A method for the quantification of bacterial protein in the presence of Jarosite

A variation on Peterson's modification of the Lowry method for microbial protein determination was developed in which 10% (w/v) oxalic acid was used to remove jarosite. This allowed the quantification of Thiobacillus ferrooxidans entrapped in solid jarosite or in aqueous suspensions containing jarosite. The quantity of protein measured was not affected by the amount of jarosite in the culture, the concentration of oxalic acid, or the time of exposure (up to 72 h) of the sample to oxalic acid. An application of this method was demonstrated in the quantification of biomass immobilized in jarosite on the surface of polystyrene beads in an inverse fluidized bed bioreactor used for the rapid microbial oxidation of ferrous iron.     

Discovery of aryl ureas and aryl amides as potent and selective histamine H3 receptor antagonists for the treatment of obesity (Part I)

A series of structurally novel aryl ureas was derived from optimization of the HTS lead as selective histamine H₃ receptor (H₃R) antagonists. The SAR was explored and the data obtained set up the starting point and foundation for further optimization. The most potent tool compounds, as exemplified by compounds 2l, 5b, 5d, and 5e, displayed antagonism potencies in the subnanomolar range in in vitro human-H₃R FLIPR assays and rhesus monkey H₃R binding assays.     

液体负平衡对感染性休克合并急性肺损伤患者早期复苏及预后的影响

目的:探讨液体负平衡对感染性休克合并急性肺损伤(ALI)患者早期复苏及预后的影响。方法:将2010年1月~2014年9月我院急诊外科收治的84例感染性休克合并ALI的患者随机分为治疗组和对照组,每组各42例患者。治疗组采用出入量负平衡方式进行液体管理,对照组采用出入量平衡方式进行液体管理。观察和比较两组治疗前后氧合指数(PaO_2/FiO_2)、中心静脉压(CVP)、平均动脉压(MAP)、心指数(CI)、血管外肺水指数(ELWI)及APACHEⅡ评分的变化,记录和比较两组的机械通气时间、ICU住院时间、多器官功能障碍综合征(MODS)的发生率及28 d的病死率。结果:入院后3d、7d,治疗组的MAP较对照组明显降低,ELWI、PaO_2/FiO_2则明显升高(P0.05);治疗前、治疗后6h,两组的APACHEⅡ评分比较均无显著性差异(P0.05),而治疗组治疗后24h、48h的APACHEⅡ评分较对照组则明显降低(P0.05);与对照组比较,治疗组的机械通气时间、ICU住院时间显著缩短,MODS发生率明显降低(P0.05)。结论:在维持循环稳定和保证器官灌注的前提下,液体负平衡有助于减轻感染性休克合并ALI患者的心肺损伤,促进患者早期复苏,改善患者的预后。     

Evaluating ecosystem-level anthropogenic impacts in a stressed transitional environment: The case of the Seine estuary

During 2002–2005, a new container terminal in the commercial harbour of Le Havre, named “Port2000”, was built on the northern flank of the Seine estuary, northern France. This extension is already known to have modified the estuary current and sediment dynamics, as well as reducing biomass of the suprabenthos assemblage, for the whole downstream part of the system. However, studies on other biotic communities were largely inconclusive, and an ecosystem-wide analysis was still lacking. Here, we performed a before/after study of ecosystem dynamics of the different habitats of the Seine estuary, using a Linear Inverse Modelling technique (LIM-MCMC) to estimate all flows occurring in the food web. Ecological Network Analysis indices were calculated, summarising ecosystem functioning traits and giving indications about the habitat health status. Results showed that the southern flank (FS, Fosse Sud) exhibits all characteristics to be considered as the least stressed habitat of the estuary: system activity and functional specialisation of flows were stable between periods, ecosystem recycling processes and detrital dynamics were also stable; an increase in trophic specialisation (decrease in system omnivory) was the only change confirming a general ecological succession. The northern flank (FN, Fosse Nord), where the actual terminal was built, showed a food web with increased importance of lower trophic levels (increased detritivory and carbon recycling), increased stability and flow efficiency, but possibly regressed to a previous step in ecological succession. In the central navigation channel (CH), patterns of network indices were overall inconclusive and the general image is one of a constantly shifting food web, a condition possibly caused by the year-round dredging activities. The functioning of the Seine estuary – especially of FN and FS – seems to have been modified by the combination of harbour construction and the related mitigation measures. Network indices partially captured this combination of changes and, although not fully operational yet, they are promising tools to comply with the European Union mandate of defining ecosystem health status.     

大鼠室旁核注射orexin-A对体重的影响

目的:探讨大鼠室旁核(PVN)注射orexin-A对体重的影响。方法:大鼠室旁核(PVN)微量注射orexin-A,用大脑置管埋管、组织化学染色等方法探讨PVN注射orexin-A对其体重的影响。结果:与安慰剂组大鼠相比,PVN注射orexin-A组大鼠体重明显减轻(P0.05),而orexin-A组和安慰剂组摄食量无明显差异(P0.05)。注射结束后6天,orexin-A处理大鼠的体重仍显著低于注射前(P0.05),而安慰剂组大鼠则比注射前显著增重(P0.05)。药物注射可显著降低机体脂肪,但并不特异存在于注射orexin-A或安慰剂的大鼠身上。Orexin-A组和安慰剂组大鼠的肌肉量和脂肪量均显著降低(P0.05),但注射orexin-A的大鼠降低更明显。与安慰剂组相比,orexin-A处理后的摄食转化率显著降低(P0.05)。结论:大鼠室旁核(PVN)注射orexin-A可通过增加活动量产生负能量平衡,引起体重减轻。     

Microbial metabolite fluxes in a model marine anoxic ecosystem

Permanently anoxic regions in the ocean are widespread and exhibit unique microbial metabolic activity exerting substantial influence on global elemental cycles and climate. Reconstructing microbial metabolic activity rates in these regions has been challenging, due to the technical difficulty of direct rate measurements. In Cariaco Basin, which is the largest permanently anoxic marine basin and an important model system for geobiology, long‐term monitoring has yielded time series for the concentrations of biologically important compounds; however, the underlying metabolite fluxes remain poorly quantified. Here, we present a computational approach for reconstructing vertical fluxes and in situ net production/consumption rates from chemical concentration data, based on a 1‐dimensional time‐dependent diffusive transport model that includes adaptive penalization of overfitting. We use this approach to estimate spatiotemporally resolved fluxes of oxygen, nitrate, hydrogen sulfide, ammonium, methane, and phosphate within the sub‐euphotic Cariaco Basin water column (depths 150–900 m, years 2001–2014) and to identify hotspots of microbial chemolithotrophic activity. Predictions of the fitted models are in excellent agreement with the data and substantially expand our knowledge of the geobiology in Cariaco Basin. In particular, we find that the diffusivity, and consequently fluxes of major reductants such as hydrogen sulfide, and methane, is about two orders of magnitude greater than previously estimated, thus resolving a long‐standing apparent conundrum between electron donor fluxes and measured dark carbon assimilation rates.     

Methods to produce marker-free transgenic plants

Selectable marker genes (SMGs) have been extraordinarily useful in enabling plant transformation because of the low efficiency of transgene integration. The most used SMGs encode proteins resistant to antibiotics or herbicides and use negative selection, i.e., by killing nontransgenic tissue. However, there are perceived risks in wide-scale deployment of SMG-transgenic plants, and therefore research has recently been performed to develop marker-free systems. In this review, transformation using markers not based on antibiotic or herbicide resistance genes, as well as different systems of marker gene deletion, are discussed.     

BAmSA: Visualising transmembrane regions in protein complexes using biotinylated amphipols and electron microscopy

Membrane protein (MP) complexes play key roles in all living cells. Their structural characterisation is hampered by difficulties in purifying and crystallising them. Recent progress in electron microscopy (EM) have revolutionised the field, not only by providing higher-resolution structures for previously characterised MPs but also by yielding first glimpses into the structure of larger and more challenging complexes, such as bacterial secretion systems. However, the resolution of pioneering EM structures may be difficult and their interpretation requires clues regarding the overall organisation of the complexes. In this context, we present BAmSA, a new method for localising transmembrane (TM) regions in MP complexes, using a general procedure that allows tagging them without resorting to neither genetic nor chemical modification. Labels bound to TM regions can be visualised directly on raw negative-stain EM images, on class averages, or on three-dimensional reconstructions, providing a novel strategy to explore the organisation of MP complexes.     

Unraveling the genetic basis of fast l-arabinose consumption on top of recombinant xylose-fermenting Saccharomyces cerevisiae

One major challenge in the bioconversion of lignocelluloses into ethanol is to develop Saccharomyces cerevisiae strains that can utilize all available sugars in biomass hydrolysates, especially the d -xylose and l -arabinose that cannot be fermented by the S. cerevisiae strain naturally. Here, we integrated an l -arabinose utilization cassette (AUC) into the genome of an efficient d -xylose fermenting industrial diploid S. cerevisiae strain CIBTS0735 to make strain CIBTS1972. After evolving on arabinose, CIBTS1974 with excellent fermentation capacity was obtained. A comparison between genome sequences of strains CIBTS1974 and CIBTS1972 revealed that the copy number of the AUC had increased from 1 to 12. We then constructed the AUC null-mutant CIBTS1975 and gradually rescued the l -arabinose utilization defect by integrating AUC iteratively. On the other hand, the parental strain CIBTS0735 was able to acquire the same performance as CIBTS1974 by the direct introduction of 12 copies of the AUC; the performance was further improved by adding two more copies. Besides, we found that not the two transporters present in the AUC were both needed during l -arabinose utilization, GAL2 was necessary and STP2 was not essential. We have described for the first time that a high copy number of AUC is sufficient for the strain to metabolize l -arabinose efficiently independent of evolution.     

Memory in coal tits: an alternative model

Jolliffe and Jolliffe (1997, Biometrics 53, 1136-1142) proposed various models for data from an experiment on memory in coal tits. This article describes an alternative model, which fits equally well and which may be simpler to interpret.