Reliability criteria of a bioassay using rats trained to detect estrus-specific odor in cow urine

An odor discrimination apparatus was used to quantify the reaction of rats against odor differences of estrous and diestrous urine of cows. Rats were trained to release impulses from a microswitch when they came into contact with estrous but not diestrous urine. After training, the discrimination ability was highly significant (P≤ 0.001) between both urine types. The reaction was specific for cow urine, and no significant discrimination occurred for estrous and diestrous urine from pigs or goats, or that of women. The discrimination of urine diluted with water was still highly significant up to 1:25. Another group of rats was trained to detect camphor. Decreasing concentrations were detected down to 0.1/ug/ml when diluted in water, and down to 1/ug/ml when diluted in estrous urine. Once trained to discriminate between estrous and diestrous urine, rats maintained this ability for 12 mo. Thus, a bioassay was set up to accompany fractionation steps in the laboratory which could lead to the isolation of estrous pheromones in cow urine.     

Responses of tabanids to Nzi traps baited with octenol,cow urine and phenols in Canada

Cow urine and the two phenols responsible for the attraction of biting flies to cow urine (4‐methylphenol, 3‐n‐propylphenol) were compared with octenol (1‐octen‐3‐ol) as baits for Tabanidae. Relative to an unbaited Nzi trap, catches of the horseflies Hybomitra lasiophthalma (Macquart), Tabanus similis Macquart and Tabanus quinquevittatus Wiedemann (Diptera: Tabanidae) were increased by 1.5–2.6, 1.4–2.0 and 1.4–1.9 times, respectively, whenever a bait included octenol released at either 0.13 mg/h or 1.5 mg/h, regardless of the presence of phenols or urine. Catches were not affected when traps were baited with phenols alone at evaporation rates of 0.38 mg/h (4‐methylphenol) and 0.022 mg/h (3‐n‐propylphenol). Catches of Hybomitra horseflies were increased by 1.5–1.9 times with cow urine and 2.6 times with cow urine + octenol. This bait combination could prove to be particularly useful for Hybomitra horseflies, the common tabanids of northern environments.     

Antibacterial activity of wine phenolic compounds and oenological extracts against potential respiratory pathogens

Aims: To investigate the effect of seven wine phenolic compounds and six oenological phenolic extracts on the growth of pathogenic bacteria associated with respiratory diseases (Pseudomonas aeruginosa, Staphylococcus aureus, Moraxella catarrhalis, Enterococcus faecalis, Streptococcus sp Group F, Streptococcus agalactiae and Streptococcus pneumoniae). Methods and Results: Antimicrobial activity was determined using a microdilution method and quantified as IC₅₀. Mor. catarrhalis was the most susceptible specie to phenolic compounds and extracts. Gallic acid and ethyl gallate were the compounds that showed the greatest antimicrobial activity. Regarding phenolic extracts, GSE (grape seed extract) and GSE‐O (oligomeric‐rich fraction from GSE) were the ones that displayed the strongest antimicrobial effects. Conclusions: Results highlight the antimicrobial properties of wine phenolic compounds and oenological extracts against potential respiratory pathogens. The antimicrobial activity of wine phenolic compounds was influenced by the type of phenolic compounds. Gram‐negative bacteria were more susceptible than Gram‐positive bacteria to the action of phenolic compounds and extracts; however, the effect was species‐dependent. Significance and Impact of Study: The ability to inhibit the growth of respiratory pathogenic bacteria as shown by several wine phenolic compounds and oenological extracts warrants further investigations to explore the use of grape and wine preparations in oral hygiene.     

Enhancement and inhibition of luminol chemiluminescence by phenolic acids

We explored the behaviour of a series of phenolic acids used as enhancers or inhibitors of luminol chemiluminescence by three different methods to determine if behaviour was associated with phenolic acid structure and redox character. All the phenolic acids inhibited chemiluminescence when hexacyanoferrate(III) was reacted with the phenolic acids before adding luminol. The redox character of these compounds was clearly related to structure. When hexacyanoferrate(III)-luminol-O₂ chemiluminescence was initiated by phenolic acid-luminol mixtures some phenolic acids behaved as enhancers of chemiluminescence, and others as inhibitors. We propose a mechanism to explain these findings. We found direct relationships between the redox character of the phenolic acids and the enhancement or inhibition of the chemiluminescence of the luminol–H₂O₂–peroxidase system and we propose mechanism to explain these phenomena.     

Potential cow milk xanthine oxidase inhibitory and antioxidant activity of selected phenolic acid derivatives

Xanthine oxidase (XO) found in all mammals and excess activity leads to urolithiasis. The cow milk XO was purified to 305‐fold with a specific activity of 8.76 EU/mg and overall yield of 87% by using DEAE‐Sepharose chromatography. The phenolics showed potent XO inhibitory effect with K_i, P1 (0.412), P2 (0.632), P3 (0.585), P4 (0.886), P5 (1.633), P6 (0.503), P7 (2.882), P8 (3.761), P9 (4.487), and P10 (5.841) μM. The phenolics P9 and P10 exhibited uncompetitive inhibition; the phenolics P1, P2, P3, P4, and P6 showed competitive inhibition, and other phenolic acids showed noncompetitive inhibition. The studied phenolic compounds showed potent antioxidant activity and expressed as EC₅₀, ranged from, DPPH (4.2–25.8 μg mL^–1), ABTS (10.2–42.5 mmol TE 100 g^–1), and FRAP (6.3–36.8 mol Fe (II) 100 g^–1). The results obtained from this study might be utilized for design of XO inhibitors and as antigout agent.     

酸枣果抗菌增敏活性成分的提取及生物学活性研究

采用不同极性的溶剂,从野生酸枣果和木枣果中由低极性到高极性依次提取获得不同极性范围的提取物,通过检测其抗菌作用和提取物与抗生素的协同抗菌作用,从中筛选具有抗菌增敏作用的活性提取物,并经活性追踪的柱层析分离纯化进一步得到活性精提物,通过GC-MS分析确定其组成成分,最后检测了该活性精提物的生物学活性。结果表明:(1)在所有的枣果提取物中仅酸枣果氯仿提取物具有广谱的抗菌作用,并能显著增强铜绿假单胞菌对氨苄青霉素的敏感性,而其他提取物均无相应的生物学活性。(2)由酸枣果氯仿提取物进一步精制得到的Fr.2a组分,经GC-MS初步分析显示它包含49.59%1,3-二氯丙醇、5.49%1,1-二氯甲醚、0.96%六氯乙烷、7.81%1,1,2,3-四氯-2-丙烯、1.33%月桂酸、1.34%十四酸、0.87%棕榈油酸、7.37%棕榈酸、9.75%邻苯二甲酸二丁酯、2.02%反式-13-十八碳烯酸、1.88%油酸酰胺、3.06%β-香树精、0.93%α-菖蒲醇、6.20%羽扇豆醇和1.42%乌索醛等成分。(3)酸枣活性提取物Fr.2a与多种抗生素联用显示出广泛的协同抗菌作用,同时Fr.2a呈剂量依赖性地促进微生物生物膜的形成,降低微生物的运动性和显著抑制牛奶中微生物的生长。该研究结果为酸枣果的药用产品和天然防腐剂的开发奠定了基础。     

Exudation of peroxidase from roots of Festuca rubra and its effects on exuded phenolic acids

Peroxidase is exuded from roots of Festuca rubra under axenic conditions. No phenolase was detected. Peroxidase can use phenolic acids. probably differentially, as hydrogen donors for the H₂O₂ substrate and could thus have an effect on the qualitative and quantitative determinations of phenolic acids also exuded by plants.     

酚酸类化感自毒物质对枸杞种子萌发的抑制作用

酚酸类化感自毒物质是导致农作物连作障碍的重要诱因之一。枸杞作为多年生木本植物,连作障碍突出,成因复杂。为揭示酚酸类物质对枸杞的化感自毒作用,研究了22种常见酚酸对枸杞种子萌发的抑制作用,并运用比较分子场分析方法(CoMFA)进行构效关系研究。结果发现,酚酸化合物对枸杞种子萌发作用的半抑制浓度IC₅₀范围为39.94—115.97 mg/L;CoMFA结果表明,运用立体场和静电场能较好解释该类化合物对枸杞种子萌发的化感自毒特征,苯环1/2/3/4位具有大体积、1位具有强负电性取代基的酚酸化合物,对枸杞种子萌发具有较强抑制作用。研究结果可为客观评价酚酸类化合物对枸杞的化感自毒作用提供基础数据支撑。     

蜜蜂巢脾抗氧化活性与抗菌活性的研究

以意蜂巢脾和中蜂巢脾为研究材料,分别采用水提法和醇提法处理,对获得的4种提取物,以DPPH.法测定自由基清除能力,福林酚法测定总酚含量,琼脂扩散法测定其对金黄色葡萄球菌和大肠杆菌的抑菌半径。结果表明,巢脾提取物具有一定的抗氧化活性和抑菌活性,且含有丰富的酚类化合物。除了对大肠杆菌的抑制作用外,巢脾水提液效果优于巢脾醇提液;除DPPH.清除能力外,意蜂巢脾效果优于中蜂巢脾效果;生物学活性呈现浓度依赖效应。本研究为巢脾在中医药领域的应用提供了一定的解释,并证明了巢脾是一种潜在的天然生物资源。     

Free and bound phenolic acids of lucerne (Medicago sativa cv europe)

The distribution of free and covalently-bound phenolic acids was studied in various fractions obtained from fresh lucerne shoots. p-Hydroxybenzoic, vanillic, p-coumaric and ferulic acids were present, both free and bound, in all the fractions. Salicylic and sinapic acids occurred only in a bound, alkali-labile state and were found almost entirely in the ‘aqueous phase’ fraction after treatment of methanol-chloroform-water extract according to Bligh and Dyer. Many other common phenolics were absent. Amounts of the phenolic acids much larger than those extracted by methanol-chloroform-water were extracted subsequently by phenol-acetic acid-water and passed into the ‘diffusate’ fraction on dialysis of this extract against 70% acetic acid. Small, though significant, quantities of phenolic acids remained with the bulk protein in the ‘bag contents’ fraction. The extent to which the phenolic acids in these last two fractions are held to protein by covalent bonds or by secondary-valence attractions is discussed, particularly in relation to the isolation of N-feruloylglycylphenylalanine after partial hydrolysis. Suggestions are made for improving analytical procedures.     

连作花生土壤中酚酸类物质的检测及其对花生的化感作用

研究了南方红壤区不同连作年限花生土壤中酚酸物质的种类、含量,及其对花生生长的影响。结果表明:连作花生土壤中对羟基苯甲酸、香草酸和香豆酸随着连作年限的增加而增加,连作10a后3种酚酸总量达11.09mg·kg-1干土,显著高于连作3a和6a的土壤;而土壤中香豆素和苯甲酸含量比较低,且变化没有规律。所有酚酸处理组对花生幼苗的株高和根长表现出抑制作用,对花生幼苗地下部的干鲜重均表现出"低促高抑"的特点。香草酸和香豆酸处理组对花生幼苗地上部的干鲜重表现出"低促高抑"的特点,其他处理组均表现出抑制作用。花生幼苗根系活力随着酚酸处理浓度的增加而降低,花生幼苗的超氧化物歧化酶活力(SOD)、过氧化物酶活力(POD)、丙二醛含量(MDA)则随着酚酸浓度的增加而增加。与只用茄腐镰刀菌孢子悬液浸泡花生种子的对照相比,加入酚酸后,花生种子的病原菌的感染率随着酚酸浓度的增加而增加,发芽率则随着酚酸浓度的增加而下降。以上结果说明,酚酸物质可以抑制花生幼苗的生长和提高花生的发病率,可能是因为酚酸物质破坏花生细胞膜的完整性而使病原菌入侵,影响花生生长,产生连作障碍。     

Quenching of fluorescence of phenolic compounds and modified humic acids by cadmium ions

The interaction of a number of phenolic compounds, being ‘model fragments’ of humic acids, with cadmium ions was investigated. The fluorescence quenching method was used to determine the complexation constants of these compounds with cadmium ions. It was established that bonding of phenolic compounds by cadmium ions at рН 7 is weak and reaches a maximum value of 15% for interaction with resorcinol. It was demonstrated that modification of humic acids by the mechanoactivation method increases by three times bonding of cadmium ions, which is caused by strengthening the acid properties of carboxyl and hydroxyl groups at the aromatic ring. Copyright © 2016 John Wiley & Sons, Ltd.     

First Report about Mineral Content,Fatty Acids Composition and Biological Activities of Four Wild Edible Mushrooms

The goal of this research was a comprehensive analysis of four wild edible mushroom species, Cantharellus cinereus, Clavariadelphus pistillaris, Clitocybe nebularis and Hygrocybe punicea, which have not been analyzed so far. Extracts of different polarities have been prepared and evaluated for their antioxidant activities by DPPH, ABTS, FRAP, TRP and CUPRAC methods. For all extracts, total phenolic content was determined. Based on the analysis, it was shown that solvent type had a significant effect on the antioxidant capacities of mushroom extracts, so water extracts showed the highest activity. Furthermore, the analysis includes determination of mineral composition, fatty acid profiles and antimicrobial activity. Unsaturated fatty acids, which are very important for human health, are dominant in the studied mushroom species. Linoleic and oleic acid consist of over 50 % of the total fatty acid composition. Seventeen biologically important and toxic elements have been analyzed by ICP‐OES and ICP‐MS and results showed that the element concentrations were species‐dependent. Also, it has been found that analyzed mushrooms did not show any antimicrobial activity. Chemometric analysis was used to understand the connection between the extracts of different polarities.     

Antimicrobial activity and stability to proteolysis of small linear cationic peptides with D-amino acid substitutions

Antimicrobial peptides contribute to innate host defense against a number of bacteria and fungal pathogens. Some of antimicrobial synthetic peptides were systemically administered in vivo; however, effective protection has so far not been obtained because the effective dose of peptides in vivo seems to be very high, often close to the toxic level against the host. Alternatively, peptides administered in vivo may be degraded by certain proteases present in serum. In this study, D-amino acids were substituted for the L-amino acids of antimicrobial peptides to circumvent these problems. Initially a peptide (L-peptide) rich in five arginine residues and consisting of an 11-amino acid peptide (residues 32-42) of human granulysin was synthesized. Subsequently, the L-amino acids of the 11-amino acid peptide were replaced partially (D-peptide) or wholly (AD-peptide) with D-amino acids. Activity and stability to proteolysis, in particular, in the serum of antimicrobial peptides with D-amino acid substitutions were examined. Peptides with D-amino acid substitutions were found to lyse bacteria as efficiently as their all-L-amino acid parent, L-peptide. In addition, the peptide composed of L-amino acids was susceptible to trypsin, whereas peptides containing D-amino acid substitutions were highly stable to trypsin treatment. Similarly, the peptide consisting of L-amino acids alone was also susceptible to fetal calf serum (FCS), however, protease inhibitors restored the lowered antimicrobial activity of the FCS-incubated peptide. Thus, D-amino acid substitutions can make antimicrobial peptides resistant to proteolysis, suggesting that the antimicrobial peptides consisting of D-amino acids are potential candidates for clinical therapeutic use.     

Analytical Profiling of Phenolic Compounds in Extracts of Three Cistus Species from Sardinia and Their Potential Antimicrobial and Antioxidant Activity

The Cistaceae family is well represented in Sardinia, and the Cistus genus is widely used in traditional medicine. Nowadays only few studies have been performed on this genus vegetating in Sardinia in spite of its ethnobotanical importance. Moreover, in the past there have been conflicting opinions among botanists for the exact assignment of the species growing in Sardinia. We started several years ago to carried out studies on this genus and in the present study was to evaluate the in vitro activity of several samples of Cistus salvifolius L., Cistus monspeliensis L., and Cistus albidus L. collected in Sardinia as antimicrobial agents against Escherichia coli, Staphylococcus aureus, and four Candida species and their antioxidant activity using DPPH, ABTS, and FRAP assays. Furthermore, the phenolic content and composition of the extracts were first evaluated. Using statistical multivariate analysis on the complete metabolomics profile of all Cistus species growing wild in Sardinia, we confirmed the botanical classification, and we observed an interesting correlation between metabolomics profile and antioxidant activity.     

Levels of 8-oxo-dGsn and 8-oxo-Gsn in random urine are consistent with 24 h urine in healthy subjects and patients with renal disease

Oxidatively generated damage to nucleic acids may play an important role in the pathophysiological processes of a variety of diseases. 8-oxo-7,8-dihydro-2′-deoxyguanosine (8-oxo-dGsn) and 8-oxo-7,8-dihydroguanosine (8-oxo-Gsn) are oxidatively generated products of DNA and RNA, respectively. Our previous studies have suggested that the amounts of 8-oxo-dGsn and 8-oxo-Gsn in urine were considerably higher than other body fluid or tissue. The aim of this study was to investigate whether 8-oxo-dGsn and 8-oxo-Gsn levels in random urine samples are consistent with those in 24?h urine samples in healthy subjects and patients with renal disease. A total of 16 healthy subjects and 104 renal disease patients were enrolled in this study, and their random and 24?h urine samples were collected. The levels of urinary 8-oxo-dGsn and 8-oxo-Gsn were quantified by LC-MS/MS and corrected by creatinine. Regardless of healthy subjects or renal disease patients, the levels of oxidised nucleosides in random urine samples were consistent with 24?h urine samples. Regardless of the age bracket, there is no significant difference between random samples and 24?h urine samples. In conclusion, 8-oxo-dGsn and 8-oxo-Gsn levels in random urine samples could replace those in 24?h urine samples, and were considered as the representative of the level of systemic oxidative stress for the whole day.     

Influence of phenolic acids on growth and inactivation of Oenococcus oeni and Lactobacillus hilgardii

Aims: To determine the effect of several wine-associated, phenolic acids on the growth and viability of strains of Oenococcus oeni and Lactobacillus hilgardii. Methods and Results: Growth was monitored in ethanol-containing medium supplemented with varying concentrations of hydroxybenzoic acids (p-hydroxybenzoic, protocatechuic, gallic, vanillic and syringic acids) and hydroxycinnamic acids (p-coumaric, caffeic and ferulic acids). Progressive inactivation was monitored in ethanol-containing phosphate buffer supplemented in a similar manner to the growth experiments. Hydroxycinnamic acids proved to be more inhibitory to the growth of O. oeni than hydroxybenzoic acids. On the other hand, some acids showed a beneficial effect on growth of Lact. hilgardii. p-Coumaric acid showed the strongest inhibitory effect on growth and survival of both bacteria. Conclusions: Most phenolic acids had a negative effect on growth of O. oeni, for Lact. hilgardii this effect was only noted for p-coumaric acid. Generally, O. oeni was more sensitive to phenolic acid inactivation than Lact. hilgardii. Significance and Impact of the Study: Eight wine-derived, phenolic acids were compared for their effects on wine lactic acid bacteria. Results indicate that phenolic acids have the capacity to influence growth and survival parameters. The differences found between phenolic compounds could be related to their different chemical structures.     

Hydrocaffeic and p-coumaric acids,natural phenolic compounds,inhibit UV-B damage in WKD human conjunctival cells in vitro and rabbit eye in vivo

This paper studied the effect on UV-B ocular damage of 10µm hydrocaffeic acid (HCAF) alone and as a mixture (MIX) (5µm HCAF+5µm p-coumaric acid). Since ocular UV-B damage is mediated by reactive oxygen species, the aim was to test if HCAF and MIX could reduce oxidation damage in human conjunctival cells (WKD) in vitro and in cornea and sclera of rabbits in vivo. After UVB irradiation (44 J/m²) of WKD cells, 8-oxodG levels in DNA were markedly increased and this effect was attenuated by HCAF and MIX. Rabbit eyes were treated by application of HCAF and MIX drops before UV-B exposure (79 J/m²). Corneal and scleral DNA oxidation damage, xanthine-oxidase (XO) activity and malondialdehyde levels (MDA) in corneal tissue and prostaglandin E₂ (PGE₂) in the aqueous humour were reduced by HCAF alone and in combination with p-coumaric acid, showing their potential as a topical treatment against UV-B damage.     

Flavonoids and phenolic acids in adenostoma, a dominant genus of the californian chaparral

The flavonoid and phenolic acid pattern of Adenostoma fasciculatum and A. sparsifolium, two dominant, endemic species of the Californian chaparral, was analysed qualitatively and quantitatively. Adenostoma sparsifolium was found to secrete large amounts of lipophilic, unusually substituted flavonoids onto the leaf surface; A. fasciculatum produces five hydrophilic flavonol 3-O-glycosides of kaempferol, quercetin and isorhamnetin. The phenolic acid pattern differed quantitatively but not qualitatively between the species. The amounts of phenolic acids that could be detected within the leaves, leaf litter and soil beneath the shrubs seem too small to explain allelopathic effects as the main reason for the dominance of the two species.