Plasmid Metagenome Reveals High Levels of Antibiotic Resistance Genes and Mobile Genetic Elements in Activated Sludge

The overuse or misuse of antibiotics has accelerated antibiotic resistance, creating a major challenge for the public health in the world. Sewage treatment plants (STPs) are considered as important reservoirs for antibiotic resistance genes (ARGs) and activated sludge characterized with high microbial density and diversity facilitates ARG horizontal gene transfer (HGT) via mobile genetic elements (MGEs). However, little is known regarding the pool of ARGs and MGEs in sludge microbiome. In this study, the transposon aided capture (TRACA) system was employed to isolate novel plasmids from activated sludge of one STP in Hong Kong, China. We also used Illumina Hiseq 2000 high-throughput sequencing and metagenomics analysis to investigate the plasmid metagenome. Two novel plasmids were acquired from the sludge microbiome by using TRACA system and one novel plasmid was identified through metagenomics analysis. Our results revealed high levels of various ARGs as well as MGEs for HGT, including integrons, transposons and plasmids. The application of the TRACA system to isolate novel plasmids from the environmental metagenome, coupled with subsequent high-throughput sequencing and metagenomic analysis, highlighted the prevalence of ARGs and MGEs in microbial community of STPs

Recent developments of acupuncture in Australia and the way forward

Almost one in ten Australians has received acupuncture treatment by acupuncturists and/or medical doctors in private clinics. The majority of Australian health insurance funds offer rebates for acupuncture. Statutory regulations for acupuncture have been implemented in the State of Victoria, Australia. Six acupuncture degree courses have been approved by the Chinese Medicine Registration Board of Victoria and/or accredited by the Australian Acupuncture and Chinese Medicine Association. Furthermore, a number of clinical trials of acupuncture on allergic rhinitis, pain and women's health were carried out in Australia. Recent developments of acupuncture in Australia indicate that through adequate and appropriate evaluation, acupuncture begins to integrate into mainstream health care in Australia

Placebo devices as effective control methods in acupuncture clinical trials : a systematic review

While the use of acupuncture has been recognised by the World Health Organisation, its efficacy for many of the common clinical conditions is still undergoing validation through randomised controlled trials (RCTs). A credible placebo control for such RCTs to enable meaningful evaluation of its efficacy is to be established. While several non-penetrating acupuncture placebo devices, namely the Streitberger, the Park and the Takakura Devices, have been developed and used in RCTs, their suitability as inert placebo controls needs to be rigorously determined. This article systematically reviews these devices as placebo interventions. Electronic searches were conducted on four English and two Chinese databases from their inceptions to July 2014; hand searches of relevant references were also conducted. RCTs, in English or Chinese language, comparing acupuncture with one of the aforementioned devices as the control intervention on human participants with any clinical condition and evaluating clinically related outcomes were included. Thirty-six studies were included for qualitative analysis while 14 were in the meta-analysis. The meta-analysis does not support the notion of either the Streitberger or the Park Device being inert control interventions while none of the studies involving the Takakura Device was included in the meta-analysis. Sixteen studies reported the occurrence of adverse events, with no significant difference between verum and placebo acupuncture. Author-reported blinding credibility showed that participant blinding was successful in most cases; however, when blinding index was calculated, only one study, which utilised the Park Device, seemed to have an ideal blinding scenario. Although the blinding index could not be calculated for the Takakura Device, it was the only device reported to enable practitioner blinding. There are limitations with each of the placebo devices and more rigorous studies are needed to further evaluate their effects and blinding credibility

Sanofi in-house medium: Exceeding expectations in cell line development as an alternative to commercially available basal medium

Our current CHO cell line development process has used commercially supplied media for all steps from post-transfection recovery through scale-up and screening of clones. Media alternatives have been developed in house to improve cell culture performance, decrease media cost, and enable modification by Sanofi. A current version of the basal medium has been utilized for cell culture production processes for some early stage development programs. With the goal of utilizing the same media for cell line development and cell culture production, we evaluated this basal medium in different steps of the cell line generation process, with the current (commercial) medium used as the control. Overall, the results demonstrated that the in-house medium met or exceeded the performance of the commercial medium in the cell line development workflow. To assess the in-house basal medium for post-transfection methotrexate (MTX) selection, cell growth and viability were measured during MTX selection, and recombinant protein titers of the MTX-selected pools were assessed. Results show that protein production is comparable between pools selected in the in-house medium versus the control. In addition, use of the in-house medium improved cell growth during selection and decreased the selection timeline by a week. A subsequent MTX kill curve study using the in-house medium showed excellent recovery of transfected cells, even at the higher MTX concentrations used, and high protein titers with less time in selection. Next, the medium was evaluated for single cell cloning in a process using flow cytometry cell sorting to deposit one cell per well in 96-well plates. The initial experiments revealed a minor limitation of the in-house medium for single cell cloning and, based on these results, a cloning version of the basal medium was prepared and tested. Several experiments demonstrated that the cloning version of the in-house medium supported robust cell growth and cloning efficiency that is similar to or better than that obtained with the control. Further studies showed that, for both pools and clones in batch cultures, the in-house medium supports more rapid growth rates and higher cell densities. Experimental results to be presented demonstrate success of the in-house medium in the MTX selection, single cell cloning and outgrowth, and pool and clone screening steps of the cell line development workflow. Furthermore, the advantages of using the in-house medium for pre-master cell banking will be discussed. Proof of concept work from recent cell line generation projects will further demonstrate the utility and advantage of using our in-house medium in the cell line development workflow for new preclinical development programs

Atmospheric nitrogen deposition in the Yangtze River basin: spatial pattern and source attribution

The Yangtze River basin is one of the world's hotspots for nitrogen (N) deposition and likely plays an important role in China's riverine N output. Here we constructed a basin-scale total dissolved inorganic N (DIN) deposition (bulk plus dry) pattern based on published data at 100 observational sites between 2000 and 2014, and assessed the relative contributions of different reactive N (Nr) emission sectors to total DIN deposition using the GEOS-Chem model. Our results show a significant spatial variation in total DIN deposition across the Yangtze River basin (33.2 kg N ha−1 yr−1 on average), with the highest fluxes occurring mainly in the central basin (e.g., Sichuan, Hubei and Hunan provinces, and Chongqing municipality). This indicates that controlling N deposition should build on mitigation strategies according to local conditions, namely, implementation of stricter control of Nr emissions in N deposition hotspots but moderate control in the areas with low N deposition levels. Total DIN deposition in approximately 82% of the basin area exceeded the critical load of N deposition for semi-natural ecosystems along the basin. On the basin scale, the dominant source of DIN deposition is fertilizer use (40%) relative to livestock (11%), industry (13%), power plant (9%), transportation (9%), and others (18%, which is the sum of contributions from human waste, residential activities, soil, lighting and biomass burning), suggesting that reducing NH3 emissions from improper fertilizer (including chemical and organic fertilizer) application should be a priority in curbing N deposition. This, together with distinct spatial variations in emission sector contributions to total DIN deposition also suggest that, in addition to fertilizer, major emission sectors in different regions of the basin should be considered when developing synergistic control measures

Dipole-field-assisted charge extraction in metal-perovskite-metal back-contact solar cells

Hybrid organic-inorganic halide perovskites are low-cost solution-processable solar cell materials with photovoltaic properties that rival those of crystalline silicon. The perovskite films are typically sandwiched between thin layers of hole and electron transport materials, which efficiently extract photogenerated charges. This affords high-energy conversion efficiencies but results in significant performance and fabrication challenges. Herein we present a simple charge transport layer-free perovskite solar cell (PSC), comprising only a perovskite layer with two interdigitated gold back-contacts. Charge extraction is achieved via self-assembled molecular monolayers (SAMs) and their associated dipole fields at the metal/perovskite interface. Photovoltages of approximately 600 mV generated by SAM-modified PSCs are equivalent to the built-in potential generated by individual dipole layers. Efficient charge extraction results in photocurrents of up to 12.1 mA/cm2 under simulated sunlight, despite a large electrode spacing.Comment: 18 pages, 5 figure

Structural Basis for Calmodulin as a Dynamic Calcium Sensor

Calmodulin is a prototypical and versatile Ca2+ sensor with EF-hands as its high-affinity Ca2+ binding domains. Calmodulin is present in all eukaryotic cells, mediating Ca2+-dependent signaling. Upon binding Ca2+, calmodulin changes its conformation to form complexes with a diverse array of target proteins. Despite a wealth of knowledge on calmodulin, little is known on how target proteins regulate calmodulin’s ability to bind Ca2+. Here, we take advantage of two splice variants of SK2 channels, which are activated by Ca2+-bound calmodulin, but show different sensitivity to Ca2+ for their activation. Protein crystal structures and other experiments show that depending on which SK2 splice variant it binds to calmodulin adopts drastically different conformations with different affinities for Ca2+ at its C-lobe. Such target protein induced conformational changes make calmodulin a dynamic Ca2+ sensor, capable of responding to different Ca2+ concentrations in cellular Ca2+ signaling

Structural Basis for Calmodulin as a Dynamic Calcium Sensor

Calmodulin is a prototypical and versatile Ca2+ sensor with EF-hands as its high-affinity Ca2+ binding domains. Calmodulin is present in all eukaryotic cells, mediating Ca2+-dependent signaling. Upon binding Ca2+, calmodulin changes its conformation to form complexes with a diverse array of target proteins. Despite a wealth of knowledge on calmodulin, little is known on how target proteins regulate calmodulin’s ability to bind Ca2+. Here, we take advantage of two splice variants of SK2 channels, which are activated by Ca2+-bound calmodulin, but show different sensitivity to Ca2+ for their activation. Protein crystal structures and other experiments show that depending on which SK2 splice variant it binds to calmodulin adopts drastically different conformations with different affinities for Ca2+ at its C-lobe. Such target protein induced conformational changes make calmodulin a dynamic Ca2+ sensor, capable of responding to different Ca2+ concentrations in cellular Ca2+ signaling