Thermodynamics of antimicrobial peptide JCpep8 binding to living Staphylococcus aureus as a pseudo-stationary phase in capillary electrochromatography and consequences for antimicrobial activity

To understand the details of the permeation pathways of antimicrobial peptide JCpep8, the antimicrobial processes were investigated step by step in this paper. First, the characterization of the initial binding process was explored by introducing the living Staphylococcus aureus cells (LSACs) into electrophoretic buffer used as pseudo-stationary phase in capillary electrochromatography (CEC), and the thermodynamic parameters were determined. The binding constants at 298, 303, and 309 K were 7.40 × 1011, 1.43 × 1012, and 2.6 × 1012 M?1, respectively, which indicated the evident interaction between JCpep8 and LSACs. This binding process was spontaneous. Both the electrostatic force and hydrophobic effect play major roles in this binding process. Second, antibacterial activity kinetics and outer membrane and inner membrane disruption assays were investigated. Data indicated that JCpep8 killed microbes principally by breaking their cell wall and membrane, followed by cell lysis. The results were confirmed by Fourier transform infrared (FTIR) spectroscopy and transmission electron microscopy (TEM). In summary, JCpep8 kills microbes mainly by wall-/membrane-targeting pore-forming mechanisms.     

Essential oil constituents and in vitro antimicrobial activity of Decalepis hamiltonii roots against foodborne pathogens

Hydrodistillation of Decalepis hamiltonii roots yielded an essential oil (0.33% v/w) that contained 2-hydroxy-4-methoxybenzaldehyde (37.45%), 2-hydroxybenzaldehyde (31.01%), 4-O-methylresorcylaldehyde (9.12%), benzyl alcohol (3.16%), and alpha-atlantone (2.06%) as major constituents, with aromatic aldehydes constituting the main fraction of this root's essential oil. The oil was tested for its antimicrobial activity against foodborne pathogens responsible for food spoilage and human pathologies using standard antimicrobial assays. It exhibited strong antimicrobial activity against Bacillus cereus, Bacillus megaterium, Candida albicans, Escherichia coli, Micrococcus luteus, Micrococcus roseus, and Staphylococcus aureus at a concentration range of 1:0 with inhibitory activities of 27, 23, 16, 19, 22, 19, and 23 mm, respectively, which are comparable to those of the standards. The roots of D. hamiltonii, therefore, may be considered as an inexpensive source of an essential oil rich in antimicrobial compounds against foodborne pathogens.     

In vitro antimicrobial activity of aminoreductone against the pathogenic bacteria methicillin-resistant Staphylococcus aureus (MRSA)

In this study, antimicrobial activity of aminoreductone (AR), a product formed during the initial stage of the Maillard reaction, against methicillin-resistant Staphylococcus aureus (MRSA) was evaluated. The significant growth inhibition of all 51 MRSA isolates irrespective of drug susceptibility by AR was observed. The minimum inhibitory concentration (MIC) of AR ranged from 13 to 26 mM. The bactericidal activity of AR was evaluated by a killing assay with multiples of MIC, and it was recognized to depend on its dose. The combined effects of AR and antibiotics frequently used for the treatment of infections caused by Gram-positive and Gram-negative bacteria, such as amikacin (AN), ciprofloxacin, imipenem and levofloxacin, were examined. As a result, AR did not interfere with these antibiotic activities against 12 MRSA isolates selected and showed the advanced effect of growth inhibition in combination with antibiotics. Moreover, the inhibitory effects of AR were similar to those of AN, an antibiotic with known adverse effects, some serious. These findings show that AR is a naturally formed antimicrobial agent present in thermally processed foods with potential health benefits in medical practice.     

Structural requirements for the antimicrobial activity of carvacrol

Carvacrol is a component of several essential oils and has been shown to exert antimicrobial activity. The structural requirements for the activity of carvacrol were determined by comparison to structurally related (nonessential oil) compounds. Removal of the aliphatic ring substituents of carvacrol slightly decreased the antimicrobial activity. The effect of the hydroxyl group of carvacrol on activity could not be determined by simply comparing it to p-cymene, because this compound is immiscible with water; therefore, 2-amino-p-cymene, the amino analogue of carvacrol, which has a similar hydrophobicity and structural characteristics, was used. 2-Amino-p-cymene had similar membrane disruption and bacterial killing characteristics as carvacrol showing that, contrary to previous reports, the hydroxyl group of carvacrol itself is not essential for the antimicrobial activity. However, the observed 3-fold lower activity for 2-amino-p-cymene as compared to carvacrol indicates special features in the antimicrobial mode of action of carvacrol due to the hydroxyl group.     

黄瓜内生细菌E1～E4防治黄瓜枯萎病研究

内生细菌是植物体自然生态系成员[1],是微生态系统的天然组成成分,作为生防微生物效果显著。有益内生细菌与植物在长期共同进化过程中建立了一种和谐的关系,从植物体内获得充分的营养,占居有利的生态位,在植物保护下不易受外界环境条件影响,并可以经受住植物防卫反应[2]。利用有益的植物内生细菌作用于植物微生态系,进行微生态调控,可达到抗病[3,5,6]、促进植物生长[6]的目的。目前国内在棉花、水稻、番茄、辣椒、花生、小麦、烟草、甘蔗、马铃薯等作物中分离到内生细菌并作了相关研究,但对黄瓜内生细菌的研究未见报道。本研究从黄瓜内生微…     

鸡传染性支气管炎病毒S1基因在大肠杆菌中的表达及其生物活性的初步鉴定

摘要：利用基因重组技术将鸡传染性支气管炎病毒S1基因分段（A段和B段）克隆到pGEX-6p-1载体中，并利用IPTG诱导表达。表达产物裂解后，经SDS-PAGE电泳和考马司亮蓝染色，有明显的融合表达蛋白条带。通过特异性抗体的Western-blot检测，两段表达产物均在相应位置出现了明显的条带，证明表达产物具有部分抗原性。用Glutathione Sepharose 4B纯化出GST融合蛋白。将纯化得到的融合蛋白免疫家兔，连续免疫3次，将得到的抗体与200TOCID50/0.1ml病毒工作液在37℃温育1h进行气管环试验，计算结果 A段抗体的50%血清中和终点为1：18，而B段抗体的50%血清中和终点为1：53，从而表明这两段的表达产物具有较好的免疫原性，且B段表达产物的免疫原性比A段高。     

Evaluation of the antimicrobial activity of citrus essences on Saccharomyces cerevisiae

The aim of this research was to assess the antimicrobial activity of nine different industrial essences used in a soft drink factory in relation to their composition, as well as to verify the role of vapor pressure on their bioactivity. The essences were tested against a Saccharomyces cerevisiae strain isolated from spoiled soft drinks. The tests were carried out by adding the essences directly to a liquid medium or into the headspace of closed systems inoculated with the yeast. The headspace composition was evaluated through a solid phase microextraction-gas chromatography technique. The use of a mass spectrometer allowed the identification of the peaks detected. The microbial growth was indirectly monitored by measuring the metabolic CO2 released by the yeast. The results obtained indicated that the most effective essences were characterized by the highest concentration of some terpenes, such as citral, beta-pinene, and p-cymene. Moreover, all of the essences were more bioactive when added directly to the liquid medium.     

牛白细胞介素18的原核表达及其生物学活性研究

将牛白细胞介素18(bovine interleukin-18,BoIL-18)的成熟蛋白基因亚克隆到原核表达载体pGEX6p-1中，并在宿主菌BL21（DE3）中进行表达；用BoIL-18在昆虫细胞中的表达产物制备的兔多克隆抗体作为一抗进行Western blot分析；采用亲和层析法，对表达产物进行纯化；利用MTT染色法研究重组BoIL-18（re BoIL-18）对牛外周血单核细胞(PBMC) 增殖的促进作用；采用双抗体夹心ELISA法研究reBoIL-18诱导脾淋巴细胞产生IFN-γ的情况；采用微量细胞病变抑制法检测reBoIL-18对VSV导致细胞病变的抑制作用。结果:得到了BoIL-18的高效表达产物，表达量为31.8％，Western blot 显示，在44KD处有一特异性条带；纯化后获得了纯度较高的reBoIL-18蛋白；纯化的BoIL-18对PBMC增殖具有明显的促进作用，并且能够促进IFN-γ的诱生和抑制VSV病毒的活性。     

胶质芽胞杆菌(Bacillus mucilaginosus)D4B1菌株生理特性研究

对胶质芽孢杆菌D4B1菌株生理特性研究表明,D4B1菌株在供试培养基A中生长良好,12 h进入对数生长期,48 h达到生长高峰,菌体浓度为6.0×108个/mL;对氨苄青霉素、卡那霉素、氯霉素、硫酸链霉素没有抗性;能分泌胞外活性物质抑制植物病原真菌的生长;具有较强的解磷解钾能力,但无植酸酶活性.其结果可以作为进一步分子改造胶质芽胞杆菌的参考.     

RP-HPLC analysis of the phenolic compounds of plant extracts. investigation of their antioxidant capacity and antimicrobial activity

Extracts of aromatic plants of Greek origin were examined as potential sources of phenolic compounds. RP-HPLC with UV detection was employed for the identification and quantification of the phenolic antioxidants, present in methanolic extracts. The most abundant phenolic acids were ferulic acid (1.1-280 mg/100 g of dry sample) and caffeic acid (1.2-60 mg/100 g of dry sample). (+)-Catechin and quercetin were the most abundant flavonoids. Apigenin and luteolin were detected in high amounts in Menta pulegium and Thymus vulgaris, respectively. The antioxidant capacity was determined, in dried ground plants and in their methanol extracts, with the Rancimat test using sunflower oil as substrate. Both pulverized plants and extracts showed antioxidant capacity. Total phenolic content in the extracts was determined spectrometrically according to the Folin-Ciocalteu assay and ranged from 1 to 21 mg of gallic acid/100 g of dry sample. Antimicrobial activity of the extracts against selected microbes was also conducted in this study.     

Composition and antimicrobial activity of the essential oils of two Origanum species

The essential oils obtained from the aerial parts of Origanum scabrum and Origaum microphyllum, both endemic species in Greece, were analyzed by means of GC and GC-MS. Forty-eight constituents were identified, representing 98.59 and 98.66% of the oils, respectively. Carvacrol, terpinen-4-ol, linalool, sabinene, alpha-terpinene, and gamma-terpinene were found as the major components. Furthermore, both samples exhibited a very interesting antimicrobial profile after they were tested against six Gram-negative and -positive bacteria and three pathogenic fungi.     

Antioxidant activity of a flavonoid-rich extract of Hypericum perforatum L. in vitro

A flavonoid-rich extract of Hypericum perforatum L. (FEHP) was prepared by adsorption on macroporous resin and desorption by ethanol. Total flavonoid content of FEHP was determined by a colorimetric method. The major constituents of FEHP, including rutin, hyperoside, isoquercitrin, avicularin, quercitrin, and quercetin, were determined by HPLC analysis and confirmed by LC-MS. Different antioxidant assays were utilized to evaluate free radical scavenging activity and antioxidant activity of FEHP. FEHP was an effective scavenger in quenching DPPH and superoxide radical with IC50 of 10.63 microg/mL and 54.3 microg/mL, respectively. A linear correlation between concentration of FEHP and reducing power was observed with a coefficient of r2 = 0.9991. Addition of 150 microg of FEHP obviously decreased the peroxidation of linoleic acid during 84 h incubation, but the amount of FEHP over 150 microg did not show statistically significant inhibitory effect of peroxidation of linoliec acid (p > 0.05). FEHP exhibited inhibitory effect of peroxidation of liposome induced both by hydroxyl radical generated with iron-ascorbic acid system and peroxyl radical and showed prominent inhibitory effect of deoxyribose degradation in a concentration-dependent manner in site-specific assay but poor effect in non-site-specific assay, which suggested that chelation of metal ion was the main antioxidant action. According to the results obtained in the present study, the antioxidant mechanism of FEHP might be attributed to its free radical scavenging activity, metal-chelation activity, and reactive oxygen quenching activity.     

Composition and antimicrobial activity of the essential oils of Micromeria cristata subsp. phrygia and the enantiomeric distribution of borneol

Water-distilled essential oils from herbal parts of Micromeria cristata (Hampe) Griseb. subsp. phrygia P. H. Davis (Endemic) (Lamiaceae) collected from three different localities were analyzed by GC-MS. The major component characterized in the three oils was borneol (27-39%). Other main components were determined as camphor (9-15%), caryophyllene oxide (4-6%), and trans-verbenol (4-6%) in the oils. Enantiomeric distributions of borneol and camphor in the oils were determined on a fused silica Lipodex-E capillary column using a multidimensional GC-MS system. The three essential oils and both enantiomers of borneol have been evaluated for their antimicrobial activity. They showed inhibitory effects on Gr (-) and Gr (+) pathogenic microorganisms.     

Influence of natural and synthetic humic substances on the activity of urease

The activity of a purified urease, obtained from Bacillus pasteurii, was inhibited by humic and fulvic acids obtained from an agricultural soil. Enzyme kinetic studies showed that the humic substances affected the affinity of the enzyme for its substrate (K_m) and the maximum velocity of the reaction (V_max). The V_max was inhibited to the same extent by both humic (HA) and fulvic (FA) acids, the precise effect depending on the pH and concentration of humic substance. At pH 4.0, HA concentrations of 25 pg cm^?3 and 10 μg cm^?3 inhibited the V_max by 38.5% and 20% respectively. HA and FA had similar effects on the K_m but in this case the lowering of the affinity of the enzyme for its substrate was not concentration dependent in the range 0–25 μg cm^?3 of humic substance. Typically, the affinity was decreased from a K_M of 50 mM in the control to 67 mM in the presence of HA and FA. The effects were not due primarily to the ash or N contents of the humic substances because de-ashed humic acid and synthetic model humic (made from catechol, guaiacol, pyrogallol, resorcinol and protocatechuic acid) and fulvic acid (made from polymaleic acid), containing virtually no ash or N, were equally as effective. The effect was not related to the phenolic monomers which, before polymerization, had no effect on urease activity.     

In vitro antimicrobial and antioxidant activities of the essential oils and various extracts of Thymus eigii M. Zohary et P.H. Davis

This study was designed to examine the in vitro antimicrobial and antioxidant activities of the essential oil and various extracts obtained from aerial parts of Thymus eigii. The essential oil was particularly found to possess stronger antimicrobial activity, whereas other nonpolar extracts and subfractions showed moderate activity and polar extracts remained almost inactive. GC-MS analysis of the oil resulted in the identification of 39 compounds, representing 93.7% of the oil; thymol (30.6%), carvacrol (26.1%), and p-cymene (13.0%) were the main components. The samples were also subjected to a screening for their possible antioxidant activity by using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and beta-carotene-linoleic acid assays. In the former case, the polar subfraction of the methanol extract was found to be superior to all extracts tested, only 16.8 microg/mL of which provided 50% inhibition, whereas all extracts, particularly the polar ones, seem to inhibit the oxidation of linoleic acid in the latter case. These data were further supported by total phenolics analysis, indicating that the antioxidative potential of the extracts was closely related to their phenolic constituents.     

Antioxidant activity of uruguayan propolis. In vitro and cellular assays

The antioxidant capacity of propolis from the southern region of Uruguay was evaluated using in vitro as well as cellular assays. Free radical scavenging capacity was assessed by ORAC, obtaining values significantly higher than those of other natural products (8000 μmol Trolox equiv/g propolis). ORAC values correlated well with total polyphenol content (determined by Folin-Ciocalteu method) and UV absorption. Total polyphenol content (150 mg gallic acid equiv/g propolis) and flavonoids (45 mg quercetin equiv/g propolis) were similar to values reported for southern Brazilian (group 3) and Argentinean propolis. Flavonoid composition determined by RP-HPLC indicates a strong poplar-tree origin. Samples high in polyphenols efficiently inhibit low-density lipoprotein lipoperoxidation and tyrosine nitration. In addition, Uruguayan propolis was found to induce the expression of endothelial nitric oxide synthase and inhibit endothelial NADPH oxidase, suggesting a potential cardiovascular benefit by increasing nitric oxide bioavailability in the endothelium.     

Tentative identification of polyphenols in Sempervivum tectorum and assessment of the antimicrobial activity of Sempervivum L

Polyphenols were isolated from sliced fresh leaves of Sempervivum tectorum. After 21 h of extraction by methanol and removal of chlorophyll, ethyl acetate was used to separate oligomeric and polymeric polyphenols: 0.07% of oligomeric and 0.13% of polymeric polyphenols were found. After acidic hydrolysis of the oligomeric polyphenols, it was established by TLC, HPLC, and FAB mass spectra that kaempferol was the unique aglycon of the three main oligomeric constituents of S. tectorum. Paper chromatography suggested delphinidol to be the only anthocyanidin detectable in the material obtained by acidic hydrolysis of the polymeric polyphenol fraction. After Haslam degradation of the same polymeric polyphenol fraction, only 4-thiobenzyl-(-)-epigallocatechin and 4-thiobenzyl-(-)-epigallocatechin-3-gallate were found and tentatively identified. We concluded that procyanidins of B2 type could be the major components of the polymeric polyphenol fraction of this plant. Antimicrobial activity of Sempervivum L. leaves against six of seven selected microorganisms was observed.     

Effect of fruiting body maturity stage on chemical composition and antimicrobial activity of Lactarius sp. mushrooms

The effects of fruiting body maturity on chemical composition and antimicrobial activity of the wild mushrooms, Lactarius deliciosus and Lactarius piperatus, were evaluated. Immature and mature samples were analyzed for proximate constituents (moisture, fat, crude protein, ash, carbohydrates), nutritional value, and individual composition in fatty acids and sugars. Protein content, MUFA (monounsaturated fatty acids), and PUFA (polyunsaturated fatty acids) percentages increased with the fruiting body maturity stage, while carbohydrate and SFA (saturated fatty acids) content decreased. The maturity stage did not significantly affect the individual sugar profile. The antimicrobial activity of the mushrooms was screened against Gram positive and Gram negative bacteria and fungi, and correlated to the amounts of phenols, flavonoids, ascorbic acid, beta-carotene, and lycopene present in the immature and mature fruiting bodies. Mature fruiting bodies with mature spores presented lower antimicrobial activity, which was in agreement with the bioactive compound content found in those samples.     

Analysis of fat-soluble vitamins. XVI. Antirachitic activity of 5,6-trans-vitamin D3 alone and in the presence of 5,6-cis-vitamin D3 resin, using chick bioassays.

Two biological assays were conducted in which the antirachitic activity in chicks of 5,6-trans-vitamin D3 added to feed is compared with that of 5,6-cis-vitamin D3. On the basis of the results obtained it is concluded that the relative potency of the trans isomer is, at the most, 5% and that the antirachitic activity of the trans isomer is not markedly enhanced (an increase to a relative potency of 16%, at the most) if the cis isomer is also included in the diet. The results are not conclusive on the inhibition or lack of inhibition of the antirachitic activity of 5,6-cis-vitamin D3 by the presence of 5,6-trans-vitamin D3 in the feed.