A novel syphilis Treponema pallidum lipoprotein peptide antigen diagnostic assay using red cell kodecytes in routine blood centre column agglutination testing platforms.

BACKGROUND AND OBJECTIVES: The detection of treponemal antibodies, which are used to make a diagnosis of syphilis, is important both for diagnostic purposes and as a mandatory blood donor test in most countries. We evaluated the feasibility of using Kode Technology to make syphilis peptide red cell kodecytes for use in column agglutination serologic platforms. MATERIALS AND METHODS: Candidate Kode Technology function-spacer-lipid (FSL) constructs were made for the Treponema pallidum lipoprotein (TmpA) of T. pallidum, using the peptide and FSL selection algorithms, and then used to make kodecytes. Developmental kodecytes were evaluated against a large range of syphilis antibody reactive and non-reactive samples in column agglutination platforms and compared against established methodologies. Overall, 150 reactive and 2072 non-reactive Syphicheck assay (a modified T. pallidum particle agglutination) blood donor samples were used to evaluate the agreement rate of the developed kodecyte assay. RESULTS: From three FSL-peptide candidate constructs, one was found to be the most suitable for diagnostics. Of 150 Syphicheck assay reactive samples, 146 were TmpA-kodecyte reactive (97.3% agreement), compared with 58.0% with the rapid plasmin reagin (RPR) assay for the same samples. Against the 2072 expected syphilis non-reactive samples the agreement rate for TmpA-kodecytes was 98.8%. CONCLUSION: TmpA-kodecytes are viable for use as cost-effective serologic reagent red cells for the detection of treponemal antibodies to diagnose syphilis with a high level of specificity in blood centres. This kodecyte methodology also potentially allows for introduction of the reverse-algorithm testing into low-volume laboratories, by utilizing existing transfusion laboratory infrastructure.

Surface Glycans of Microvesicles Derived from Endothelial Cells, as Probed Using Plant Lectins.

Glycans of MVs are proposed to be candidates for mediating targeting specificity or at least promoting it. In contrast to exosomes, glycomic studies of MVs are largely absent. We studied the glycoprofile of endothelial cell-derived MVs using 21 plant lectins, and the results show the dominance of oligolactosamines and their α2-6-sialylated forms as N-glycans and low levels of α2-3-sialylated glycans. The low levels of α2-3-sialosides could not be explained by the action of extracellular glycosidases. Additionally, the level of some Man-containing glycans was also decreased in MVs. Spatial masking as the causative relationship between these low level glycans (as glycosphingolipids) by integral proteins or proteoglycans (thus, their lack of interaction with lectins) seems unlikely. The results suggest that integral proteins do not pass randomly into MVs, but instead only some types, differing in terms of their specific glycosylation, are integrated into MVs.

Chemical Resolution of an Epitope Recognized by Blood Group Antibodies Capable of Binding Both A and B Red Blood Cells.

The high specificity of human antibodies to blood group A and B antigens is impressive, especially when considering the structural difference between these antigens (tetrasaccharides) is a NHAc versus a OH-group on the terminal monosaccharide residue. It is well established that in addition to anti-A and anti-B there is a third antibody, anti-A,B capable of recognizing both A and B antigens. To analyze this AB specificity, we synthesized a tetrasaccharide, where the NHAc of the A antigen was replaced with NH2. This NH2-group was used to attach the glycan to an affinity resin, creating an AB-epitope (ABep) adsorbent where the critical site for recognition by A and B antibodies was not accessible, while the rest of the (conformationally compact) tetrasaccharide remained accessible. Anti-ABep antibodies were isolated from blood group O donors and found to have expected A,B-specificity against immobilized and red cell bound synthetic antigens, including ABep, and were able to agglutinate both A and B red cells. The amount of these anti-ABep (anti-A,B) antibodies found in the blood of group O donors was comparable to levels of anti-A and anti-B found in group B and A individuals. Using STD-NMR the location for the AB-epitope on the tetrasaccharide was found.

Comparison of record linkage software for deduplicating patient identities in California's Prescription Drug Monitoring Program.

BACKGROUND: To help prevent overdose deaths involving prescription drugs, accurate linkage of prescription drug monitoring program (PDMP) records for individual patients is essential. OBJECTIVES: To compare the accuracy of the linkage program used by California's PDMP against various record linkage programs with respect to accuracy in deduplicating patient identities in the PDMP, with implications for identifying high-risk opioid use and outlier behaviors. RESEARCH DESIGN: We evaluated California's program, Link Plus, LinkSolv, and The Link King on 557 861 PDMP identity records with addresses in two 3-digit zip code areas for patients who filled a controlled substance prescription in 2013. Manual review was performed on a stratified sample of 720 paired records identified as matches by at least one program. MEASURES: We estimated sensitivity and positive predictive value, and computed PDMP patient alerts for the patient entities identified by each program. RESULTS: Sensitivity was 95% for LinkSolv and The Link King, 84% for Link Plus, and 73% for California's program; positive predictive value was ≥93% for all programs. The number of patient entities prompting a PDMP alert was similar among the programs for all alerts except multiple provider episodes (obtaining prescriptions from ≥6 prescribers or ≥6 pharmacies in the last 6 months), which were 10.9%, 26.6%, and 16.9% greater using The Link King, Link Plus, and LinkSolv, respectively, compared to California's program. CONCLUSIONS: PDMPs should assess the accuracy of record linkage algorithms and the impacts of these algorithms on patient safety alerts and develop national best practices for PDMP record linkage.

Impact of 30-day prescribed opioid dose trajectory on fatal overdose risk: A population-based, statewide cohort study.

BACKGROUND: Both increases and decreases in patients' prescribed daily opioid dose have been linked to increased overdose risk, but associations between 30-day dose trajectories and subsequent overdose risk have not been systematically examined. OBJECTIVE: To examine the associations between 30-day prescribed opioid dose trajectories and fatal opioid overdose risk during the subsequent 15 days. DESIGN: Statewide cohort study using linked prescription drug monitoring program and death certificate data. We constructed a multivariable Cox proportional hazards model that accounted for time-varying prescription-, prescriber-, and pharmacy-level factors. PARTICIPANTS: All patients prescribed an opioid analgesic in California from March to December, 2013 (5,326,392 patients). MAIN MEASURES: Dependent variable: fatal drug overdose involving opioids. Primary independent variable: a 16-level variable denoting all possible opioid dose trajectories using the following categories for current and 30-day previously prescribed daily dose: 0-29, 30-59, 60-89, or ≥90 milligram morphine equivalents (MME). KEY RESULTS: Relative to patients prescribed a stable daily dose of 0-29 MME, large (≥2 categories) dose increases and having a previous or current dose ≥60 MME per day were associated with significantly greater 15-day overdose risk. Patients whose dose decreased from ≥90 to 0-29 MME per day had significantly greater overdose risk compared to both patients prescribed a stable daily dose of ≥90 MME (aHR 3.56, 95%CI 2.24-5.67) and to patients prescribed a stable daily dose of 0-29 MME (aHR 7.87, 95%CI 5.49-11.28). Patients prescribed benzodiazepines also had significantly greater overdose risk; being prescribed Z-drugs, carisoprodol, or psychostimulants was not associated with overdose risk. CONCLUSIONS: Large (≥2 categories) 30-day dose increases and decreases were both associated with increased risk of fatal opioid overdose, particularly for patients taking ≥90 MME whose opioids were abruptly stopped. Results align with 2022 CDC guidelines that urge caution when reducing opioid doses for patients taking long-term opioid for chronic pain.

Stigma Against Patients With Substance Use Disorders Among Health Care Professionals and Trainees and Stigma-Reducing Interventions: A Systematic Review.

PURPOSE: In this systematic review, the authors examine the prevalence and extent of stigmatizing attitudes among health care professionals (HCPs) and trainees against patients with substance use disorders (SUDs), including research on interventions to reduce stigma. METHOD: The authors searched 7 databases for articles published from January 1, 2011, through February 15, 2023, that quantified SUD stigma among HCPs or trainees. Inclusion criteria allowed both observational and intervention studies from the United States or Canada to be included in this review. Quality assessment was applied to all included studies; studies were not excluded based on quality. RESULTS: A total of 1,992 unique articles were identified of which 32 articles (17 observational studies and 15 intervention studies), all conducted in the United States, met the inclusion criteria. Half of the included studies (16 of 32) were published in 2020 or later. Most of the intervention studies (13 of 15) used a single-group pre-post design; interventions involved didactics and/or interactions with persons with SUDs. The 32 included studies used a total of 19 different measures of stigma. All 17 observational studies showed some degree of HCP or trainee stigma against patients with SUDs. Most intervention studies (12 of 15) found small but statistically significant reductions in stigma after intervention. CONCLUSIONS: SUD stigma exists among HCPs and trainees. Some interventions to reduce this stigma had positive impacts, but future studies with larger, diverse participants and comparison groups are needed. Heterogeneity among studies and stigma measures limits the ability to interpret results across studies. Future rigorous research is needed to determine validated, consensus measures of SUD stigma among HCPs and trainees, identify stigma scores that are associated with clinical outcomes, and develop effective antistigma interventions for HCPs and trainees.

Communicative Practices Clinicians Use to Correct Patient Misconceptions in Primary Care Visits.

To investigate how clinicians correct patient misconceptions, we analyzed 23 video recordings of primary care visits. Analysis focused on operationalizing, identifying, and characterizing clinician corrections, integrating two inductive approaches: microanalysis of clinical interaction and conversation analysis. According to our definition, patient misconception-clinician correction episodes met three essential criteria: (1) the clinician refuted something the patient had said, (2) which the patient had presented without uncertainty, and (3) which contained a proposition that was factually incorrect. We identified 59 such episodes; the patient misconceptions most commonly related to medication issues; fewer than half had foreseeable implications for patients' future actions. We identified seven clinician correction practices: Three direct practices (displaying surprise, marking disagreement, contradicting the patient) and four indirect practices (presenting the correct proposition, providing explanations, invoking an outside authority, demonstrating with evidence). We found an almost equal distribution of these direct and indirect practices.

Paediatric sand toe: radiographic, ultrasound and MRI findings of dorsomedial capsular injury.

Hyperflexion injury to the metatarsophalangeal joint of the great toe, referred to as sand toe, can cause significant functional impairment. To our knowledge, there have been no radiological descriptions of this injury in the paediatric age group. Here, we report radiographic, sonographic and MRI findings in a male paediatric patient who sustained a sand toe injury, highlighting structural damage to the dorsomedial capsule and medial sagittal band, and discuss sand toe's favourable prognosis with conservative management.

Galectin-9 as a Potential Modulator of Lymphocyte Adhesion to Endothelium via Binding to Blood Group H Glycan.

The recruitment of leukocytes from blood is one of the most important cellular processes in response to tissue damage and inflammation. This multi-step process includes rolling leukocytes and their adhesion to endothelial cells (EC), culminating in crossing the EC barrier to reach the inflamed tissue. Galectin-8 and galectin-9 expressed on the immune system cells are part of this process and can induce cell adhesion via binding to oligolactosamine glycans. Similarly, these galectins have an order of magnitude higher affinity towards glycans of the ABH blood group system, widely represented on ECs. However, the roles of gal-8 and gal-9 as mediators of adhesion to endothelial ABH antigens are practically unknown. In this work, we investigated whether H antigen-gal-9-mediated adhesion occurred between Jurkat cells (of lymphocytic origin and known to have gal-9) and EA.hy 926 cells (immortalized endothelial cells and known to have blood group H antigen). Baseline experiments showed that Jurkat cells adhered to EA.hy 926 cells; however when these EA.hy 926 cells were defucosylated (despite the unmasking of lactosamine chains), adherence was abolished. Restoration of fucosylation by insertion of synthetic glycolipids in the form of H (type 2) trisaccharide Fucα1-2Galß1-4GlcNAc restored adhesion. The degree of lymphocyte adhesion to native and the "H-restored" (glycolipid-loaded) EA.hy 926 cells was comparable. If this gal-9/H (type 2) interaction is similar to processes that occur in vivo, this suggests that only the short (trisaccharide) H glycan on ECs is required.

Professional knowledge acquisition of dental students employed part-time as dental assistants.

INTRODUCTION: Whilst admission to dental school in many jurisdictions requires applicants to shadow dentists or undertake work experience at a dental practice, little is known about the impacts on professional learning when dental students work part-time as dental assistants whilst concurrently studying dentistry. This paper explores what, how and from whom dental students who work as dental assistants acquire professional knowledges during employment. METHODS: This study draws on a qualitative analysis of interviews with sixteen senior dental students who have extra-curricular part-time employment as dental assistants in private dental practices. RESULTS: Analysis produced four themes that relate to students learning in the professional environment: students learn about the responsibilities, rhythms and routines of practice, as well as patient communication and interactions. Students embedded in the dental team noticed and related to the dispositions and the work of dentistry. Students learned from all members of the dental team including clinical and non-clinical staff (reception, administration, laboratory and sterilisation). DISCUSSION: Students used their experiences in a practice setting to further their professional education. The ability to "read" a situation and formulate an appropriate response requires the integration of complex and actionable professional knowledges. CONCLUSION: This research study presents insight into the ways dental students employed as dental assistants are embedded in and learn from the dental team in a critically evaluative manner. Students professionally notice and make sense of complex practice environments whilst undertaking university studies to learn about practice routines, rhythms and responsibilities as well as advancing confidence in relating with patients. This study provides a stimulus for further research about the contribution of workplace experiences to dental education.

Development and Testing of a Communication Intervention to Improve Chronic Pain Management in Primary Care: A Pilot Randomized Clinical Trial.

OBJECTIVES: Effective communication skills are essential for optimally managing chronic pain and opioids. This exploratory, sequential mixed methods study tested the effect of a novel framework designed to improve pain-related communication and outcomes. METHODS: Study 1 developed a novel 5-step framework for helping primary care clinicians discuss chronic pain and opioids with patients. Study 2 pilot tested an intervention for teaching this framework using standardized patient instructors-actors trained to portray patients and provide immediate clinician feedback-deployed during regular clinic hours. Primary care physicians were randomized to receive either the intervention or pain management recommendations from the Centers for Disease Control and Prevention. Primary outcomes were pain-related interference at 2 months and clinician use of targeted communication skills (coded from transcripts of audio-recorded visits); secondary outcomes were pain intensity at 2 months, clinician self-efficacy for communicating about chronic pain, patient experience, and clinician-reported visit difficulty. RESULTS: We enrolled 47 primary care physicians from 2 academic teaching clinics and recorded visits with 48 patients taking opioids for chronic pain who had an appointment scheduled with an enrolled physician. The intervention was not associated with significant changes in primary or secondary outcomes other than clinician self-efficacy, which was significantly greater in the intervention group. DISCUSSION: This study developed a novel framework and intervention for teaching clinician pain-related communications skills. Although the intervention showed promise, more intensive or multicomponent interventions may be needed to have a significant impact on clinicians' pain-related communication and pain outcomes.

The impact of California wildfires on patient access to prescription opioids.

BACKGROUND: Patients on long-term opioid therapy are particularly vulnerable to disruptions in medication access, especially during traumatic and chaotic events such as wildfires and other natural disasters. OBJECTIVES: To determine whether past highly destructive California wildfires were associated with disrupted access to prescription opioids for patients receiving long-term, and therefore physically dependent on, opioid medications. METHODS: Using California prescription drug monitoring program data, this retrospective study selected patients with long-term prescription opioid use episodes residing in ZIP code tabulation areas impacted by either the Camp Fire or Tubbs Fire. Autoregressive integrated moving average time series models were fit to pre-fire data to forecast post-fire expected values and then compared with observed post-fire data, specifically for weekly proportions of long-term episodes with early fills, late fills, changes in patients' prescriber and pharmacy, and fills within a different ZIP code tabulation area than the patient's residence. RESULTS: After the Camp Fire, there were significant spikes in the proportions of early fills (peak at 56% of total, week 1 after fire), late fills (peak at 29%, week 6), and immediate significant increases in prescriber (peak at 37%, week 3) and pharmacy changes (peak at 71%, week 1) in high-impact ZIP code tabulation areas. Low-impact ZIP code tabulation areas experienced no similar disruptions. Disruptions due to the Tubbs Fire were far less severe. CONCLUSION: Access to prescription opioids was greatly disrupted for patients living in areas most impacted by the Camp Fire. Future research should explore effectiveness of current state and federal controlled substance prescribing policies to determine what improvements are needed to minimize disruptions in medication access due to wildfires and other natural disasters.

Procedural Approach to Ultrasound-Guided Geniculate Nerve Blockade for Knee Pain in Patients with OA.

ABSTRACT: Ultrasound-guided genicular nerve block can provide excellent pain control for patients with knee OA. This procedure has the advantage of providing sensory blockade with concomitant sparing of motor compromise, which is observed when the femoral and its lateral femoral cutaneous branches are blocked. Once the geniculate nerve of interest is identified, the operator can use ultrasound guidance to surround nerve fascicles with an injectate mixture of anesthetic and corticosteroid, yielding decreased pain sensation at the joint capsule. Given the role of the geniculate nerve in providing sensory innervation to the joint capsule and knee ligaments, blockade of this nerve can serve as a useful tool for managing patients with acute knee pain secondary to OA.

Simulated RBC antibody identification training panels created using SARS-CoV-2 kodecytes and immune plasma.

BACKGROUND: Training is essential to develop and maintain skills required to be a competent serologist, yet samples required to achieve this are often difficult to obtain. We evaluated the feasibility of SARS-CoV-2 peptide modified RBCs (1144-kodecytes) to develop simulated antibody screening and identification panels of reagent RBCs suitable for practical training, recognition, and grading of serologic reactions. STUDY DESIGN AND METHODS: RBCs from a single donor were modified into kodecytes using Kode Technology function-spacer-lipid constructs bearing a short SARS-CoV-2 peptide. Kodecytes and unmodified cells were then arranged in patterns representative of RBC antibody profiles as simulated antibody screening and identification reagent cell panels (SASID), and then tested against immune donor plasma samples containing SARS-CoV-2 antibodies. Manual tube and two different gel card serologic platforms were evaluated by routine techniques. SASID exemplars were created for antibodies including D, Cw , f (ce), Jka (strong, weak, dosing), mixtures of D + E, Jka + K, Fya + E, high and low frequency antibodies and a warm IgG autoantibody. RESULTS: Kodecytes (positive reactions) and unmodified cells (negative) when arranged and tested in appropriate patterns in SASID panels were able to mimic IgG antibody reactions, and were capable of measuring both accuracy and precision in reaction grading. CONCLUSIONS: Kodecytes can be used to rapidly create in-house simulated yet realistic antibody screening and identification panels suitable for large scale training in the recognition and grading of serologic reactions.

Erytra blood group analyser and kode technology testing of SARS-CoV-2 antibodies among convalescent patients and vaccinated individuals.

Surveillance of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic requires tests to monitor antibody formation and prevalence. We detected SARS-CoV-2 antibodies using red cells coated by Kode technology with short peptides derived from the SARS-CoV-2 spike protein (SP). Such modified red cells, called C19-kodecytes, can be used as reagent cells in any manual or automated column agglutination assay. We investigated the presence of SARS-CoV-2 antibodies in 130 samples from COVID-19 convalescent plasma donors using standard manual technique, two FDA-authorized enzyme-linked immunosorbent assay (ELISA) assays and a virus neutralisation assay. The sensitivity of the C19-kodecyte assay was 88%, comparable to the anti-SP and anti-nucleocapsid protein (NCP) ELISAs (86% and 83%) and the virus neutralisation assay (88%). The specificity of the C19-kodecyte assay was 90% (anti-SP 100% and anti-NCP 97%). Likewise, 231 samples from 73 vaccinated individuals were tested with an automated analyser, and we monitored the appearance and persistence of SARS-CoV-2 antibodies. The C19-kodecyte assay is a robust tool for SARS-CoV-2 antibody detection. Automated blood group analyser use enables large-scale SARS-CoV-2 antibody testing for vaccination monitoring in population surveys.

Visit Linearity in Primary Care Visits for Patients with Chronic Pain on Long-term Opioid Therapy.

BACKGROUND: Physicians and patients report frustration after primary care visits for chronic pain. The need to shift between multiple clinical topics to address competing demands during visits may contribute to this frustration. OBJECTIVE: This study creates a novel measure, "visit linearity," to assess visit organization and examines whether visits that require less shifting back and forth between topics are associated with better patient and physician visit experiences. It also explores whether visit linearity differs depending on the following: (1) whether or not pain is a major topic of the visit and (2) whether or not pain is the first topic raised. DESIGN: This study analyzed 41 video-recorded visits using inductive, qualitative analysis informed by conversation analysis. We used linear regression to evaluate associations between visit organization and post-visit measures of participant experience. PARTICIPANTS: Patients were established adult patients planning to discuss pain management during routine primary care. Physicians were internal or family medicine residents. MAIN MEASURES: Visit linearity, total topics, return topics, topic shifts, time per topic, visit duration, pain main topic, pain first topic, patient experience, and physician difficulty. KEY RESULTS: Visits had a mean of 8.1 total topics (standard deviation (SD)=3.46), 14.5 topic shifts (SD=6.28), and 1.9 topic shifts per topic (SD=0.62). Less linear visits (higher topic shifts to topic ratio) were associated with greater physician visit difficulty (ß=7.28, p<0.001) and worse patient experience (ß= -0.62, p=0.03). Visit linearity was not significantly impacted by pain as a major or first topic raised. CONCLUSIONS: In primary care visits for patients with chronic pain taking opioids, more linear visits were associated with better physician and patient experience. Frequent topic shifts may be disruptive. If confirmed in future research, this finding implies that reducing shifts between topics could help decrease mutual frustration related to discussions about pain.

COVID-19 Antibody Detection and Assay Performance Using Red Cell Agglutination.

Red cells can be labeled with peptides from the SARS-CoV-2 spike protein (C-19 kodecytes) and used as reagent cells for serologic screening of SARS-CoV-2 antibodies. We evaluated 140 convalescent COVID-19 donors and 275 healthy controls using C19-kodecytes. The analytical performance of the C19-kodecyte assay was compared with a virus neutralizing assay and two commercial chemiluminescent antibody tests (Total assay and IgG assay, Ortho). The C19-kodecyte assay detected SARS-CoV-2 antibodies with a sensitivity of 92.8% and specificity of 96.3%, well within the minimum performance range required by FDA for EUA authorization of serologic tests. The Cohen's kappa coefficient was 0.90 indicating an almost perfect agreement with the Total assay. The Spearman's correlation coefficient was 0.20 with the neutralizing assay (0.49 with IgG, and 0.41 with Total assays). The limited correlation in assay reaction strengths suggested that the assays may be influenced by different antibody specificities. The C19-kodecyte assay is easily scalable and may vastly improve test capacity in any blood typing laboratory using its routine column agglutination platforms. IMPORTANCE We recently developed a red cell based assay to detect SARS-CoV-2 antibodies in human plasma. In the current study, we show the hands-on application of this assay in a group of COVID-19 convalescent plasma donors and healthy individuals. We compared our assay against three published assays, including two that are widely used for patient care in the United States. Our assay compared well with all three assays. Our easily scalable assay can be used for population-wide screening of SARS-CoV-2 antibody status. It can be readily established in any hospital blood bank worldwide using its routine equipment.