Depolymerized products of lambda-carrageenan as a potent angiogenesis inhibitor

Since angiogenesis is involved in initiating and promoting several diseases such as cancer and cardiovascular events, this study was designed to evaluate the anti-angiogenesis of low-molecular-weight (LMW), highly sulfated lambda-carrageenan oligosaccharides (lambda-CO) obtained by carrageenan depolymerization, by CAM (chick chorioallantoic membrane) model and human umbilical vein endothelial cells (HUVECs). Significant inhibition of vessel growth was observed at 200 microg/pellet. A histochemistry assay also revealed a decrease of capillary plexus and connective tissue in lambda-CO treated samples. lambda-CO inhibited the viability of cells at the high concentration of 1 mg/mL, whereas it affected the cell survival slightly (>95%) at a low concentration (<250 microg/mL), and HUVEC is the most sensitive to lambda-CO among three kinds of cells. Furthermore, the inhibitory action of lambda-CO was also observed in the endothelial cell invasion and migration at relatively low concentration (150-300 microg/mL), through down-regulation of intracellular matrix metalloproteinases (MMP-2) expression on endothelial cells. Taken together, these findings demonstrate that lambda-CO is a potential angiogenesis inhibitor with combined effects of inhibiting invasion, migration, and proliferation.     

Resveratrol is a potent inhibitor of the dioxygenase activity of lipoxygenase

Resveratrol is a naturally occurring phytoalexin, present in grapes and other food products, with important antioxidant properties. Although still under debate, it is generally assumed that resveratrol has protective effects against heart diseases and probably tumor development. Lipoxygenase is a dioxygenase with peroxidase activity involved in the synthesis of mediators in inflammatory, atherosclerotic, and carcinogenic processes. Lipoxygenase activity is also involved in the generation of flavors and aromas in foods from animal or vegetal sources. The results presented here show that resveratrol was a potent inhibitor of the dioxygenase activity of lipoxygenase, with an IC(50) = 13 microM. Simultaneously, resveratrol was oxidized by the peroxidase activity of lipoxygenase with a V(max) = 0.28 microM min(-1) and a k(M) = 16.6 microM. Furthermore, oxidized resveratrol was as efficient a lipoxygenase inhibitor as in its reduced form. From the data obtained it can be concluded that both resveratrol and its oxidized form can act as inhibitors of the dioxygenase activity of lipoxygenase. In contrast, the hydroperoxidase activity of lipoxygenase was not inhibited by resveratrol. These results suggest that resveratrol may be used as an antioxidant food additive and as a pharmacological agent to prevent the generation of eicosanoids involved in pathological processes.     

C-phycocyanin, a very potent and novel platelet aggregation inhibitor from Spirulina platensis

The aim of this study was to systematically examine the inhibitory mechanisms of C-phycocyanin (C-PC), one of the major phycobiliproteins of Spirulina platensis (a blue-green alga), in platelet activation. In this study, C-PC concentration-dependently (0.5-10 nM) inhibited platelet aggregation stimulated by agonists. C-PC (4 and 8 nM) inhibited intracellular Ca2+ mobilization and thromboxane A2 formation but not phosphoinositide breakdown stimulated by collagen (1 microg/mL) in human platelets. In addition, C-PC (4 and 8 nM) markedly increased levels of cyclic GMP and cyclic GMP-induced vasodilator-stimulated phosphoprotein (VASP) Ser(157) phosphorylation. Rapid phosphorylation of a platelet protein of Mw 47,000 (P47), a marker of protein kinase C activation, was triggered by phorbol-12,13-dibutyrate (150 nM). This phosphorylation was markedly inhibited by C-PC (4 and 8 nM). In addition, C-PC (4 and 8 nM) markedly reduced the electron spin resonance (ESR) signal intensity of hydroxyl radicals in collagen (1 microg/mL)-activated platelets. The present study reports on a novel and very potent (in nanomolar concentrations) antiplatelet agent, C-PC, which is involved in the following inhibitory pathways: (1) C-phycocyanin increases cyclic GMP/VASP Ser157 phosphorylation and subsequently inhibits protein kinase C activity, resulting in inhibition of both P47 phosphorylation and intracellular Ca2+ mobilization, and (2) C-PC may inhibit free radicals (such as hydroxyl radicals) released from activated platelets, which ultimately inhibits platelet aggregation. These results strongly indicate that C-PC appears to represent a novel and potential antiplatelet agent for treatment of arterial thromboembolism.     

Dinoseb as a specific inhibitor of nitrogen fixation in soil

N₂ fixation, estimated by reduction of C₂H₂, by a loamy soil amended with glucose was highly sensitive to the pesticide, 4,6-dinitro-o-sec-butylphenol (DNBP).At a concentration of 6 parts/10⁶ DNBP inhibited reduction of C₂H₂ by 90% in an anaerobic artificial soil system. Studies of the nitrogen-fixing enzyme system isolated from Azotobacter suggested that DNBP may compete with ATP for the same site on the enzyme.     

Phenylboronic acid--a potent inhibitor of lipase from Oryza sativa

The kinetics of inhibition of rice bran lipase (RBL) by phenylboronic acid (PBA) was studied to elucidate the nature of inhibition and the effect of the inhibitor on the structure-function of RBL. The effectiveness of an inhibitor is normally expressed by the constant K(i), which is calculated from the Lineweaver-Burk plot and found to be 1.7 mM at pH 7.4. The kinetics of inhibition by PBA was competitive, indicating the presence of serine in the active site of the enzyme. The loss of activity of RBL was concentration dependent on the inhibitor (PBA), and the inactivation followed a pseudo-first-order kinetics. Fluorescence emission measurements indicated a decrease in the fluorescence emission intensity and a red shift in the emission maximum as the inhibitor concentration was increased. The inhibition of the enzyme by PBA was also confirmed by thermal denaturation measurements, which indicated a shift in the thermal denaturation temperature of the enzyme toward lower temperatures. The far-UV-CD data suggest that there were no significant changes in the conformation of the enzyme as a result of binding of PBA. These results indicate that PBA is a potential inhibitor of RBL and binds to the enzyme in bringing about inhibition without any structural alterations.     

A growth inhibitor specific to upland rice produced by Pyrenochaeta SP.

An attempt was made to isolate a growth inhibitor of upland rice produced by Pyrenochaeta sp. Partially purified pink substance having absorption maxima at 481, 502, 512,537 and 548 nm was obtained from mycelia of Pyrenochaeta sp. This substance inhibited the growth of upland rice at a lower concentration than p-coumaric acid. It inhibited the growth of upland rice and sorghum, but stimulated the growth of Chinese cabbage and cucumber.     

Isolation of a new phlorotannin, a potent inhibitor of carbohydrate-hydrolyzing enzymes, from the brown alga Sargassum patens

Ethanol extracts from 15 kinds of marine algae collected from the coast of the Noto Peninsula in Japan were examined for their inhibitory effects on human salivary α-amylase. Four extracts significantly suppressed the enzyme activity. An inhibitor was purified from the extract of Sargassum patens . The compound was a new phloroglucinol derivative, 2-(4-(3,5-dihydroxyphenoxy)-3,5-dihydroxyphenoxy) benzene-1,3,5-triol (DDBT), which strongly suppressed the hydrolysis of amylopectin by human salivary and pancreatic α-amylases. The 50% inhibitory activity (IC(50)) for α-amylase inhibition of DDBT (3.2 μg/mL) was much lower than that of commercially available α-amylase inhibitors, acarbose (26.3 μg/mL), quercetagetin (764 μg/mL), and α-amylase inhibitor from Triticum aestivum (88.3 μg/mL). A kinetic study indicated that DDBT was a competitive α-amylase inhibitor with a K(i) of 1.8 μg/mL. DDBT also inhibited rat intestinal α-glucosidase with an IC(50) value of 25.4 μg/mL for sucrase activity and 114 μg/mL for maltase activity. These results suggest that DDBT, a potent inhibitor of carbohydrate-hydrolyzing enzymes, may be useful as a natural nutraceutical to prevent diabetes.     

Activity of octylthiotrifluoropropan-2-one,a potent esterase inhibitor,on growth,development, and intraspecific communication in Spodoptera littoralis and Sesamia nonagrioides

A series of experiments were conducted to examine the effect of 3-octylthio-1,1,1-trifluoro-2-propanone (OTFP) on growth, development, and behavior of the cotton leafworm, Spodoptera littoralis (Lepidoptera: Noctuidae), and the corn stalk borer, Sesamia nonagrioides (Lepidoptera: Noctuidae). The chemical behaved as an oviposition deterrent and, when added to the diet of the second-instar larvae of both insects, reduced diet consumption and growth, pupation, and adult emergence. Treatment of 100-5000 ng of the compound on fourth-instar larvae for 3-24 h, however, did not produce significant differences in the amount of diet ingested. Our results suggest that the effect of OTFP is long-lasting and that the inhibitor is not fully detoxified by the detoxification enzymes of the digestive tract of the insects. In behavioral assays, adult males which had been treated with the chemical at the larval stage were less attracted to the pheromone source than regular untreated males. When Sp. littoralis untreated females were used as the attractant source, treated males also displayed significantly fewer contacts with the cage-containing females than untreated or solvent-treated males. In the presence of treated females, only 27% of treated males successfully completed the flight in comparison to animals responding to solvent-treated females (54.5%). By contrast, when Se. nonagrioides females, whether they had been subjected or not to the treatment, were used as the attractant source, males were similarly attracted to them regardless of whether they had been treated or not at the larval stage. Analyses of gland extracts of Sp. littoralis treated females showed no difference from control insects in the qualitative or quantitative composition of the pheromone blend. The results obtained, in combination with other results previously reported by us (Riba, M.; Sans, A.; Bau, P.; Grolleau, G.; Renou, M.; Guerrero, A. J. Chem. Ecol. 2001, 27, 1879-1897), provide new and relevant information about the possible utility of these chemicals in future studies directed to the development of new approaches for pest control.     

Extremely potent antifeedant neo-clerodane derivatives of scutecyprol A

Two known neo-clerodane diterpenoids, scutecyprol A (1) and scutalbin C (2), have been isolated from the acetone extract of the aerial parts of Scutellaria sieberi. The antifeedant activity of scutecyprol A (1), of its 15-oxo derivative (3), and of several halohydrins (4-9), synthesized starting from compounds 1 and 3, against Spodoptera littoralis have been determined and structure-antifeedant relationships are discussed.     

树木年轮信息解析系统研制与试验

树木年轮信息在树木年代学、气候学、生态学、灾害学和环境学等领域研究中有重要的作用。为准确、快速、地获取树木年轮信息,降低工作强度和操作复杂度,该研究设计了一种树木年轮信息解析系统。该系统在硬件上进行了机电一体化设计,主要包含滑台模组、旋转编码器、步进电机、置物台、手轮、工业相机、控制箱、光学显微镜、电脑等组件,基于C#语言和OpenCvSharp工具包设计上位机,所设计的上位机软件通过与工业相机、采集器和控制器通信来实现对年轮信息采集、处理和存储,实现位置计算、电机控制和图像识别。系统设计了2种方法进行年轮信息解析：1） 电子显微定位法。在常规显微定位法的基础上,基于旋转编码器和螺旋测距法感知滑台线性位移进而实现年轮宽度测量,并使用工业相机和显示器代替传统显微镜来判读年轮线,最后可生成年轮解析图表;2）图机辅助法。在电子显微法的基础上,结合图像识别与电机控制算法进行改进。为验证系统的年轮宽度测量精度、年轮线识别准确率和解析效率,以传统显微镜和光学标定板作为参照工具,采集油松、落叶松、杨树、白桦4个树种的木芯样品进行试验。试验结果表明：在年轮宽度测量精度上,电子显微定位法和图机辅助法的平均绝对误差分别为5.68和7.84 μm,均方根误差分别为6.31和8.62 μm,满足国家标准;在年轮线识别上,油松、落叶松、杨树、白桦4个树种的准确率分别为83.3%、93.7%、55.0%和68.3%,针叶树种组、阔叶树种组的平均召回率分别为96.4%和72.6%,能够辅助判读年轮线;在解析效率上,平均每个年轮采集耗时为电子显微定位法1.58 s、图机辅助定位法1.24 s,分别比显微定位法提高了约19%和36%。综合而言,该系统实用性强、精度高、效率高,工作疲劳度低、成本低(<1万元),在多个学科领域中具有推广应用前景。     

Response of ammonia oxidizers and denitrifiers to repeated applications of a nitrification inhibitor and a urease inhibitor in two pasture soils

Purpose

The nitrification inhibitor 3,4-dimethylpyrazol-phosphate (DMPP) and the urease inhibitor N-(n-butyl) thiophosphoric triamide (nBTPT) can mitigate N losses through reducing nitrification and ammonia volatilization, respectively. However, the impact of repeated applications of these inhibitors on nitrogen cycling microorganisms is not well documented. This study aimed to investigate the changes in the abundance and community structure of the functional microorganisms involved in nitrification and denitrification in Australian pasture soils after repeated applications of DMPP and nBTPT.

Materials and methods

Soil was collected in autumn and spring, 2014 from two pasture sites where control, urea, urea ammonium nitrate, and urea-coated inhibitors had been repeatedly applied over 2 year. Soil samples were analyzed to determine the potential nitrification rates (PNRs), the abundances of amoA, narG, nirK and bacterial 16S rRNA genes, and the community structure of ammonia oxidizers.

Results and discussion

Two years of urea application resulted in a significantly lower soil pH at Terang and a significant decrease in total bacterial 16S rRNA gene abundance at Glenormiston and led to significantly higher PNRs and abundances of ammonia oxidizers compared to the control. Amendment with either DMPP or nBTPT significantly decreased PNRs and the abundance of amoA and narG genes. However, there was no fertilizer- or inhibitor-induced change in the community structure of ammonia oxidizers.

Conclusions

These results suggest that there were inhibitory effects of DMPP and nBTPT on the functional groups mediating nitrification and denitrification, while no significant impact on the community structure of ammonia oxidizers was observed. The application of nitrification or urease inhibitor appears to be an effective approach targeting specific microbial groups with minimal effects on soil pH and the total bacterial abundance.

     

Tetrahydrofolic Acid is a potent suicide substrate of mushroom tyrosinase

The coenzyme tetrahydrofolic acid is the most rapid suicide substrate of tyrosinase that has been characterized to date. A kinetic study of the suicide inactivation process provides the kinetic constants that characterize it: λ(max), the maximum apparent inactivation constant; r, the partition ratio or the number of turnovers made by one enzyme molecule before inactivation; and k(cat) and K(m), the catalytic and Michaelis constants, respectively. From these values, it is possible to establish the ratio λ(max)/K(m), which represents the potency of the inactivation process. Besides acting as a suicide substrate of tyrosinase, tetrahydrofolic acid reduces o-quinones generated by the enzyme in its action on substrates, such as l-tyrosine and l-DOPA (o-dopaquinone), thus inhibiting enzymatic browning.     

Nitrogen assimilation studies using 15N in soybean plants treated with imazethapyr, an inhibitor of branched-chain amino acid biosynthesis

The pattern of nitrogen assimilation in soybean plants treated with a herbicide that inhibits branched-chain amino acid biosynthesis was evaluated by (15)N isotopic analysis. The herbicide imazethapyr caused a strong decrease in nitrate uptake by roots, partly due to a reduced stomatal conductance. The inhibition of (15)N uptake was accompanied by a decrease in the (15)N content in the plant and, concomitantly, an inhibition of translocation to the shoot. Imazethapyr inhibited nitrate reductase activity in leaves and roots. Among all parameters studied, "de novo" synthesis of proteins was the first parameter of the N assimilation metabolism affected by the herbicide. These results show that this class of herbicides totally damages N metabolism and indicates a regulatory effect on N uptake and translocation that would be mediated by the increase in free amino acid pool provoked by the inhibition of branched-chain amino acid biosynthesis.     

Alterations of flavonoid biosynthesis in young grapevine (Vitis vinifera L.) leaves, flowers, and berries induced by the dioxygenase inhibitor prohexadione-Ca

Prohexadione-Ca is a structural mimic of 2-oxoglutarate, and according to this property, it is able to inhibit dioxygenase enzymes, which require 2-oxoglutarate as a cosubstrate. Such enzymes are involved in flavonoid biosynthesis; therefore, prohexadione-Ca treatment leads to alterations in the flavonoid metabolism in grapevine tissues. Because of the fact that phenolic compounds often are responsible for enhanced plant resistance, modification of phenylpropanoid metabolism using elicitation can be considered as a new potential strategy in plant protection. The phenolic compounds were analyzed by high-performance liquid chromatography combined with chemical reaction detection. Tissue treatment induced the accumulation of unusual flavonoids, which were identified as derivatives of pentahydroxyflavanone, eriodictyol, and luteoliflavan. Concentrations of constitutive flavonoids were also affected by the bioregulator treatment. The alterations of the flavonoid profiles are discussed with respect to substrate preferences of relevant enzymes.     

Purification and characterization of a trypsin inhibitor from Plathymenia foliolosa seeds

A novel trypsin inhibitor (PFTI) was isolated from Plathymenia foliolosa (Benth.) seeds by gel filtration chromatography on a Sephadex G-100, DEAE-Sepharose, and trypsin-Sepharose columns. By SDSPAGE, PFTI yielded a single band with a M(r) of 19 kDa. PFTI inhibited bovine trypsin and bovine chymotrypsin with equilibrium dissociation constants (K(i)) of 4 x 10(-8) and 1.4 x 10(-6) M, respectively. PFTI retained more than 50% of activity at up to 50 degrees C for 30 min, but there were 80 and 100% losses of activity at 60 and 70 degrees C, respectively. DTT affected the activity or stability of PFTI. The N-terminal amino acid sequence of PFTI showed a high degree of homology with various members of the Kunitz family of inhibitors. Anagasta kuehniella is found worldwide; this insect attacks stored grains and products of rice, oat, rye, corn, and wheat. The velvet bean caterpillar (Anticarsia gemmatalis) is considered the main defoliator pest of soybean in Brazil. Diatraea saccharalis, the sugar cane borer, is the major pest of sugar cane crops, and its caterpillar-feeding behavior, inside the stems, hampers control. PFTI showed significant inhibitory activity against trypsin-like proteases present in the larval midguts on A. kuehniella and D. saccharalis and could suppress the growth of larvae.     

Research on dicyandiamide as a nitrification inhibitor and future outlook

Abstract

The action and decomposition of dicyandiamide (DCD), a nitrification inhibitor, is discussed.

DCD is especially efficient when used with animal manure slurries or potato starch waste water. As a consequence, nitrate leaching can be reduced, yields and N uptake increased. DCD‐amended mineral N fertilizers applied once can substitute for split N applications, thus reducing labor costs without any loss in crop yield and quality. With wheat and sugar beets, use of a DCD‐containing product ("Alzon 22") reduced the requirement of N for maximum yield. New formulations, such as DCD plus a reducing substance, ammonium thiosulfate (ATS), or new inhibitors, such as guanylthiourea (GTU), will receive more attention in the future.     

结构参数对增压器浮环轴承润滑特性和环速比的影响

基于以往对增压器的浮环轴承润滑分析中大都忽略浮环的环速比影响,或将润滑性能和环速比独立分析。该文采用数值分析方法研究了增压器浮环轴承的润滑特性和环速比,分析中考虑了转轴、浮环、轴承座之间的传热因素,基于Reynolds方程和浮环平衡方程,建立了浮环轴承润滑模型,对比分析了浮环内、外层间隙,内、外圆半径4个结构参数对浮环轴承润滑特性和环速比的影响。结果表明,实际设计浮环时,需综合考虑结构参数对浮环润滑特性和环速比的影响及影响程度;浮环内层间隙增加,环速比降低,与内层间隙0.02 mm时相比,转速60 000 r/min时,内层间隙0.04 mm时的环速比减幅达23%,内层间隙增加,内、外膜温度减小,摩擦功耗略有增加,内层间隙0.03 mm时,浮环具有较理想的润滑性能和环速比;外层间隙0.06 mm的环速比均比外层间隙0.04 mm的环速比增加30%以上,外层间隙增加,外膜温度减小,且转速越高,外膜温度减幅越大;浮环内圆半径越小,环速比越小,内、外膜温度和摩擦功耗越小,浮环润滑性能越好;浮环外圆半径增加,环速比降低,但内膜温度、外膜温度、总摩擦功耗和总端泄流量变化幅度均在5%以内,外圆半径对浮环润滑性能影响不显著;浮环实际设计时,调整内圆半径比调整外圆半径对改善浮环润滑性能更有效。     

Laboratory evaluation of dicyandiamide as a soil nitrification inhibitor

Abstract

Laboratory studies to evaluate dicyandiamide (DCD) as a soil nitrification inhibitor showed that it is considerably more effective than several compounds that have been patented or proposed as fertilizer amendments for retarding nitrification of fertilizer nitrogen (N) in soil, but is considerably less effective than 2‐ethynylpyridine, nitrapyrin (N‐Serve), etridiazole (Dwell), 3‐methylpyrazole‐l‐carboxamide (MPC), or 4‐amino‐l,2,4‐triazole (ATC). Other findings in studies reported were as follows: a) DCD is more effective for inhibiting nitrification of ammonium‐N than of urea‐N; b) the effectiveness of DCD as a nitrification inhibitor is markedly affected by soil temperature and soil type and is limited by the susceptibility of DCD to leaching; c) DCD has very little, if any, effect on urea hydrolysis, denitrification, or seed germination in soil; d) products of DCD decomposition in soil (guanylurea and guanidine) have little, if any, effect on nitrification compared with DCD; e) in the absence of leaching, the persistence of the inhibitory effect of DCD on nitrification decreases with increase in soil temperature from 10 to 30°C, but the inhibitory effect of 50 μg DCD g^‐1 soil is substantial even after incubation of DCD‐treated soils at 20 or 30°C for 24 weeks.     

Isolation and identification of 2-methoxy-3,5-dimethylpyrazine, a potent musty compound from wine corks

The compound responsible for a "fungal must" taint evident in industry assessments of wine corks was identified as 2-methoxy-3,5-dimethylpyrazine. The identification was made on the basis of gas chromatography/odor analyses, collection of material using micropreparative techniques, determination of chemical properties of collected material, and comparison by gas chromatography/mass spectrometry with an authentic sample, synthesized from 2-hydroxy-3,5-dimethylpyrazine. 2-Methoxy-3,5-dimethylpyrazine is an extremely potent compound with an unpleasant, musty, moldy aroma and an aroma threshold in a white wine of 2.1 ng/L. While its contribution to the frequency and intensity of cork taint in bottled wine is yet to be established, it has been assessed by some wine industry personnel as second only to 2,4,6-trichloroanisole as a cause of cork taint in Australian wine.     

干旱胁迫下喷施14-羟基芸苔素甾醇对冬小麦穗花发育及碳氮代谢的调控

[目的]明确外源芸苔素甾醇类化合物(brassinosteroids,BRs)中14-羟基芸苔素甾醇(14-hydroxylated brassinosteroid,14-HBR)对干旱胁迫下冬小麦穗花发育成粒的调控效果,为小麦大田生产减轻干旱胁迫危害提供技术支撑.[方法]以大穗型品种周麦16(ZM16)和多穗型品种豫...