Open-source electrochemical cell for in situ X-ray absorption spectroscopy in transmission and fluorescence modes.

X-ray spectroscopy is a valuable technique for the study of many materials systems. Characterizing reactions in situ and operando can reveal complex reaction kinetics, which is crucial to understanding active site composition and reaction mechanisms. In this project, the design, fabrication and testing of an open-source and easy-to-fabricate electrochemical cell for in situ electrochemistry compatible with X-ray absorption spectroscopy in both transmission and fluorescence modes are accomplished via windows with large opening angles on both the upstream and downstream sides of the cell. Using a hobbyist computer numerical control machine and free 3D CAD software, anyone can make a reliable electrochemical cell using this design. Onion-like carbon nanoparticles, with a 1:3 iron-to-cobalt ratio, were drop-coated onto carbon paper for testing in situ X-ray absorption spectroscopy. Cyclic voltammetry of the carbon paper showed the expected behavior, with no increased ohmic drop, even in sandwiched cells. Chronoamperometry was used to apply 0.4 V versus reversible hydrogen electrode, with and without 15 min of oxygen purging to ensure that the electrochemical cell does not provide any artefacts due to gas purging. The XANES and EXAFS spectra showed no differences with and without oxygen, as expected at 0.4 V, without any artefacts due to gas purging. The development of this open-source electrochemical cell design allows for improved collection of in situ X-ray absorption spectroscopy data and enables researchers to perform both transmission and fluorescence simultaneously. It additionally addresses key practical considerations including gas purging, reduced ionic resistance and leak prevention.

Promotores' perspectives on the virtual adaptation of a hereditary breast cancer education program.

Breast cancer is the most common cancer in women in the United States (U.S.) and the leading cause of cancer related death among U.S. Hispanics/Latinas (H/Ls). H/Ls have lower rates of screening and longer time to follow up after an abnormal mammogram. We developed a comprehensive community health educator (promotores)-led education and risk identification program for Spanish-speaking H/Ls in California to increase mammography screening, genetic testing, and the understanding of the impact of family history on cancer risk. Due to COVID-19, we adapted the program to a virtual platform. The experience of transforming the program to a virtual platform provided unique opportunities for collaboration between researchers, community partners, and participants. Promotores are major partners in community based participatory research and in the provision of health care services, but their voices are often excluded from scientific reports. This commentary is an effort to provide a platform for promotores to share their experiences and for the readers to understand their approach in bridging the gap between health care services and communities.

Cysteine Oxidation in Proteins: Structure, Biophysics, and Simulation.

Cysteine side chains can exist in distinct oxidation states depending on the pH and redox potential of the environment, and cysteine oxidation plays important yet complex regulatory roles. Compared with the effects of post-translational modifications such as phosphorylation, the effects of oxidation of cysteine to sulfenic, sulfinic, and sulfonic acid on protein structure and function remain relatively poorly characterized. We present an analysis of the role of cysteine reactivity as a regulatory factor in proteins, emphasizing the interplay between electrostatics and redox potential as key determinants of the resulting oxidation state. A review of current computational approaches suggests underdeveloped areas of research for studying cysteine reactivity through molecular simulations.

Thermodynamic stabilization of von Willebrand factor A1 domain induces protein loss of function.

The von Willebrand disease (vWD) is the most common hereditary bleeding disorder caused by defects of the von Willebrand Factor (vWF), a large extracellular protein in charge of adhering platelets to sites of vascular lesions. vWF performs this essential homeostatic task via specific protein-protein interactions between the vWF A1 domain and the platelet receptor, the glycoprotein Ib alpha (GPIBα). The two naturally occurring vWF A1 domain mutations G1324A and G1324S, near the GPIBα binding site, induce a dramatic decrease in platelet adhesion, resulting in a bleeding disorder classified as type 2M vWD. However, the reason for the drastic phenotypic response induced by these two supposedly minor modifications remains unclear. We addressed this question using a combination of equilibrium-molecular dynamics (MD) and nonequilibrium MD-based free energy simulations. Our data confirms that both mutations maintain the highly stable Rossmann fold of the vWF A1 domain. G1324A and G1324S mutations hardly changed the per-residue flexibility of the A1 domain but induced a global conformational change affecting the region near the binding site to GPIBα. Furthermore, we observed two significant changes in the vWF A1 domain upon mutation, the global redistribution of the internal mechanical stress and the increased thermodynamic stability of the A1 domain. These observations are consistent with previously reported mutations increasing the melting temperature. Overall, our results support the idea of thermodynamic conformational restriction of A1-before the binding to GPIBα-as a crucial factor determining the loss-of-function of the G1324A(S) vWD mutants.

DNA binds to a specific site of the adhesive blood-protein von Willebrand factor guided by electrostatic interactions.

Neutrophils release their intracellular content, DNA included, into the bloodstream to form neutrophil extracellular traps (NETs) that confine and kill circulating pathogens. The mechanosensitive adhesive blood protein, von Willebrand Factor (vWF), interacts with the extracellular DNA of NETs to potentially immobilize them during inflammatory and coagulatory conditions. Here, we elucidate the previously unknown molecular mechanism governing the DNA-vWF interaction by integrating atomistic, coarse-grained, and Brownian dynamics simulations, with thermophoresis, gel electrophoresis, fluorescence correlation spectroscopy (FCS), and microfluidic experiments. We demonstrate that, independently of its nucleotide sequence, double-stranded DNA binds to a specific helix of the vWF A1 domain, via three arginines. This interaction is attenuated by increasing the ionic strength. Our FCS and microfluidic measurements also highlight the key role shear-stress has in enabling this interaction. Our simulations attribute the previously-observed platelet-recruitment reduction and heparin-size modulation, upon establishment of DNA-vWF interactions, to indirect steric hindrance and partial overlap of the binding sites, respectively. Overall, we suggest electrostatics-guiding DNA to a specific protein binding site-as the main driving force defining DNA-vWF recognition. The molecular picture of a key shear-mediated DNA-protein interaction is provided here and it constitutes the basis for understanding NETs-mediated immune and hemostatic responses.

Metodologías para la priorización en investigación en salud: una revisión sistemática de la literatura.

OBJETIVO: Identificar elementos metodológicos clave para la priorización en investigación en salud, a partir de las metodologías reportadas en la literatura científica. MÉTODOS: Se realizó una búsqueda sistemática en Medline, Embase, LILACS, y fuentes complementarias de literatura gris. Se utilizaron las palabras clave: research, methods y health priorities, en combinación con términos libres. Dos revisores independientes, de acuerdo con criterios previamente definidos, seleccionaron revisiones de la literatura o documentos metodológicos que presentaran metodologías para priorización en investigación en salud. Se extrajeron las principales características de las metodologías reportadas y se identificaron elementos comunes. RESULTADOS: Se incluyeron siete revisiones y cinco documentos metodológicos, que reportaron cuatro metodologías estructuradas específicas y múltiples aproximaciones metodológicas que combinan elementos diversos. En general, estas metodologías integran la perspectiva de actores clave con información objetiva, mediante la aplicación de técnicas estandarizadas de participación, para establecer un ranking de prioridades, con base en criterios previamente definidos. Se identificaron elementos metodológicos comunes relacionados con pasos del proceso, mecanismos de participación, criterios para priorizar y análisis de resultados. CONCLUSIÓN: La priorización en investigación en salud requiere el empleo de una metodología definida a priori, que debe contener como mínimo cuatro elementos clave: pasos claros del proceso, criterios para priorizar, técnicas formales de participación y métodos de análisis de resultados. Estos elementos deben ajustarse a las condiciones y necesidades del contexto de aplicación.

Mesenchymal stromal cells express GARP/LRRC32 on their surface: effects on their biology and immunomodulatory capacity.

Mesenchymal stromal cells (MSCs) represent a promising tool for therapy in regenerative medicine, transplantation, and autoimmune disease due to their trophic and immunomodulatory activities. However, we are still far from understanding the mechanisms of action of MSCs in these processes. Transforming growth factor (TGF)-ß1 is a pleiotropic cytokine involved in MSC migration, differentiation, and immunomodulation. Recently, glycoprotein A repetitions predominant (GARP) was shown to bind latency-associated peptide (LAP)/TGF-ß1 to the cell surface of activated Foxp3(+) regulatory T cells (Tregs) and megakaryocytes/platelets. In this manuscript, we show that human and mouse MSCs express GARP which presents LAP/TGF-ß1 on their cell surface. Silencing GARP expression in MSCs increased their secretion and activation of TGF-ß1 and reduced their proliferative capacity in a TGF-ß1-independent manner. Importantly, we showed that GARP expression on MSCs contributed to their ability to inhibit T-cell responses in vitro. In summary, we have found that GARP is an essential molecule for MSC biology, regulating their immunomodulatory and proliferative activities. We envision GARP as a new target for improving the therapeutic efficacy of MSCs and also as a novel MSC marker.

Quantifying efficacy and limits of unmanned aerial vehicle (UAV) technology for weed seedling detection as affected by sensor resolution.

In order to optimize the application of herbicides in weed-crop systems, accurate and timely weed maps of the crop-field are required. In this context, this investigation quantified the efficacy and limitations of remote images collected with an unmanned aerial vehicle (UAV) for early detection of weed seedlings. The ability to discriminate weeds was significantly affected by the imagery spectral (type of camera), spatial (flight altitude) and temporal (the date of the study) resolutions. The colour-infrared images captured at 40 m and 50 days after sowing (date 2), when plants had 5-6 true leaves, had the highest weed detection accuracy (up to 91%). At this flight altitude, the images captured before date 2 had slightly better results than the images captured later. However, this trend changed in the visible-light images captured at 60 m and higher, which had notably better results on date 3 (57 days after sowing) because of the larger size of the weed plants. Our results showed the requirements on spectral and spatial resolutions needed to generate a suitable weed map early in the growing season, as well as the best moment for the UAV image acquisition, with the ultimate objective of applying site-specific weed management operations.

Nicotinamide inhibits the lysosomal cathepsin b-like protease and kills African trypanosomes.

Nicotinamide, a soluble compound of the vitamin B3 group, has antimicrobial activity against several microorganisms ranging from viruses to parasite protozoans. However, the mode of action of this antimicrobial activity is unknown. Here, we investigate the trypanocidal activity of nicotinamide on Trypanosoma brucei, the causative agent of African trypanosomiasis. Incubation of trypanosomes with nicotinamide causes deleterious defects in endocytic traffic, disruption of the lysosome, failure of cytokinesis, and, ultimately, cell death. At the same concentrations there was no effect on a cultured mammalian cell line. The effects on endocytosis and vesicle traffic were visible within 3 h and can be attributed to inhibition of lysosomal cathepsin b-like protease activity. The inhibitory effect of nicotinamide was confirmed by a direct activity assay of recombinant cathepsin b-like protein. Taken together, these data demonstrate that inhibition of the lysosomal protease cathepsin b-like blocks endocytosis, causing cell death. In addition, these results demonstrate for the first time the inhibitory effect of nicotinamide on a protease.

Clinical outcomes of a toric continuous range of vision presbyopia-correcting intraocular lens.

PURPOSE: To evaluate the clinical outcomes of the toric version of a presbyopia-correcting intraocular lens (IOL) based on the combination of a diffractive-based extended depth-of-focus (EDOF) pattern and a diffractive multifocal platform. SETTING: Miranza COI Bilbao, Bilbao, Spain. DESIGN: Prospective case series. METHODS: 35 patients (51 to 84 years) with corneal astigmatism ranging from 0.75 to 2.19 diopters (D) undergoing bilateral cataract surgery with implantation of the Synergy Toric II IOL were evaluated during a 3-month follow-up. Visual acuity, refraction, defocus curve, and patient-reported outcomes with the Catquest-9SF questionnaire were analyzed. A vectorial analysis was used to analyze the accuracy of astigmatic correction. RESULTS: Mean 3-month monocular postoperative uncorrected distance, intermediate (80 cm) and near (40 cm) visual acuities were 0.06 ± 0.11 logMAR, 0.13 ± 0.12 logMAR, and 0.13 ± 0.09 logMAR, respectively. Mean monocular distance-corrected intermediate (80 cm) and near visual acuity (40 cm) were 0.11 ± 0.12 logMAR and 0.10 ± 0.10 logMAR, respectively. Mean binocular defocus curve showed visual acuities of 0.10 logMAR or better for defocus levels from +0.50 to -2.50 D. Residual cylinder was within ±0.50 D in 97.0% of eyes. The surgically induced astigmatism prediction error ranged between -0.49 D and 0.50 D, with a mean value of 0.04 ± 0.16 D. Mean absolute IOL rotation was 3.79 ± 2.94 degrees. Significant improvements were found in all Rasch-calibrated scores obtained with Catquest-9SF ( P < .001). CONCLUSIONS: The implantation of the toric presbyopia-correcting IOL evaluated provides an efficacious astigmatic correction while providing a fully restoration of the visual function across different distances.

Building a High-Potential Silver-Sulfur Redox Reaction Based on the Hard-Soft Acid-Base Theory.

Sulfur holds immense promise for battery applications owing to its abundant availability, low cost, and high capacity. Currently, sulfur is commonly combined with alkali or alkaline earth metals in metal-sulfur batteries. However, these batteries universally face challenges in cycling stability due to the inevitable issue of polysulfide dissolution and shuttling. Additionally, the inferior stability of metal sulfide discharge compounds results in low S0/S2- redox potentials (<-0.41 V vs SHE). Herein, we leverage the principle of the hard-soft acid-base theory to introduce a novel silver-sulfur (Ag-S) battery system, which operates on the reaction between the soft acid of Ag+ and the soft base of S2-. Due to their high reaction affinity, the discharge compound of silver sulfide (Ag2S) is intrinsically insoluble and fundamentally stable. This not only resolves the polysulfide dissolution issue but also leads to a predominantly high S0/S2- redox potential (+1.0 V vs. SHE). We thus exploit the Ag-S reaction for a primary zinc battery application, which exhibits a high capacity of ∼620 mAh g-1 and a high voltage of ∼1.45 V. This work offers valuable insights into the application of classic chemistry theories in the development of innovative energy storage devices.

An Integrated Characterization Strategy on Board for Recycling of poly(vinyl butyral) (PVB) from Laminated Glass Wastes.

Polyvinyl butyral (PVB) is widely used as an interlayer material in laminated glass applications, mainly in the automotive industry, but also for construction and photovoltaic applications. Post-consumed laminated glass is a waste that is mainly landfilled; nevertheless, it can be revalorized upon efficient separation and removal of adhered glass. PVB interlayers in laminated glass are always plasticized with a significant fraction in the 20-40% w/w range of plasticizer, and they are protected from the environment by two sheets of glass. In this work, the aim is to develop a thorough characterization strategy for PVB films. Neat reference PVB grades intended for interlayer use are compared with properly processed (delaminated) post-consumed PVB grades from the automotive and construction sectors. Methods are developed to open opportunities for recycling and reuse of the latter. The plasticizer content and chemical nature are determined by applying well-known analytical techniques, namely, FT-IR, TGA, NMR. The issue of potential aging during the life cycle of the original laminated material is also addressed through NMR. Based on the findings, a sensor capable of directly sorting PVB post-consumer materials will be developed and calibrated at a later stage.

Mutations in α-synuclein, TDP-43 and tau prolong protein half-life through diminished degradation by lysosomal proteases.

BACKGROUND: Autosomal dominant mutations in α-synuclein, TDP-43 and tau are thought to predispose to neurodegeneration by enhancing protein aggregation. While a subset of α-synuclein, TDP-43 and tau mutations has been shown to increase the structural propensity of these proteins toward self-association, rates of aggregation are also highly dependent on protein steady state concentrations, which are in large part regulated by their rates of lysosomal degradation. Previous studies have shown that lysosomal proteases operate precisely and not indiscriminately, cleaving their substrates at very specific linear amino acid sequences. With this knowledge, we hypothesized that certain coding mutations in α-synuclein, TDP-43 and tau may lead to increased protein steady state concentrations and eventual aggregation by an alternative mechanism, that is, through disrupting lysosomal protease cleavage recognition motifs and subsequently conferring protease resistance to these proteins. RESULTS: To test this possibility, we first generated comprehensive proteolysis maps containing all of the potential lysosomal protease cleavage sites for α-synuclein, TDP-43 and tau. In silico analyses of these maps indicated that certain mutations would diminish cathepsin cleavage, a prediction we confirmed utilizing in vitro protease assays. We then validated these findings in cell models and induced neurons, demonstrating that mutant forms of α-synuclein, TDP-43 and tau are degraded less efficiently than wild type despite being imported into lysosomes at similar rates. CONCLUSIONS: Together, this study provides evidence that pathogenic mutations in the N-terminal domain of α-synuclein (G51D, A53T), low complexity domain of TDP-43 (A315T, Q331K, M337V) and R1 and R2 domains of tau (K257T, N279K, S305N) directly impair their own lysosomal degradation, altering protein homeostasis and increasing cellular protein concentrations by extending the degradation half-lives of these proteins. These results also point to novel, shared, alternative mechanism by which different forms of neurodegeneration, including synucleinopathies, TDP-43 proteinopathies and tauopathies, may arise. Importantly, they also provide a roadmap for how the upregulation of particular lysosomal proteases could be targeted as potential therapeutics for human neurodegenerative disease.

Efficient generation of a self-organizing neuromuscular junction model from human pluripotent stem cells.

The complex neuromuscular network that controls body movements is the target of severe diseases that result in paralysis and death. Here, we report the development of a robust and efficient self-organizing neuromuscular junction (soNMJ) model from human pluripotent stem cells that can be maintained long-term in simple adherent conditions. The timely application of specific patterning signals instructs the simultaneous development and differentiation of position-specific brachial spinal neurons, skeletal muscles, and terminal Schwann cells. High-content imaging reveals self-organized bundles of aligned muscle fibers surrounded by innervating motor neurons that form functional neuromuscular junctions. Optogenetic activation and pharmacological interventions show that the spinal neurons actively instruct the synchronous skeletal muscle contraction. The generation of a soNMJ model from spinal muscular atrophy patient-specific iPSCs reveals that the number of NMJs and muscle contraction is severely affected, resembling the patient's pathology. In the future, the soNMJ model could be used for high-throughput studies in disease modeling and drug development. Thus, this model will allow us to address unmet needs in the neuromuscular disease field.

Integration of Genomic Sequencing Drives Therapeutic Targeting of PDGFRA in T-Cell Acute Lymphoblastic Leukemia/Lymphoblastic Lymphoma.

PURPOSE: Patients with relapsed or refractory T-cell acute lymphoblastic leukemia (T-ALL) or lymphoblastic lymphoma (T-LBL) have limited therapeutic options. Clinical use of genomic profiling provides an opportunity to identify targetable alterations to inform therapy. EXPERIMENTAL DESIGN: We describe a cohort of 14 pediatric patients with relapsed or refractory T-ALL enrolled on the Leukemia Precision-based Therapy (LEAP) Consortium trial (NCT02670525) and a patient with T-LBL, discovering alterations in platelet-derived growth factor receptor-α (PDGFRA) in 3 of these patients. We identified a novel mutation in PDGFRA, p.D842N, and used an integrated structural modeling and molecular biology approach to characterize mutations at D842 to guide therapeutic targeting. We conducted a preclinical study of avapritinib in a mouse patient-derived xenograft (PDX) model of FIP1L1-PDGFRA and PDGFRA p.D842N leukemia. RESULTS: Two patients with T-ALL in the LEAP cohort (14%) had targetable genomic alterations affecting PDGFRA, a FIP1-like 1 protein/PDGFRA (FIP1L1-PDGFRA) fusion and a novel mutation in PDGFRA, p.D842N. The D842N mutation resulted in PDGFRA activation and sensitivity to tested PDGFRA inhibitors. In a T-ALL PDX model, avapritinib treatment led to decreased leukemia burden, significantly prolonged survival, and even cured a subset of mice. Avapritinib treatment was well tolerated and yielded clinical benefit in a patient with refractory T-ALL. CONCLUSIONS: Refractory T-ALL has not been fully characterized. Alterations in PDGFRA or other targetable kinases may inform therapy for patients with refractory T-ALL who otherwise have limited treatment options. Clinical genomic profiling, in real time, is needed for fully informed therapeutic decision making.

Equine-assisted services for individuals with substance use disorders: a scoping review.

The implementation of equine-assisted services (EAS) during treatment for psychological disorders has been frequently documented; however, little is known about the effect of EAS on outcomes for populations with substance use disorder (SUD). The purpose of this scoping review was to synthesize existing literature reporting the effects of EAS when incorporated into SUD treatment. This review followed guidelines in the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) checklist for scoping reviews. A search of four databases (MEDLINE/PubMed, PsycINFO, the Cumulative Index to Nursing and Allied Health Literature (CINAHL), and Academic OneFile) identified studies reporting quantitative and/or qualitative data for an EAS intervention that was implemented among a population receiving treatment for and/or diagnosed with SUD. Of 188 titles, 71 underwent full-text assessment and six met inclusion criteria. Three additional articles were identified through other sources. Quantitative data suggested a potential positive effect of EAS on SUD treatment retention and completion as well as participants' mental health; however, studies were limited by small sample sizes. Qualitative data indicated that in addition to participants developing bonds with the horses and perceiving EAS as welcomed breaks from traditional treatment, they experienced increased self-efficacy, positive emotional affect, and SUD treatment motivation. Due to methodological limitations and an overall dearth of available studies, the effectiveness of EAS as adjuncts to SUD treatment is inconclusive. Additional research is necessary before supporting broad implementation of EAS as adjuncts to SUD treatment.

Assessing the Activation of Tyrosine Kinase KIT through Free Energy Calculations.

KIT is a type 3 receptor tyrosine kinase that plays a crucial role in cellular growth and proliferation. Mutations in KIT can dysregulate its active-inactive equilibrium. Activating mutations drive cancer growth, while deactivating mutations result in the loss of skin and hair pigmentation in a disease known as piebaldism. Here, we propose a method based on molecular dynamics and free energy calculations to predict the functional effect of KIT mutations. Our calculations may have important clinical implications by defining the functional significance of previously uncharacterized KIT mutations and guiding targeted therapy.

[Vitamin D deficiency/insufficiency on healthy infants receiving standard supplementation]. / Deficiencia e insuficiencia de vitamina D en lactantes sanos recibiendo suplementación estándar.

Vitamin D (VD) is essential for calcium and phosphorus metabolism. Its deficiency can cause rickets. In Chile, newborns receive 400UI/day supplementation from the first day of age until the first year. OBJECTIVE: To describe the VD plasma levels in healthy infants who received supplementation and secondarily to correlate this with seasonality and nutritional status. SUBJECTS AND METHOD: Cross sectional study. Infants on exclusive or mixed breastfeeding, with monthly pediatric checkups recei ving 400 UI VD supplementation were evaluated, measuring VD plasma levels at 6 months of age, weight, and length, and their nutritional status was classified according to the WHO growth referen ces (weight/age and weight/length). The VD cut-off concentration values were < 20 ng/ml, 21- 29 ng/ ml, and ≥ 30 ng/ml considered as deficiency, insufficiency, and sufficiency, respectively. RESULTS: 40 infants were studied, 40% had insufficient levels and 40% presented deficiency. Season and nutritio nal status were variables significantly related to lower VD values (Winter-Spring p = 0.007; at risk of malnutrition p = 0.038). CONCLUSIONS: The population who received supplementation presented a high frequency of VD deficiency and insufficiency which increases during winter and spring and in subjects at risk of malnutrition.

Rationale, design, and methodology of a randomized pilot trial of an integrated intervention combining computerized behavioral therapy and recovery coaching for people with opioid use disorder: The OVERCOME study.

Background: Opioid use disorder (OUD) has led to a staggering death toll in terms of drug-related overdoses. Despite the demonstrated benefits and effectiveness of buprenorphine, retention is suboptimal, and patients typically present with high rates of ongoing polysubstance use during treatment. A pilot trial provided preliminary support for the efficacy of computer-based cognitive-behavioral therapy (CBT4CBT) as an add-on to buprenorphine in reducing substance use. Recovery coaching services provided by individuals with substance use experience and successful recovery have also shown to positively influence recovery outcomes for people with OUD by increasing buprenorphine initiation and reducing opioid use. Methods: The OVERCOME study is a randomized clinical trial (RCT) aimed to tests an integrated intervention combining CBT4CBT and Recovery Coaching relative to treatment-as-usual (TAU) among individuals with OUD on buprenorphine. The primary outcome measure is the percentage of samples with any drug tested as positive at each research visit conducted during treatment (visits 1 to 8). Secondary outcomes include the percentage of samples with any drug tested as positive at 1- and 3- month follow-up and retention to buprenorphine at 3- month follow-up. Results: We describe the rationale, design, and methodology of the OVERCOME Study. Conclusion: This trial will provide evidence of the efficacy of an integrated intervention combining CBT4CBT and Recovery Coaching for reducing substance use and increasing buprenorphine adherence which has the potential to reduce mortality among people with OUD.