In this work, we assessed the in-vitro effects of eicosapentaenoic acid (EPA; C20:5n-3) and docosahexaenoic acid (DHA; C22:6n-3) (final concentration, 15 microM) on T cell blastogenesis, interleukin-2 and -4 (IL-2, IL-4) secretion, fatty acid composition and intracellular oxidative status in type I diabetic patients with or without complications. Con A stimulated lymphocyte proliferation, glucose uptake, intracellular reduced glutathione levels and catalase activity were lower in diabetics as compared to controls, regardless to the presence of complications. EPA and DHA diminished T-lymphocyte proliferation and IL-2 production but enhanced IL-4 secretion in both diabetic and control groups. No changes in the levels of reduced glutathione and in the activities of catalase and SOD were observed in control T cells cultured in the presence of EPA and DHA. However, in diabetic patients, addition of n-3 PUFA to culture induced an increase in T cell levels of reduced glutathione and hydroperoxide, and in activities of catalase and SOD. Low levels of arachidonic acid (C20:4n-6) were found in plasma membrane phospholipids of lymphocytes from diabetic patients compared to controls. Incubation of lymphocytes with EPA and DHA was associated with an incorporation of these fatty acids in membrane phospholipids. In conclusion, the beneficial effects of n-3 PUFA on T cell functions in type I diabetes could be attributed to their suppressive action and modulation of cytokine secretion, and to the improvement of intracellular oxidative status. 相似文献
Reconstitution of the biosynthetic machinery for fungal secondary metabolites in Aspergillus oryzae provides an opportunity both for stepwise determination of the biosynthetic pathways and the total biosynthesis of fungal natural products. However, to maximize the utility of the reconstitution system, a simple and rapid strategy for the introduction of heterologous genes into A. oryzae is required. In this study, we demonstrated an effective method for introducing multiple genes involved in the biosynthesis of fungal metabolites by using the expression vectors pUARA2 and pUSA2, each of which contains two cloning sites. The successful introduction of all the aflatrem biosynthetic genes (seven genes in total) after two rounds of transformation enabled the total biosynthesis of aflatrem. This rapid reconstitution strategy will facilitate the functional analysis of the biosynthetic machinery of fungal metabolites. 相似文献
Isoprenoids form the largest family of compounds found in nature. Isoprenoids are often attached to other moieties such as aromatic compounds, indoles/tryptophan, and flavonoids. These reactions are catalyzed by three phylogenetically distinct prenyltransferases: soluble aromatic prenyltransferases identified mainly in actinobacteria, soluble indole prenyltransferases mostly in fungi, and membrane‐bound prenyltransferases in various organisms. Fusicoccin A (FC A) is a diterpene glycoside produced by the plant‐pathogenic fungus Phomopsis amygdali and has a unique O‐prenylated glucose moiety. In this study, we identified for the first time, from a genome database of P. amygdali, a gene (papt) encoding a prenyltransferase that reversibly transfers dimethylallyl diphosphate (DMAPP) to the 6′‐hydroxy group of the glucose moiety of FC A to yield an O‐prenylated sugar. An in vitro assay with a recombinant enzyme was also developed. Detailed analyses with recombinant PAPT showed that the enzyme is likely to be a monomer and requires no divalent cations. The optimum pH and temperature were 8.0 and 50 °C, respectively. Km values were calculated as 0.49±0.037 μM for FC P (a plausible intermediate of FC A biosynthesis) and 8.3±0.63 μM for DMAPP, with a kcat of 55.3±3.3×10?3 s. The enzyme did not act on representative substrates of the above‐mentioned three types of prenyltransferase, but showed a weak transfer activity of geranyl diphosphate to FC P. 相似文献
Two series samples of Iron Oxide nanoparticles doped with nickel and cobalt with different doping values (x?=?0.01; 0.03; 0.05 and 0.07), were successfully synthesized by using sol–gel method, and then they were characterized by X-ray diffraction, scanning electron and vibrating sample magnetometer (VSM). X-ray diffraction analysis of two series samples showed the formation α-Fe2O3 nanoparticles, accompanied by two phases iron spinels, CoFe2O4 and NiFe2O4. In addition, the variations in grain size were observed for both two series. The observation by scanning electron microscopy reveals a change in the morphology of the grains of all the samples doped, which confirm the cobalt and nickel effect on the morphology of iron oxide nanoparticles. Magnetic measurements which were measured by VSM showed significant magnetic parameters such as coercivity and magnetization besides the ferromagnetic behavior of both two series doped with Cobalt and Nickel.
Antioxidant activities of Myrtus communis leaf phenolic compounds (McPCs) were investigated on 2,2′‐9‐azino‐bis‐3‐ethylbenzothiazoline‐6‐sulfonic acid (ABTS+?) and oxygen radical absorbance capacity (ORAC) tests or on oxidation of biological models, human low‐density lipoprotein (LDL) and phospholipid aqueous dispersion (l ‐α‐phosphatidylcholine stabilized by bile salts). Two extraction techniques, microwave‐assisted (MAE) and conventional (CE), were used to isolate McPCs, producing similar results of phenolic compound content. ABTS+? assay showed clearly that myrtle extracts exhibited a stronger scavenging effect than butylated hydroxyanisole and α‐tocopherol, with a slight advantage for myrtle CE extract. In ORAC assay, the both McPC extracts were similarly less effective than the pure compounds as caffeic acid and myricitrin (myricetin 3‐O‐rhamnoside) but stronger than butylated hydroxytoluene. Moreover, myrtle CE and MAE extracts, and myricitrin were able to inhibit similarly the production of conjugated dienes and to prolong the lag phase (Tlag) during Cu2+‐induced LDL oxidation with a dose‐response effect. The cryo‐electron microscopy observations on studied phospholipid dispersion stabilized by bile salts (BS) revealed the presence of bilayer vesicles and micelles. In 2,2′‐azobis (2‐amidinopropane) hydrochloride–induced phospholipid/BS oxidation, myrtle CE and MAE extracts gave similar effects to α‐tocopherol and caffeic acid but myricitrin showed a higher protective effect than myrtle extracts. We showed also that no synergic or additive effect between α‐tocopherol and myrtle extracts or caffeic acid in α‐tocopherol–enriched phospholipid/BS dispersion, but myricitrin showed an additive effect and thus promoted the total antioxidant activity. These data showed that myrtle extract could be used as potential natural antioxidants, food stabilizers, or natural health products. 相似文献
BACKGROUND: This study was designed to determine the prognostic value of positive surgical resection margin or highest nodal station sampled at thoracotomy in patients with non-small cell lung cancer. METHODS: Two reviewers independently examined the surgical records and pathologic reports from a randomized trial comparing computed tomography versus mediastinoscopy for staging of lung cancer. They recorded pathologic findings at the surgical resection margin, the highest mediastinal nodal station sampled at thoracotomy, histologic type, tumor size, N status, and evidence of vascular or lymphatic invasion. These variables formed the independent variables in logistic regression models to predict recurrence. RESULTS: Except for 1 patient, follow-up at 3 years for 399 included patients was complete. Significant predictors of recurrence were tumor size (odds ratio [OR], 1.2 (per centimeter); 99% CI [confidence interval], 1.1 to 1.4), and N status (compared with N0, N1: OR, 1.6; CI, 0.8 to 3.1; N2: OR, 3.2; CI, 1.4 to 7.5). Other variables, including positive surgical resection margin, did not predict early recurrence or death. CONCLUSIONS: In patients with non-small cell lung cancer, surgical resection margin or highest nodal station sampled at thoracotomy that are involved by carcinoma do not predict recurrence. The current definition of incomplete resection has limited prognostic significance. 相似文献
Silica-supported nickel catalysts have been prepared by a solventless technique involving the deposition of the [Ni(tmen)(acac)2] precursor from the gas phase. According to UV–vis and IR spectroscopy and to elemental analysis, the initial deposition step results in the grafting of Ni(II) complexes on silanol groups to give predominantly [Ni(acac)2 (Si-OH)2]0 monomeric inner-sphere complexes, with elimination of the neutral (tmen) ligands into the gas phase. Subsequent thermal treatments in oxidising atmospheres causes an oxidative decomposition of the (acac) ligands without nickel desorption or coalescence into oxide crystallites. The ensuing coordinatively unsaturated Ni(II) centres are reduced to Ni(0) by flowing hydrogen at low temperatures (300 °C), yielding nanosized Ni particles as evidenced by TEM and O2 titration. Thus, the gas-phase deposition technique represents an interesting alternative to conventional wet deposition procedures since it allows one to obtain well-dispersed supported nickel catalysts by reduction at low-temperatures. 相似文献