All of whom? Limitations encountered using All of Us Researcher Workbench in a Primary Care residents secondary data analysis research training block.

OBJECTIVES: The goal of this case report is to detail experiences and challenges experienced in the training of Primary Care residents in secondary analysis using All of Us Researcher Workbench. At our large, urban safety net hospital, Primary Care/Internal Medicine residents in their third year undergo a research intensive block, the Research Practicum, where they work as a team to conduct secondary data analysis on a dataset with faculty facilitation. In 2023, this research block focused on use of the All of Us Researcher Workbench for secondary data analysis. MATERIALS AND METHODS: Two groups of 5 residents underwent training to access the All of Us Researcher Workbench, and each group explored available data with a faculty facilitator and generated original research questions. Two blocks of residents successfully completed their research blocks and created original presentations on "social isolation and A1C" levels and "medical discrimination and diabetes management." RESULTS: Departmental faculty were satisfied with the depth of learning and data exploration. In focus groups, some residents noted that for those without interest in performing research, the activity felt extraneous to their career goals, while others were glad for the opportunity to publish. In both blocks, residents highlighted dissatisfaction with the degree to which the All of Us Researcher Workbench was representative of patients they encounter in a large safety net hospital. DISCUSSION: Using the All of Us Researcher Workbench provided residents with an opportunity to explore novel questions in a massive data source. Many residents however noted that because the population described in the All of Us Researcher Workbench appeared to be more highly educated and less racially diverse than patients they encounter in their practice, research may be hard to generalize in a community health context. Additionally, given that the data required knowledge of 1 of 2 code-based data analysis languages (R or Python) and work within an idiosyncratic coding environment, residents were heavily reliant on a faculty facilitator to assist with analysis. CONCLUSION: Using the All of Us Researcher Workbench for research training allowed residents to explore novel questions and gain first-hand exposure to opportunities and challenges in secondary data analysis.

Fentanyl Test Strips for Harm Reduction: A Scoping Review.

BACKGROUND: High potency synthetic opioids like fentanyl have continued to replace or contaminate the supply of illicit drugs in North America, with fentanyl test strips (FTSs) often used as a harm reduction tool for overdose prevention. The available evidence to support FTS for harm reduction has yet to be summarized. METHODS: A search of PubMed, Ovid Embase, and Web of Science was conducted in March 2023. A 2-stage review was conducted to screen by title and abstract and then by full text by 2 reviewers. Data were extracted from each study using a standardized template. RESULTS: A total of 91 articles were included, mostly from North America, predominantly reporting on FTS along with other harm reduction tools, and all conducted after 2016. No randomized controlled trials are reported. Robust evidence exists supporting the sensitivity and specificity of FTS, along with their acceptability and feasibility of use for people who use drugs and as a public health intervention. However, limited research is available on the efficacy of FTS as a harm reduction tool for behavior change, engagement in care, or overdose prevention. CONCLUSIONS: Though FTSs are highly sensitive and specific for point of care testing, further research is needed to assess the association of FTS use with overdose prevention. Differences in FTS efficacy likely exist between people who use opioids and nonopioid drugs, with additional investigation strongly needed. As drug testing with point-of-care immunoassays is embraced for nonfentanyl contaminants such as xylazine and benzodiazepines, increased investment in examining overdose prevention is necessary.

Efficient and Accurate Analog Voltage Measurement Using a Direct Sensor-to-Digital Port Interface for Microcontrollers and Field-Programmable Gate Arrays.

Portable sensor systems are usually based on microcontrollers and/or Field-Programmable Gate Arrays (FPGAs) that are interfaced with sensors by means of an Analog-to-Digital converter (ADC), either integrated in the computing device or external. An alternative solution is based on the direct connection of the sensors to the digital input port of the microcontroller or FPGA. This solution is particularly interesting in the case of devices not integrating an internal ADC or featuring a small number of ADC channels. In this paper, a technique is presented to directly interface sensors with analog voltage output to the digital input port of a microcontroller or FPGA. The proposed method requires only a few passive components and is based on the measurements of the duty cycle of a digital square-wave signal. This technique was investigated by means of circuit simulations using LTSpice and was implemented in a commercial low-cost FPGA device (Gowin GW1NR-9). The duty cycle of the square-wave signal features a good linear correlation with the analog voltage to be measured. Thus, a look-up table to map the analog voltage values to the measured duty cycle is not required with benefits in terms of memory occupation. The experimental results on the FPGA device have shown that the analog voltage can be measured with a maximum accuracy of 1.09 mV and a sampling rate of 9.75 Hz. The sampling rate can be increased to 31.35 Hz and 128.31 Hz with an accuracy of 1.61 mV and 2.68 mV, respectively.

Field-Deployable Determinations of Peroxide Index and Total Phenolic Content in Olive Oil Using a Promising Portable Sensor System.

Useful information about the oxidative stability of a virgin olive oil in terms of oxidation products and antioxidant compounds can be obtained by analyzing the peroxide index (PI) and total phenolic content (TPC), respectively. These quality parameters are usually determined in a chemical laboratory using expensive equipment, toxic solvents, and well-trained personnel. This paper presents a novel portable sensor system for in the field and rapid determination of PI and TPC that is particularly suited in the case of small production environments that cannot afford an internal laboratory for quality control analysis. The system is small, can be powered by both USB ports and batteries, is easy to operate, and integrates a Bluetooth module for wireless data transmission. It estimates the PI and TPC in olive oil from the measurement of the optical attenuation of an emulsion between a reagent and the sample under test. The system has been tested on a set of 12 olive oil samples (eight for calibration and four for validation), and the results have shown how the considered parameters can be estimated with good accuracy. The maximum deviation from the results obtained with the reference analytical techniques is 4.7 meq O2/kg in the case of PI and 45.3 ppm in the case of TPC for the calibration set, while it is 14.8 meq O2/kg in the case of PI and 55 ppm in the case of TPC for the validation set.

Computer Vision Approach for the Determination of Microbial Concentration and Growth Kinetics Using a Low Cost Sensor System.

The measurement of microbial contamination is of primary importance in different fields, from environmental monitoring to food safety and clinical analysis. Today, almost all microbiology laboratories make microbial concentration measurements using the standard Plate Count Technique (PCT), a manual method that must be performed by trained personnel. Since manual PCT analysis can result in eye fatigue and errors, in particular when hundreds of samples are processed every day, automatic colony counters have been built and are commercially available. While quick and reliable, these instruments are generally expensive, thus, portable colony counters based on smartphones have been developed and are of low cost but also not accurate as the commercial benchtop instruments. In this paper, a novel computer vision sensor system is presented that can measure the microbial concentration of a sample under test and also estimate the microbial growth kinetics by monitoring the colonies grown on a Petri dish at regular time intervals. The proposed method has been in-house validated by performing PCT analysis in parallel under the same conditions and using these results as a reference. All the measurements have been carried out in a laboratory using benchtop instruments, however, such a system can also be realized as an embedded sensor system to be deployed for microbial analysis outside a laboratory environment.

RGD conjugated cell uptake off to on responsive NIR-AZA fluorophores: applications toward intraoperative fluorescence guided surgery.

The use of NIR-fluorescence imaging to demarcate tumour boundaries for real-time guidance of their surgical resection has a huge untapped potential. However, fluorescence imaging using molecular fluorophores, even with a targeting biomolecule attached, has a major shortcoming of signal interference from non-specific background fluorescence outside the region of interest. This poor selectivity necessitates prolonged time delays to allow clearance of background fluorophore and retention within the tumour prior to image acquisition. In this report, an innovative approach to overcome this issue is described in which cancer targeted off to on bio-responsive NIR-fluorophores are utilised to switch-on first within the tumour. Bio-responsive cRGD, iRGD and PEG conjugates have been synthesised using activated ester/amine or maleimide/thiol couplings to link targeting and fluorophore components. Their off to on emission responses were measured and compared with an always-on non-responsive control with each bio-responsive derivative showing large fluorescence enhancement values. Live cell imaging experiments using metastatic breast cancer cells confirmed in vitro bio-responsive capabilities. An in vivo assessment of MDA-MB 231 tumour imaging performance for bio-responsive and always-on fluorophores was conducted with monitoring of fluorescence distributions over 96 h. As anticipated, the always-on fluorophore gave an immediate, non-specific and very strong emission throughout whereas the bio-responsive derivatives initially displayed very low fluorescence. All three bio-responsive derivatives switched on within tumours at time points consistent with their conjugated targeting groups. cRGD and iRGD conjugates both had effective tumour turn-on in the first hour, though the cRGD derivative had superior specificity for tumour over the iRGD conjugate. The pegylated derivative had similar switch-on characteristics but over a much longer period, taking 9 h before a significant emission was observable from the tumour. Evidence for in vivo active tumour targeting was obtained for the best performing cRGD bio-responsive NIR-AZA derivative from competitive binding studies. Overall, this cRGD-conjugate has the potential to overcome the inherent drawback of targeted always-on fluorophores requiring prolonged clearance times and shows excellent potential for clinical translation for intraoperative use in fluorescence guided tumour resections.

Lysosome triggered near-infrared fluorescence imaging of cellular trafficking processes in real time.

Bioresponsive NIR-fluorophores offer the possibility for continual visualization of dynamic cellular processes with added potential for direct translation to in vivo imaging. Here we show the design, synthesis and lysosome-responsive emission properties of a new NIR fluorophore. The NIR fluorescent probe design differs from typical amine functionalized lysosomotropic stains with off/on fluorescence switching controlled by a reversible phenol/phenolate interconversion. Emission from the probe is shown to be highly selective for the lysosomes in co-imaging experiments using a HeLa cell line expressing the lysosomal-associated membrane protein 1 fused to green fluorescent protein. The responsive probe is capable of real-time continuous imaging of fundamental cellular processes such as endocytosis, lysosomal trafficking and efflux in 3D and 4D. The advantage of the NIR emission allows for direct translation to in vivo tumour imaging, which is successfully demonstrated using an MDA-MB-231 subcutaneous tumour model. This bioresponsive NIR fluorophore offers significant potential for use in live cellular and in vivo imaging, for which currently there is a deficit of suitable molecular fluorescent tools.

NIR fluorescence labelled carbon nano-onions: synthesis, analysis and cellular imaging.

The preparation of novel NIR fluorescent carbon based nanomaterials, consisting of boron difluoride azadipyrromethene fluorophores covalently attached to carbon nano-onions, is demonstrated. In addition, the analysis of the new nanomaterial is presented. The fluorescent nano-derivative properties are customized such that their emission can be reversibly on/off modulated in response to pH, which is demonstrated in solution and in cells. The in vitro imaging of HeLa Kyoto cells is carried out and the cellular uptake of the carbon nano-onion NIR fluorophore conjugates is verified.

Mechanistic insight into the formation of tetraarylazadipyrromethenes.

The tetraarylazadipyrromethene chromophore class has gained increasing attention in the past decade for a diverse set of scientific interests and applications. The most direct synthetic route available for their generation is heating of 4-nitro-1,3-diarylbutan-1-ones with an ammonium source in an alcohol solvent. Despite the practical simplicity, the reaction pathway(s) for these conversions are lengthy and unclear. To gain insight into the steps involved, (15)N labeling experiments with MS and NMR analysis were utilized for conversion of 4-nitro-1,3-diphenylbutan-1-one 1 into tetraphenylazadipyrromethene 2 with (15)NH(4)OAc. To permit examination of later stages of the reaction sequence to 2, the (15)N-labeled potential intermediate 3,5-diphenyl-1H-pyrrol-2-amine 10 was synthesized. A study of the dimerization pathway utilizing (15)N-labeled 10 revealed an unprecedented nitrogen rearrangement in the final stages of the pathway involving a ring-opening/closing of a pyrrole ring. Utilizing (15)N labeling experiments we have shown that 2,4-diphenylpyrrole 8 can also react under the reaction conditions with 3,5-diphenyl-2H-pyrrol-2-imine 7 (from oxidation of 10) to produce 2. Overall in the conversion of 1 into 2, two related pathways are ongoing concurrently; the first involves a dimerization of 3,5-diphenyl-2H-pyrrol-2-imine 7, and the other a reaction of 7 with 2,4-diphenylpyrrole 8.

Cellular uptake mediated off/on responsive near-infrared fluorescent nanoparticles.

Fluorescence imaging, utilizing molecular fluorophores, often acts as a central tool for the investigation of fundamental biological processes and offers huge future potential for human imaging coupled to therapeutic procedures. An often encountered limitation with fluorescence imaging is the difficulty in discriminating nonspecific background fluorophore emission from a fluorophore localized at a specific region of interest. This limits imaging to individual time points at which background fluorescence has been minimized. It would be of significant advantage if the fluorescence output could be modulated from off to on in response to specific biological events as this would permit imaging of such events in real time without background interference. Here we report our approach to achieve this for the most fundamental of cellular processes, i.e. endocytosis. We describe a new near-infrared off to on fluorescence switchable nanoparticle construct that is capable of switching its fluorescence on following cellular uptake but remains switched off in extracellular environments. This permits continuous real-time imaging of the uptake process as extracellular particles are nonfluorescent. The principles behind the fluorescence off/on switch can be understood by encapsulation of particles in cellular organelles which effect a microenvironmental change establishing a fluorescence signal.

Light induced antimicrobial properties of a brominated boron difluoride (BF(2)) chelated tetraarylazadipyrromethene photosensitizer.

Herein we describe a new antimicrobial photodynamic therapeutic (PDT) agent based upon the brominated BF(2) chelated tetraarylazadipyrromethene photosensitizer class. Bis-ammonium salt substitution of the photosensitizer promoted a rapid 10 min uptake into Gram-positive and -negative bacterial strains and pathogenic yeasts. A photosensitizer and light dose response analysis for methicillin-sensitive S. aureus showed an impressive antibacterial efficacy with 1, 2, and 5 µg/mL 6. Specifically, light activation with a dose of 16 J/cm(2) and 5 µg/mL 6 resulted in a 6.8 and 3.4 log(10) reduction of S. aureus and a clinically defined methicillin-resistant Staphylococcus aureus (MRSA) strain, respectively. Encouragingly, a broad spectrum pathogen response (using 5 µg/mL 6 and 75 J/cm(2)) was observed with 3.6 and 5.7 log(10) decreases in viable cell numbers achievable for Gram-negative bacterium E. coli and the pathogenic yeast C. albicans, respectively. The photophysical and cell eradicating characteristics of this bis-cationic PDT agent suggest that it has broad potential in antimicrobial therapeutics.