Alpha-glucosidase inhibition from a Chinese medical herb (Ramulus mori) in normal and diabetic rats and mice

Alpha-glucosidase inhibitors are oral antidiabetic drugs. A traditional Chinese medical herb, Sangzhi (Ramulus mori), appears to have properties similar to those of alpha-glucosidase inhibitors. The effects of an aqueous extract of Shangzhi (SZ) were studied in normal and alloxan diabetic rats and mice, and these results compared with those for acarbose, an alpha-glucosidase inhibitor. In our grade-dose studies, SZ was found to lower and prolong the zenith of blood glucose concentration (ZBG) after sucrose or starch loading and stabilize blood glucose levels in fasting normal and alloxan diabetic mice. After 2 weeks of SZ administration with high-calorie chow or a normal diet, the fasting and non-fasting blood glucose concentrations in alloxan diabetic mice and rats were decreased. In alloxan rats, the blood fructosamine concentration was lowered. Results for acarbose and SZ were similar. These indicate that SZ has alpha-glucosidase inhibitory effects.     

Towards robust evolutionary inference with integral projection models

Integral projection models (IPMs) are extremely flexible tools for ecological and evolutionary inference. IPMs track the distribution of phenotype in populations through time, using functions describing phenotype‐dependent development, inheritance, survival and fecundity. For evolutionary inference, two important features of any model are the ability to (i) characterize relationships among traits (including values of the same traits across ages) within individuals, and (ii) characterize similarity between individuals and their descendants. In IPM analyses, the former depends on regressions of observed trait values at each age on values at the previous age (development functions), and the latter on regressions of offspring values at birth on parent values as adults (inheritance functions). We show analytically that development functions, characterized this way, will typically underestimate covariances of trait values across ages, due to compounding of regression to the mean across projection steps. Similarly, we show that inheritance, characterized this way, is inconsistent with a modern understanding of inheritance, and underestimates the degree to which relatives are phenotypically similar. Additionally, we show that the use of a constant biometric inheritance function, particularly with a constant intercept, is incompatible with evolution. Consequently, current implementations of IPMs will predict little or no phenotypic evolution, purely as artefacts of their construction. We present alternative approaches to constructing development and inheritance functions, based on a quantitative genetic approach, and show analytically and through an empirical example on a population of bighorn sheep how they can potentially recover patterns that are critical to evolutionary inference.     

Influence of micronization (fine grinding) of soya bean meal and fullfat soya bean on productive performance and digestive traits in young pigs

Two trials were conducted to test the effect of micronization (very fine grinding) of soya bean meal (SBM) and fullfat soya bean (FFSB) on productive performance and digestive traits of piglets. The experimental design was completely randomized with four treatments arranged factorially (SBM and FFSB, micronized and ground). The mean particle size (MPS) was 47 and 881 μm for the SBM and 41 and 778 μm for the FFSB, micronized and ground, respectively. In trial 1 growth traits from 23 to 45 days of age and the coefficient of total tract apparent digestibility (CTTAD) of dietary components at 33 and 45 days of age were assessed. In trial 2 the coefficient of ileal apparent digestibility (CIAD) of dietary components, the pH of the gastro intestinal tract (GIT) and the weight of digestive organs and spleen were measured at 45 days of age. From 23 to 33 days of age pigs fed SBM grew faster (253 g/d versus 213 g/day; P<0.05) and were more efficient (0.87 g/g versus 0.98 g/g; P<0.01) than pigs fed FFSB. For the entire experiment (23–45 days of age) pigs fed SBM tended to grow more (360 g/day versus 324 g/day) and to eat more feed (414 g/day versus 380 g/day) than pigs fed FFSB (P<0.10). The CTTAD of crude protein (0.798 g/kg versus 0.778 g/kg), organic matter (0.864 g/kg versus 0.839 g/kg) and gross energy (0.849 g/kg versus 0.830 g/kg) were higher for pigs fed SBM than for pigs fed FFSB (P<0.001). In addition, CIAD of organic matter (0.765 g/kg versus 0.705 g/kg) and gross energy (0.761 g/kg versus 0.711 g/kg) were higher for SBM than for FFSB diets (P<0.001). The pH of the different segments of the GIT was not affected by the protein source (P>0.10). Particle size did not affect any trait studied (P>0.10). The poor performance and digestibility of pigs fed FFSB as compared to pigs fed SBM might be related to the conditions applied during processing. It is concluded that pigs fed soya bean meal perform better than pigs fed FFSB and that micronization of the soya protein sources does not affect any trait studied.     

Cell‐mediated immunity and multi‐locus heterozygosity in bluethroat nestlings

Recent evidence suggests that marker‐based heterozygosity‐fitness correlations may be driven by only one or a few markers, indicating local heterozygosity effects caused by linkage disequilibrium with functional genes. In this study, we investigated the relationship between microsatellite heterozygosity and a measure of cell‐mediated immunity (phytohaemagglutinin; PHA) in bluethroat (Luscinia s. svecica) nestlings using a full‐sibling design. We found significant positive associations between PHA response and two different indices of microsatellite heterozygosity, i.e. multi‐locus heterozygosity and mean d². However, model comparisons disclosed that both associations were more likely caused by local effects rather than general effects and that the two local effects appeared to be realized through two different genetic mechanisms. Our results indicate that both the random assortment of parental chromosomes during meiosis as well as inbreeding can drive heterozygosity‐fitness correlations.     

High temperatures limit plant growth but hasten flowering in root chicory (Cichorium intybus) independently of vernalisation

An increase in mean and extreme summer temperatures is expected as a consequence of climate changes and this might have an impact on plant development in numerous species. Root chicory (Cichorium intybus L.) is a major crop in northern Europe, and it is cultivated as a source of inulin. This polysaccharide is stored in the tap root during the first growing season when the plant grows as a leafy rosette, whereas bolting and flowering occur in the second year after winter vernalisation. The impact of heat stress on plant phenology, water status, photosynthesis-related parameters, and inulin content was studied in the field and under controlled phytotron conditions. In the field, plants of the Crescendo cultivar were cultivated under a closed plastic-panelled greenhouse to investigate heat-stress conditions, while the control plants were shielded with a similar, but open, structure. In the phytotrons, the Crescendo and Fredonia cultivars were exposed to high temperatures (35 °C day/28 °C night) and compared to control conditions (17 °C) over 10 weeks. In the field, heat reduced the root weight, the inulin content of the root and its degree of polymerisation in non-bolting plants. Flowering was observed in 12% of the heat stressed plants during the first growing season in the field. In the phytotron, the heat stress increased the total number of leaves per plant, but reduced the mean leaf area. Photosynthesis efficiency was increased in these plants, whereas osmotic potential was decreased. High temperature was also found to induced flowering of up to 50% of these plants, especially for the Fredonia cultivar. In conclusion, high temperatures induced a reduction in the growth of root chicory, although photosynthesis is not affected. Flowering was also induced, which indicates that high temperatures can partly substitute for the vernalisation requirement for the flowering of root chicory.     

Modulation of calcium signalling by mitochondria

In this review we will attempt to summarise the complex and sometimes contradictory effects that mitochondria have on different forms of calcium signalling. Mitochondria can influence Ca²⁺ signalling indirectly by changing the concentration of ATP, NAD(P)H, pyruvate and reactive oxygen species — which in turn modulate components of the Ca²⁺ signalling machinery i.e. buffering, release from internal stores, influx from the extracellular solution, uptake into cellular organelles and extrusion by plasma membrane Ca²⁺ pumps. Mitochondria can directly influence the calcium concentration in the cytosol of the cell by importing Ca²⁺ via the mitochondrial Ca²⁺ uniporter or transporting Ca²⁺ from the interior of the organelle into the cytosol by means of Na⁺/Ca²⁺ or H⁺/Ca²⁺ exchangers. Considerable progress in understanding the relationship between Ca²⁺ signalling cascades and mitochondrial physiology has been accumulated over the last few years due to the development of more advanced optical techniques and electrophysiological approaches.     

Coupling Molecular Dynamics Simulations with Experiments for the Rational Design of Indolicidin-Analogous Antimicrobial Peptides

Antimicrobial peptides (AMPs) have attracted much interest in recent years because of their potential use as new-generation antibiotics. Indolicidin (IL) is a 13-residue cationic AMP that is effective against a broad spectrum of bacteria, fungi, and even viruses. Unfortunately, its high hemolytic activity retards its clinical applications. In this study, we adopted molecular dynamics (MD) simulations as an aid toward the rational design of IL analogues exhibiting high antimicrobial activity but low hemolysis. We employed long-timescale, multi-trajectory all-atom MD simulations to investigate the interactions of the peptide IL with model membranes. The lipid bilayer formed by the zwitterionic 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) was chosen as the model erythrocyte membrane; lipid bilayers formed from a mixture of POPC and the negatively charged 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol were chosen to model bacterial membranes. MD simulations with a total simulation time of up to 4 μs revealed the mechanisms of the processes of IL adsorption onto and insertion into the membranes. The packing order of these lipid bilayers presumably correlated to the membrane stability upon IL adsorption and insertion. We used the degree of local membrane thinning and the reduction in the order parameter of the acyl chains of the lipids to characterize the membrane stability. The order of the mixed 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol/POPC lipid bilayer reduced significantly upon the adsorption of IL. On the other hand, although the order of the pure-POPC lipid bilayer was perturbed slightly during the adsorption stage, the value was reduced more dramatically upon the insertion of IL into the membrane's hydrophobic region. The results imply that enhancing IL adsorption on the microbial membrane may amplify its antimicrobial activity, while the degree of hemolysis may be reduced through inhibition of IL insertion into the hydrophobic region of the erythrocyte membrane. In addition, through simulations, we identified the amino acids that are most responsible for the adsorption onto or insertion into the two model membranes. Positive charges are critical to the peptide's adsorption, whereas the presence of hydrophobic Trp8 and Trp9 leads to its deeper insertion. Combining the hypothetical relationships between the membrane disordering and the antimicrobial and hemolytical activities with the simulated results, we designed three new IL-analogous peptides: IL-K7 (Pro7 → Lys), IL-F89 (Trp8 and Trp9 → Phe), and IL-K7F89 (Pro7 → Lys; Trp8 and Trp9 → Phe). The hemolytic activity of IL-F89 is considerably lower than that of IL, whereas the antimicrobial activity of IL-K7 is greatly enhanced. In particular, the de novo peptide IL-K7F89 exhibits higher antimicrobial activity against Escherichia coli; its hemolytic activity decreased to only 10% of that of IL. Our simulated and experimental results correlated well. This approach—coupling MD simulations with experimental design—is a useful strategy toward the rational design of AMPs for potential therapeutic use.     

植物光合CO2响应模型对光下(暗)呼吸速率拟合的探讨

运用LI-6400便携式光合作用系统测定了不同光强(2000、1500、1000和500 μmol·m^-2·s^-1)和两种O₂浓度(21%和2%的O₂)下冬小麦(Triticum aestivum)灌浆期旗叶的CO₂响应曲线, 比较了现有CO₂响应模型(生化模型、直角双曲线模型和直角双曲线修正模型)拟合给出光下(暗)呼吸与测量值之间的差异。结果显示, 直角双曲线修正模型所给出的光下呼吸速率拟合值与测量值最为接近。植物光合作用对大气CO₂响应(A/C_a)的拟合结果优于光合作用对胞间CO₂浓度(A/C_i)的拟合。然而, 所有模型基于A/C_a拟合的光下(暗)呼吸在整体上与测量值存在显著差异(p < 0.05), 推测与现有模型没有考虑CO₂浓度对光呼吸和光下暗呼吸速率的影响有关。对小麦的试验结果表明, CO₂浓度对光呼吸和光下暗呼吸均有显著影响: 随着CO₂浓度的增加(0-1400 μmol·mol^-1), 不同光强下的表观光呼吸变化范围分别为5.035-11.670、4.222-11.650、4.330-10.999和3.263-9.094 μmol CO_2·m^-2·s^-1; 光下暗呼吸的变化范围分别为0.491-2.987、0.457-2.955、0.545-3.139和0.448-3.139 μmol CO₂·m^-2·s^-1。回归分析发现, 表观光呼吸和光下暗呼吸与CO₂浓度之间均存在较好的相关性。然而, 将该回归关系整合到现有模型中, 是否会优化模型, 从而提高模型对相关光合参数估算的准确性尚有待于进一步研究。     

Functional comparison of hemoglobin purified by different methods and their biophysical implications

Hemoglobin (Hb) that is purified from red blood cells (RBCs) is commonly subjected to harsh processing conditions, such as high temperatures and extensive column separation, which may damage the Hb by altering the heme prosthetic group and/or the Hb protein structure. In this study, bovine and human Hb purified by tangential flow filtration (TFF) was compared to commercial preparations of human Hb (Hemosol, Inc., Toronto, Canada) and bovine Hb (Biopure, Inc., Cambridge, MA). Purified Hbs were characterized by measuring their overall purity (SDS–PAGE, SEC, and ESI‐MS), susceptibility to oxidation (k_ox), responses to physiological conditions (pH, [Cl^?], [IHP], and T), and ligand binding kinetics (O₂, NO, and CO). All Hbs evaluated possessed comparable biophysical properties, however, a small amount of catalase was detected in the TFF‐purified Hbs that reduced the rate of autoxidation. Mass changes observed by mass spectrometry suggest that structural alterations may be introduced into Hbs that are purified by extensive chromatographic separations. These results demonstrate that TFF is a suitable process for the purification of Hb from RBCs with a quality equivalent to that of commercial Hb preparations that employ more extensive purification strategies. This work also shows that TFF can yield highly pure Hb which can be used for Hb‐based O₂ carrier synthesis. Biotechnol. Bioeng. 2010; 106: 76–85. © 2010 Wiley Periodicals, Inc.