Assessment Of Potency Research Articles

Assessment of Shelf-Life Stability and Bioactive Potency of HCA-Rich <i>Garcinia Atroviridis</i> Powder to Address Obesity and Metabolic Health

Assessment of Shelf-Life Stability and Bioactive Potency of HCA-Rich <i>Garcinia Atroviridis</i> Powder to Address Obesity and Metabolic Health

Label-free assessment of complement-dependent cytotoxicity of therapeutic antibodies via a whole-cell MALDI mass spectrometry bioassay

Potency assessment of monoclonal antibodies or corresponding biosimilars in cell-based assays is an essential prerequisite in biopharmaceutical research and development. However, cellular bioassays are still subject to limitations in sample throughput, speed, and often need costly reagents or labels as they are based on an indirect readout by luminescence or fluorescence. In contrast, whole-cell Matrix-Assisted Laser Desorption/Ionization Time-of-Flight (MALDI-TOF) Mass Spectrometry (MS) has emerged as a direct, fast and label-free technology for functional drug screening being able to unravel the molecular complexity of cellular response to pharmaceutical reagents. However, this approach has not yet been used for cellular testing of biologicals. In this study, we have conceived, developed and benchmarked a label-free MALDI-MS based cell bioassay workflow for the functional assessment of complement-dependent cytotoxicity (CDC) of Rituximab antibody. By computational evaluation of response profiles followed by subsequent m/z feature annotation via fragmentation analysis and trapped ion mobility MS, we identified adenosine triphosphate and glutathione as readily MS-assessable metabolite markers for CDC and demonstrate that robust concentration–response characteristics can be obtained by MALDI-TOF MS. Statistical assay performance indicators suggest that whole-cell MALDI-TOF MS could complement the toolbox for functional cellular testing of biopharmaceuticals.

Predicting points of departure and potency categories for fragrance ingredients by integrating OECD in vitro models

Continuous potency assessment is crucial for conducting quantitative risk assessment (QRA) of sensitizers. Quantitative regression models, based on in vitro methods, have been developed to calculate points of departure for use in skin sensitization QRA. These models calculate a point of departure as a predicted value for Local Lymph Node Assay (LLNA) EC3 or potency value (PV), integrating data from the kinetic Direct Peptide Reactivity Assay (kDPRA), KeratinoSens (KS) assay, and human Cell Line Activation Test (h-CLAT). The goal of this study was to determine how in vitro predicted EC3s and PVs compare to published reference data. In vitro data were combined in point of departure regression models to predict EC3s and PVs. These points of departure were then grouped into sensitization potency categories, such as extreme, strong, moderate, weak, very weak, or non-sensitizer, as previously described. Trends in potency distribution and high concordance between predicted EC3 and PV categories and published potency categories were observed. Furthermore, the median absolute fold-misprediction ranged between 1.8 and 2.5 for models predicting EC3 and between 1.7 and 3.4 for those predicting PVs. These regression models are a promising animal alternative for determining sensitization quantitative potency for fragrance ingredients, thereby facilitating QRA.

Increasing Accessibility of Bayesian Network-Based Defined Approaches for Skin Sensitisation Potency Assessment.

Skin sensitisation is a critical adverse effect assessed to ensure the safety of compounds and materials exposed to the skin. Alongside the development of new approach methodologies (NAMs), defined approaches (DAs) have been established to promote skin sensitisation potency assessment by adopting and integrating standardised in vitro, in chemico, and in silico methods with specified data analysis procedures to achieve reliable and reproducible predictions. The incorporation of additional NAMs could help increase accessibility and flexibility. Using superior algorithms may help improve the accuracy of hazard and potency assessment and build confidence in the results. Here, we introduce two new DA models, with the aim to build DAs on freely available software and the newly developed kDPRA for covalent binding of a chemical to skin peptides and proteins. The new DA models are built on an existing Bayesian network (BN) modelling approach and expand on it. The new DA models include kDPRA data as one of the in vitro parameters and utilise in silico inputs from open-source QSAR models. Both approaches perform at least on par with the existing BN DA and show 63% and 68% accuracy when predicting four LLNA potency classes, respectively. We demonstrate the value of the Bayesian network's confidence indications for predictions, as they provide a measure for differentiating between highly accurate and reliable predictions (accuracies up to 87%) in contrast to low-reliability predictions associated with inaccurate predictions.

P19-57 Improved Confidence of Quantitative Sensitizing Potency Assessment for Point of Departure Using GARDskin Dose-Response

P19-57 Improved Confidence of Quantitative Sensitizing Potency Assessment for Point of Departure Using GARDskin Dose-Response

Quantitative Phytochemical Investigation, Antibacterial Potency, and Drug-ability Assessment of Three Indian Medicinal Plants Leaf Extracts Using Bioinformatics Tools

Natural regimens have long-held ethnomedicinal values, serving as primary sources for mainstream medicine. Therefore, scientists are paying more attention to studying the biological activity of existing plant species in organized ways to select potent bioactive metabolites to use for specific therapeutic purposes. This study used the same approach to find potent antibacterial phytoconstituents in three well-known Indian medicinal plants: Psidium guajava L., Syzygium cumini L. and Punica granatum L. In the earlier study, methanolic leaf extracts of the above plant extracts were more effective than n-hexane extracts against biofilm and drug-resistant pathogenic bacteria. Accordingly, we selected methanolic crude extracts for gas chromatography-mass spectrometry (GC-MS) to identify the phytoconstituents presented. In addition, we added a few reported candidates from the above plant extracts for molecular docking studies against four bacterial targets. For molecular docking studies, we retrieved all phytoconstituents or ligands from the PubChem database and bacterial target proteins from the protein data bank using PyRx 0.8-AutoDock 4.2 software. Furthermore, we used various bioinformatics and chemoinformatics tools to examine the investigated phytoconstituents physicochemical properties, toxicity, and drug-ability profiles. Out of the 30 GC-MS report-derived candidates from three plants, P5 from P. guajava, P18 from S. cumini, and P21 from P. granatum had the potent binding ability with bacterial targets. In the same way, out of the 30 reported candidates, P39, P43, and P56 from three plants, along with amikacin, showed strong binding against the same bacterial target. Both sets of candidates showed favorable physicochemical and toxicity profiles; however, all GC-MS-derived and few reported candidates exhibited negative drug-likeness. The study reveals that these crude extracts have antibacterial properties because they contain both GC-MS and existing phytoconstituents. The study starts with a crude extraction and then uses bioinformatics to choose two possible antibacterial candidates, ursolic acid and punicacortein A. This platform could be useful for finding an antibacterial agent that works specifically on a specific target. To sum up, the study encourages the isolation of more bioactive candidates from different Indian medicinal plants and uses bioinformatics tools to speed up the selection of strong leads that can speed up the process of making antibacterial drugs within limited resources.

Assessment of the Antimicrobial Potency of Unripe Plantain Fruits and Peelings Extracts against Escherichia coli, Staphylococcus aureus and Salmonella typhi

Assessment of the Antimicrobial Potency of Unripe Plantain Fruits and Peelings Extracts against Escherichia coli, Staphylococcus aureus and Salmonella typhi

Supporting an integrated QTc risk assessment using the hERG margin distributions for three positive control agents derived from multiple laboratories and on multiple occasions.

BackgroundDetermination of a drug's potency in blocking the hERG channel is an established safety pharmacology study. Best practice guidelines have been published for reliable assessment of hERG potency. In addition, a set of plasma concentration and plasma protein binding fraction data were provided as denominators for margin calculations.The aims of the current analysis were five-fold: provide data allowing creation of consistent denominators for the hERG margin distributions of the key reference agents, explore the variation in hERG margins within and across laboratories, provide a hERG margin to 10 ms QTc prolongation based on several newer studies, provide information to use these analyses for reference purposes, and provide recommended hERG margin ‘cut-off’ values. MethodsThe analyses used 12 hERG IC50 ‘best practice’ data sets (for the 3 reference agents). A group of 5 data sets came from a single laboratory. The other 7 data sets were collected by 6 different laboratories. ResultsThe denominator exposure distributions were consistent with the ICH E14/S7B Training Materials. The inter-occasion and inter-laboratory variability in hERG IC50 values were comparable. Inter-drug differences were most important in determining the pooled margin variability. The combined data provided a robust hERG margin reference based on best practice guidelines and consistent exposure denominators. The sensitivity of hERG margin thresholds were consistent with the sensitivity described over the course of the last two decades. ConclusionThe current data provide further insight into the sensitivity of the 30-fold hERG margin ‘cut-off’ used for two decades. Using similar hERG assessments and these analyses, a future researcher can use a hERG margin threshold to support a negative QTc integrated risk assessment.

Exploring the Nutritional and Sensorial Enhancement of Traditional Indian Laddu through Flaxseed Fortification

Abstract: The purpose of the current study was to cultivate laddu by adding different amounts of flaxseed. to evaluate laddu, including its microbiological, tactile, and textural qualities, as well as its chemical and fatty acid composition and antioxidant activity. The primary objective of this research endeavor was to explore the potential enhancement of laddu, a traditional Indian sweet, through the incorporation of varied proportions of flaxseed. This exploration encompassed a multifaceted evaluation, including microbiological scrutiny, tactile discernment, textural analysis, chemical profiling, fatty acid characterization, and assessment of antioxidant potency. The procedural standardization entailed the utilization of sesame seeds, jaggery, and whole flaxseed in the formulation of Nutra Laddu. Distinct formulations denoted as T1, T2, T3, T4 were devised, featuring incremental adjustments in the proportion of whole flaxseed, ranging from 0% to 50%. Subsequent to the preparation process, the laddu specimens underwent meticulous evaluation of their organoleptic attributes, encompassing considerations such as color and visual appeal, flavor profile, textural attributes, taste perception, and overall palatability. This comprehensive assessment was facilitated through the application of a 9-point hedonic scale, enabling nuanced appraisal. Furthermore, it emerged from the analysis that Nutra Laddu fortified with flaxseed presents a notable nutritional profile, characterized by heightened levels of dietary fiber and protein. This underscores the potential of flaxseed as a valuable adjunct in laddu formulation, given its recognized nutritional and therapeutic attributes.

Quantum Mechanical Assessment of Nitrosamine Potency.

Nitrosamines are in the cohort of concern (CoC) as determined by regulatory guidance. CoC compounds are considered highly potent carcinogens that need to be limited below the threshold of toxicological concern, 1.5 μg/day. Nitrosamines like NDMA and NDEA require strict control, while novel nitrosamine drug substance-related impurities (NDSRIs) may or may not be characterized as potent carcinogens. A risk assessment based on the structural features of NDSRIs is important in order to predict potency because they lack substance-specific carcinogenicity. Herein, we present a quantum mechanical (QM)-based analysis on structurally diverse sets of nitrosamines to better understand how structure influences the reactivity that could result in carcinogenicity. We describe the potency trend through activation energies corresponding to α-hydroxylation, aldehyde formation, diazonium intermediate formation, reaction with DNA base, and hydrolysis reactions, and other probable metabolic pathways associated with the carcinogenicity of nitrosamines. We evaluated activation energies for selected cases such as N-nitroso pyrrolidines, N-nitroso piperidines, N-nitroso piperazines, N-nitroso morpholines, N-nitroso thiomorpholine, N-methyl nitroso aromatic, fluorine-substituted nitrosamines, and substituted aliphatic nitrosamines. We compare these results to the recent framework of the carcinogenic potency characterization approach (CPCA) proposed by health authorities which is meant to give guidance on acceptable intakes (AI) for NDSRIs lacking substance-specific carcinogenicity data. We show examples where QM modeling and CPCA are aligned and examples where CPCA both underestimates and overestimates the AI. In cases where CPCA predicts high potency for NDSRIs, QM modeling can help better estimate an AI. Our results suggest that a combined mechanistic understanding of α-hydroxylation, aldehyde formation, hydrolysis, and reaction with DNA bases could help identify the structural features that underpin the potency of nitrosamines. We anticipate this work will be a valuable addition to the CPCA and provide a more analytical way to estimate AI for novel NDSRIs.

Uveal melanoma immunogenomics predict immunotherapy resistance and susceptibility

Immune checkpoint inhibition has shown success in treating metastatic cutaneous melanoma but has limited efficacy against metastatic uveal melanoma, a rare variant arising from the immune privileged eye. To better understand this resistance, we comprehensively profile 100 human uveal melanoma metastases using clinicogenomics, transcriptomics, and tumor infiltrating lymphocyte potency assessment. We find that over half of these metastases harbor tumor infiltrating lymphocytes with potent autologous tumor specificity, despite low mutational burden and resistance to prior immunotherapies. However, we observe strikingly low intratumoral T cell receptor clonality within the tumor microenvironment even after prior immunotherapies. To harness these quiescent tumor infiltrating lymphocytes, we develop a transcriptomic biomarker to enable in vivo identification and ex vivo liberation to counter their growth suppression. Finally, we demonstrate that adoptive transfer of these transcriptomically selected tumor infiltrating lymphocytes can promote tumor immunity in patients with metastatic uveal melanoma when other immunotherapies are incapable.

Establishment and Validation of an Automated System for the Antifactor IIa Assay: A Case Study of Potency Assessment of a Pharmaceutical Gel Formulation.

Antithrombotic agents and anticoagulant drugs, such as those from the heparin family, are employed in clinical settings for the prevention and treatment of clotting, thromboembolism, and wound healing. The potency assessment of antithrombotic agents is typically conducted using antifactor IIa assay with manual systems which are time-consuming and often lack repeatability. Here, we present a novel automated system that significantly enhances assay repeatability, attaining an outstandingly low relative standard deviation (RSD) % of only 0.6% for repeatability. This system has been applied to a pharmaceutical gel formulation for wound healing developed by Abdi Ibrahim Pharmaceuticals R&D Center as a case study for validation. The automated system demonstrated substantial improvements over manual systems in linearity (R2 = 0.9927), precision, accuracy, specificity, and robustness. The system aligns with the European Pharmacopoeia specifications, promising to enhance quality control across pharmaceutical formulations and conduct absorbance-based end-point assays within the pharmaceutical industry while offering increased throughput and cost-effectiveness.

Assessment of Cinnamaldehyde's Potency on Heat Stress-induced Testicular Impairments in Wistar Rats.

Male sterility results from high testicular temperatures, which affect mammalian spermatogenesis. High testicular temperatures affect sperm motility, morphology and fertility according to their magnitude and duration. The aim of the current study is to examine the effects of heat-induced oxidative stress and cinnamaldehyde on Wistar rat testicular structure and function. The rats used in this experiment were Wistar albino rats. This research has six animals per group. Male Wistar albino rats of 2.5-3 months old and 275-300 g. (I) control, (II) heat stress (HS) in a closed chamber at 41°C for 14 days and (III) HS with cinnamaldehyde (CA) 50 mg/kg body weight for 14 days. (IV) CA alone. After the study, the animals were euthanised, and test samples were taken for sperm count, morphology, haematoxylin and eosin stain for normal cellular morphology, antioxidants and DNA integrity assessments. The data were analysed statistically using one- and two-way ANOVA tests for comparisons between groups. The stress group had significantly lower sperm counts and poor sperm morphology. The stress group's antioxidant capacity is much lower than that of the control group. Animals under stress have fragmented DNA. Treatment with cinnamaldehyde increased overall antioxidant capacity and seminal parameters, and rats behaved most like controls. CA restores malondialdehyde levels, total antioxidant capacity, sperm characteristics and mitigates testicular damage in rats exposed to experimental HS.

Development of a Potency Assay for Nous-209, a Multivalent Neoantigens-Based Genetic Cancer Vaccine.

Quality control testing of vaccines, including potency assessment, is critical to ensure equivalence of clinical lots. We developed a potency assay to support the clinical advancement of Nous-209, a cancer vaccine based on heterologous prime/boost administration of two multivalent viral vector products: GAd-209 and MVA-209. These consist of a mix of four Adeno (Great Ape Adenovirus; GAd) and four Modified Vaccinia Ankara (MVA) vectors respectively, each containing a different transgene encoding a synthetic polypeptide composed of antigenic peptide fragments joined one after the other. The potency assay employs quantitative Reverse Transcription PCR (RT-Q-PCR) to quantitatively measure the transcripts from the four transgenes encoded by each product in in vitro infected cells, enabling simultaneous detection. Results showcase the assay's robustness and biological relevance, as it effectively detects potency loss in one component of the mixture comparably to in vivo immunogenicity testing. This report details the assay's setup and validation, offering valuable insights for the clinical development of similar genetic vaccines, particularly those encoding synthetic polypeptides.

Integrated Electrochemical Aptasensor Array toward Monitoring Anticancer Drugs in Sweat.

In the realm of clinical practice, the concurrent utilization of anticancer medications can enhance their overall therapeutic efficacy. However, it is crucial to acknowledge that the interactions among these anticancer drugs can potentially yield detrimental consequences on their intended outcomes. Consequently, the assessment of both anticancer potency and potential toxic side effects is greatly refined when multiple anticancer drugs are simultaneously detected and evaluated. Here, we designed a wearable electrochemical aptasensor array for monitoring multiple anticancer drugs in sweat. The integrated sensor array consists of three working electrodes modified with three different aptamers (Apt1, Apt2, and Apt3), a Au counter electrode, and a Ag/AgCl reference electrode. Molecular docking simulations were performed to show the binding affinities between three anticancer drugs and their corresponding aptamers. Various eigenvalues were derived from the square-wave voltammetry electrochemical signals, and these data sets were subjected to rigorous analysis through multivariate data analysis techniques. This analytical approach demonstrated exceptional performance by achieving flawless 100% accuracy in the precise identification of nine anticancer drugs consistently at uniform concentrations. Furthermore, the integrated wearable sensor array exhibited impressive capabilities, correctly recognizing all nine anticancer drugs with 100% accuracy and successfully distinguishing between these drugs in artificial sweat samples. The proposed sensor array presents good stability for 15 days. Flexibility tests showed stable device performance after 500 twisting cycles. This innovative wearable sensing array represents a novel approach for achieving real-time monitoring and precise adjustment of drug dosages. It offers invaluable insights for tailoring the treatment of anticancer drugs to individual patients, predicting both drug efficacy and potential adverse reactions within the field of clinical medicine.

Assessment of myogenic potency in patient-derived fibroblasts with c.1289-2A>G Desmin mutation

Abstract Objectives The ultra-rare DES c.1289-2A>G mutation, resulting in a 48-base pair insertion in the Desmin tail domain, is associated with late-onset MFM1 (myofibrillar myopathy-1; OMIM number; 601419) and exhibits distinctive pathological features. Despite sustained expression and cytoskeletal integrity, muscle biopsies reveal dystrophic characteristics through an unidentified mechanism. A deeper understanding of the molecular mechanisms underlying Desmin-related MFM1 could enhance our perspective and comprehension of the disease’s pathophysiology. In this study, we aimed to investigate the pathological phenotype by assessing the myogenic potency of MyoD-induced patient-derived fibroblasts. Methods Following the immortalization and myoconversion of unaffected and patient-derived fibroblast cells, we analyzed the myogenic potency of the mutant and control groups on day 5 post-differentiation. This analysis involved staining cells with MF20 antibody and DAPI after MyoD induction. Results Employing six parameters to quantify extra nuclei and myotube properties, we unveil impaired myogenic differentiation in c.1289-2A>G mutant cells, as evidenced by a compromised fusion index and distinctive myogenic features. In summary, our preliminary findings indicate phenotypic abnormalities and suggest an association between the DES c.1289-2A>G mutation and delayed maturation and MFM in affected individuals. Conclusions Our results indicate a significant involvement of Desmin in the myogenic maturation of muscle cells. Further investigation is required to understand the changes in the transcriptome during the myoconversion of patient-derived fibroblasts.

Synthesis, characterization and assessment of anticancer potency of oxcarbazepine with folic acid conjugated Fe2O3 nanostructures as nano-drugs

The well-known chemical drug oxcarbazepine (OXZ) is used to treat partial and secondary generalized tonic-clonic seizures. Because extended-release formulations of antiepileptic drug provide adequate drug delivery, they can help many patients significantly in achieving the main goals of their therapy. To improve the effectiveness of drug administration, OXZ was encapsulated with Fe2O3 nanoparticles (FeNPs) conjugated with folic acid and N-hydroxysuccinimide (FeNPs-FA-NHS-OXZ) in the current work. The nano-drug FeNPs-FA-NHS-OXZ was thoroughly investigated, and its structural and morphological characteristics were confirmed, using a range of sophisticated techniques such as UV–vis spectra, FTIR, DLS, SEM-EDAX, and XRD. X-ray diffraction analyses validated the pure Fe2O3 NPs' hexagonal structure, with an estimated crystallite size of 83 nm. Moreover, the thermograms of FeNPs-FA-NHS-OXZ showed a weight decrease of around 30–40 %, which supported the conjugation of a biomolecule with Fe2O3 NPs. The SEM scans revealed that the morphology of the materials as-prepared showed a combination of hexagonal rod-like shape. The particle size analyzer revealed an observed particle size of 82.2 nm (DLS). Additionally effective for drug delivery are the FeNPs-FA-NHS-OXZ drug loading and drug-releasing capabilities. Additionally, using the SRB test, the cytotoxicity effects of the produced substance were assessed on rat neuroblastoma cell lines. To further investigate the nanodrug's in vivo toxicity, drosophila melanogaster was used in the experiment. When FeNPs-FA-NHS-OXZ was administered to drosophila melanogaster, the results demonstrated little toxicity, as well as enhanced lifespan and reproductive capacity.

High-Throughput Bacteriophage Testing with Potency Determination: Validation of an Automated Pipetting and Phage Drop-Off Method.

The development of bacteriophages (phages) as active pharmaceutical ingredients for the treatment of patients is on its way and regulatory agencies are calling for reliable methods to assess phage potency. As the number of phage banks is increasing, so is the number of phages that need to be tested to identify therapeutic candidates. Currently, assessment of phage potency on a semi-solid medium to observe plaque-forming units is unavoidable and proves to be labor intensive when considering dozens of phage candidates. Here, we present a method based on automated pipetting and phage drop-off performed by a liquid-handling robot, allowing high-throughput testing and phage potency determination (based on phage titer and efficiency of plaquing). Ten phages were tested, individually and assembled into one cocktail, against 126 Escherichia coli strains. This automated method was compared to the reference one (manual assay) and validated in terms of reproducibility and concordance (ratio of results according to the Bland and Altman method: 0.99; Lin's concordance correlation coefficient: 0.86). We found that coefficients of variation were lower with automated pipetting (mean CV: 13.3% vs. 24.5%). Beyond speeding up the process of phage screening, this method could be used to standardize phage potency evaluation.

Identification of Piperazinyl-Difluoro-indene Derivatives Containing Pyridyl Groups as Potent FGFR Inhibitors against FGFR Mutant Tumor: Design, Synthesis, and Biological Evaluation.

The fibroblast growth factor receptor (FGFR) signaling pathway plays important roles in cellular processes such as proliferation, differentiation, and migration. In this study, we highlighted the potential of FGFR inhibitors bearing the (S)-3,3-difluoro-1-(4-methylpiperazin-1-yl)-2,3-dihydro-1H-indene scaffold containing a crucial 3-pyridyl group for the treatment of FGFR mutant cancers. The representative compound (S)-23, which was identified through comprehensive evaluation, exhibited potent antiproliferative activity with GI50 in the range of 6.4-10.4 nM against FGFR1 fusion protein-carrying, FGFR2-amplified, and FGFR2 mutant cancer cell lines and good antiproliferative activity against FGFR3 translocation and mutant FGFR4 cancer cell lines, as well as potency assessment against FGFR1-4 kinases. Moreover, compound (S)-23 exhibited favorable pharmacokinetic properties, low potential for drug-drug interactions, and very potent antitumor activity in MFE-296 xenograft mouse models with a TGI of 99.1% at the dose of 10 mg/kg. These findings demonstrate that compound (S)-23 is a potential therapeutic agent for FGFR mutant tumors.

Assessment of simultaneous alcohol and cannabis use and its related consequences and cognitions in college students: Anarrative review.

As rates of students using cannabis continue to rise, simultaneoususe of alcohol and cannabis (such that their effects overlap; commonly referred to as simultaneous alcohol and marijuana [SAM] use)is prevalent among college students who use both substances.Although research focusing on SAM use and related cognitions and consequences continues to grow,there are nocommon established measures,as approaches varyacross studies.This narrative reviewidentifiescurrent methods for assessingSAM use and measures of SAM-related consequences and cognitions (motives and expectancies) among college students, evaluateshow they were developed, identifiesgaps in the literature, and providesrecommendations for future directions of assessment. We conclude that the assessment of SAM use is limited by difficulties in the assessment of cannabis quantity and potency. However, and the lack of a psychometrically validated measure of SAM consequences. However, measures of SAM motives and expectancies have been published with support from psychometric examinations such as exploratory factor analysis, confirmatory factor analysis, and measurement invariance. Research is needed that incorporates qualitative approaches in the development of SAM use measures so that unique items specific to SAM use rather than single-substance use can be identified. Additionally, validation of these measures is needed across different samples that varydemographically, such asby race and gender or sex. Future research should consider the development of ameasure of protective behavioral strategiesspecific to SAM use to inform interventionsthat target thereduction of negative consequences of SAM use.