Purification to homogeneity of human placental acid sphingomyelinase

Acid sphingomyelinase was purified to homogeneity from human placenta in the presence of a dialyzable detergent, n-octyl-beta-D-glucopyranoside. The major steps in the procedure included column chromatographies with Con A-Sepharose, sphingosylphosphorylcholine-Sepharose 4B, hexyl-agarose, and Mono P. The purified enzyme with pI 7.4 had a specific activity of approx 170,000 units/mg protein with a yield of 3.6%. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed a single protein band of Mr 62,000. Gel filtration with a Superose 12 column gave a single peak, and the enzyme in the presence 50 mM n-octyl-beta-D-glucopyranoside was of Mr 123,000, indicating that the native enzyme occurs in a dimeric form. The optimal pH was 5.5 with both sphingomyelin and an artificial substrate, 2-N-hexadecanoylamino-4-nitrophenylphosphorylcholine. The Km values were 55 microM with sphingomyelin and 340 microM with the artificial substrate. The enzyme activity was not affected by Mg2+ (1-5 mM), confirming that the enzyme is acid sphingomyelinase. The enzyme was stable at -80 degrees C for more than 4 months. In addition to the enzyme with pI 7.4, the Mono P chromatofocusing gave two peaks (pI 7.0 and 6.7) possessing the enzymatic activity.     

“秦油2号”油菜植株内含物与萝卜蚜有翅率的关系

通过对Ｎ、Ｐ、Ｋ不同施肥条件下的“秦油２号”油菜植株内含物等与萝卜蚜有翅率关系的研究,建立了８个数学模型,结果表明可溶性糖、总Ｎ、糖与蛋白质之比和丝氨酸与萝卜蚜有翅率有密切关系,其次是天门冬氨酸、苏氨酸、谷氨酸、甘氨酸、丙氨酸、蛋氨酸、异亮氨酸、亮氨酸、酪氨酸、含水量、水溶性蛋白、酰胺氮和含Ｐ量等．     

山楂叶螨种群空间格局及其应用的研究Ⅱ．成螨种群密度综合估计及其抽样技术

本文结合Ｉｗａｏ零频率法和Ｇｅｒｒａｒｄ阈限密度法,提出一种改进的种群密度估计方法──“综合阈限密度估计法”,并由此探讨了苹果树上山楂叶螨成螨的密度估计及其抽样技术．采用零样频率来估计成螨的平均密度,并得到用概率保证的理论抽样数模型．比较结果表明,零频率法所需的理论抽样数少于直接计数法．综合阈限密度估计法的拟合效果更为显著．     

The legacy effect of biochar application on soil nitrous oxide emissions

Existing studies suggest that biochar application can reduce soil nitrous oxide (N₂O) emissions, mainly based on short-term results. However, it remains unclear what the effects (i.e., legacy effects) and underlying mechanisms are on N₂O emissions after many years of a single application of biochar. Here, we collected intact soil columns from plots without and with biochar application in a subtropical tea plantation 7 years ago for an incubation experiment. We used the N₂O isotopocule analysis combined with ammonia oxidizer-specific inhibitors and molecular biology approaches to investigate how the legacy effect of biochar affected soil N₂O emissions. Results showed that the soil in the presence of biochar had lower N₂O emissions than the control albeit statistically insignificant. The legacy effect of biochar in decreasing N₂O emissions may be attributed to the reduced effectiveness of the soil substrate, nitrification and denitrification activities, and the promotion of the further reduction of N₂O. The legacy effect of biochar reduced the relative contribution of nitrifier denitrification/bacterial denitrification, nitrification-related N₂O production, and the relative abundance of several microorganisms involved in the nitrogen cycle. Our global meta-analysis also showed that the reduction of N₂O by biochar increased with increasing application rate but diminished and possibly even reversed with increasing experimental time. In conclusion, our findings suggest that the abatement capacity of biochar on soil N₂O emissions may weaken over time after biochar application, but this remains under further investigation.     

Alteration of seed fatty acid composition by an ethyl methanesulfonate-induced mutation in Arabidopsis thaliana affecting diacylglycerol acyltransferase activity.

In characterizing the enzymes involved in the formation of very long-chain fatty acids (VLCFAs) in the Brassicaceae, we have generated a series of mutants of Arabidopsis thaliana that have reduced VLCFA content. Here we report the characterization of a seed lipid mutant, AS11, which, in comparison to wild type (WT), has reduced levels of 20:1 and 18:1 and accumulates 18:3 as the major fatty acid in triacylglycerols. Proportions of 18:2 remain similar to WT. Genetic analyses indicate that the fatty acid phenotype is caused by a semidominant mutation in a single nuclear gene, designated TAG1, located on chromosome 2. Biochemical analyses have shown that the AS11 phenotype is not due to a deficiency in the capacity to elongate 18:1 or to an increase in the relative delta 15 or delta 12 desaturase activities. Indeed, the ratio of desaturase/elongase activities measured in vitro is virtually identical in developing WT and AS11 seed homogenates. Rather, the fatty acid phenotype of AS11 is the result of reduced diacylglycerol acyltransferase activity throughout development, such that triacylglycerol biosynthesis is reduced. This leads to a reduction in 20:1 biosynthesis during seed development, leaving more 18:1 available for desaturation. Thus, we have demonstrated that changes to triacylglycerol biosynthesis can result in dramatic changes in fatty acid composition and, in particular, in the accumulation of VLCFAs in seed storage lipids.     

Regulation of GABAB receptors by histamine and neuronal activity in the isolated spinal cord of neonatal opossum in culture.

The aim of these experiments has been to analyse the properties of receptors for the transmitter gamma-aminobutyric acid (GABA) in developing mammalian nervous system. Changes in responses of GABAB receptors have been measured after alterations of the chemical environment and the level of electrical activity. We have previously shown that when the central nervous system (CNS) of the new-born opossum, Monodelphis domestica, is cultured for three to five days in the presence of histidine, inhibition by baclofen, a GABAB agonist, disappears (Stewart et al. 1991). We have now investigated whether histidine acts indirectly by way of conversion to histamine. As with histidine, culture with 150 microM histamine for five days virtually abolished the inhibition by baclofen. The effects of histidine, as well as histamine, were blocked by mepyramine, a histamine H1-receptor antagonist, and by ranitidine, an H2-antagonist. Tetrodotoxin (TTX), which blocks all electrical activity, protected preparations from the action of histidine but not histamine. Our results suggest that histidine is converted to histamine, which reduces the efficacy of GABAB agonists. We conclude that, in the developing mammalian CNS, transmitter levels and electrical activity can selectively influence the properties of receptors.