Baicalein induction of hydroxyl radical formation via 12-lipoxygenase in human platelets: an ESR study

The pro-oxidant activities of baicalein, morin, myricetin, quercetin, and rutin were examined in various cell-containing systems including human platelets, rat vascular smooth muscle cells, human umbilical vein endothelial cells (HUVECs), human THP-1 cells, and fibroblast cells. Electron spin resonance (ESR) results showed that only baicalein generated hydroxyl radicals in a resting human platelet suspension, whereas the other flavonoids showed no effects on any of the resting cell systems. A low concentration of arachidonic acid (AA) increased the intensity of hydroxyl radicals, but a high concentration inhibited it. Collagen and thrombin, platelet aggregatory agents that can cause the release of AA by platelets, enhanced baicalein-induced hydroxyl radical formation, whereas ADP and U44619 showed no significant effects. Quinacrine and 5,8,11,14-eicosatetraenoic trifluoromethyl ketone, both PLA2 inhibitors, significantly attenuated baicalein-induced hydroxyl radical formation. These results suggest that baicalein-induced hydroxyl radical formation is associated with AA metabolite enzymes in human platelets. The formation of hydroxyl radicals was significantly inhibited by lipoxygenase inhibitors including nordihydroguaiaretic acid, (-)-epicatechin, (-)-epicatechin gallate, and hinokitiol, but was not affected by desferroxamine or the heme protein inhibitors KCN and NaN3. On the other hand, semiquinone free radicals were generated when baicalein was incubated with horseradish peroxidase/H2O2 or platelets/AA. The semiquinone radicals formed in the platelets/AA system could be extensively inhibited by desferroxamine, diethylenetriaminepentaacetic acid, KCN, and NaN3, indicating that prostaglandin H synthase (PGHS)-peroxidase may be involved. The results of this study led to the proposal that baicalein induces hydroxyl radical formation via 12-lipoxygenase and induces semiquinone radical formation via PGHS-peroxidase in human platelets.     

Long-term tillage and crop rotation effects on weed seedbank size and composition

Size and composition of the weed seedbank was assessed after 12 years of application of four tillage systems in two crop rotations. Mouldboard and chisel ploughing at 45 cm, minimum tillage at 15 cm and no tillage were compared in continuous winter wheat and a pigeon bean/winter wheat 2-year rotation. Weed control was based upon post-emergence herbicide application. Weed seedling emergence from soil samples taken at 0–15, 15–30 and 30–45 cm depths was assessed in a non-heated glasshouse for 12 months. The tillage system influenced weed seedbank size and composition to a much greater extent than crop rotation. Total weed seedling density was higher in no tillage, minimum tillage and chisel ploughing plots in the 0–15, 15–30 and 30–45 cm layers respectively. Density in the whole (0–45 cm) layer did not differ significantly among tillage systems. With no tillage, more than 60% of the total seedlings emerged from the surface layer, compared with an average 43% in the other tillage systems. Crop rotation did not influence either weed seedbank size or seedling distribution among soil layers, and only had a small influence on major species abundance. The weed seedbank was dominated (>66%) by Conyza canadensis (L.) Cronq. and Amaranthus retroflexus (L.), which thrived in chisel ploughing and no tillage respectively. Results suggested that crop rotation and substitution of mouldboard ploughing by non-inversion tillage (especially by minimum tillage) would not result in increased weed problems, whereas use of no tillage might increase weed infestations because of higher seedling recruitment from the topsoil.     

Seed phosphorus remobilization is not a major limiting step for phosphorus nutrition during early growth of maize

Phosphorus (P) is the least mobile nutrient in the soil as compared to other macronutrients and therefore frequently limits crop growth. During germination and early growth, seed‐phytate hydrolysis and seed‐P remobilization is the major P source for developing seedlings. The objective of this paper was to investigate whether seed‐P hydrolysis and remobilization of nonphytate P are sufficient for seedling P nutrition during early growth stages of maize. A large part of initial maize endogenous seed P reserves are mainly in the form of phytate. Till 70 cumulated degree days after sowing, nearly all the phytate (98%) was hydrolyzed and caused an increase in nonphytate P in seeds. Phytate hydrolysis and remobilization of nonphytate P was the main source of P supply for the newly growing seedlings and was not a limiting step for seedling P nutrition during the first four weeks of early growth.     

Hibiscus protocatechuic acid or esculetin can inhibit oxidative LDL induced by either copper ion or nitric oxide donor

Oxidation of low-density lipoprotein (LDL) could increase the incidence of atherosclerosis. Previous studies have shown that copper and sodium nitroprusside (SNP) possess the ability to oxidize LDL in a dose-dependent condition. They increase the existing negative charge in LDL and increase the electrophoretic mobility. In this study, we used protocatechuic acid (PCA) and/or esculetin (ECT) to define the antioxidative activity in oxidative LDL by relative electrophoretic mobility (REM) and thiobarbituric acid-relative substances (TBARS). The data showed that ECT and PCA possessed stronger antioxidative activity than vitamin E in oxidative LDL. A previous study showed that the level of oxidative LDL can be determined by the cholesterol degradation and fragmentation of Apo B. Our results showed that Cu(2+)-mediated oxidative LDL can induce 31% cholesterol degradation and significant fragmentation of Apo B. Both PCA and ECT exhibited remarkable ability to rescue the cholesterol degradation and Apo B fragmentation. Taken together, both PCA and ECT showed strong potency to inhibit oxidative LDL induced by copper or an NO donor. Additionally, their nontoxic characteristics elevated the possibility for their use in the daily diet; and should further prevent atherosclerosis effectively.     

Computed tomography of proximal metatarsal and concurrent third tarsal bone fractures in a Thoroughbred racehorse

A 2‐year‐old Thoroughbred filly presented to the William R. Pritchard Veterinary Medical Teaching Hospital at the University of California at Davis for whole body nuclear scintigraphic evaluation after an acute onset of nonweightbearing right hindlimb lameness post race. The use of computed tomography (CT) enabled visualisation of additional pathology that was not initially appreciable using combined imaging modalities of nuclear scintigraphy and digital radiography, which ultimately altered the selected course of treatment. Computed tomography in horses with fractures of the cuboidal bones of the hock can provide valuable additional information regarding fracture configuration and radiographically occult pathology, which may help to guide clinical decisions about treatment.     

High molecular weight glutenin subunits in some durum wheat cultivars investigated by means of mass spectrometric techniques

The primary structures of high molecular weight glutenin subunits (HMW-GS) of 5 Triticum durum Desf. cultivars (Simeto, Svevo, Duilio, Bronte, and Sant'Agata), largely cultivated in the south of Italy, and of 13 populations of the old spring Sicilian durum wheat landrace Timilia (Triticum durum Desf.) (accession nos. 1, 2, 3, 4, 7, 8, 9, 13, 14, 15, SG1, SG2, and SG3) were investigated using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) and reversed-phase high performance liquid chromatography/nanoelectrospray ionization mass spectrometry (RP-HPLC/nESI-MS/MS). M(r) of the intact proteins determined by MALDI mass spectrometry showed that all the 13 populations of Timilia contained the same two HMW-GS with 75.2 kDa and 86.4 kDa, whereas the other durum wheat cultivars showed the presence of the expected HMW-GS 1By8 and 1Bx7 at 75.1 kDa and 83.1 kDa, respectively. By MALDI mass spectrometry of the tryptic digestion peptides of the isolated HMW-GS of Timilia, the 1Bx and 1By subunits were identified as the NCBInr Acc. No AAQ93629, and AAQ93633, respectively. Sequence verification for HMW-GS 1Bx and 1By both in Simeto and Timilia was obtained by MALDI mass mapping and HPLC/nESI-MSMS of the tryptic peptides. The Bx subunit of Timila presents a sequence similarity of 96% with respect to Simeto, with differences in the insertion of 3 peptides of 5, 9, and 15 amino acids, for a total insertion of 29 amino acids and 25 amino acid substitutions. These differences in the amino acidic sequence account for the determined Δm of 3294 Da between the M(r) of the 1Bx subunits in Timilia and Simeto. Sequence alignment between the two By subunits shows 10 amino acid substitutions and is consistent with the Δm of 148 Da found in the MALDI mass spectra of the intact subunits.     

5-Hydroxy-3,6,7,8,3',4'-hexamethoxyflavone induces apoptosis through reactive oxygen species production, growth arrest and DNA damage-inducible gene 153 expression, and caspase activation in human leukemia cells

This study examined the growth inhibitory effects of structurally related polymethoxylated flavones in human cancer cells. Here, we report that 5-hydroxy-3,6,7,8,3',4'-hexamethoxyflavone (5-OH-HxMF) induces growth inhibition of human cancer cells and induction of apoptosis in HL-60 cells through modulation of mitochondrial functions regulated by reactive oxygen species (ROS). ROS generation occurs in the early stages of 5-OH-HxMF-induced apoptosis, preceding cytochrome c release, caspase activation, and DNA fragmentation. The changes occurred after single breaks in DNA were detected, suggesting that 5-OH-HxMF induced irreparable DNA damage, which in turn triggered the process of apoptosis. Up-regulation of Bax was found in 5-OH-HxMF-treated HL-60 cells. In addition, a caspase-independent pathway indicated by endonuclease G also contributed to apoptosis caused by 5-OH-HxMF. Antioxidants suppress 5-OH-HxMF-induced apoptosis. 5-OH-HxMF markedly enhanced growth arrest DNA damage-inducible gene 153 (GADD153) protein in a time-dependent manner. N-acetylcysteine (NAC) and catalase prevented up-regulation of GADD153 expression caused by 5-OH-HxMF. These findings suggest that 5-OH-HxMF creates an oxidative cellular environment that induces DNA damage and GADD153 gene activation, which in turn helps trigger apoptosis in HL-60 cells. Meanwhile, ROS were proven an important inducer in this apoptotic process. The C-5 hydroxyl on the ring of 5-OH-HxMF was found to be essential for the antiproliferative and apoptosis-inducing activity. Our study identified the novel mechanisms of 5-OH-HxMF-induced apoptosis and indicated that these results have significant applications as potential chemopreventive and chemotherapeutic agents.     

桑苗圃和桑园的化学除草

用氟乐灵和精稳杀得或高效盖草能进行桑苗圃除草，用草甘磷和乙草胺混合进行桑园除草，可有效地防除杂草。