Synergistic effects between silibinin and antibiotics on methicillin-resistant Staphylococcus aureus isolated from clinical specimens

Methicillin-resistant Staphylococcus aureus (MRSA) is a dangerous microorganism, and creates serious medical problems. It causes many types of infections in humans and often acquires multi-drug resistance. In this study, silibinin was evaluated against 20 clinical isolates of MRSA, either alone or in combination with ampicillin or oxacillin, using a checkerboard assay. The silibinin exhibited good activity against isolates of MRSA, and MRSA ATCC33952 and MSSA ATCC25923, with minimum inhibitory concentrations/minimum bactericidal concentrations (MICs/MBCs) ranging between 2-8/4-16 μg/mL, for ampicillin 2-1024/2-2048 μg/mL, and for oxacillin 0.25-32/0.5-64 μg/mL. The range of MIC(50) and MIC(90) were 0.5-4 μg/mL and 2-8 μg/mL, respectively. The MICs/MBCs for the combination of silibinin plus oxacillin or ampicillin were reduced by ≥4-fold against the MRSA isolates tested, demonstrating a synergistic effect, as defined by a fractional inhibitory concentration index (FICI) of ≤0.5. Furthermore, a time-kill study evaluating the growth of the tested bacteria showed that growth was completely attenuated after 2-5 h of treatment with the 1/2 MIC of silibinin, regardless of whether it was administered alone or with oxacillin (1/2 MIC) or ampicillin (1/2 MIC). In conclusion, silibinin exerted synergistic effects when administered with oxacillin or ampicillin and the antibacterial activity and resistant regulation of silibinin against clinical isolates of MRSA might be useful in controlling MRSA infections.     

Identification of metabolic fluxes in hepatic cells from transient 13C-labeling experiments: Part I. Experimental observations

An experimental set-up for acquiring metabolite and transient (13)C-labeling data in mammalian cells is presented. An efficient sampling procedure was established for hepatic cells cultured in six-well plates as a monolayer attached to collagen, which allowed simultaneous quenching of metabolism and extraction of the intracellular intermediates of interest. Extracellular concentrations of glucose, amino acids, lactate, pyruvate, and urea were determined by GC-MS procedures and were used for estimation of metabolic uptake and excretion rates. Sensitive LC-MS and GC-MS methods were used to quantify the intracellular intermediates of tricarboxylic acid cycle, glycolysis, and pentose phosphate pathway and for the determination of isotopomer fractions of the respective metabolites. Mass isotopomer fractions were determined in a transient (13)C-labeling experiment using (13)C-labeled glucose as substrate. The absolute amounts of intracellular metabolites were obtained from a non-labeled experiment carried out in exactly the same way as the (13)C-labeling experiment, except that the media contained naturally labeled glucose only. Estimation of intracellular metabolic fluxes from the presented data is addressed in part II of this contribution.     

Sublethal effects of hexaflumuron on development and reproduction of the diamondback moth,Plutella xylostella (Lepidoptera: Yponomeutidae)

Abstract Effects of hexaflumuron at 10% lethal concentration (LC₁₀) and LC₂₅ on development and reproduction parameters of the diamondback moth, Plutella xylostella (Linnaeus, 1753) (Lep.: Yponomeutidae) were investigated. Estimated LC₅₀, LC₁₀ and LC₂₅ values of leaf dip bioassay of hexaflumuron on the third instar larvae of the P. xylostella were 1.48, 0.59 and 0.91 mg/L, respectively. Hexaflumuron decreased pupal weight in the parent generation at sublethal concentrations but in the offspring generation, this effect was not observed. Sublethal concentrations increased egg, first and second larval instar and pupa developmental time and shortened life span of adults, but did not change the third and fourth larval instars and pre‐pupa developmental period. Also fecundity of females reduced significantly but hatchability of treatments and control were similar. Survival rate of pre‐adult stages declined significantly at LC₂₅ concentration. Reproduction parameters such as reproductive rate (R₀) and intrinsic rate of increase in sublethal concentrations were significantly lower compared with control, but gross reproduction rate (GRR) at the LC₁₀ concentration was increased and it could be hormoligosis. Also hexaflumuron significantly increased doubling time (Dt). We conclude that the sublethal effects of hexaflumuron might exhibit significant effects on the population dynamics of P. xylostella.     

Chlorogenic acid modifies plasma and liver concentrations of: cholesterol,triacylglycerol, and minerals in (fa/fa) Zucker rats

Chlorogenic acid, a phenolic compound found ubiquitously in plants, is an in vitro antioxidant and metal chelator. Some derivatives of chlorogenic acid are hypoglycemic agents and may affect lipid metabolism. Concentrations of cholesterol and triacylglycerols are of interest due to their association with diseases such as non-insulin-dependent-diabetes- mellitus and obese insulin resistance. As little is known about the effects of chlorogenic acid in vivo, studies using obese, hyperlipidemic, and insulin resistant (fa/fa) Zucker rats were conducted to test the effect of chlorogenic acid on fasting plasma glucose, plasma and liver triacylglycerols and cholesterol concentrations. Aditionally, the effects of chlorogenic acid on selected mineral concentrations in plasma, spleen, and liver were determined. Rats were implanted with jugular vein catheters. Chlorogenic acid was infused (5 mg/Kg body weight/day) for 3 weeks via intravenous infusion. Chlorogenic acid did not promote sustained hypoglycemia and significantly lowered the postprandial peak response to a glucose challenge when compared to the same group of rats before Chlorogenic acid treatment. In Chlorogenic acid-treated rats, fasting plasma cholesterol and triacylglycerols concentrations significantly decreased by 44% and 58% respectively, as did in liver triacylglycerols concentrations (24%). We did not find differences (p > 0.05) in adipose triacylglycerols concentration. Significant differences (p < 0.05) in the plasma, liver, and spleen concentration of selected minerals were found in chlorogenic acid-treated rats. In vivo, chlorogenic acid was found to improve glucose tolerance, decreased some plasma and liver lipids, and improve mineral pool distribution under the conditions of this study.     

光强和氮源及其浓度对缺刻缘绿藻生长、油脂和花生四烯酸积累的影响

对缺刻缘绿藻（Parietochloris incisa（Reisigl） S.Watanabe）在不同光强和氮源及其浓度条件下的生长状况及油脂和花生四烯酸（AA）的积累规律进行了研究。结果显示,缺刻缘绿藻在3种氮源条件下均能较好地生长。在高氮浓度条件下,增大光强能显著提高缺刻缘绿藻的生物量并促进油脂和AA的积累。缺刻缘绿藻在300 μmol·m^-2·s^-1光强、8.8 mmol·L^-1NaNO₃条件下生物量达到最大（4.17 g·L^-1）。油脂含量在100 μmol·m^-2·s^-1光强、1.0 mmol·L^-1氮浓度下达到最高,分别为41.17%（NaNO₃）、42.04%（NH₄HCO₃）和39.96%（CO（NH₂）₂）。AA绝对含量在300 μmol·m^-2·s^-1光强、2.9 mmol·L^-1 NaNO₃条件下达到最高,占细胞干重的16.44%。油脂和AA产率,在300 μmol·m^-2·s^-1光强、以NaNO₃为氮源的条件下达到最大,分别为134.6 mg·L^-1·d^-1（1.0 mmol·L^-1）和35.85 mg·L^-1·d^-1（2.9 mmol·L^-1）。综合考虑成本等因素,选择NH₄HCO₃（5.9 mmol·L^-1）和CO（NH₂）₂（2.9 mmol·L^-1）为氮源、在300 μmol·m^-2·s^-1高光强下培养缺刻缘绿藻进行AA的生产为最优方案。     

不同生活型被子植物功能性状与基因组大小的关系

基因组大小在被子植物物种之间存在着巨大的变异, 但目前对不同生活型被子植物功能性状与基因组大小的关系缺乏统一的认识。本研究基于被子植物245科2,226属11,215个物种的基因组大小数据, 探讨了不同生活型物种种子重量、最大植株高度和叶片氮、磷含量4个功能性状与基因组大小之间的关系。结果表明, 被子植物最大植株高度和种子重量与基因组大小间的关系在草本和木本植物中存在显著差异。草本植物最大植株高度与基因组大小的关系不显著, 但种子重量与其呈极显著的正相关关系。木本植物最大植株高度与基因组大小显著负相关, 但种子重量与其关系不显著。木本植物叶片氮含量与基因组大小呈显著正相关, 但其他生活型植物的叶片氮、磷含量与基因组大小均无显著相关性。本研究表明被子植物功能性状与基因组大小的相关性在不同生活型间存在差异, 这为深入研究植物多种功能性状和植物生活型与基因组大小的权衡关系在植物演化和生态适应中的作用提供了重要依据。     

Response of bacterial communities to cyanobacterial harmful algal blooms in Lake Taihu,China

Cyanobacterial harmful algal blooms are prevalent around the world, influencing aquatic organisms and altering the physico-chemical properties in freshwater systems. However, the response of bacterial communities to toxic cyanobacterial blooms and associated microcystins (MC) remain poorly understood even though global concentrations of MC have increased dramatically in the past few decades. To address this issue, the dynamics of bacterial community composition (BCC) in the water column and how BCC is influenced by both harmful cyanobacterial blooms and environmental factors were investigated on a monthly basis from August 2013 to July 2014 in Lake Taihu, China. Non-metric multidimensional scaling (NMDS) revealed that seasonal variation in BCC was significant, and that the succession of BCC greatly depends on changes in environmental conditions. Redundancy analysis (RDA) results showed that the overall variation of BCC was explained mainly by dissolved oxygen (DO), nitrate nitrogen (NO₃⁻-N), and Microcystis. The alpha biodiversity of the bacterial community was different among months with the highest diversity in February and the lowest diversity in October. Furthermore, significant negative relationships were found between alpha biodiversity indices and Microcystis abundance as well as with intracellular MC concentrations, indicating that Microcystis and associated MC may influence the bacterial community structure by reducing its biodiversity. This study shows that potential associations exist between toxic cyanobacterial blooms and bacterial communities but more investigations are needed to obtain a mechanistic understanding of their complex relationships.     

Functional response of suspension feeding anuran larvae to different particle sizes at low concentrations (Amphibia)

The influence of particle size, initial particle concentration and larval stage on the ingestion rate, ‘retention efficiency’, and filtering rate of anuran larvae with varying filter apparatus anatomy and different life histories was investigated for four species. Larvae of premetamorphic Stages 28 and 32 and prometamorphic Stage 40 were selected for filtering experiments on the basis of their different growth rates. Three different sizes of silica gel particles were offered as mock food. Particle concentration was measured photometrically. The Michaelis-Menten model was used to describe the dependency of ingestion rate, filtering rate, and ‘retention efficiency’ upon initial particle concentration, and to calculate maximum ingestion rate, threshold concentration, and the half-saturation constant. (1) The highest ingestion rates, filtering rates and ‘retention efficiencies’ were achieved by Xenopus laevis larvae, followed by Bufo calamita larvae. Bufo bufo larvae lay at the opposite end of the scale. Rana temporaria larvae were placed between B. calamita and B. bufo larvae. This order is attributed to differences in life histories, especially the different breeding environments in which these larvae occur. (2) The larger the particle size and the older the stage, the greater the tendency toward saturation of the ingestion rate, filtering rate and ‘retention efficiency’. These filtration parameters are graded according to particle size. The ingestion rate (number of particles), filtration rate and ‘retention efficiency’ are greatest for PS3. Ingestion volume is greatest for PS 1. The difference between PS3 and PS2 on the one hand, and PS1 on the other, is often great; for Stage 28 X. laevis it is very great. This shows that larvae ingest large particles more effectively, and that the most effective ingestion takes place at Stages 28 and 32, owing to the growth function of these stages. The ability of larvae to ingest large particles effectively is possibly a very basic phylogenetic characteristic. (3) The threshold concentration is lowest when the particles are at their largest. In accordance with conclusions drawn by other authors, threshold feeding is attributed to regulation by buccal pumping and mucus production. Considerable importance is attributed to threshold feeding with respect to larval adaptation to oligotrophic environments.     

Photoluminescent properties of Eu3+‐Eu2+ activated MAl2SixO2x + 4 (M = Mg,Ca, Sr,Ba) phosphors prepared in air

In this paper, MAl₂Si_xO_2x+4:Eu²⁺/Eu³⁺ (Eu²⁺ + Eu³⁺ = 2%, molar ratio; M = Mg, Ca, Sr, Ba; x = 0, 0.5, 1, 1.5, 2) phosphors with different SiO₂ concentrations (the ratio of SiO₂ to MAl₂O₄ is n%, n = 0, 50, 100, 150, 200, respectively) were prepared by high‐temperature solid‐state reaction under atmospheric air conditions. Their structures and photoluminescent properties were systematically researched. The results indicate that Eu³⁺ ions have been reduced and Eu²⁺ ions are obtained in air through the self‐reduction mechanism. The alkaline earth metal ions and doping SiO₂ strongly affect the crystalline phase and photoluminescent properties of samples, including microstructures, relative intensity of Eu²⁺ to Eu³⁺, location of emission lines/bands. It is interesting and important that the emission color and intensities of europium‐doped various phosphors which consist of aluminosilicate matrices prepared under atmospheric air conditions could be modulated by changing the kinds of alkaline earth metal and the content of SiO₂.     

Steady state peripheral blood provides cells with functional and metabolic characteristics of real hematopoietic stem cells

Hematopoietic stem cells (HSCs), which are located in the bone marrow, also circulate in cord and peripheral blood. Despite high availability, HSCs from steady state peripheral blood (SSPB) are little known and not used for research or cell therapy. We thus aimed to characterize and select HSCs from SSPB by a direct approach with a view to delineating their main functional and metabolic properties and the mechanisms responsible for their maintenance. We chose to work on Side Population (SP) cells which are highly enriched in HSCs in mouse, human bone marrow, and cord blood. However, no SP cells from SSBP have as yet been characterized. Here we showed that SP cells from SSPB exhibited a higher proliferative capacity and generated more clonogenic progenitors than non‐SP cells in vitro. Furthermore, xenotransplantation studies on immunodeficient mice demonstrated that SP cells are up to 45 times more enriched in cells with engraftment capacity than non‐SP cells. From a cell regulation point of view, we showed that SP activity depended on O₂ concentrations close to those found in HSC niches, an effect which is dependent on both hypoxia‐induced factors HIF‐1α and HIF‐2α. Moreover SP cells displayed a reduced mitochondrial mass and, in particular, a lower mitochondrial activity compared to non‐SP cells, while they exhibited a similar level of glucose incorporation. These results provided evidence that SP cells from SSPB displayed properties of very primitive cells and HSC, thus rendering them an interesting model for research and cell therapy.