Antibiotic resistance, driven by antibiotic consumption, is a growing global health threat. Our report on antibiotic use in 76 countries over 16 years provides an up-to-date comprehensive assessment of global trends in antibiotic consumption. We find that the antibiotic consumption rate in low- and middle-income countries (LMICs) has been converging to (and in some countries surpassing) levels typically observed in high-income countries. However, inequities in drug access persist, as many LMICs continue to be burdened with high rates of infectious disease-related mortality and low rates of antibiotic consumption. Our findings emphasize the need for global surveillance of antibiotic consumption to support policies to reduce antibiotic consumption and resistance while providing access to these lifesaving drugs. P = 0.004). In high-income countries (HICs), although overall consumption increased modestly, DDDs per 1,000 inhabitants per day fell 4%, and there was no correlation with GDPPC. Of particular concern was the rapid increase in the use of last-resort compounds, both in HICs and LMICs, such as glycylcyclines, oxazolidinones, carbapenems, and polymyxins. Projections of global antibiotic consumption in 2030, assuming no policy changes, were up to 200% higher than the 42 billion DDDs estimated in 2015. Although antibiotic consumption rates in most LMICs remain lower than in HICs despite higher bacterial disease burden, consumption in LMICs is rapidly converging to rates similar to HICs. Reducing global consumption is critical for reducing the threat of antibiotic resistance, but reduction efforts must balance access limitations in LMICs and take account of local and global resistance patterns.Tracking antibiotic consumption patterns over time and across countries could inform policies to optimize antibiotic prescribing and minimize antibiotic resistance, such as setting and enforcing per capita consumption targets or aiding investments in alternatives to antibiotics. In this study, we analyzed the trends and drivers of antibiotic consumption from 2000 to 2015 in 76 countries and projected total global antibiotic consumption through 2030. Between 2000 and 2015, antibiotic consumption, expressed in defined daily doses (DDD), increased 65% (21.1–34.8 billion DDDs), and the antibiotic consumption rate increased 39% . The increase was driven by low- and middle-income countries (LMICs), where rising consumption was correlated with gross domestic product per capita (GDPPC) growth ( Antibiotic resistance—the ability of microbes to evolve and withstand the effects of antibiotics—is a significant cause of morbidity and mortality globally –3. Antibi) monitor national and global trends over time; (ii) compare antibiotic use among countries; (iii) provide a baseline for the evaluation of future efforts to reduce antibiotic use; (iv) enable epidemiological analysis of the association between antibiotic use and resistance over time (v) support policies that aim to reduce antibiotic resistance. Given the urgency of the threat posed by rising AMR levels  and over a longer time period (2000–2015). In addition, we assess differences in consumption between high-income countries (HICs) and low- and middle-income countries (LMICs), identify drivers of antibiotic use in each income group from a set of socioeconomic and medical indicators, and project future growth in global antibiotic consumption.There have been few attempts to assess antibiotic consumption globally or in multiple countries –17, noneSI Appendix, Table S1). In countries where both hospital and retail data were reported for some but not all years (2000–2015), consumption in the missing sector was estimated by interpolation, using the ratio of antibiotic consumption in the hospital and retail sectors for the years for which data had been reported.We estimated global antibiotic consumption using the IQVIA MIDAS database. IQVIA uses national sample surveys of antibiotic sales to develop estimates of the total volume of sales of each antibiotic molecule (or combination of molecules). For each country, antibiotic consumption was reported by month or quarter and broken down between the retail and hospital sectors. We obtained data for 76 countries from 2000 through 2015. Central America  and French West Africa  were included as two individual groups of countries with aggregated sales for these regions. Sixty-six countries had data available for every year between 2000 and 2015, while data on the remaining countries covered partial time periods . Data for SUs were available for all years, whereas kilogram data were available only for the period 2005–2015. The ratio of SUs to kilograms for 2005–2015 was used to estimate kilograms and DDDs for 2000–2004. A country’s annual antibiotic consumption rate in DDDs per 1,000 inhabitants per day was calculated using population estimates from the World Bank DataBank. Consumption rates were subsequently compared between groups of countries based on their World Bank income classification in 2007.Data on antibiotic sales in standard units (SUs) and kilograms were purchased under license from IQVIA. An SU is an IQVIA designation that represents a single-dose unit such as a pill, capsule, or equal amount of liquid. Sales expressed in kilograms were converted into DDDs using the Anatomical Therapeutic Chemical Classification System  developed by the WHO Collaborating Centre for Drug Statistics Methodology. For molecules not included in the ATC/DDD index, DDD values were estimated from other sources or as the average of DDD unit values by class ; imports of goods and services as a percentage of GDP (as a measure of trade); measles vaccination coverage in children between the ages of 12 and 23 mo [lack of pneumococcal conjugate vaccination (PCV) coverage information limited our ability to include PCV coverage as a variable]; physician density per 1,000 population; and percentage of the population living in urban areas . All explanatory variables were pooled by country/year. Regression models analyzed data for HICs and LMICs separately. Health and economic factors were obtained from the World Development Indicators in the World Bank DataBank . Serial i) no policy changes, where countries’ antibiotic consumption rates for 2016 through 2030 were assumed to continue to change based on their compounded growth rate between 2010 and 2015 and (ii) a target policy in which countries were assumed to converge to the 2015 global median antibiotic consumption rate by 2020 . Population projections were retrieved from the World Bank DataBank  DDDs per 1,000 inhabitants per day to 20.9 (SD 9.8), and the median antibiotic consumption rate increased 25% from 15.5 to 19.5 DDDs per 1,000 inhabitants per day.SI Appendix, Fig. S1). In HICs, although the total amount of antibiotics consumed increased 6% between 2000 and 2015, from 9.7 to 10.3 billion DDDs, the antibiotic consumption rate decreased by a modest 4%, from 26.8 to 25.7 DDDs per 1,000 inhabitants per day  in India, from 2.3 to 4.2 billion DDDs (79%) in China, and from 0.8 to 1.3 billion DDDs (65%) in Pakistan. The antibiotic consumption rate increased from 8.2 to 13.6 DDDs per 1,000 inhabitants per day (63%) in India, from 5.1 to 8.4 DDDs per 1,000 inhabitants per day (65%) in China, and from 16.2 to 19.6 DDDs per 1,000 inhabitants per day (21%) in Pakistan.The antibiotic consumption rate of broad-spectrum penicillins, the most commonly consumed class of antibiotics , increased 36% between 2000 and 2015 globally. The greatest increase was in LMICs (56%), although the antibiotic consumption rate in HICs increased 15% A. While Consumption of newer and last-resort antibiotic classes increased across all country income groups between 2000 and 2015. The United States was the largest consumer of glycylcyclines (tigecycline) and oxazolidinones (primarily linezolid as tedizolid was not introduced until 2014) through the late 2000s. However, the antibiotic consumption rate of these drugs in the United States began declining in 2009 A and B, P = 0.004), although no statistically significant association was found between these factors in HICs (P = 0.52). Other indicators, including the measles vaccination rate , imports as a percentage of GDP, and physician density, were not correlated with changes in per capita antibiotic use across countries, irrespective of income group . Total global antibiotic consumption in 2015 was estimated to be 42.3 billion DDDs —10.7 billion DDDs in HICs and 31.6 billion DDDs in LMICs. In our baseline condition, where we assumed no policy changes and constant antibiotic consumption rates set at current levels of use, global antibiotic use is projected to increase 15% between 2015 and 2030. If all countries continue to increase their antibiotic consumption rates at their compounded annual growth rates, we estimate that total consumption would increase 202% to 128 billion DDDs, while the antibiotic consumption rate would increase 161% to 41.1 DDDs per 1,000 inhabitants per day. Finally, if all countries converge on the global median antibiotic consumption rate in 2015 of 17.9 DDDs per 1,000 inhabitants per day by 2020, we estimate global antibiotic consumption would increase 32% to 55.6 billion DDDs  decreased slightly in HICs from 27.0 to 25.7 DDDs per 1,000 inhabitants per day in HICs and increased from 8.6 to 13.9 DDDs per 1,000 inhabitants per day in LMICs , as well as others that have reported global antibiotic consumption  (sumption , 16. Whisumption , 45. Indsumption  and indusumption . As partsumption . HoweverWith antibiotic consumption increasing worldwide, the challenge posed by antibiotic resistance is likely to get worse. As with climate change, there may be an unknown tipping point , and thiSupplementary File


E. coli O157:H7, S. Typhimurium, and L. monocytogenes) while maintaining higher system performance efficiency.Control of foodborne pathogens is an important issue for the fruit juice industry and ohmic heating treatment has been considered as one of the promising antimicrobial interventions. However, to date, evaluation of the relationship between inactivation of foodborne pathogens and system performance efficiency based on differing soluble solids content of apple juice during ohmic heating treatment has not been well studied. This study aims to investigate effective voltage gradients of an ohmic heating system and corresponding sugar concentrations (°Brix) of apple juice for inactivating major foodborne pathogens  under two treatment conditions (30 V/cm in 36 °Brix or 60 V/cm in 48 °Brix juice) were relatively greater than for other combinations. Meanwhile, 5-log reductions of the three foodborne pathogens were achieved after treatment for 60 s in 36 °Brix at 30 V/cm, but this same reduction was observed in 48 °Brix juice at 60 V/cm within 20 s without affecting product quality.With respect to both bactericidal efficiency and SPC values, 60 V/cm in 48 °Brix was the most effective ohmic heating treatment combination for decontaminating apple juice concentrates. Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium .For enumeration of bacteria, each treated 25 ml sample was immediately transferred into a sterile stomacher bag  containing 225 ml of iced 0.2% PW (maintained on crushed ice) and homogenized for 2 min with a stomacher . One ml aliquots of homogenized samples were tenfold serially diluted in 9 ml of 0.2% PW, and 0.1 ml of sample or diluent was spread-plated onto each selective medium. For the enumeration of The system performance coefficient (SPC) of ohmic heating was determined from temperature, voltage, and current data  and calcm is mass (g), Cp is specific heat capacity (J/g K), ∆T is difference between final temperature and initial temperature (K), ∆V is voltage applied (V), I is electric current (A), and t is time (s). ∑ΔVIt is the energy given to the system, mCpΔT is energy given to the system minus energy loss during ohmic heating. The ratio of mCpΔT to ∑ΔVIt indicates the system performance coefficient [Where fficient .To assess color changes of treated apple juice, a Minolta colorimeter  was used in this study. Color of apple juice were expressed by values of L*, a*, and b*  . A pH meP value of <0.05 was used to indicate significant differences.All experiments were conducted three times with duplicate samples. Data were analyzed by the ANOVA procedure of SAS , and mean values were separated using Tukey-Kramer’s multiple range test. A There are various factors affecting electrical conductivity of liquids. Electrical conductivity relies on chemical components, ion activity, and viscosity of liquids. Such an electrical characteristic, along with juice concentration, could have an influence on temperature rise and microbial inactivation , 21. A sThe system performance coefficient (SPC), which affects processing cost, was considered as an important factor in this study. Icier and Ilicali  reportedE. coli O157:H7, S. Typhimurium, and L. monocytogenes resulting from ohmic heating was significantly higher (P < 0.05) than that resulting from conventional heating at equal temperatures of 55, 58, and 60 °C in apple juice. These results showed that electric field-induced ohmic heating led to additional bacterial inactivation due not only to thermal effect but also to electroporation-caused cell damage [Control of foodborne pathogens is an important issue for the fruit juice industry and ohmic heating treatment has been considered as one of the promising antimicrobial interventions. In our previous study , reductil damage . As the E. coli O157:H7, S. Typhimurium, and L. monocytogenes in different apple juice concentrations during ohmic heating, respectively. At 30 V/cm, ohmic heating for 60 s achieved 0.95, 2.59, 6.78, 5.21, and 2.71 log reductions of E. coli O157:H7 in 72, 48, 36, 24, and 18 °Brix apple juice, respectively. Also, reductions of 1.40, 2.88, 6.71, 6.70, and 3.27 log CFU/ml in concentrations of 72, 48, 36, 24, and 18 °Brix, respectively, were observed in S. Typhimurium. In the case of L. monocytogenes, levels of log reduction following ohmic heating were 0.47, 1.74, 5.01, 3.91, and 1.13, respectively, in juice concentrations of 72, 48, 36, 24, and 18 °Brix. From these results at 30 V/cm, maximum log reductions of the three foodborne pathogens were observed in 36 °Brix apple juice. Dramatic levels of inactivation were achieved in 18–48 °Brix apple juice during ohmic heating at 60 V/cm. Reductions of E. coli O157:H7 were 6.32, 6.58, 6.88, and 6.93 log CFU/ml in 48, 36, 24, and 18 °Brix juice, respectively, after ohmic heating for 20 s. Similarly, ohmic heating for 20 s accomplished 5.80, 6.10, 6.60, and 6.68 log reductions of S. Typhimurium in 48, 36, 24, and 18 °Brix juice, respectively. Log reductions of 5.71, 5.70, 5.82, and 5.93 in 48, 36, 24, and 18 °Brix apple juice, respectively, were observed for L. monocytogenes. Thus, the time duration required for 5-log reduction at 30 V/cm in 36 °Brix apple juice was three times longer than for 60 V/cm at all apple juice concentrations with the exception of 72 °Brix. Also, commercial processing of higher concentration apple juice has the advantage of greater production yield (of 18 °Brix juice). Therefore, with respect to bactericidal efficiency, SPC values, and treatment time, ohmic heating application of 60 V/cm in 48 °Brix apple juice could be more efficient than that of 30 V/cm in 36°Brix.Tables P > 0.05) different. The pH values of treated samples did not significantly differ from those of non-treated samples. Thus, the proposed parameters for optimal ohmic heating did not significantly affect the quality of apple juice product (Table Additionally, ohmic heating is a suitable technology for minimizing degradation of juice quality due to the fundamental property of ohmic heating, which generates internal heat in food materials . Color act Table .Table 4CAlthough ohmic heating is no longer regarded as a new technology, target microbe reductions have to be assessed in new application environments which include product type and production setting. In this study, optimized voltage gradient and juice concentration for ohmic heating gave a distinct advantage in terms of both bactericidal and economic aspects but also ensured minimal quality loss. However, since ohmic heating was performed in a small-scale batch system, energy and performance criteria have limited significance relative to larger-scale processing units. Therefore, further research incorporating more sophisticated experimental conditions to industrial-scale continuous systems is needed.Novel thermal processing interventions employed by the fruit juice industry for controlling foodborne pathogens involve the utilization of sophisticated systems, which enable reduced processing times and temperatures to prevent loss of nutritional and sensory quality while still securing outstanding bactericidal efficacy. Ohmic heating is one of the most promising thermal technologies for effectively inactivating foodborne pathogens in this respect. In the present study, the optimum processing parameters of ohmic heating treatment such as applied voltage gradients and °Brix of apple juice concentrates were investigated to provide benefits with regard to bactericidal, sensory, and economic aspects. These results can be utilized by the apple juice industry for effective application of ohmic heating.


Additionally, Portland cement made from the tested fly ash concrete was tested to confirm recyclability, with the cement quality meeting the Japanese classification of ordinary Portland cement. Limestone-based recyclable fly ash concrete pavement is, thus, a preferred material in terms of sustainability.Recyclable concrete pavement was made from fly ash and crushed limestone sand and gravel as aggregates so that the concrete pavement could be recycled to raw materials for cement production. With the aim to use as much fly ash as possible for the sustainable development of society, while achieving adequate strength development, pavement concrete having a cement-replacement ratio of 40% by mass was experimentally investigated, focusing on the strength development at an early age. Limestone powder was added to improve the early strength; flexural strength at two days reached 3.5 MPa, the minimum strength for traffic service in Japan. The matured fly ash concrete made with a cement content of 200 kg/m Asphalt concrete pavement accounts for approximately 95% of the total length of road in Japan. Cement concrete pavement accounts for less than 5% of the total length because of the concerns of construction costs and traffic noise, yet it has adequate durability and superior life-cycle cost. Most concrete pavement requires a relatively long curing term before traffic service, which negatively affects its use as a pavement material.Fly ash collected from coal-burning power plants has remarkable properties that can improve the performance of cement concrete when added, and the powder material is, thus, often used as a supplemental binder of Portland cement in concrete (fly ash concrete). The greatest concern about fly ash concrete is the gradual strength development at an early age. Thus, when fly ash concrete is employed as a pavement material, the traffic service must be further delayed. et al. [et al. [et al. [Limestone powder can be added to improve strength development via the micro-filler effect and help accelerate the reaction at an early age. Bentz et al.  reported [et al.  used lim [et al. ,4 examinFundamental properties and applications of HVFA concrete were summarized in high-performance and high-volume fly ash concrete . Additioet al. [et al. [et al. [w/cm = 0.40, 0.34, 0.30) and 20%–60% cement replacement by fly ash. The test results showed that the concrete containing 40% fly ash developed the maximum strength. Rashad et al. [et al. [Naik et al.  reported [et al.  conducte [et al.  investig [et al.  examined [et al.  performed et al.  prepared [et al.  reportedFA/cm = 0.4) by referring to previous investigations [According to the definition of HVFA concrete , fly ashigations ,11. To iigations ,4. Concrigations ,15. In p3) and crushed limestone (2.68 g/cm3) so as to ensure the recyclability for cement production. To eliminate the influence of temperature on the fresh properties of concrete, all materials had been stored in a temperature-controlling room (20 °C) for 24 h or more. Powder materials in the recyclable concrete were ordinary Portland cement, limestone powder and fly ash. w/cm = 0.33). The HRWRA (1.06 g/cm3) used in the study was a commercial air entraining (AE) water-reducing agent of polycarboxylic acid.3) was designed through mixing-tests of fresh concrete. The M-1 mixture was fly ash concrete incorporating limestone powder, while M-2 was fly ash concrete made with more fly ash (40 kg/m3) instead of limestone powder. The additional fly ash is regarded for convenience as a mineral admixture like limestone powder. The accurate cementitious material ratios are given in the parenthesis in Concrete pavement having three mixture proportions were prepared for the experimental study . All conThirty concrete beam specimens (100 mm × 100 mm × 400 mm) for each mixture proportion were prepared in a laboratory. All specimens were cured in a temperature-controlled room (20 °C) for 1 day and then stored in a water tank at 20 °C after demolding. To examine the strength development of the pavement concretes, four-point loading tests were conducted at ages of 1, 2, 3, 5, 7, 28, 56, 91, 182 and 364 days. The flexural strength at each test age was taken as the average strength of three beam specimens at the same test age.i.e., the micro-filler effect) and the acceleration of the reaction at an early age. Concrete pavement is passable for traffic when the flexural strength is equal to or higher than 3.5 MPa [ 3.5 MPa . The flyFA/cm = 0.4, w/cm = 0.34). All concretes tested in the present study had higher strength than the referenced strength data [gth data . The difgth data .3 of Portland cement. The fly ash concrete tested in the present study demonstrated adequate strength development for application as pavement concrete.To confirm the recyclability of the pavement concrete, a Portland cement (recycled cement) was made using test specimens of the fly ash concrete. The age of the concrete specimens was half a year or more. The tested concrete was a raw material for manufacturing cement as an alternative material to limestone. The study examined the chemical and physical properties of the recycled cement.The hardened concrete was dried in an oven (105 °C) for 24 h and crushed using a ball mill. Chemical compositions of the concrete were examined employing a Japanese standard test  and X-raA cement clinker, having components similar to those of a reference clinker  given inThe cement clinker was made with the above materials in an electric furnace. The temperature of the furnace increased from 1000 to 1200 °C over 20 min and from 1200 to 1450 °C over 10 min. The temperature was maintained at 1450 °C for 30 min and then reduced to 1350 °C. Afterward, the clinker was removed from the furnace and quenched to a room temperature of 20 °C. The chemical properties of the cement clinker made from the fly-ash concrete were then examined. The chemical compositions were examined employing the test method given in JIS R5202 .The cement clinker and gypsum were mixed and ground in a ball mill to produce the recycled cement. Additionally, the chemical compositions of the recycled cement were examined, employing a Japanese standard test  and X-ra3) of ordinary Portland cement in Japan. The strengths of the cement were higher than the minimum strength given in JIS R5210 [In addition, the study examined fundamental physical properties given in JIS R5201 . Table 9IS R5210 . It was IS R5210 .w/cm = 0.33) to improve strength development at an early age. The study examined the development of flexural strength up to age of 364 days, and examined physical and chemical properties of the recycled cement to confirm recyclability. The conclusions drawn from the results of the study are as follows.The present study investigated recyclable pavement concrete made with a cement-replacement ratio of 40% by mass. The early-age strength is the most concern relating to the practical use of fly ash concrete, and the study thus aimed to develop a concrete pavement having adequate early strength. Limestone aggregate was used to ensure recyclability for cement production, and limestone powder was added to the concrete of very low water-cementitious material ratio (3.The fly ash concrete with limestone powder achieved the minimum strength (3.5 MPa) for passable pavement at an age of two days, and the specified strength (4.5 MPa) of most concrete pavement at an age of seven days. Furthermore, the matured fly ash concrete achieved a strength almost equal to that of the control concrete without fly ash despite the concrete being made with a unit cement weight of 200 kg/mThe fly ash concrete was used in cement manufacturing as a primary calcium material. Fundamental examinations revealed that the chemical and physical properties of the recycled cement meet the quality of ordinary Portland cement defined in Japanese industrial standards. It is concluded that the recyclable fly ash concrete pavement is a preferred material in terms of sustainability.Although the recycled cement made in the test furnace has shown promising test results, further investigations are necessary to confirm the practical use of the cement, such as production process and cost performance.


We extracted a dataset from Kudos of 830,565 unique publications claimed by authors, for which 20,775 had actions taken to explain or share via Kudos, and for 4,867 of these full text download data from publishers was available. Findings show that researchers are most likely to share their work on Facebook, but links shared on Twitter are more likely to be clicked on. A Mann-Whitney U test revealed that a treatment group (publications having actions in Kudos) had a significantly higher median average of 149 full text downloads (23.1% more) per publication as compared to a control group (having no actions in Kudos) with a median average of 121 full text downloads per publication. These findings suggest that performing actions on publications, such as sharing, explaining, or enriching, could help to increase the number of full text downloads of a publication.With the growth of scholarly collaboration networks and social communication platforms, members of the scholarly community are experimenting with their approach to disseminating research outputs, in an effort to increase their audience and outreach. However, from a researcher’s point of view, it is difficult to determine whether efforts to make work more visible are worthwhile (in terms of the association with publication metrics) and within that, difficult to assess which platform or network is most effective for sharing work and connecting to a wider audience. We undertook a case study of Kudos ( Unlike traditional print and broadcast media, new web-based platforms have made it possible for audiences to step out of their passive roles and connect with the communicators themselves . Hence, l impact .altmetrics is still very controversial, as it could also be seen as just a new form of informetrics [Altmetrics can be described as new or alternative metrics based on activities on social media for measuring scholarly impact , 4. Howermetrics , and thermetrics . Many atrmetrics . Downloarmetrics .Traditional metrics such as citation counts or the h-index are slow indications of impact, for instance, citations could take years to appear. They also represent a narrow academic audience . In conthttps://www.altmetric.com), Plum Analytics , and Impactstory (https://impactstory.org) compile a range of altmetrics from different data sources and provide them to researchers to view and track [https://www.scopus.com) introduced “Altmetric for Scopus”, a third party web application that integrates into the sidebar of pages in Scopus, where researchers can track the mentions of different social media metrics and reference managers. Furthermore, editors and authors find it challenging to engage online with different audiences, and many readers find it difficult to locate and give feedback to publications, as these have been shared on different social media sites [Since traditional metrics have been criticized for not comprehensively representing scholarly impact, not only researchers but publishers and service providers are starting to consider alternative tools to track the online presence of authors, articles and journals. Altmetric aggregators such as Altmetric.com  as a case study in this paper. In the next section, we deliberate on the significance of new media in academia and discuss several features and services of social media platforms and altmetric services that provide support for researchers in their efforts to promote their research outputs and increase their outreach. We then look at related studies that investigate these systems. We present our reasons for choosing Kudos as a case study for this paper, we introduce the system and features provided to researchers. We then present the research questions we aim to address and the methodology used for the analysis. After this, the results of the analysis are presented. We then discuss our findings, limitations, and future work.The above arguments and challenges motivate us to study the sources  that are commonly used to share publications, and to analyse the effectiveness of the sources for sharing a publication  using Kudos , LinkedIn (https://www.linkedin.com), ResearchGate, Academia.edu (https://www.academia.edu), Facebook (https://www.facebook.com), Mendeley (https://www.mendeley.com), and Twitter with varying degrees of usage intensity, and for diverse purposes, including maintaining their scholarly profiles, and sharing their research outputs [A comprehensive literature review on the usage of social media in academia and in scholarly communication shows the interest and popularity amongst researchers in using social media platforms . Social  outputs .twimpact factor, have been proposed to complement traditional citation metrics [Specifically, scholars have cited information dissemination as a major benefit of using Twitter . In a st metrics .https://www.youtube.com), Slideshare (http://www.slideshare.net), and Flickr (https://www.flickr.com) to disseminate a wider range of research outputs, such as videos, presentation slides, or photos [Also, a survey of 2,414 researchers shows that researchers use YouTube , could be associated with an increase in a publication’s metrics . A study in the clinical pain sciences, albeit with a small sample of 16 PLOS ONE articles, does show that sharing via Facebook, Twitter, LinkedIn and ResearchBlogging.org is associated with an increase in the number of views and downloads of a publication, although the absolute effect size might not be considered substantial . Thus, thttps://www.growkudos.com, launched in May 2014) is a web-based service that claims to help researchers, institutions and funders maximise the visibility and outreach of published articles. Kudos enables analysis not only of the current outreach of a publication, but also how that outreach has been achieved.Kudos , 1,485 had share referrals, and 10,521 had Altmetric Scores available. The metrics were available on a daily count from the time of claiming to the date of extraction from Kudos. Kudos tracks daily full text downloads of publications from participating publishers’ websites, and share referrals . Some publications were shared multiple times across the different channels, and sometimes multiple times via the same channel. When this occurred, the multiple records of a publication being shared were aggregated. Publication dates are not available in the metadata for all publications in Kudos. The dates on which publications were claimed by authors in Kudos ranged from 17 September 2013 to 08 January 2016. The datasets used for the analysis have been made available online .For our analysis, a metric increase was defined as an increase in a metric  between the date the publication was first claimed in Kudos and the date of extraction from Kudos (30 January 2016) as shown in The assumption was made that the metrics are cumulative scores, thus the metric on the extraction date could only be greater or equal to the metric on the claim date. Unfortunately, the Altmetric Score from Altmetric.com does not respect this assumption as sometimes the Altmetric Score experienced a decrease in value due to changes in the calculation of the Altmetric Score by Altmetric.com. Furthermore, Kudos “trackable URLs” were only recognised by Altmetric.com following a change made part way through the time period represented in the dataset for this project. Thus, the Altmetric Score could not be considered further in the analysis.https://www.oecd.org/science/inno/38235147.pdf) classification scheme, which provides broader disciplines: natural sciences, engineering and technology, medical and health sciences, agricultural sciences, social sciences, humanities, and other disciplines . In this analysis, the self-reported career levels and disciplines of the first authors who claimed their publications on Kudos are considered. Data records on first authors not having a career level or a discipline were excluded from the analysis, consequently the total number of authors analysed per career level  and discipline  varies. A Chi-Square test is applied to determine statistical significance among the differences in career levels and disciplines.r = .10 to .29 , r = .30 to .49 (medium), and r = .50 to 1.0 (large), regardless whether the values are positive or negative. The statistical significance level is shown at **p < .01 and *p < .05.RQ2 aims to investigate which online channels are most effective for sharing a publication. For this analysis, we investigate the self-reported career and discipline levels of the authors who claimed the publications in Kudos. We apply the Spearman correlation to determine the association between the number of links shared on the different social media channels and an increase in share referrals during the time between the claim date and extraction date. The Spearman correlation is applied since the underlying distribution is highly skewed. The analysis is performed on the 4,610 publications with share actions. Following ’s guidelFor RQ3, which aims to investigate if any of the actions offered by Kudos have an association with the full text downloads of a publication, two groups were compared with each other, a treatment group and a control group. An increase in metric (in full text downloads) was calculated in the same way as explained previously. An increase in full text downloads of a publication was measured between the first claim date in Kudos and the extraction date selected. In order to create two groups that are comparable, a data sampling procedure was performed. From the 830,565 unique publications claimed in Kudos, those having full text downloads available were extracted. This resulted in 71,081 publications with full text download information. The 71,081 publications were filtered into two parts: those with actions  and those without actions . For this analysis, the 4,867 publications with actions were taken to form the Treatment group. To form the Control group, 4,867 publications were randomly selected from the 66,214 publications without actions.A correlation analysis was initially performed to measure the association between publications from the Treatment group (having actions) and an increase in full text downloads and the publications from the control group (having no actions) and an increase in full text downloads. The results did not offer much insight into the association between publications having actions or having no actions and an increase in full text downloads. We therefore looked into the characteristics of the publications considered for the analysis. We noticed that many of the publications in Kudos were only recently published, most between 2015 and 2011  had been shared, and 20,775 (93.7%) had been explained. Of those explained, 4,499 (21.7%) had been further enriched. As shown in As seen in p < .01), followed by LinkedIn , and Facebook . The results are based on n = 1,485 shared links that had been clicked.The findings in p < .01). For faculty, a small correlation is found for LinkedIn , and a medium correlation for Twitter . A small correlation is also found between the number of shares made by professionals and an increase in share referrals on Twitter .The correlations between the number of shares on social media channels and an increase in share referrals by career level and discipline are also investigated. As shown in Small to large Spearman correlations between the number of shares made on Twitter and an increase in share referrals are seen across all disciplines in p < .001). The results were not remarkably affected by filtering by document type. When considering only document types: article, article in press, and conference paper, the Treatment group  still had a significantly higher median average of 158 increase in full text downloads (23.4% more) per publication as compared to the Control group  with a median average of 128 increase in full text downloads per publication . The results are shown in Overall for all publications, the Treatment group  had a significantly higher median average of 149 increase in full text downloads (23.1% more) per publication in comparison to the median average of 121 of the Control group . The Mann-Whitney U value was found to be significant at , in 2012 , and in 2011 . 2016 and the other earlier years (including 2008) had too small sample sizes of n < 40 for the Treatment group  being faculty. Also, findings for the discipline Agricultural Sciences must be interpreted with care, as the sample sizes were small for sharing on Twitter and LinkedIn with n < 40.RQ2 aimed to investigate how effective researchers on Kudos were in sharing their publications on social media. Overall, the results show that sharing a Kudos generated URL on any of the popular social media channels  is associated with an increase in the clicks on this link . Although Facebook might be the most commonly used platform, it seems that links shared on Twitter are more likely to be clicked on across all disciplines, and almost all career levels (except for students). In addition, researchers in the fields of medical and health sciences were moderately effective in sharing their research across all social media channels. This could be a motivation for the scholarly community to share their publications more on social media. The results are in line with other studies such as , which aRQ3 aimed to analyse if any actions offered by Kudos had an association with the increase in full text downloads of a publication. Our study does not aim to generally compare the full text downloads of publications, but rather to analyse an increase in full text downloads for a publication from the time the author first claimed it in Kudos up till the data extraction date for this study. The aim is to analyse whether publications in Kudos having actions performed on them have a higher increase in full text downloads in this period when compared to publications in Kudos without any actions performed on them. The calculation of an increase in full text downloads in such a period is thus dependent on the publication having been claimed in Kudos and Kudos having tracked the full text downloads over this period. Thus, we are unable to expand this analysis to include papers that have not been claimed by authors in Kudos in the Control group as neither the increase in full text downloads can be calculated for them, nor does the period in Kudos for the analysis exist in such a case (there would be no claim date).Several different data analysis approaches were tested on the dataset, but they did not yield conclusive results. For instance, individual actions were analysed using a correlation test to examine the association between actions and an increase in full text downloads, and between actions and an increase in share referrals. It was considered if a publication had an action or not, but the number of times the action was performed on the publication was not considered for the analysis. The increase in metric was calculated in the same way as previously mentioned. The findings however did not show that any specific actions could be associated with an increase in any of the metrics, and as such these analyses are not presented here.The results of Mann-Whitney U tests show that overall the Treatment group had a significantly higher median average than the Control group, independent of document type. This finding was partially supported when analysed per publication year and journal. However, since most publication years and journals had too small sample sizes, the results have to be interpreted cautiously. Moreover, as most publications in the Treatment group and Control group had been published in journals belonging to the Royal Society of Chemistry, this means the Treatment group and Control group are rather homogeneous in discipline, with most publications coming from the broader field of chemistry. The Treatment group however had a large number of publications published in the journal the “Analyst”. But still in an attempt to drill down by discipline, we used the self-reported disciplines of the first author who claimed the publication in Kudos. It is however important to note that social behaviours and expectations change in reaction to social statistics, rankings and public evaluation . As schoThere are some limitations to this study. This study by no means aims to generalize the findings made here to the entire research community. The dataset extracted from Kudos represents only a small fraction of researchers and published papers worldwide. The profile information about authors who have claimed publications in Kudos is self-reported, and we only analyse authors who claimed and shared in Kudos, thus, some results might be biased and the results cannot be generalized to all researchers and faculty. A number of authors and publications had to be excluded from the analysis due to insufficient data, such as missing dates of publication, or missing career level or discipline information for authors.Moreover, this dataset could be criticised as being biased and represent only a narrow group of researchers as the services offered by Kudos might potentially attract a certain profile of users . This can only be investigated with a user survey of Kudos’ users, which is out of scope for this study. However, it should be noted, that the services offered by Kudos are not restricted to any specific group of users, nor has any group of users been specifically targeted as potential users. Kudos is freely accessible to all who wish to use its services and we have had no influence over the users of Kudos, nor the publications they claim in the system. We also have had no influence over the actions they took on Kudos.Furthermore, the treatment group could be criticised as having a potential selection bias as the users of Kudos might have chosen their most promising papers  to further perform actions on in Kudos to boost these papers’ metrics. This can also only be resolved by investigating the motivations of the users of Kudos in a user study.Due to the limited sample size of the Treatment group , the possible variations of analyses were limited. When broken down into smaller samples for example, per year, per journal, per discipline, most of the sample sizes got too small for analysis. We could also not compare publications on a per month basis as most publications did not have this information available. Also, due to the small sample sizes, drilling down per month led to even smaller sample sizes, in most cases too small for a comparison.Some characteristics of the publications and authors were not considered in this study, such as geographical location of the author, open access status of the publication, and daily citation counts for publications. Thus there remain unanswered questions such as, does the geographical location of the author affect the sharing of publications in Kudos? Do open access publications receive more actions and full text downloads? Do actions in Kudos correlate to increased citation counts? Further work could be undertaken to explore other sources for metadata and metrics to overcome limitations in data availability.In this paper, we investigate the most effective actions that researchers could take, and channels that they could use to communicate their research outputs, in terms of the increase in full text downloads of the publications. The results show that Facebook is the most popular channel, but that links shared on Twitter are more likely to be clicked on. Moreover, we also find that actions performed in Kudos, such as sharing via social media and other channels, are associated with 23.1% more full text downloads. Citation counts were not analysed in this study so its findings do not claim that an increased rate of full text downloads is associated with a higher citation count in future.We believe the findings from this study will be useful for individuals and institutions to improve and update understanding of researchers’ outreach efforts and the potential increase in a publication’s metrics. The findings may encourage researchers to use communications media more strategically for building the readership of their work. The findings could also motivate improved provision by publishers and service providers of metadata, metrics and services to support further analysis.In the future, it would be interesting to gain more understanding of the scholarly community’s strategies of promoting their research, their opinion of disseminating research outputs on social media, and how they keep track of their research outputs. We could also study the information sharing behaviour, habits, and experiences of users on different social media channels, considering their demographics, such as the age group of the authors, career level, discipline, geographical location, etc. Furthermore, sharing via other channels such as wikis, blogs, forums, etc., could also be studied further.S1 FigPercentages of publications on Kudos with share actions  that were shared via Facebook , and Twitter , as well as via other channels (661), before 07 July 2015. LinkedIn was introduced by Kudos as an integrated sharing option in July 2015. Some publications were shared via multiple channels and on different share dates. Multiple shares on the same channel on the same share date were counted only once in this analysis.(TIFF)Click here for additional data file.S2 FigPercentages of publications on Kudos with share actions  that were shared via the three social media channels: Facebook , Twitter (877), and LinkedIn (969), as well as via other channels (111), after 07 July 2015. LinkedIn was introduced by Kudos as an integrated sharing option in July 2015. Some publications were shared via multiple channels and on different share dates. Multiple shares on the same channel on the same share date were counted only once in this analysis.(TIFF)Click here for additional data file.S3 FigThe publication years for the publications with publication dates available, and restricted to publications with document types: article, article in press, and conference paper for the Treatment group  and the Control group .(TIFF)Click here for additional data file.S4 FigThe month and year the publications were first claimed by an author in Kudos for publications with publication dates available, and restricted to publications with document types: article, article in press, and conference paper for the Treatment Group  and the Control Group .(TIFF)Click here for additional data file.S5 FigPercentage distribution of disciplines of the first authors who claimed the publications in Kudos. For publications with publication dates available and restricted to publications with document types: article, article in press, and conference paper in the Treatment Group : Natural Sciences , Engineering and Technology (313), Medical and Health Sciences (185), Agricultural Sciences (23), Social Sciences (87), Humanities (14), and other disciplines (36).(TIFF)Click here for additional data file.S6 FigPercentage distribution of disciplines of the first authors who claimed the publications in Kudos. For publications with publication dates available and restricted to publications with document types: article, article in press, and conference paper in the Control Group .: Natural Sciences , Engineering and Technology (678), Medical and Health Sciences (455), Agricultural Sciences (44), Social Sciences (102), Humanities (15), and other disciplines (49).(TIFF)Click here for additional data file.S1 TableFor career levels: professionals (n = 506), students (n = 256), researchers (n = 689), faculty , and other career levels (n = 241) who shared their publications via Kudos on Facebook.(PDF)Click here for additional data file.S2 TableFor career levels: professionals (n = 506), students (n = 256), researchers (n = 689), faculty , and other career levels (n = 241) who shared their publications via Kudos on Twitter.(PDF)Click here for additional data file.S3 TableFor career levels: professionals (n = 506), students (n = 256), researchers (n = 689), faculty , and other career levels (n = 241) who shared their publications via Kudos on LinkedIn.(PDF)Click here for additional data file.S4 TableFor career levels: professionals (n = 506), students (n = 256), researchers (n = 689), faculty , and other career levels (n = 241) who shared their publications via Kudos on other channels. * Due to small sample sizes (n < 40), results should be interpreted with caution.(PDF)Click here for additional data file.S5 TableAuthors from Social Sciences , Natural Sciences , Medical and Health Sciences (759), Engineering and Technology (590), Humanities (304), Agricultural Sciences (65), and other disciplines (102), who shared their publications via Kudos on Facebook. * Due to small sample sizes (n < 40), results should be interpreted with caution.(PDF)Click here for additional data file.S6 TableAuthors from Social Sciences , Natural Sciences , Medical and Health Sciences (759), Engineering and Technology (590), Humanities (304), Agricultural Sciences (65), and other disciplines (102), who shared their publications via Kudos on Twitter. * Due to small sample sizes (n < 40), results should be interpreted with caution.(PDF)Click here for additional data file.S7 TableAuthors from Social Sciences , Natural Sciences , Medical and Health Sciences (759), Engineering and Technology (590), Humanities (304), Agricultural Sciences (65), and other disciplines (102), who shared their publications via Kudos on LinkedIn. * Due to small sample sizes (n < 40), results should be interpreted with caution.(PDF)Click here for additional data file.S8 TableAuthors from Social Sciences , Natural Sciences , Medical and Health Sciences (759), Engineering and Technology (590), Humanities (304), Agricultural Sciences (65), and other disciplines (102), who shared their publications via Kudos on other channels. * Due to small sample sizes (n < 40), results should be interpreted with caution.(PDF)Click here for additional data file.S9 Tablehttps://www.oecd.org/science/inno/38235147.pdf) career categories: Professionals, students, researchers, faculty, and other career levels.The career levels of the first authors who claimed publications on Kudos were re-coded using the five broad OECD Click here for additional data file.S10 Tablehttps://www.oecd.org/science/inno/38235147.pdf) classification scheme, which provides seven broad disciplines: Natural Sciences, Engineering and Technology, Medical and Health Sciences, Agricultural Sciences, Social Sciences, Humanities, and other disciplines.The disciplines of the first authors who claimed publications in Kudos were recoded using the OECD Click here for additional data file.S11 TableThe document type was retrieved from Scopus for the publications in the Treatment group  and the Control group . Only 4,208 had document types available in the Treatment group, and only 4,155 had document types available in the Control group. The document types: article, article in press, conference paper, editorial, erratum, letter, note, review, and short survey were available.(PDF)Click here for additional data file.S12 TableFull text downloads for publications on the day they were first claimed by an author in Kudos, on the extraction date from Kudos for this study (30 January 2016), and the calculated increase in full text downloads between the claim date and extraction date, for publications with publication dates available and restricted to publications with document types: article, article in press, and conference paper for the Treatment group  and the Control group .(PDF)Click here for additional data file.


Then, with the OLR increased, the removal efficiencies of BOD5, COD, and TSS increased as long as the organic loading reached 0.44 gCOD/L.d. But with the OLR increased more, the removal efficiency of these parameters decreased. The ANOVA statistical test results showed that the mean difference of removal efficiency in organic loadings for BOD5 (p ≤ 0.001) and COD (p = 0.003) was significant, while it was insignificant for TSS (p = 0.23). The best removal efficiencies of Total Nitrogen (TN) and Total Phosphorus (TP) were 89.60 and 86.67%, respectively, which were obtained at an OLR of 0.44 gCOD/L.d. By increasing OLR up to 0.44 gCOD/L.d, removal efficiencies of TN and TP increased, while the removal efficiency decreased with the OLR increased more, and this difference was statistically significant (p = 0.021). Finally, the results showed that the IFAS system provided a proper efficiency in treatment of the synthetic greywater and it could be used in a full scale.In this study, the efficiency of Integrated Fixed-film Activated Sludge (IFAS) system in synthetic greywater treatment and nutrients removal was studied in duration of 105 days according to different Organic Loadings Rates (OLRs). The study was operated in pilot-scale and OLRs of 0.11–1.3 gCOD/L.d. Scanning Electron Microscope (SEM) image showed that the biofilm with a proper thickness was formed on IFAS reactor’s media. The results indicated that the best removal efficiency of BOD Nowadays, the shortage of water resources, growing population and increasing water demand, inappropriate distribution of water resources, mismanagement of the available water, and climate changes have steered most of the countries toward the research for new water resources  with a flow rate of 20 L/day. Organic loading rate in IFAS system at 5 loadings was 0.11–1.3 gCOD/L.d, hydraulic retention time was 10.8 h, and Input COD in 5 OLRs was 50–600 mg/L according to a previously reported study  = 5500 mg/L, Mixed Liquor Volatile Suspended Solids (MLVSS) = 4510 mg/L, and the Sludge Volume Index (SVI) = 160 mL/gr was used. After 25 days and formation of fixed-biofilm on aeration reactor, firstly the greywater with COD 50 mg/L and OLR of 0.11 gCOD/L.d was injected into the system, and then different greywater samples with CODs of 100, 200, 400, and 600 mg/L and OLRs of 0.22, 0.44, 0.88, and 1.3 gCOD/L.d were logged in IFAS system.5, COD, Total Suspended Solids (TSS), Total Nitrogen (TN), and Total Phosphorous (TP) parameters as well as pH, DO, temperature, and MLSS were measured. Composite Sampling was done on input and output from the system, and at least five samples were measured for each parameter in each loading. Measurement of the parameters was carried out via diverse methods as follows: COD by dichromate method , BOD5 by Winkler method with standard Number of 2510, TSS by spectrophotometry with standard Number of 2540 D, total nitrogen (TN) by the Kjeldahl method, total phosphorus (TP) by digestion and Conversion of the various forms of phosphorus to orthophosphate, and determination of the orthophosphate by spectrophotometry. Also, temperature, pH, and DO were measured by portable YSI and MLSS using filter paper of 0.45 micron according to the standard . The highest MLSS was 3900 mg/L at the OLR of 0.44 gCOD/L.d. The results showed that the IFAS system provides a high efficiency in synthetic greywater treatment.In the present study, the efficiency of IFAS system in synthetic greywater treatment from different organic loadings’ rates was studied within a duration of 105 days. SEM image showed that the biofilm was formed on the media of IFAS reactor. The best removal efficiencies of BOD


PAE) refers to the temporary suppression of bacterial growth following transient antibiotic treatment. This effect has been observed for decades for a wide variety of antibiotics and microbial species. However, despite empirical observations, a mechanistic understanding of this phenomenon is lacking. Using a combination of modeling and quantitative experiments, we show that the PAE can be explained by the temporal dynamics of drug detoxification in individual cells after an antibiotic is removed from the extracellular environment. These dynamics are dictated by both the export of the antibiotic and the intracellular titration of the antibiotic by its target. This mechanism is generally applicable for antibiotics with different modes of action. We further show that efflux inhibition is effective against certain antibiotic motifs, which may help explain mixed cotreatment success.The postantibiotic effect ( Pneumococcus and Streptococcus in the 1940s. Even after the antibiotic had been degraded by a penicillinase, the target populations exhibited a significant lag before resuming growth  refers to the temporary suppression of bacterial growth following transient exposure to antibiotics. This transient inhibition has been observed since the first studies of penicillin against  Bigger, . Subsequ Bigger, , β‐lactaet al, et al, Despite widespread observations of PAE, its underlying mechanisms are not well established. Previous studies have speculated on a number of possible explanations, including nonspecific binding and nonlethal damage induced by antibiotic treatment  as the doubling time relative to the end of antibiotic treatment . We found that the fluorescence signal serves as a reliable surrogate measure of the population density over the range of dosing conditions used . Therefore, although these cells contributed to total fluorescence, they did not contribute to population recovery. To account for this, we measured population viability as a function of streptomycin concentration and duration in the absence of growth . We adjuet al, Ain or Aout), synthesis of the ribosome (C) through a positive feedback loop, and antibiotic‐mediated ribosome inhibition and potential degradation . In terms of the core structure, our model is similar to one developed by zur Wiesch et al =kinkoutAout(1−e−koutD)≈kinAoutD . That is, at the end of an antibiotic pulse, the intracellular antibiotic concentration is proportional to total amount of the antibiotic used  . Moreover, this correlation is robust to changes in the antibiotic‐mediated ribosome degradation rate  will induce a significant RTn. However, regardless of the frequency of antibiotic dosing, population recovery time remains a function of total antibiotic exposure , which does not induce HSR , a phosphorylation inhibitor that inhibits a variety of efflux pumps that rely on the proton motive force (PMF), including resistance nodulation division pumps and others , and antibiotic motifs, to changes in efflux. We reasoned that the induction of PAE by antibiotic accumulation and detoxification dynamics Fig A is applet al, To test this hypothesis, we developed simplified kinetic models of intracellular dynamics to investigate three motifs of antibiotic action that encompass a wide variety of common antibiotics  factors including availability, toxicity, and plasma vs total drug concentrations 567, lacZ4787(del)::rrnB‐3, LAM−, rph‐1, DE(rhaD‐rhaB)568, hsdR514) was used throughout this study. Unless otherwise noted, all experiments were conducted in M9 media supplemented with 0.4% w/v glucose and 0.1% w/v casamino acids, and at 37°C unless otherwise indicated. BW25113 cells were transformed with a constitutively expressed GFP plasmid (kanamycin resistant) for microfluidic and viability experiments; all cultures were supplemented with 50 μg/ml kanamycin for selection purposes. For luminescence assays, BW25113 was transformed with a constitutively expressed luminescence reporter plasmid . Cells were grown in 3 ml aliquots in the presence of increasing streptomycin concentrations  for 2 h at 37°C and 250 rpm shaking. Following treatment, cells were serially diluted and plated on selective agar plates to measure CFU counts; GFP fluorescence and luminescence were measured in 96‐well black‐walled plates (Corning) using a Tecan InfinitePro M200 plate reader (GFP emission/excitation 485/535 nm). Plates were incubated overnight at 37°C and CFUs counted the following morning. All measurements were done in quadruplicate.et al, Transient antibiotic dosing experiments were carried out using a previously published microfluidic platform , followed by antibiotic treatment (500 μl/h with applicable antibiotic concentrations) and a recovery phase . These flow rates were selected to satisfy two physical limits: Excessively high flow rates resulted in cell washout, and insufficiently low flow rates resulted in minimal antibiotic effect over the range of dose durations used. The entire device was kept at 37°C. Five replicates were used per experimental condition. Culturing chambers were imaged every 5 min using a DeltaVision Elite deconvolution microscope.Overnight cultures were diluted 100‐fold into 3 ml M9 and grown for 2 h at 37°C with 250 rpm shaking, reaching an ODAll image data from microfluidic experiments were analyzed using custom MATLAB scripts. Following background subtraction, total fluorescence was quantified and normalized to the initial time point following antibiotic treatment. Figures show mean and standard deviation for five replicates.7‐fold and plated on selecting agar plates (50 μg/ml kanamycin). Plates were incubated at 37°C overnight and CFUs counted the next day. Cell density was also measured prior to antibiotic treatment (~7E8 CFU/ml). All data points show the mean and standard deviation of six replicates. Viability curves for each streptomycin concentration were normalized to initial densities and fit exponential decay curves. The time constants of these fits were used to account for population death.Overnight cultures were regrown to mid‐exponential phase by 100‐fold dilution in 3 ml M9 media and incubating for 2 h at 37°C with 250 rpm shaking. Cells were then washed and resuspended in a standard 1× M9 buffer (M9 not containing glucose or casamino acids); 500 μl aliquots in 1.5‐ml Eppendorf tubes were incubated with appropriate antibiotic concentrations at 37°C with 250 rpm shaking. Samples were then serially diluted to 10X) of cells. Then, the population fluorescence immediately following antibiotic treatment can be expressed as a combination of dead and viable cells: XF0+1−XF0, assuming dead cells contribute equally to the fluorescence signal. Therefore, when the viable fraction doubles, the total fluorescence would be XF0+21−XF0=2−XF0.Viability curves were used to adjust the twofold cutoff used to calculate recovery time. As an example, suppose a particular antibiotic treatment killed a fraction (600) data were collected every 10 min using a Tecan M200 Infinite Pro plate reader at 37°C. IC50 values were determined by first calculating growth rates by iteratively finding the linear region of increase; these growth rates were then fit to a Hill equation, where μmax, n, and A are the maximum growth rates, Hill coefficient, and antibiotic concentration, respectively  with 200 μl liquid per well; wells were covered with 50 μl mineral oil  to prevent evaporation and density ; stock solutions were prepared by dissolving CCCP in dimethyl sulfoxide (DMSO) to a final concentration of 1 mg/ml and stored at 4°C. Cells were plated and covered with 50 μl mineral oil to prevent evaporation. Intracellular EtBr signal and cell density (OD600) were measured every 10 min at 30°C using a Tecan InifintePro M200 plate reader (EtBr excitation/emission 530/585 nm). Fluorescence time courses were normalized to OD600. Efflux rates were calculated assuming that intracellular EtBr concentration is dictated by the following equation:Efflux rates were measured by adapting an ethidium bromide (EtBr)‐based assay for efflux pump activity , and efflux rates were determined, assuming et al, We modeled aminoglycoside action similarly to a previously published framework  and intracellular ([Ain]) environment according to the rates of influx and efflux, kin and kout, respectively. Ribosomes reversibly bind to intracellular antibiotic (rates kf and kb) yielding the complex [CA]. This complex undergoes a two‐step degradation process, releasing antibiotic to the intracellular concentration  will remain effective.Periodic dosing simulations were specified by the following parameters: the antibiotic concentration A0 and duration D, we assumed a symmetric profile, with a maximum value of A0 at t = D/2 For exponentially decaying doses, we assumed a rate constant such that AoutD=0.01A0.To demonstrate the generality of the dependence between recovery time and total antibiotic, implemented both triangular and exponentially decaying dose profiles. For triangular doses, given the initial antibiotic concentration T] and [A] represent the drug target and intracellular antibiotic concentration, respectively; [P] represents the antibiotic–target binding product. k0 represents the basal target synthesis rate, kf and kb represent the binding and dissociation rates between antibiotic and target, kp and KT represent the maximum synthesis and half‐maximal values of [T], dT and dp represent the degradation rates of target and antibiotic–target complex, respectively. Finally, Aout represents the extracellular antibiotic concentration during a dose, which we assume to be constant; kin and kout represent the influx and efflux rates across the cell membrane, respectively. By setting various parameters to zero, these equations can be used to compare diverse modes of action. Setting k0 = 0 results in target synthesis by positive feedback with rapid antibiotic‐mediated degradation; setting dp = 0 corresponds to antibiotic‐inducing minimal target degradation; and setting dp = 0 = kt corresponds to antibiotics whose targets are not subject to positive feedback. As with the full model, intracellular recovery time was calculated as the time to achieve 0.5∗maxdTdt during recovery.We adopted a minimal set of ODEs capable of representing various mechanisms of action (equations 10–12, https://fairdomhub.org/investigations/155 and an SED‐ML simulation reproducing Fig https://jjj.bio.vu.nl/models/experiments/srimani2017_fig2cii/simulate.MATLAB codes for all main figure simulation results are provided as JKS conceived the work, designed and carried out experimental and modeling studies and data analysis, and wrote the manuscript. SH designed and fabricated the microfluidic platform, assisted in experimental studies, and contributed to manuscript revisions. AJL fabricated microfluidic chips, assisted in carrying out experiments and data analysis, and contributed to manuscript revisions. LY conceived the work and assisted in research design, data interpretation, and writing and revising the manuscript.The authors declare that they have no conflict of interest.AppendixClick here for additional data file.Code EV1Click here for additional data file.Review Process FileClick here for additional data file.


MET amplification. Whereas T790M mutation confers prolonged survival and sensitivity to 3rd generation TKIs, data are lacking on clinical features and outcome of MET-driven resistant EGFR-mutated NSCLC patients.Several mechanisms of acquired resistance to EGFR tyrosine kinase inhibitors (TKIs) in EGFR-mutated NSCLC have been described including the T790M mutation and MET amplification, detected on a biopsy performed after progression on EGFR TKI, were identified in 15 centers. Clinical and molecular data were retrospectively collected.Patients with metastatic EGFR-mutated NSCLC displaying high MET overexpression or MET amplification. A T790M mutation was found in 11/41 (26.8%) patients. T790M-positive patients had a better OS than T790M-negative patients (p=0.0224). Nineteen patients received a MET TKI. Objective response was reported in 1 out of 12 evaluable patients treated with a MET inhibitor as a single agent and in 1 of 2 patients treated with a combination of MET and EGFR TKIs.Forty two patients were included. The median overall survival (OS), and the median post EGFR TKI progression overall survival (PPOS) were 36.2 months [95%CI 27.3-66.5] and 18.5 months [95%CI 10.6-27.4] respectively. Nineteen out of 36 tumors tested for MET FISH had MET-driven resistance to EGFR TKI defines a specific pattern of resistance characterized by low objective response rate to MET inhibitors given alone and overlapping with T790M mutations. Further studies are warranted to define adequate therapeutic strategies for MET-driven resistance to EGFR TKI. EGFR mutations are found in 10% of non small cell lung cancer (NSCLC) in Caucasians and 40% in Asians . Mo. MoEGFR pression  and may pression . BecauseEGFR mutated NSCLC with a MET-driven resistance to EGFR TKI did not display specific clinical features compared to those observed in previous studies. Median PPOS (18.5 months) was also in the range of what has been reported in previous studies focusing on patients with acquired resistance to EGFR TKIs (14.3-20 months) [MET GCN alterations and MET overexpression have been associated with poor prognosis in resected NSCLC [EGFR mutated MET amplified NSCLC responding to combined MET and EGFR TKIs have been reported [EGFR mutated NSCLC patients who progressed on EGFR TKI, reported an ORR of 19% in patients with MET IHC3+ or MET IHC≥2+ and MET GCN≥5 and raised to 30% in the subgroup of patients with MET GCN≥6 [In our cohort, metastatic  months) , 32, 33.ed NSCLC , 35, we ed NSCLC , 36. Casian PPOS .5 monthsreported . This duET GCN≥6 .MET amplification and a T790M mutation on the same sample. The coexistence of these mechanisms of resistance has been previously described [MET amplification and overexpression. Of note, Gou et al. found that patients with a T790M mutation and MET overexpression had a worse PPOS than patients with only MET overexpression or the T790M mutation alone. This discrepancy with our results might be due to the small size of both cohorts, and variations in the characteristics of the study population (higher rate of never smokers and caucasians in our study). We cannot also exclude the influence of confounding factors such as the younger age and the use of 3rd generation EGFR TKI in T790M positive patients. Indeed in our study, 9 T790M-positive patients received a 3rd generation EGFR TKI and 3 partial responses were achieved. Very few data are available regarding the efficacy of 3rd generation TKIs in the context of multiple resistance mechanisms. Our results suggest that 3rd generation TKIs may retain activity against T790M-positive tumors in some patients, even in the presence of MET activation, which may be due to the spatial heterogeneity of resistance mechanisms rather than co-existence of 2 resistance mechanisms in the same tumor cells.In our study, 11 patients were T790M positive. Three had both a escribed . Compareescribed , 32, 38.rd generation EGFR TKIs and prolonged survival. Further studies are warranted to define adequate therapeutic strategies for MET-driven resistance to EGFR TKI.Finally, MET-driven resistance to EGFR TKI defines a specific pattern of resistance characterized by low objective response rate to MET TKIs given alone and overlapping with T790M mutations. Even when associated with MET dysregulation, the T790M mutation was still associated with relative efficacy of 3EGFR mutation on tumor sample at diagnosis, treatment with at least one EGFR TKI and detection of MET overexpression or MET amplification after the time of clinical or radiological progression on EGFR TKI. Post-progression re-biopsy and MET status assessment were routinely performed in participating centers during the study period. Clinical and pathological data were retrospectively collected in each center for all included patients. Best overall response, defined as the best response from the start of treatment until disease progression, was assessed by investigators from available follow-up exams in each center using Response Evaluation Criteria in Solid Tumor (RECIST) v1.1. Crizotinib, gefinitib, and osimertinib were respectively prescribed with the following doses and schedules : 250mg bid, 250mg daily, 80mg daily. The study was approved by a national ethic committee (CEPRO 2016-001).We constituted a multicenter retrospective observational cohort of patients identified among 15 French centers. Inclusion criteria were documented diagnosis of metastatic NSCLC, detection of an MET FISH analyses were realized according to locally certified and nationally approved procedures. MET overexpression was defined as a 3+ MET Immunoscore (≥ 50% of tumor cells showing high-intensity staining) on MET immunochemistry using MET monoclonal antibody (clone SP44 Ventana). MET amplification was defined by Fluorescence In Situ Hybridization (FISH) as a mean MET GNC per cell ≥ 6, or a Ratio MET/CEP7 ≥ 2, or the presence of MET clusters [EGFR T790M mutation was detected in post-progression circulating tumor DNA, on the re-biopsy on which MET overexpression or MET amplification was found, or on a tumor sample collected before the re-biopsy.Histological and molecular analyses of formalin-fixed paraffin-embedded (FFPE) samples were prospectively assessed by local pathologists in accredited and quality controlled laboratories, as part of the routine procedure. MET immunochemistry and clusters . PatientMET amplified vs. MET non-amplified and T790M+ vs. T790M-). Chi-Squared tests or Fisher's exact test (when expected cell frequency <5) were used to study the association between categorical variables and different groups. A Mann-Withney U test was used to compare age, time between EGFR TKI initiation and rebiopsy, and duration of TKI EGFR therapy between groups. We estimated and compared overall survival (OS), post progression overall survival (PPOS) and progression free survival (PFS) between the study groups (MET amplified vs. MET non amplified and T790M+ vs. T790M-) using the Kaplan-Meier Method and log-rank test. OS was measured from the date of metastatic NSCLC diagnosis to the date of death from any cause or last follow-up. The PPOS was measured as the time from EGFR TKI RECIST progression to death from any cause. The PFS was defined as the time from treatment start to disease progression or death from any cause. Objective response rate (ORR) was defined as the percentage of patients with partial or complete response to the indicated treatment. Statistical testing was conducted at the 2-tailed α level of 0.05. Data were analyzed with SAS software version 9.3 .Categorical variables are expressed as frequencies and percentages. Quantitative variables are expressed as medians (range). Normality of distribution was assessed graphically and by using the Shapiro–Wilk test. Bivariate analyses were realized to assess the sub-group comparability (


The Central Sensitization Inventory (CSI) is a new patient-reported instrument, which measures symptoms related to Central Sensitivity Syndromes and Central Sensitization. The aim of this study was to translate the CSI into Spanish, and then to perform a psychometric validation, including a factor analysis to reveal the underlying structure.In this two-stage psychometric study participated 395 subjects with various chronic pain conditions and that were recruited from two Primary Care Centres. The CSI was cross-culturally adapted to Spanish through double forward and backward translations. The psychometric properties were then evaluated with analyses of construct validity, factor structure and internal consistency. One subgroup (n = 45) determined test-retest reliability at 7 days.The Spanish Version of CSI demonstrated high internal consistency (α = 0.872) and test-retest reliability (r = 0.91). Factor structure was one-dimensional and supported construct validity.The psychometric properties of the Spanish version were found to be strong, with high test-retest reliability and internal consistency, with similar psychometric properties to the English language version. Unlike the English version, however, a one factor solution was found to be a best fit for the Spanish version. Patient-reported outcome (PRO) measures Garratt  are commThe Central Sensitization Inventory (CSI) was designed as a tool to identify when a patient’s symptoms may be related to CS/CSSs  were recruited consecutively from the community-based Physiotherapy Program at the Malaga University. Exclusion criteria were; Chronic musculoskeletal pain for less than 3 months; diagnosis of specific medical conditions that can negatively affect the central nervous system, including cancer, brain or spinal cord injury, neurological disease or injury; Aged <18 years old and Poor Spanish language comprehension. Diagnoses were made by a physician in two primary care centres in Torremolinos, Malaga, Spanish National Health Service. All eligible participants completed the three Spanish language versions of the self-administered questionnaire CSI-Sp.2.1) test–retest methodology in a randomly selected subgroup of the full sample determined at 7 days .Descriptive analyses were applied to calculate means and standard deviations of demographic variables. Distribution and normality were determined by one-sample Kolmogorov–Smirnov tests (significance <0.05). Construct validity and factor structure were determined through the use of questionnaire principal component analysis with Maximum Likelihood Extraction (MLE), with the requirements for extraction being the satisfaction of all three points: screeplot inflection point, Eigen value >1.0 and accounting for >10 % of variance  with an individual range from 0.87 to 0.95. Measurement error was determined from SEM and MDC90, being at 2.52 and 7.83 %, respectively. No significance differences were found between genders in item responses.The CSI-Sp showed no missing responses and it showed a high degree of internal consistency (Cronbach’s α = 0.872) with an individual item range from 0.851 to 0.891. The test–retest reliability was high at (ICCp < 0.001). This indicated that the correlation matrix was unlikely to be an identity matrix and, therefore, was suitable for MLE. The factor analysis revealed a satisfactory percentage of total variance explained by the one factor at 25.9 %. However, the items with an Eigenvalue >1.0 each accounted for <10 % of variance and were shown to be after the screeplot initial inflection point and consequently not extracted. The screeplot .The correlation matrix for the CSI-Sp was determined suitable from the Kaiser-Meyer-Oklin values (0.864) and Barlett’s Test of Sphericity p < 0.00. This inIn the present study, a cross-cultural adaptation of the CSI, from English to the Spanish, was completed, resulting in a CSI-Sp version of this Inventory. Construct validity and internal consistency of the CSI-Sp were determined independently, and were both found to be strong. The single factor structure from this psychometric properties indicated that a single summated score could be used , which was in-line with the test–retest results of the English (0.82)  across the different countries/languages .The psychometric properties of the CSI-Sp are reported for the first time. The determined values were satisfactory and supportive of the validation of the CSI-Sp, particularly in the areas of internal consistency, factor structure and reliability. Consequently, the CSI-Sp may be useful in Spanish-speaking populations and for making cross-cultural comparisons in other English-speaking countries with a high Spanish-speaking population.


Advances from pharmacogenetics (PGx) have not been implemented into health care to the expected extent. One gap that will be addressed in this study is a lack of reporting on clinical validity and clinical utility of PGx-tests. A systematic review of current reporting in scientific literature was conducted on publications addressing PGx in the context of statins and muscle toxicity. Eighty-nine publications were included and information was selected on reported measures of effect, arguments, and accompanying conclusions. Most authors report associations to quantify the relationship between a genetic variation an outcome, such as adverse drug responses. Conclusions on the implementation of a PGx-test are generally based on these associations, without explicit mention of other measures relevant to evaluate the test's clinical validity and clinical utility. To gain insight in the clinical impact and select useful tests, additional outcomes are needed to estimate the clinical validity and utility, such as cost-effectiveness. Health care professionals, scientists, and policy makers have recognized the potential of precision medicine to optimize therapeutic outcomes. Appropriate therapeutic choices could be made by prospectively identifying patients through pharmacogenetic (PGx) tests. A PGx test can identify patients at high risk of treatment failure, for example due to drug toxicity or inferior treatment efficacy. However, predictive biomarkers are not used to the expected extent in health care practice Teng, . NumerouSeveral initiatives exist to translate knowledge on PGx to the broader clinical practice  independently extracted data following a predefined template of relevant topics. The topics included in this template are summarized in Table From the extracted data two researchers (MJ and TR) independently analyzed the following items: 1) measures of effect; (2) arguments used for or against eligibility; and (3) implementation advice. In the data on measures of effect and arguments for eligibility we specifically analyzed: parameters for potential clinical validity, clinical validity, and clinical utility  and arguments about eligibility (item 2) it was evaluated if the implementation advice was inconclusive.In the next analysis step, the consistency within publications was analyzed between the three items. For example, when a publication reported In total 132 publications were identified in the search, of which 89 were included for review (Supplementary Table n = 45), the ratio of positive outcomes reported compared to the total number of studies was the largest with 35 studies that reported a positive outcome. In the review publications, a ratio of 21 to 36 was found.The reviewed publications included 45 original studies , 36 reviews, and eight expert opinions . A range of statins was studied in the publications  clinical validity. However, none of these papers gave a positive implementation advice: three gave inconclusive advice, and one did not give an implementation advice.Some authors commented on the research and translation process in a more general manner. These comments suggest general steps for research and translation, which apply to ensure evidence-based guidelines for PGx for statins, such as (Needham and Mastaglia, “It is essential to first establish a valid association between a genetic variant and a drug response, which has been done for some genetic variations and muscle toxicity as reviewed above, and to then demonstrate a clinically significant outcome that results in improved patient management.”While many scientific publications describe promising PGx results, implementation appears to lag behind. Our results suggest an explanation can be found in the type of evidence reported in literature, because solely associations are often reported. Other types of evidence are needed for implementation, including guideline development and coverage decisions. To review arguments for the eligibility of PGx testing and what evidence is used to substantiate this, 89 scientific publications were evaluated on PGx for statins in cardiovascular disease. In 64 of the 89 publications, positive findings were reported on the research aims. Only in nine of these 64 publications a positive implementation advice was given. As multiple authors also discuss (Ong et al., Though our study is limited to published research and is inherently influenced by publication bias, it is a unique approach to reviewing scientific publications. The broad review of arguments in publications on the topic of PGx for statins, enabled a clear identification of the gap between scientific efforts and evidence needs for implementation. The call for reporting on a level of clinical utility finds resonance in other publications that discuss the gap between reported measures and necessary measures for implementation (Voora and Ginsburg, The context in which a test should take place and be piloted remains unclear in research suggestions from current publications. The timing of a PGx test influences the impact of the test greatly, for example there are significant differences between pre-emptively testing for a broad panel or targeted testing at first prescription. One of the few international initiatives on translation of PGx information into practice—CPIC—only offers information on dosing advice (Ramsey et al., A bridge is needed between the necessary and highly relevant work from basic science to relevant implementation research for clinical practice. Only through following the steps from clinical validity to clinical utility in a well-defined health care context Box , scientiResults from PGx studies on statins for cardiovascular disease are often summarized as associations between the genotype and phenotype: from effective drug levels to ADRs. For the next steps in translation, evidence is needed on clinical validity and clinical utility through implementation studies. It needs to be clearly defined what drug is studied, what the target population is, and when they should be tested. To support this contextual thinking, experts from health technology assessment could be included in early phases of research. This will offer opportunities to make recommendations on the steps from clinical validity to clinical utility, and for example include recommendation to study cost per QALY. Furthermore, to have an incentive for fundamental researchers to think about these steps, journals and peer reviewers can easily ask for measures that can directly be derived from measures for association. As Tonk et al.  describeMJ contributed to the analysis and interpretation of data, drafting the work and revising it critically. TR, WR, and MC contributed to the conception of the work, analysis and interpretation of data, and revising the work critically. TF, MW contributed to the analysis and interpretation of data, and revising the work critically. EH contributed to the conception of the work, and the data analysis. All authors approved the version to be published.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.