Content of trans- and cis-resveratrol in Galician white and red wines

Content in resveratrol (trans and cis) present in commercial white and red wines of the five Galician Controlled Denomination of Origin (CDO Monterrei, CDO Rías Baixas, CDO Ribeira Sacra, CDO Ribeiro and CDO Valdeorras) has been determined by reversed-phase high-performance liquid chromatography (HPLC) with fluorimetric detection. The wines were filtered and directly injected, and also the samples were analyzed after the addition of two known amounts of the two standard isomers. The results obtained showed that the content in cis-resveratrol found in white and red wines ranged from 0.12 to 0.06 mg/L, respectively, whereas the concentration of trans-resveratrol was highest in the red wines (mean=12.68 mg/L), elaborated mainly from varieties such as Mencía, and practically null in the white wines (mean=2.51 mg/L), except in Godello and Albariño wines.     

Effects of five rice herbicides on the growth of two threatened aquatic ferns

The effects of five rice herbicides bensulfuron methyl, mefenacet, quinoclamine, simetryn, and thiobencarb on the growth of two threatened aquatic ferns Azolla japonica and Salvinia natans were tested using 12-day exposure experiments at 0.1-100 nM which are expected to be present in drainages and rivers in Japan. As a reference species, Lenma minor was also used to examine the toxicity of bensulfuron methyl. Bensulfuron methyl had the most pronounced effect on the relative growth rate (RGR) of A. japonica, S. natans, and L. minor with an EC50 of 5.0, 0.54, and 10 nM, respectively. The other herbicides reduced the RGR of the aquatic ferns only at the highest concentration (100 nM) or not at all. S. natans showed the highest susceptibility to bensulfuron methyl among the three species, and the EC50 for this species was comparable to or below the maximum concentration (0.49-5.6 nM) that had been previously detected in 7 of 11 rivers in Japan. These results suggest that bensulfuron methyl runoff in drainages and rivers in Japan is expected to have adverse effects on the growth of threatened aquatic ferns in some cases, and that no or small effects occur for the other four herbicides tested.     

Synthesis and cytotoxic activity of the heptaphylline and 7-methoxyheptaphylline series

Nineteen carbazole alkaloids modified from heptaphylline (I) and 7-methoxyheptaphylline (II) isolated from Clausena harmandiana were synthesized. Among these derivatives, Ih and IIi showed cytotoxicity against the NCI-H187 cell line with IC₅₀ values of 0.02 and 0.66 μM, respectively, which are about 138 and 4 fold stronger than the ellipticine standard. In addition, oxime Ih displayed cytotoxicity against KB cells with an IC₅₀ value of 0.17 μM which is about 10 times stronger than the ellipticine. This compound demonstrated weak cytotoxicity against Vero cells (IC₅₀ = 66.01 μM). The results show convincingly that Ih may be a promising lead for the development of cytotoxic agents.     

L-Proline anchored multicomponent synthesis of novel pyrido[2,3-a]carbazoles; investigation of in vitro antimicrobial, antioxidant, cytotoxicity and structure activity relationship studies

The newly synthesized pyrido[2,3-a]carbazoles were prepared in good yields by multicomponent reactions under l-proline as promoter and structurally characterized. Few compounds showed significant activity toward both gram-positive, gram-negative bacterial strains. All compounds exerted negative efficacy for antifungal activity except compounds 5f and 7f which showed moderate activity. All compounds showed weak to moderate capacity for scavenging DPPH. The cytotoxicity was evaluated by Sulforhodamine B assay against A-549, B16F10, HCT-15, SKMel2 and SKOV3 cell lines and compared with standard drug cisplatin. Compound 5f outperformed cisplatin against A-549, HCT-15, SKMel2 and B16F10 cell lines. Compound 5e outranged cisplatin against A-549, HCT-15, and SKMel2 cell lines. 5b outperformed cisplatin specifically against B16F10. The preliminary structure activity relationships were carried out.     

Antioxidant properties of trans-3,3',5,5'-tetrahydroxy-4'-methoxystilbene against modification of variety of biomolecules in human blood cells treated with platinum compounds

OBJECTIVE: A diet rich in natural antioxidants in combination with anticancer therapy is important in reducing morbidity and mortality in addition to diminishing toxicity and side effects of chemotherapeutic agents. Cisplatin (cis-diamminedichloroplatinum II, cis-Pt) is a common chemotherapeutic agent, but it causes side effects, including hematologic toxicity with changes in the biological function of blood cells. METHODS: We compared the action of two phenolic compounds isolated from the bark of Yucca schidigera: trans-3,3',5,5'-tetrahydroxy-4'-methoxystilbene and resveratrol (trans-3,4',5 - trihydroxystilbene, present also in grapes and wine) on oxidative stress induced by cisplatin (used in chemotherapy) and selenium-cisplatin conjugate ([NH(3)](2)Pt(SeO(3) [Se-Pt], with a slight toxic effect on blood cells) in human blood platelets and peripheral blood lymphocytes. RESULTS: The trans-3,3',5,5'-tetrahydroxy-4'-methoxystilbene, like resveratrol, significantly inhibited protein carbonylation (measured by enzyme-linked immunosorbent assay and western blot analysis) in blood platelets treated with platinum compounds (10 microg/mL) and markedly reduced oxidation of thiol groups of proteins in these cells. The trans-3,3',5,5'-tetrahydroxy-4'-methoxystilbene, like resveratrol, caused a distinct reduction of platelet lipid peroxidation induced by platinum compounds. The combined action of the tested phenolic compounds with Se-Pt evoked a significant decrease in DNA damage (measured by the comet assay) in lymphocytes compared with cells treated with Se-Pt only. CONCLUSION: We conclude that one promising natural product may be trans-3,3',5,5'-tetrahydroxy-4'-methoxystilbene, because it is a stronger antioxidant in the tested models in vitro compared with resveratrol (P < 0.05). The trans-3,3',5,5'-tetrahydroxy-4'-methoxystilbene can also be useful as a protective factor against platinum compounds during chemotherapy or cancer prophylaxis.     

温石棉与岩棉及硅灰石纤维体外细胞毒性的比较

目的 研究温石棉(CA)与岩棉(RW)和硅灰石(WS)纤维的细胞毒性.方法 将V79细胞分为CA组、RW组及WS组3个实验及1个阴性对照组(200 μl PBS),CA、RW及WS粉尘终质量浓度为100 mg/L,噻唑蓝(MTT)法观察V79细胞存活率,用动力学法测乳酸脱氢酶(LDH)活力,用扫描电子显微镜观察粉尘对V79细胞形态的影响.结果 SiO2均为3种纤维的主要成分,WS中的SiO2含量最高,为50.83%.CA组、RW组、WS的细胞存活率分别为54.5%、64.8%、65.7%,RW组和WS 组的存活率明[显高于CA组,差异有统计学意义(P＜0.01),RW组和WS组的LDH活力[分别为(15.7±0.9)、(12.3±3.7)U/L]明显低于CA组[(20.2±0.9)U/L].差异有统计学意义(P＜0.05).CA组的V79细胞两端堆积大量颗粒状残余体,RW组和WS组V79细胞形态基本正常.结论 RW和WS对V79细胞的毒性低于CA.

Abstract：

Objective To study the cytotoxicity induced by chrysotile asbestos (CA), rock wool (RW)and wollastonite (WS). Methods V79 cells were divided into 4 groups. i.e. CA group, WS group, RW group and control group (200 μ1 PBS). The exposure concentration of dusts was 100 mg/L, The cell viability was detected by MTT and lactate dehydrogenate (LDH) activity assays. The technique of scanning electron microscopy was used to examine the change of V79 cells. Results SiO2 was main constituent for 3 kinds of dusts. In MTT assay, the cell viability of RW and WS groups was 64.8% and 65.7%, respectively, which were significantly higher than that (54.5%) of CA group (P＜0.01). In LDH assay, the LDH activity of RW and WS groups [(15.7±50.9),( 12.3±3.7)U/L, respectively] was significantly lower than that[( 20.2±0.9) U/L]of CA group (P＜0.05). In scanning electron microscopy examination, it was found that the two ends of V79 cells in CA group contained a great deal of fibers remaining bodies, but the V79 cell appearance in RW and WS groups was normal. Conclusion The cytotoxicity induced by RW and WS is significantly lower than that induced by CA for V79 cell.     

Synthesis and cytotoxicity of 2-phenylquinazolin-4(3H)-one derivatives

Thirty 2-phenylquinazolin-4(3H)-one derivatives were prepared and their cytotoxic activities were tested in five human tumor cell lines. Some compounds (5e, 5k, 5t, 6c and 6f) showed relatively high cytotoxic activity. Especially, compound 6c showed the most cytotoxicity against all cell lines tested among the synthesized derivatives, and the inhibitory activity of 6c against HeLa cell was higher than that of adriamycin. The putative mechanism of antitumor action in apoptotic cell death was cell cycle arrest in the G0/G1 phase by compounds 5k, 5v, 5m, 6c, and 6f in HeLa cells. These compounds showed relatively high cytotoxicity in this cell type.     

Soil <Emphasis Type="Italic">Pseudomonas</Emphasis> Community Structure and Its Antagonism Towards <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> Under the Stress of Acetochlor

In a microcosm experiment, the amplified ribosomal DNA restriction analysis was adopted to investigate the Pseudomonas community structure in soils applied with different concentrations (0, 50, 150, and 250 mg/kg) of acetochlor, and an in vitro assay was made to examine the antagonistic activity of isolated Pseudomonas strains acting on soil-borne pathogen Rhizoctonia solani. The results showed that acetochlor application changed the community structure of Pseudomonas in aquic brown soil. The diversity of Pseudomonas and the amount of isolated Pseudomonas strains with antagonistic activity decreased with an increasing acetochlor concentration, and the toxic effect of acetochlor reached to a steady level at 150–250 mg/kg.     

Toxicity assessment of 40 herbicides to the green alga Raphidocelis subcapitata

The effects of 40 herbicides with nine modes of action on the green alga Raphidocelis subcapitata were studied by 96-h acute toxicity tests. Results showed that the EC50 of the herbicides with respect to the photosynthetic processes of R. subcapitata ranged from 0.0007 to 4.2286 mgL(-1). Photosynthesis was the process of the green alga most sensitive to the tested herbicides. The most toxic herbicides were atrazine, ametryme, simazine, prometryne, cyanazine, isoproturon, chlorotoluron, diuron, methabenzthiazuron, and paraquat. The EC50 of the protoporphyrinogen oxidase inhibitor oxadiargyl, which was the parameter least sensitive to the herbicides tested in this study, was 42.5 mgL(-1). The descending order of the average acute toxicity to R. subcapitata of herbicides with regard to the nine modes of action was as follows: photosynthetic process>cell division>lipid synthesis, acetyl-coenzyme A carboxylase>acetolactate synthase> 5-enolpyruvyl-shikimate-3-phosphate synthase, glutamine synthase, hormone synthesis>protoporphyrinogen oxidase.     

Investigation of animal and algal bioassays for reliable saxitoxin ecotoxicity and cytotoxicity risk evaluation

Contamination of water bodies by saxitoxin can result in various toxic effects in aquatic organisms. Saxitoxin contamination has also been shown to be a threat to human health in several reported cases, even resulting in death. In this study, we evaluated the sensitivity of animal (Neuro-2A) and algal (Chlamydomonas reinhardtii) bioassays to saxitoxin effect. Neuro-2A cells were found to be sensitive to saxitoxin, as shown by a 24 h EC50 value of 1.5 nM, which was obtained using a cell viability assay. Conversely, no saxitoxin effect was found in any of the algal biomarkers evaluated, for the concentration range tested (2-128 nM). These results indicate that saxitoxin may induce toxic effects in animal and human populations at concentrations where phytoplankton communities are not affected. Therefore, when evaluating STX risk of toxicity, algal bioassays do not appear to be reliable indicators and should always be conducted in combination with animal bioassays.     

Impact of silver and copper on the survival of amoebae and ciliated protozoa in vitro

The efficacy of 1:10 silver/copper combinations for inactivation of Hartmannella vermiformis amoebas and the ciliated protozoan Tetrahymena pyriformis in vitro was studied. Tetrahymena and Hartmannella/isolate 19 were inactivated for 2 log steps by 100 + 1000 micrograms/l Ag + Cu. Hartmannella/isolate 21 was more resistant. 500 + 5000 micrograms/l produced only a 0.6 log reduction. The investigations clearly showed that levels within the limit of the German drinking water regulation (10 + 100 micrograms/l Ag + Cu) could not inactivate these protozoas in vitro.     

Synthesis of 3-heteroarylthioquinoline derivatives and their in vitro antituberculosis and cytotoxicity studies

A series of 3-heteroarylthioquinoline derivatives has been synthesized by the Friedlander annulation of 2-[(5-methyl-1,3,4-thiadiazol-2-yl)sulfanyl]-1-aryl-1-ethanone/2-(1,3-benzothiazol-2-ylsulfanyl)-1-aryl-1-ethanone/1-aryl-2-[(2-phenyl-2H-1,2,3,4-tetraazol-5-yl)sulfanyl]-1-ethanone with 2-aminobenzophenone in good yields using YbCl₃ as the catalyst. These compounds have been screened for their in vitro activity against Mycobacterium tuberculosis H37Rv (MTB) and among the 21 compounds screened, 2-[2-(4-bromophenyl)-4-phenyl-3-quinolyl]sulfanyl-5-methyl-1,3,4-thiadiazole (5d) and 2-[2-(4-chlorophenyl)-4-phenyl-3-quinolyl]sulfanyl-5-methyl-1,3,4-thiadiazole (5c) were found to be the most active compounds with MIC of 3.2 and 3.5 μM respectively against MTB. The cytotoxic effects against mouse fibroblasts (NIH 3T3) in vitro were evaluated for 5c and 5d, which displayed no toxic effects (IC₅₀ > 1000 μM) against the mouse fibroblast cell line NIH 3T3.     

Relationship between the Pfcrt T76 and the Pfmdr-1 Y86 mutations in Plasmodium falciparum and in vitro/in vivo chloroquine resistance in Burkina Faso, West Africa

The relationship between Pfcrt T76 and Pfmdr-1 Y86 mutations in Plasmodium falciparum was explored in samples from patients with uncomplicated malaria and tested in vitro and in vivo with chloroquine (CQ) in Burkina Faso. The two mutations were strongly related. The Pfcrt T76 mutation was found in 82% of the samples having the Pfmdr-1 Y86 mutation too (odds ratio (OR)=4.8 [95% CI: 1.7-13.3]; P=0.002). However, only half (16/34) of samples with Pfcrt T76 mutation had also the Pfmdr-1 Y86 mutation. The latter was apparently associated with in vitro resistance (OR=4.8 [95% CI: 1.4-16.5]; P=0.01) but such association disappeared (P=0.77) after adjusting for the presence of the Pfcrt T76 mutation. This suggests that the occurrence of the Pfmdr-1 Y86 mutation is dependent on that of Pfcrt T76 mutation and could explain previous reports linking the Pfmdr-1 Y86 mutation with CQ resistance (CQR). The isolates carrying both the Pfcrt K76 and Pfmdr-1 N86 alleles (wild/wild (WW)) and the single mutant Pfmdr-1 Y86 (WM) had the lowest IC50 geometric mean (GMIC50) values, while those carrying both Pfcrt T76/Pfmdr-1 Y86 alleles (mutant/mutant (MM)), and the single mutant Pfcrt T76 (MW) had the highest. Among pre-treatment samples there was a strong linkage disequilibrium with an excess of MM and WW and a deficit of single mutants (MW and WM), suggesting that parasite fitness is higher for the former and lower for the latter.     

Bioaccumulation and toxicity of sediment associated herbicides (ioxynil, pendimethalin, and bentazone) in Lumbriculus variegatus (Oligochaeta) and Chironomus riparius (Insecta)

The benthic macroinvertebrates Lumbriculus variegatus and Chironomus riparius were used in toxicity and bioaccumulation tests to determine the toxic concentrations and accumulation potential of sediment associated herbicides. The tested chemicals were ioxynil, bentazone, and pendimethalin. The bioaccumulation tests with L. variegatus were performed in four different sediments, each having different characteristics. Water-only LC₅₀ tests were performed with both L. variegatus and C. riparius. A sublethal effect of model compounds in sediments was assessed by a C. riparius larvae growth-inhibition test. Of the model compounds, ioxynil appeared to be the most toxic, with LC₅₀ values 1.79 and 2.79 mg L⁻¹ for L. variegatus and C. riparius, respectively. The LC₅₀ water concentrations for bentazone were 79.11 and 62.31 mg L⁻¹ for L. variegatus and C. riparius, respectively. Similarly, ioxynil revealed the highest bioaccumulation potential in bioaccumulation tests. The most important characters affecting chemical fate in the sediment seemed to be the organic matter content and the particle size fraction. The sediments with low organic material and coarse particle size consistently showed high bioaccumulation potential and vice versa. In C. riparius growth tests bentazone had a statistically significant effect on larval growth at sediment concentrations of 1160 and 4650 mg kg⁻¹ (P<0.05). It is noteworthy that standard deviations tend to be greater at high chemical concentrations, which addresses the fact that part of the individuals started to suffer. Ioxynil had an effect on the larval growth in other test sediment at the highest concentration (15.46 mg kg⁻¹ dw), in which head capsule length correlated with larval weight, decreasing toward higher exposure concentrations. The current results show the importance of sediment organic matter as a binding site of xenobiotics.     

粪肠球菌临床株对泰利唑胺体外抗菌活性的研究

目的评价泰利唑胺体外抗粪肠球菌的活性,探讨泰利唑胺不敏感粪肠球菌的耐药机制及其多位点序列分型分布情况。方法收集2011年1月1日—2016年6月30日深圳市南山区人民医院临床分离的粪肠球菌菌株,使用自动化仪器法及微量肉汤稀释法对分离菌株的耐药性进行检测。应用聚合酶链式反应(PCR)方法检测恶唑烷酮类抗生素耐药基因的携带情况,并采用多位点序列分型(MLST)对分离菌株进行分型。结果共获得289株粪肠球菌,来源科室主要为外科(57.4%),标本来源主要为中段尿(126株,43.6%)。289株粪肠球菌对泰利唑胺敏感率为94.1%,对氨苄西林、呋喃西林和万古霉素有较高的敏感性(敏感率为97.9%～99.7%)。MLST结果显示,共分为47个ST型,优势ST分型为ST16和ST179,分别占29.1%(84株)和24.9%(72株),在泰利唑胺不敏感粪肠球菌中,ST16的比例高于ST179(P0.05)。共检出泰利唑胺不敏感粪肠球菌17株,其携带optrA基因比例高于敏感株。结论泰利唑胺对粪肠球菌的抗菌活性整体优于利奈唑胺,但对携带optrA基因的粪肠球菌则未显示出良好的抗菌活性。     

Salvia leriifolia Benth (Lamiaceae) extract demonstrates in vitro antioxidant properties and cholinesterase inhibitory activity

The object of the present study was to investigate the in vitro antioxidant properties and cholinesterase inhibitory activity of Salvia leriifolia Benth extracts and fractions. The functional role of herbs and spices and their constituents is a hot topic in food-related plant research. Salvia species have been used since ancient times in folk medicine for cognitive brain function and have been subjected to extensive research. Thus, we hypothesize that S leriifolia, because of its functional properties, would be a good candidate to use as a nutraceutical product for improving memory in the elderly or patients affected by Alzheimer disease (ad). To test this hypothesis, we examined the cholinesterase inhibitory activity using the modified colorimetric Ellman's method against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE). The n-hexane exhibited the highest activity, with inhibitory concentration 50% (IC₅₀) values of 0.59 and 0.21 mg/mL, for AChE and BChE, respectively. This extract was fractionated, and 9 of these fractions (A-I) were obtained and tested. Fraction G, characterized by the presence of sesquiterpenes as major components, was the most active against AChE (IC₅₀ = 0.05 mg/mL). Because oxidative stress is a critical event in the pathogenesis of AD, we decided to screen the antioxidant activity (AA) using 2,2-diphenyl-1-picrylhydrazyl test, β-carotene bleaching test, and bovine brain peroxidation (thiobarbituric acid) assay. The ethyl acetate extract showed the highest activity, with IC₅₀ values of 2 and 33 μg/mL on β-carotene bleaching test and thiobarbituric acid test, respectively. These results suggest potential health benefits of S leriifolia extracts. However, this finding requires additional investigation in vivo.     

Expression of cereolysine AB genes in Bacillus anthracis vaccine strain ensures protection against experimental hemolytic anthrax infection

The cereolysin AB genes from Bacillus cereus VKM-B164 have been expressed in Bacillus anthracis strains: virulent H-7 (PXO1, PXO2), vaccine STI-1 (PXO1), 221 (without its own plasmids). Expression was achieved by cloning the genes in a high copy number plasmid pE194. This construct was integrated with host genomes in amplified form. Gold hamsters were vaccinated with parental and recombinant B. anthracis STI-1 and 221 strains and challenged with virulent ones subcutaneously. Gold hamsters vaccinated with 221 strains showed absence of protection. STI-1 immunisation protected against the H-7 strain, but did not protect against the recombinant strain. STI-1 recombinant strain protected gold hamsters against the H-7 as well as the recombinant H-7 strains. The results describe the modulation of immunopathogenic properties of B. anthracis due to expression of cereolysin AB genes.     

The role of humic fractions from soil and compost in controlling the growth in vitro of phytopathogenic and antagonistic soil-borne fungi

The regulation capacity of four humic substance (HS) samples, a soil humic acid (HA) and two HAs and one fulvic acid (FA) isolated from a composting substrate, was evaluated at two concentrations on the growth in vitro of one plant pathogenic, Sclerotinia sclerotiorum, and two antagonistic, Trichoderma viride and T. harzianum, soil-borne fungi. The presence of any HS sample in the growing medium, especially those from the composting substrate, caused a relevant inhibition of the mycelial growth of S. sclerotiorum and a marked stimulation of sclerotial formation that was exhibited as early appearance and numerical increase. On the contrary, the same HS treatments generally did not inhibit the growth of the two Trichoderma species. In particular, T. viride was significantly stimulated by any HS sample at any concentration, with the only exception of HA from fresh-composting substrate, whereas T. harzianum appeared to be stimulated only slightly or not significantly influenced. Only S. sclerotiorum showed evident high correlations of both the extent of the inhibitory action on mycelial growth and the final number of sclerotia with some chemical and functional properties of HS, such as total acidity, COOH group content, and elemental composition.     

Detection of antibodies against Rhodococcus equi in Alpaca (Lama pacos) in Italy

After isolating the two virulent strains of Rhodococcus equi from Alpaca, a serological survey of Rhodococcus equi infection was carried out on 57 blood samples of Alpaca collected in Central Italy. The survey was performed with an ELISA test using a reference R. equi strain as antigen (ATCC 33701). Four (7.0%) sera (OD greater or equal to 0.3) tested positive, while five (8.77%) were considered doubtful (OD between 0.2 and 0.3). This is the first serological survey of R. equi infection in Alpaca in Italy. The results indicate that besides the horse R. equi infection could also affect some ruminant species. The ELISA test was recently introduced as a reliable diagnostic method in horses and was adapted to alpaca.