A blueprint for success in real-world evidence: "glocal" approach to building capabilities and generating impactful evidence.

The past decade has seen the increasing influence and relevance of real-world data (RWD) and real-world evidence (RWE) in healthcare decision making. The value added by RWD/RWE has prompted the pharmaceutical industry to develop high performing systems and practices to harness the power of evidence generated at the global level. However, this worldwide transformation provides outstanding opportunities to support capability building within local affiliates and to impact key country-level stakeholders through resulting evidence. Therefore, we present an Evidence Blueprint Initiative, which links the global and local ("glocal") skills, and furthermore addresses the opportunities and gaps in evidence generation capabilities at the local level. Cross-functional experts were recruited at the local, regional, and global level to define best practices. A framework was developed to characterize the foundational expertise needed and to assess markets' existing capabilities. Subsequently, targeted roadmaps were developed and implemented to build capabilities in specific areas within each affiliate. The impact from the Blueprint is encouraging, resulting in improved local evidence plans, established evidence teams, enhanced RWD use and strategic implementation of patient centric science in local affiliates. The success of the Blueprint resides in empowering affiliates to realise their local evidence generation ambitions and to match them to their local context. It strengthens and expands the ties between various parts of the organisation and the external environment while building fit-for-future evidence capabilities from local affiliates.

Miniaturized Microscale Solid-Phase Extraction-Based Module for Highly Efficient DNA Extraction and Purification from Grapevine Samples.

Several developments over the last few years are being directed toward improving DNA-based analysis to simplify, miniaturize, and reduce the time and cost of analysis, with the objective to allow its use in decentralized settings. One of the most interesting fields is DNA extraction and purification, a key step for ensuring good analytical performance. In this sense, microscale solid phase extraction (µSPE) offers paramount advantages for an improved DNA yield. In this work, we have developed a miniaturized module for DNA purification based on µSPE using a borosilicate glass microfiber filter as the solid phase. We also established a protocol for highly efficient DNA purification from vegetable samples, including leaves and grapes from four different varieties from the PDO Douro and two varieties from the Minho wine regions. The protocol demonstrated excellent performance when compared with a commercial kit with a DNA recovery yield of around 50%.

Dual colorimetric strategy for specific DNA detection by nicking endonuclease-assisted gold nanoparticle signal amplification.

The continuous spread of invasive alien species, as zebra mussel (Dreissena polymorpha), is a major global concern and it is urgent to stop it. Early stages of an invasion are crucial and challenging; however, detection tools based on environmental DNA analysis are promising alternatives. We present an alternative DNA target amplification strategy for signal enhancement followed by dual-mode colorimetric naked eye and optical smartphone analysis for the early detection of zebra mussel environmental DNA. Target amplification was designed based on the nicking endonuclease probe cleavage upon probe and complementary target hybridization. The cleaved/intact probe interacts with DNA-modified nanoparticles for colorimetric detection. We have demonstrated that enzyme amplification strategy enhanced 12-fold the sensitivity by naked eye detection, achieving a detection limit of ~8 nM (4.48×1010 copies) in controlled conditions, whereas target in complex environmental samples allowed the detection of 22.5 nM (1.26×1011 copies). Competitive assays also showed that the system can discriminate specific zebra mussel DNA sequences from other DNA sequences. Additionally, smartphone analysis for DNA quantification further improved the sensitivity of its detection by 130-fold, more than 2 orders of magnitude, when applied to environmental samples. The limit of detection to 0.17 nM (9.52×108 copies) is based on RGB coordinates, which is especially relevant to monitor early aggregation stages, being more accurate and reducing naked eye detection subjectivity. DNA extracted from zebra mussel meat, zebra mussel contaminated river water, and non-contaminated river water samples were successfully tested. Dual-mode colorimetric detection is useful in field analysis without the need for expensive laboratory equipment.

The 7+ Habits of Highly Effective Medical Directors.

Medical Directors in the pharmaceutical industry are responsible for fulfilling multiple roles, including scientific expert, conscience keeper and people leader, as well as being the external face of the organisation. The capabilities described in the book The 7 Habits of Highly Effective People® are not just the integral qualities of a successful leader, they are important capabilities that are critical to becoming an impactful medical leader too. As scientific, functional and enterprise leaders, country and cluster Medical Directors must possess these seven qualities. Each of the seven habits can be adapted and utilised to shape the specific areas that Medical Directors are responsible for: intuitive creativity, patient centricity, prioritisation, enterprise leadership, communication and behavioural change, building a team, partnering and scientific and technological leadership. To excel in their function, current and future Medical Directors need to sharpen their existing skills and build specific capabilities. Energy management in physical, mental, emotional and spiritual domains is the way forward.

Amplified plasmonic and microfluidic setup for DNA monitoring.

Plasmonic nanosensors for label-free detection of DNA require excellent sensing resolution, which is crucial when monitoring short DNA sequences, as these induce tiny peak shifts, compared to large biomolecules. We report a versatile and simple strategy for plasmonic sensor signal enhancement by assembling multiple (four) plasmonic sensors in series. This approach provided a fourfold signal enhancement, increased signal-to-noise ratio, and improved sensitivity for DNA detection. The response of multiple sensors based on AuNSpheres was also compared with  AuNRods, the latter showing better sensing resolution. The amplification system based on AuNR was integrated into  a microfluidic sequential injection platform and applied to the monitoring of DNA, specifically from environmental invasive species-zebra mussels. DNA from zebra mussels was log concentration-dependent from 1 to 1 × 106 pM, reaching a detection limit of 2.0 pM. In situ tests were also successfully applied to real samples, within less than 45 min, using DNA extracted from zebra mussel meat. The plasmonic nanosensors' signal can be used as a binary output (yes/no) to assess the presence of those invasive species. Even though these genosensors were applied to the monitoring of DNA in environmental samples, they potentially offer advantage in a wide range of fields, such as disease diagnostics.

Single-use microfluidic device for purification and concentration of environmental DNA from river water.

Purification and concentration of DNA is a critical step on DNA-based analysis, which should ensure efficient DNA isolation and effective removal of contaminants that may interfere with downstream DNA amplification. Complexity of samples, minute content of target analyte, or high DNA fragmentation greatly entangles the success of this step. To overcome this issue, we designed and fabricated a novel miniaturized disposable device for a highly efficient DNA purification. The microfluidic device showed binding efficiency and elution yield of 90.1% and 86.7%, respectively. Moreover, the effect of DNA fragmentation, a parameter that has not been previously addressed, showed a great impact in the recovery step. The microfluidic system integrated micropillars with chitosan being used as the solid-phase for a pH-dependent DNA capture and release. We have showed the potential of the device in the successful purification of environmental DNA (eDNA) from river water samples contaminated with Dreissena polymorpha, an invasive alien species responsible for unquestionable economic and environmental consequences in river water basins. Additionally, the device was also able to concentrate the DNA extract from highly diluted samples, showing promising results for the early detection of such invasive species, which may allow prompt measures for a more efficient control in affected areas. Suitability for integration with downstream DNA analysis was also demonstrated through qPCR analysis of the samples purified with the microfluidic device, allowing detection of the target species even if highly diluted.

Influence of the Electrolyte Salt Concentration on DNA Detection with Graphene Transistors.

Liquid-gated Graphene Field-Effect Transistors (GFET) are ultrasensitive bio-detection platforms carrying out the graphene's exceptional intrinsic functionalities. Buffer and dilution factor are prevalent strategies towards the optimum performance of the GFETs. However, beyond the Debye length (λD), the role of the graphene-electrolytes' ionic species interactions on the DNA behavior at the nanoscale interface is complicated. We studied the characteristics of the GFETs under different ionic strength, pH, and electrolyte type, e.g., phosphate buffer (PB), and phosphate buffer saline (PBS), in an automatic portable built-in system. The electrostatic gating and charge transfer phenomena were inferred from the field-effect measurements of the Dirac point position in single-layer graphene (SLG) transistors transfer curves. Results denote that λD is not the main factor governing the effective nanoscale screening environment. We observed that the longer λD was not the determining characteristic for sensitivity increment and limit of detection (LoD) as demonstrated by different types and ionic strengths of measuring buffers. In the DNA hybridization study, our findings show the role of the additional salts present in PBS, as compared to PB, in increasing graphene electron mobility, electrostatic shielding, intermolecular forces and DNA adsorption kinetics leading to an improved sensitivity.

Plasmon Resonance in Photoabsorption of Colloidal Highly Doped ZnO Nanocrystals.

A new type of dipole plasmon excitations in colloidal highly doped ZnO nanocrystals has been studied by means of many-body quantum mechanical approach. We demonstrate that in photodoped ZnO nanocrystals, the conduction band electrons are localized close to the surface and the plasmon oscillations are induced by their angular motion. The transition of this plasmon mode from classical to quantum regime is defined by the nanocrystal size. The size dependence of the resonance frequency which results from quantum effects is in remarkable agreement with experimental observations.

An impedimetric chemical sensor for determination of detergents residues.

A new impedimetric sensor based on an interdigitated electrode array with electrode digits located at the bottom of microcapillaries formed in silicon dioxide is presented. Microcapillaries are opened at the top, so that in contact with an electrolyte solution the ac current flows close to the surface of the capillary wall from one electrode to another and is significantly affected by changes in the surface conductance at the SiO2/electrolyte interface. Adsorption of detergents on the sensor surface affects the charge distribution in the electrical double layer and thus the surface conductance. These changes are registered by measuring impedance. Effect of surface adsorption of ionic and non-ionic surfactants on the sensor impedance is studied. The sensor is shown to be able to measure commercial detergents residues in a tap water starting from 5 ppm even in solutions with high electrolyte conductivity.

Application of an e-tongue to the analysis of monovarietal and blends of white wines.

This work presents a multiparametric system capable of characterizing and classifying white wines according to the grape variety and geographical origin. Besides, it quantifies specific parameters of interest for quality control in wine. The system, known as a hybrid electronic tongue, consists of an array of electrochemical microsensors-six ISFET based sensors, a conductivity sensor, a redox potential sensor and two amperometric electrodes, a gold microelectrode and a microelectrode for sensing electrochemical oxygen demand--and a miniaturized optofluidic system. The test sample set comprised eighteen Catalan monovarietal white wines from four different grape varieties, two Croatian monovarietal white wines and seven bi- and trivarietal mixtures prepared from the Catalan varieties. Different chemometric tools were used to characterize (i.e., Principal Component Analysis), classify (i.e., Soft Independent Modeling Class Analogy) and quantify (i.e., Partial-Least Squares) some parameters of interest. The results demonstrate the usefulness of the multisensor system for analysis of wine.

Integrated multi-sensor chip with photocured polymer membranes containing copolymerized plasticizer for direct pH, potassium, sodium and chloride ions determination in blood serum.

An analytical system based on a sensor array with ion-selective field effect transistors (ISFETs) monolithically integrated in one chip covered with photocured polymer membranes containing copolymerized plasticizer and a sequential injection analysis (SIA) is shown to offer an automation of the analysis of blood serum components. For sequential injection system a custom made dual channel flow cell for the sensor array was developed. Optimisation of ion-sensitive membrane characteristics and calibration solution compositions were carried out. The system was used to analyze sodium, potassium, chloride ion contents in blood serum samples. The precision of the ion determination in samples was typical for potentiometric method with standard deviation of about 3-5%.

Qualitatively Correct Charge-Transfer Excitation Energies in HeH(+) by Time-Dependent Density-Functional Theory Due to Exact Exchange Kohn-Sham Eigenvalue Differences.

Time-dependent density-functional theory in the response regime is shown to yield qualitatively correct charge-transfer excitation energies in the system HeH(+) if the exact Kohn-Sham exchange potential is employed to determine the Kohn-Sham orbitals and eigenvalues entering the time-dependent density-functional calculation. The employed exact-exchange kernel is frequency-independent and, like conventional kernels in the local density approximation or in generalized gradient approximations, does not contribute to the charge-transfer excitation energy. This shows that it can be that not the exchange-correlation kernel, as generally believed, but the exchange-correlation potential plays the crucial role in the description of charge-transfer excitations.

Electronic integrated multisensor tongue applied to grape juice and wine analysis.

An integrated multisensor composed by six ISFET devices selective to common ions and heavy metals combined with a flow injection analysis (FIA) system has been applied as an electronic tongue to grape juice and wine sample analysis. The data obtained for several grape and wine variety samples analysis have been treated using multiparametric tools like principal component analysis (PCA) and soft independent modelling class analogy technique (SIMCA) for the patterning recognition and classification of samples and partial least squares (PLS) regression for quantification of several parameters of interest in wine production. The results obtained have demonstrated the potential of using those multisensors as electronic tongues not only for distinguishing the samples according to the grape variety and the vintage year but also for quantitative prediction of several sample parameters.

Determination of cyanide using flow-injection multisensor system.

A flow-injection multisensor system (FIMS) comprising potentiometric sensors of different types for determination of free cyanide activity in basic solutions for extraction of noble metals was developed. The solvent polymeric membrane sensors based on metalloporphyrin and crystalline sensors were combined in the sensor system. The system allowed determination of cyanide activity in the range 10(-4)-1 mol l(-1) with an error less than 5% in individual cyanide solutions and acceptable precision (about 20%) in process liquids. The system was able to analyse up to 20 samples per h. The FIMS was also applied to detecting of silver ions in the presence of cyanide. Chalcogenide glass sensor was used as the detector that ensured the precision of 20%.