β-Amino acid residues: Conformational characterization of an N- and C-protected homo-β-(S)-leucine

Summary An N- and C-protected derivative ofhomo-β-leucine, Fmoc-homo-β-(S)-leucine methyl ester, synthesized from the corresponding proteinogenic parent α-amino acid in enantiopure form has been fully characterized in the solid state by X-ray diffraction analysis. The crystal conformation of this new residue indicates and extended conformation for thishomo-β-residue, with the ϕ torsion angle being more constrained than the μ and ψ angles.     

Phosphonate-based irreversible inhibitors of human γ-glutamyl transpeptidase (GGT). GGsTop is a non-toxic and highly selective inhibitor with critical electrostatic interaction with an active-site residue Lys562 for enhanced inhibitory activity

γ-Glutamyl transpeptidase (GGT, EC 2.3.2.2) that catalyzes the hydrolysis and transpeptidation of glutathione and its S-conjugates is involved in a number of physiological and pathological processes through glutathione metabolism and is an attractive pharmaceutical target. We report here the evaluation of a phosphonate-based irreversible inhibitor, 2-amino-4-{[3-(carboxymethyl)phenoxy](methoyl)phosphoryl}butanoic acid (GGsTop) and its analogues as a mechanism-based inhibitor of human GGT. GGsTop is a stable compound, but inactivated the human enzyme significantly faster than the other phosphonates, and importantly did not inhibit a glutamine amidotransferase. The structure–activity relationships, X-ray crystallography with Escherichia coli GGT, sequence alignment and site-directed mutagenesis of human GGT revealed a critical electrostatic interaction between the terminal carboxylate of GGsTop and the active-site residue Lys562 of human GGT for potent inhibition. GGsTop showed no cytotoxicity toward human fibroblasts and hepatic stellate cells up to 1 mM. GGsTop serves as a non-toxic, selective and highly potent irreversible GGT inhibitor that could be used for various in vivo as well as in vitro biochemical studies.     

Concentrations of organophosphorus pesticides in rice (Oryza sativa) and human health risk assessment from Songhua River,Northeast China

The aim of this study was to investigate organophosphorus pesticides (OPs) concentrations in rice grains and estimate potential health risk to inhabitants. A total of 102 rice samples were collected from the Irrigated Area of Songhua River, located in Jilin province, one of the most important rice production areas and a key agricultural area in China. The twelve OPs were determined by gas chromatography coupled with flame photometric detector. Results showed that OPs concentrations in rice grains did not exceed maximum residue limit. The mean concentrations obtained for the detected twelve OPs in µg kg^?1 were as follows: omethoate (0.60), diazinon (0.56), parathion-methyl (0.54), dimethoate (0.38), isocarbophos (0.38), phorate (0.31), fenitrothion (0.24), parathion (0.23), fenthion (0.23), malathion (0.13), dichlorvos (0.13), and methamidophos (0.08). Diazinon (39.2%), omethoate (31.4%), and parathion-methyl (25.5%) had the highest frequency in the detected twelve OPs. 69.6% of samples contained more than one OP, 24.5% of samples detected one OP, and only 5.9% samples did not find any OPs. The average target hazard quotients were all less than one and average hazard index for adults and children were 0.273 and 0.238, respectively. Overall, results indicated that the intake of rice may be safe for consumers.     

Health risks of consuming apples with carbendazim,imidacloprid, and thiophanate-methyl in the Chinese population: Risk assessment based on a nonparametric probabilistic evaluation model

The pesticides carbendazim (CA), imidacloprid (IM), and thiophanate-methyl (TH) are used extensively in apple production. However, the presence of pesticide residues in apples has been associated with a wide range of human health hazards. To assess whether apples are safe to consume, evaluating their potential health risk is of great significance. We tested apples from two dominant apple-producing areas for the presence of pesticide residues and constructed a statistical model to evaluate their health risks. Of the three commonly used pesticides, thirty-two samples (11.3% of all tested samples) were residue-free and 231 (81.9%) contained CA residues, which was the most frequently detectable pesticide, followed by TH (52.1%) and IM (39.0%). All of the samples were below the maximum residue limits. The probabilistic assessment results indicated that the pesticide intake risks for kids (aged 2–6 years) and children (aged 7–13 years) were significantly higher than those of other groups, so that they were the vital monitoring objects. Our findings indicate that consumption of apples with these three pesticides does not pose a health threat for the population. Nevertheless, we recommend an investigation into continuous monitoring and stricter regulations on fruits' quality and safety throughout China.     

The management of rice straw,fertilisers and leaf litters in rice cropping systems in Northeast Thailand.

The maintenance of soil organic matter (SOM) and the balancing of nutrient flows into and out of the rainfed rice cropping systems in Northeast Thailand is of paramount importance to arresting the decline in soil fertility and crop yields. A system where small applications of leaf litters from locally grown trees are applied annually to rice paddy soils prior to transplanting is described. The annual application of 1500 kg/ha of Cajanus cajan, Acacia auriculiformis, Phyllanthus taxodifolius and Samanea saman for five seasons resulted in increases in rice grain yield of 48, 35, 32 and 23% above the no-leaf litter control, respectively. Average annual nutrient inputs from the leaf litters, in kg/ha, ranged from 62.7 N, 3.9 P, 17.9 K, and 3.5 S for Cajanus cajan to 24.5 N, 1.5 P, 8.1 K and 2.0 S for Acacia auriculiformis. Nutrient balances, determined by the difference between the inputs (fertiliser and added leaf litters) and outputs (grain and straw) indicated net positive N and P balances of up to 457 and 60 kg/ha. respectively, after five seasons of leaf litter applications. Sulfur and potassium balances resulted in net deficits of up to −3 and −52 kg S and K/ha, respectively, where no leaf litter was applied and rice straw was removed following harvest. Calculated apparent nutrient recoveries reflected the decomposition rate of the added residues and were highest for P and K, reflecting their higher soil residual value than mobile nutrients such as N and S. Sustainable farming systems will require that crop yields are stable through the maintenance of soil fertility and the balance of nutrients in the system. This revised version was published online in June 2006 with corrections to the Cover Date.     

Site-Directed Mutagenesis Evidence for Arginine-384 Residue at the Active Site of Maize Branching Enzyme II

Essential arginine residues are suggested to be located at the active sites of maize branching enzymes (BE) based on the evidence that two arginine residues are conserved in all BE from various species and that as little as one arginine residue is located at the active site of maize BE by phenylglyoxal (PGO) modification. To determine the exact location of the active-site arginine residue in BE, we employed peptide mapping and site-directed mutagenesis approaches. A single trypsin-digested, [¹⁴C]PGO-labeled peptide was purified from maize BEII by two rounds of HPLC separation, but we failed to obtain amino acid sequencing information. Site-directed mutagenesis was then used to create one mutant (arginine-384 to alanine-384), R384A. Immunoblotting result showed that BEII protein was expressed at a similar level in the wild type and the R384A mutant. However, BE activity in the R384A mutant was only 1.4% of the wild type. These results support the conclusion that the conserved arginine-384 residue is important in BEII catalysis.     

Entropy capacity determines protein folding

Search and study of the general principles that govern kinetics and thermodynamics of protein folding generate a new insight into the factors controlling this process. Here, based on the known experimental data and using theoretical modeling of protein folding, we demonstrate that there exists an optimal relationship between the average conformational entropy and the average energy of contacts per residue-that is, an entropy capacity-for fast protein folding. Statistical analysis of conformational entropy and number of contacts per residue for 5829 protein structures from four general structural classes (all-alpha, all-beta, alpha/beta, alpha+beta) demonstrates that each class of proteins has its own class-specific average number of contacts (class alpha/beta has the largest number of contacts) and average conformational entropy per residue (class all-alpha has the largest number of rotatable angles phi, psi, and chi per residue). These class-specific features determine the folding rates: alpha proteins are the fastest folding proteins, then follow beta and alpha+beta proteins, and finally alpha/beta proteins are the slowest ones. Our result is in agreement with the experimental folding rates for 60 proteins. This suggests that structural and sequence properties are important determinants of protein folding rates.     

微生物菌剂在农业废弃物堆肥腐熟过程中的应用及其田间试验效果

以农村生活垃圾中的可堆肥腐熟成分和蘑菇渣为堆肥原料,通过添加微生物菌剂进行堆肥试验,研究其在农业废弃物堆肥腐熟过程中的作用,并通过田间试验研究堆肥腐熟后肥料样品对黄瓜和青椒的增产效果,以验证其肥效。结果表明添加微生物菌剂有助于堆肥腐熟后样品的氮、磷、钾的保全和有机质的增加,促进养分均衡。添加微生物菌剂的堆肥腐熟肥料样品在田间试验中对黄瓜和青椒的增产效果最为显著,分别为22.21%和19.87%。     

The S1 split signal of photosystem II; a tyrosine-manganese coupled interaction

Detailed optical and EPR analyses of states induced in dark-adapted PS II membranes by cryogenic illumination permit characterization and quantification of all pigment derived donors and acceptors, as well as optically silent (in the visible, near infrared) species which are EPR active. Near complete turnover formation of Q_A⁻ is seen in all centers, but with variable efficiency, depending on the donor species. In minimally detergent-exposed PS II membranes, negligible (< 5%) oxidation of chlorophyll or carotenoid centers occurs for illumination temperatures 5-20 K. An optically silent electron donor to P680⁺ is observed with the same decay kinetics as the S₁ split signal. Cryogenic donors to P680⁺ seen are: (i) transient (t_1/2 ∼ 150 s) tyrosine related species, including ‘split signals’ (∼ 15% total centers), (ii) reduced cytochrome b₅₅₉ (∼ 30-50% centers), and (iii) an organic donor, possibly an amino acid side chain, (∼ 30% centers).