7 mol/l, ghrelin caused a significant increase in the number of migrated cells compared with the control group (126±9 compared with 98±7, P=0.02; 142±6 compared with 98±7, P<0.01), whereas no such change could be observed in the presence of 20 μmol/l of the PI3K/Akt inhibitor LY294002 (103±7 compared with 98±7, P=0.32). Ghrelin treatment significantly enhanced NO production in a dose‐dependent fashion compared with the untreated control group [(39.93±2.12) μmol/l compared with (30.27±2.71) μmol/l, P=0.02; (56.80±1.98) μmol/l compared with (30.27±2.71) μmol/l, P<0.01]. However, pretreatment with 20 μmol/l LY294002 inhibited the ghrelin‐stimulated increase in NO secretion [(28.97±1.64) μmol/l compared with (30.27±2.71) μmol/l, P=0.37]. In summary, we have found that ghrelin treatment promotes the proliferation, migration and NO secretion of CMECs through activation of PI3K/AKT signalling pathway.     

Heteromerous Bistricyclic Aromatic Enes: Dynamic Stereochemistry of Xanthylidene‐Anthrones

The heteromerous bistricyclic aromatic ene (BAE) 2,2'‐dimethyl‐10‐(9H‐xanthylidene)‐9(10H)‐anthrone (DMXA) was synthesized by a condensation of 10,10‐dichloro‐2‐methylxanthene with 2‐methylanthrone. X‐ray crystallography of (E)‐DMXA and xanthylidene‐anthrone (XA) indicated that the molecules adopt anti‐folded conformations with folding dihedral angles of 44°/44° and 39°/41°, respectively . The crystal structure of anti‐folded (E)‐DMXA does not indicate any xanthenylium–anthracenolate push–pull effect. E,Z‐diastereomerization of DMXA was studied by ¹H‐NMR coalescence‐temperature measurements at different magnetic field strengths and by kinetic equilibration experiments . Free energy of activation for this process was 81.5 (±1.3) kJ/mol. B3LYP/6‐311+G(d,p) calculations showed that anti‐folded conformers of XA, (E)‐DMXA, bianthrone (AA), and dixanthylene (XX) were global minima. The twisted conformers of XA, AA, and XX were local minima (ΔG₂₉₈ = 16, 18, and 24 kJ/mol) with a substantial dipolar xanthenylium–anthracenolate dipolar contribution for XA. Chirality 27:919–928, 2015. © 2015 Wiley Periodicals, Inc.