The anti-oxidative,anti-cell proliferative and anti-microbial efficacies of cold-adapted Crepis flexuosa: HPTLC and GC/MS analyses

The genus Crepis constitutes cold-adapted plant spp., of these some are traditionally used in folk medicine against inflammation or fungal infections without scientific validations. Here, we report the biological activities of Crepis flexuosa total ethanol-extract (CF-EtOH) and its hexane (CF-Hex), ethyl acetate (CF-EtOA), butanol (CF-ButOH), and aqueous (CF-Aqua) fractions. Our in vitro DPPH and ABTS radical-scavenging assays showed CF-EtOH, CF-ButOH and CF-Aqua with maximal, CF-EtOA with moderate, and CF-Hex with mild anti-oxidant activities. When tested on human cancer cell lines, high cytotoxicity was demonstrated by CF-EtOH (IC₅₀: 42.45 μg/ml) and CF-Aqua (IC₅₀: 46.37 μg/ml) on HepG2, followed by CF-Hex (IC₅₀: 63.24 μg/ml) and CF-ButOH (IC₅₀: 65.32 μg/ml) on MCF7 cells. The human primary cell line (HUVEC) had comparatively lower cytotoxicity for the tested samples. Moreover, when assessed for anti-microbial efficacy, CF-ButOH and CF-Aqua exhibited the strongest activity (MIC: 156.25 μg/ml) against S. aureus, E. faecalis and C. albicans. Further, while the developed RP-HPTLC identified the bioactive flavonoid luteolin-7-O-glucoside (17.58 mg/g), GS/MS analysis revealed sixteen compounds in C. flexuosa extract. In conclusion, we for the first time show the promising anti-oxidative, anti-cell proliferative and anti-microbial efficacies of C. flexuosa. This warrants further phytochemical and bio-efficacy studies towards isolations and identifications of active principles.     

Anti-microbial action of melanocortin peptides and identification of a novel X-Pro-D/L-Val sequence in Gram-positive and Gram-negative bacteria

The melanocortin peptides alpha-MSH, Lys-Pro-Val and Lys-Pro-D-Val are known to be potent anti-inflammatory agents; however their role as antibacterial peptides is less clear. The aim of this study was to determine whether these peptides displayed antibacterial properties, and specifically whether the Lys-Pro-D-Val tripeptide was more potent than Lys-Pro-Val, consistent with their anti-inflammatory actions. alpha-MSH, Ac-Lys-Pro-D-Val-NH2 and Ac-Lys-Pro-Val-NH2 were found to be antibacterial against both Gram-positive and Gram-negative bacteria (Staphylococcus aureus and Escherichia coli) over a broad range of concentrations compared to a control peptide, Ac-Ala-Ala-Ala-NH2. However, the relative potency of alpha-MSH, Ac-Lys-Pro-D-Val-NH2, Ac-Lys-Pro-Val-NH2 did not differ. Furthermore, it was found that the cationic charge on the lysine residue was not required for activity as a variant peptide Ac-Ala-Pro-D-Val-NH2 was also antibacterial. We therefore describe a novel X-Pro-D/L-Val peptide sequence with similarity to the short melanocortin peptides, which possess antibacterial activity. The combined anti-inflammatory and antibacterial action of such peptides may also have potential value therapeutically.     

Antioxidant and antibacterial capacity of stingless bee honey from Borneo (Sarawak)

Stingless honey bees form a large group of bees that lack of a sting and are found among Meliponinae species indigenous to various tropical and subtropical regions. They are able to produce “stingless bee honey” that contains divergent categories of phenolic and flavonoid compounds and have been associated with antioxidant and antibacterial activity. This study examines the physicochemical properties, antioxidant-activity and anti-microbial activity of stingless bee honey from Malaysia that was produced by Geniotrigona thoracica, Heterotrigona itama and Heterotrigona erythrogastra. The results show that G. thoracica honey has the highest concentration of the total phenolic context (99.04?±?5.14?mg/ml) and the greatest reducing power (19.05?±?0.79%), while flavonoids (17.67?±?0.75?mg/ml), reducing power (18.10?±?0.35%), DPPH (47.40?±?3.18%) and FRAP (50.66?±?5.77?mM of Fe²⁺/100?g) of H. itama honey is significantly higher than those of the other honeys. In addition, G. thoracica honey has the highest antibacterial activity against Staphylococcus xylosus (2.10?±?0.10?cm), which is Gram-positive bacterium, and against Pseudomonas aeruginosa (1.60?±?0.10?cm) and Vibrio parahaemolyticus (2.03?±?0.06?cm), which are Gram-negative bacteria. These results suggest that stingless bee honeys possess useful amounts of phenolic and flavonoid compounds that are able to act as natural anti-oxidants and also have significant anti-microbial activity.     

Short communication: In vitro evaluation of a Bacillus sp. for the biological control of the coffee phytopathogen Mycena citricolor

A cell-free supernatant and an ethanolic extract of a 3-day-old culture of Bacillus UCR-236 inhibited the growth of Mycena citricolor, as determined by the Oxford cylinder method. A 3-day-old culture of the same bacterium also decreased leaf infection by the pathogen in a moisture-chamber test.     

Binding and insertion of alpha-helical anti-microbial peptides in POPC bilayers studied by molecular dynamics simulations

We have performed molecular dynamics simulations of the interactions of two alpha-helical anti-microbial peptides, magainin2 and its synthetic analog of MSI-78, with palmitoyl-oleoyl-phosphatidylcholine (POPC) lipid bilayers. We used various initial positions and orientations of the peptide with respect to the lipid bilayer, including a surface-bound state parallel to the interface, a trans-membrane state, and a partially inserted state. Our 20 ns long simulations show that both magainin2 and MSI-78 are most stable in the lipid environment, with the peptide destabilized to different extents in both aqueous and lipid/water interfacial environments. We found that there are strong specific interactions between the lysine residues of the peptides and the lipid head-group regions. MSI-78, owing to its large number of lysines, shows better binding characteristics and overall stability when compared to magainin2. We also find that both peptides destabilize the bilayer environment, as observed by the increase in lipid tail disorder and the induction of local curvature on the lipid head-groups by the peptides. From all the simulations, we conclude that the hydrogen bonding interactions between the lysines of the peptides and the oxygens of the polar lipid head-groups are the strongest and determine the overall peptide binding characteristics to the lipids.     

Trypanosoma cruzi: Synergistic cytotoxicity of multiple amphipathic anti-microbial peptides to T. cruzi and potential bacterial hosts

The parasite Trypanasoma cruzi is responsible for Chagas disease and its triatomine vector, Rhodnius prolixus, has a symbiotic relationship with the soil bacterium, Rhodococcus rhodnii.R. rhodnii that was previously genetically engineered to produce the anti-microbial peptide, cecropin A was co-infected with T. cruzi into R. prolixus resulting in clearance of the infectious T. cruzi in 65% of the vectors. Similar anti-microbial peptides have been isolated elsewhere and were studied for differential toxicity against T. cruzi and R. rhodnii. Of the six anti-microbial peptides tested, apidaecin, magainin II, melittin, and cecropin A were deemed potential candidates for the Chagas paratransgenic system as they were capable of killing T.cruzi at concentrations that exhibit little or no toxic effects on R. rhodnii. Subsequent treatments of T. cruzi with these peptides in pair-wise combinations resulted in synergistic killing, indicating that improvement of the 65% parasite clearance seen in previous experiments may be possible utilizing combinations of different anti-microbial peptides.     

2016—2018年某医院肺炎克雷伯菌的临床分布及耐药性分析

目的分析某院肺炎克雷伯菌(Klebsiella pneumoniae, KPN)的临床分布,产超广谱β-内酰胺酶(extended-spectrum β-lactamase, ESBLs)的情况以及综合耐药特点,为临床经验性选用抗生素提供参考。方法回顾性分析某院2016-2018年临床分离培养的KPN耐药数据及临床资料,菌株均采用全自动分析仪进行菌种鉴定和药敏分析。结果 3年间共检出非重复KPN 2 211株,样本分布结果显示,KPN最多的是痰液1 268株,占57.35%;脑脊液最少12株,仅占0.54%。临床科室分布来看,ICU病房分离KPN最多274株,占12.39%;乳甲外科最少5株,仅占0.23%。2 211株KPN共检出ESBLs阳性708株,检出率为32.02%。KPN对头孢唑林、头孢呋辛、头孢曲松和复方新诺明的耐药率均已超过40%,对亚胺培南、阿米卡星和美罗培南耐药率仅为5.20%。与胆汁、血液和痰液样本中KPN的耐药性相比,尿液分离株的耐药性普遍较高(P<0.05)。结论 ESBLs检出率较高,耐药情况较为严峻,并且不同样本来源菌株的耐药性有较大差异,以尿液分离株的耐药性更为严重。医院需加强对重点科室,不同样本中KPN的耐药性监测,合理使用抗生素。     

Use of a continuous culture fermentation system to investigate the effect of GanedenBC30 (Bacillus coagulans GBI-30, 6086) supplementation on pathogen survival in the human gut microbiota

Single-stage continuous fermentation systems were employed to examine the effects of GanedenBC(30) supplementation on the human gastrointestinal microbiota in relation to pathogen challenge in vitro. Denaturing gradient gel electrophoresis analysis demonstrated that GanedenBC(30) supplementation modified the microbial profiles in the fermentation systems compared with controls, with profiles clustering according to treatment. Overall, GanedenBC(30) supplementation did not elicit major changes in bacterial population counts in vitro, although notably higher Bcoa191 counts were seen following probiotic supplementation (compared to the controls). Pathogen challenge did not elicit significant modification of the microbial counts in vitro, although notably higher Clit135 counts were seen in the control system post-Clostridium difficile challenge than in the corresponding GanedenBC(30)-supplemented systems. Sporulation appears to be associated with the anti-microbial activity of GanedenBC(30), suggesting that a bi-modal lifecycle of GanedenBC(30)in vivo may lead to anti-microbial activity in distal regions of the gastrointestinal tract.     

Click-tailed benzenesulfonamides as potent bacterial carbonic anhydrase inhibitors for targeting Mycobacterium tuberculosis and Vibrio cholerae

A series of 1,2,3-triazole-bearing benzenesulfonamides was assessed for the inhibition of carbonic anhydrases (CA, EC 4.2.1.1) from bacteria Vibrio cholerae (VchCAα and VchCAβ) and Mycobacterium tuberculosis (β-mtCA3). Growing resistance phenomena against existing antimicrobial drugs are globally spreading and highlight a urgent need of agents endowed with alternative mechanisms of action. Two global WHO strategies aim to reduce cholera deaths by 90% and eradicate the tuberculosis epidemic by 2030. The derivatives here reported represent interesting leads towards the optimization of new antibiotic agents showing excellent inhibitory efficiency and selectivity for the target CAs over the human (h) off-target isoform hCA I. In detail, the first subset of derivatives potently inhibits VchCAα in a low nanomolar range (K_Is between 0.72 and 22.6 nM). Compounds of a second subset, differing from the first one for the position of the spacer between benzenesulfonamide and triazole, preferentially inhibit VchCAβ (K_Is in the range 54.8–102.4 nM) and β-mtCA3 (K_Is in the range 28.2–192.5 nM) even more than the clinically used AAZ, used as the standard.     

Dihydroisocoumarins and a tetralone from Cytospora eucalypticola

Two dihydroisocoumarins, 3,5-dimethyl-8-hydroxy-7-methoxy-3,4-dihydroisocoumarin and 3,5-dimethyl-8-methoxy-3,4-dihydroisocoumarin were isolated from a culture filtrate of Cytospora eucalypticola, together with three known dihydroisocoumarins and a tetralone derivative. Their structures were determined by spectroscopic methods. These isocoumarins are mildly antifungal, and antibacterial towards gram positive bacteria. A known compound, 5-hydroxymethylmellein, showed mild antifeedant activity towards Spodoptera littoralis.