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Transfusion

Wiley

Preprints posted in the last 90 days, ranked by how well they match Transfusion's content profile, based on 18 papers previously published here. The average preprint has a 0.01% match score for this journal, so anything above that is already an above-average fit.

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Association of social media-sourced blood donors with transfusion delay and donor-related irregularities: A multicentre study in Bangladesh

Hoque, A.; Rahman, M.; Basak, S. K.; Mamun, A. A.

2026-04-17 health systems and quality improvement 10.64898/2026.04.08.26350439 medRxiv
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BackgroundIn the absence of structured donor registries, social media platforms have become a dominant mechanism for blood donor recruitment in many low-resource settings. However, the implications of this shift for transfusion timeliness and system reliability remain unclear. ObjectiveTo evaluate the impact of social media-sourced donors on transfusion delay, donor reliability, and hemovigilance-related outcomes compared with conventional donor pathways. MethodsThis prospective analytical study included 400 transfusion episodes across tertiary hospitals in Bangladesh. Donor sources were categorized as social media (SM) or conventional (CON). The primary outcome was delay-to-transfusion. Secondary outcomes included donor-related irregularities, documentation completeness, near-miss events, and acute transfusion reactions. Multivariable logistic regression identified predictors of delay [&ge;]4 hours. ResultsSocial media-sourced donors were associated with significantly longer transfusion delays (5.98 vs 2.97 hours; p<0.001). Delay [&ge;]4 hours occurred in 83.6% of SM cases versus 17.6% of CON cases (OR 23.78). Donor-related irregularities were observed in 85% of SM episodes and absent in CON donors. Safety outcomes did not differ significantly between groups. Social media donor sourcing remained the strongest independent predictor of delay (adjusted OR 18.09). ConclusionUnregulated social media-based donor recruitment introduces substantial delays and undermines system reliability without improving access. Integration of digital tools into regulated donor systems is essential to strengthen transfusion timeliness and hemovigilance in resource-limited settings.

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Opportunities for Vein-to-Vein Datasets from a Blood Establishment Perspective: towards a 'Pan-European Transfusion Research InfrAstructure' (PETRA)

Wehrens, S. M.; Arvas, M.; Fustolo-Gunnink, S. F.; Vinkovic Vlah, M.; Waters, A.; Erikstrup, C.; Drechsler, L. O.; Stanworth, S. J.; van den Hurk, K.

2026-03-26 hematology 10.64898/2026.03.24.26348611 medRxiv
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iii.Background and ObjectivesThe "Pan-European Transfusion Research InfrAstructure" (PETRA) project was established to advance the use of donor, blood product, and patient datasets in Europe, aiming to benefit both patient and donor health. Here, the initial PETRA objective was to describe the landscape of existing donor and blood establishment (BE) databases. Materials and MethodsAn online survey was circulated to the European Blood Alliances BE members. The survey collected information on the feasibility of accessing donor data, and challenges and possibilities for linking these datasets with information on the associated blood products and transfusion recipients, and donors own health records. ResultsSeventeen BEs across 16 countries completed the survey. The majority could, in principle, link their donor data to product data (13 BEs (76%)) and recipient data (10 BEs (59%)), for research purposes. However, capabilities were limited and in only 29% of the BEs was the donor to recipients linkage an automated process. BEs reported significant challenges to achieve full vein-to-vein linkage, including legal constraints and lack of consent (11 BEs) and resources (10-14 BEs). IT and data issues as well as lack of knowledge and training were cited as obstacles by a minority of BEs. ConclusionWhilst the survey results suggest considerable interest in developing linkages between blood donors, their products, and recipients, many challenges remain due to a variety of obstacles. First steps in working towards a PETRA may be assistance to navigate legal frameworks as well as investing in resources and quality and harmonisation of data collections. iv. HighlightsO_LI17 blood establishments (BEs) in 16 countries responded to a survey on obstacles and opportunities for achieving vein-to-vein datasets. C_LIO_LIIn 59% of the BEs donor-to-recipient links can be established for research improving transfusion outcomes, but only in 29% this is an automated process. C_LIO_LIIn order to work towards a "Pan-European Transfusion Research InfrAstructure" (PETRA), legal frameworks, adequate donor consent and (financial and human) resources are the most common obstacles that require addressing. C_LI

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Variation in Haemostasis and VTE Prophylaxis in Elective Adult Cranial Neurosurgery: A Global Survey of Perioperative Practice

Pandit, A. S.; Chaudri, T.; Chaudri, Z.; Vasilica, A. M.; Dhaliwal, J.; Sayar, Z.; Cohen, H.; Westwood, J. P.; Toma, A. K.

2026-04-16 surgery 10.64898/2026.04.14.26350905 medRxiv
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BackgroundVenous thromboembolism (VTE) remains a major cause of perioperative morbidity in cranial neurosurgery, yet clinical practice varies widely, and formal guidelines are inconsistent. Understanding internationally sampled neurosurgical practice is essential for informing consensus and future trials. MethodsAn international, 2-stage cross-sectional, internet-based survey was conducted. Practising neurosurgeons performing elective adult cranial surgery were eligible. Descriptive statistics were used to summarise practice. Responses covered patterns of pre-operative haemostasis decision making, use and timing of mechanical and/or chemical prophylaxis, use of perioperative imaging prior to anticoagulation, and frequency of clinical assessment for VTE. Associations with geographical income status, subspecialty, and years post-certification were statistically tested. Practice heterogeneity was quantified and contextual influence was summarised using mean effect sizes across stratifying variables in order to determine domains of true equipoise. ResultsOf 585 responses, 456 (78%) met criteria for inclusion: representing 322 units across 78 countries (71% high-income). Thirteen per cent reported no departmental VTE plan; 23% followed no guidelines and 12% used multiple. Routine pre-operative testing almost universally included haemoglobin/platelets/haematocrit, with fibrinogen more common in high-income settings. Compared with high-income country respondents, low- and middle-income respondents reported higher haemoglobin transfusion thresholds (>90 g/dL; p<0.001) and shorter antiplatelet interruption (p[&le;]0.03), and less frequent outpatient VTE assessment (p<0.001). Mechanical prophylaxis was common (TEDs 81%, IPC 62%), typically started pre-or intra-operatively. Among those completing the chemoprophylaxis section (n=310), 57% required a CT or MRI scan before LMWH which was then initiated on average 31.4 hours after surgery. 1% of respondents did not routinely use LMWH. Many clinical decisions demonstrated statistical equipoise ie. high heterogeneity with low contextual influence. ConclusionPeri-operative haemostasis and VTE prophylaxis practices in adult elective cranial neurosurgery vary substantially worldwide, with some decisions reflecting geographical or socioeconomic differences and many others reflecting true clinical equipoise rather than contextual determinants. By mapping contemporary real-world practice across diverse health-system contexts, this study provides a necessary empirical foundation for rational trial design and future guideline development.

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Structured Error Analysis and Corrective Actions in Clinical Laboratory Practice: An Analysis of 7226 External Quality Assurance Participations

Strasser, B.; Mustafa, S.; Holly, M.; Grünberger, M.; Anita, S.

2026-04-04 health systems and quality improvement 10.64898/2026.04.02.26350023 medRxiv
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Background: External Quality Assurance (EQA) is an essential component of modern laboratory medicine. Current scientific evidence on EQA focuses primarily on the analyses carried out by EQA providers while relatively little research has been conducted in individual clinical laboratories. Methods: In this retrospective single-center observational study in a clinical laboratory, EQA results were analyzed over a period of four years (2021-2024). The evaluation was based on EQA action reports documented in the institutes internal quality management system. Deviations were classified according to department, type of discrepancy, root cause category (analytical, preanalytical, systemic, unidentifiable), and measures taken. Results: A total of 7226 EQA participations were evaluated during the observation period. The overall error rate remained consistently low, ranging between 0.8% and 1.6%, with no significant change over time (p = 0.87). Most deviations occurred in the departments of clinical chemistry and immuno/autoimmune diagnostics (p < 0.001). These were predominantly quantitative discrepancies (false low/false negative or false high/false positive). Root cause analysis showed a clear dominance of analytical causes (p < 0.001), while preanalytical and systemic causes were identified less frequently. In most cases, corrective measures, such as re-analyses, recalibrations, process adjustments, or staff training, were implemented promptly. Hard structural measures, such as changing methods or discontinuing tests, were rarely necessary. Conclusion: In a clinical laboratory, EQA is an important tool for structured error analysis and continuous quality improvement. Consistent processing of deviating EQA results goes hand in hand with stable analytical performance and a low error rate.

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Chyme reinfusion using the Insides System to reduce parenteral nutrition dependence in Type 2 intestinal failure: multicentre randomised controlled trial (REINFUSE)

Milne, T.; Lal, S.; Farrer, K.; Evans, J.; Thomas, K.; Randhawa, N.; Rogers, J.; Mercer, D.; Carroll, R.; Rosenthal, M.; Jafri, S.-M.; Vaizey, C.; Gabe, S.; layfield, D.; Burke, D.; Frampton, C.

2026-05-04 surgery 10.64898/2026.04.26.26351226 medRxiv
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BackgroundPatients with type 2 intestinal failure (T2IF) due to double enterostomy or enterocutaneous fistula (DES/ECF) require parenteral nutrition (PN), carrying risks of catheter sepsis, venous thrombosis, and liver disease. Chyme reinfusion therapy (CRT) may reduce PN dependence but has not been evaluated in a randomised controlled trial (RCT). This study assessed whether device-assisted CRT using The Insides System reduces PN requirements. MethodsThis multicentre, open-label RCT enrolled PN-dependent adults with T2IF due to DES/ECF across 12 centres in the UK and USA. Patients with insufficient distal limb length, proximal bowel obstruction, active sepsis, or severe hepatorenal failure were excluded. Participants were block randomised 2:1 to device-assisted CRT plus standard care (active) or standard care (control). The primary endpoint was 50% or greater reduction in PN caloric intake at 30 days, using an intention-to-treat analysis, for a comparison between randomised groups using a two tailed p-value of 0.025 to allow for a single interim analysis. Secondary outcomes were rate of PN cessation at 30 and 60 days, quality of life, and adverse events. ResultsThe population comprised 39 (26 active, 13 control) participants. At Day 30, 8/26 (31%) active participants achieved the primary endpoint versus no controls (p=0.035). By Day 60, 10/23 (43%) active participants had completely ceased PN versus no controls (p=0.008), with median intestinal losses reduced by 1,344 mL/day at Day 30 (p=0.005) and 1,450 mL/day at Day 60 (p=0.026 between group). Device-related adverse events were predominantly mild; one death unrelated to the device occurred. ConclusionCRT with the Insides System demonstrated substantial therapeutic advantages in patients with T2IF from DES/ECF, with 31% of participants reducing PN calories by 50% at 30 days and >40% of participants achieving complete PN cessation by 60 days, and an acceptable safety profile. Trial registrationClinicalTrials.gov NCT04577456 FundingThis trial was sponsored by The Insides Company Ltd. Surgical RelevanceWhat is already known: Patients with type 2 intestinal failure due to double enterostomy or enterocutaneous fistula depend on parenteral nutrition, which carries significant risks including central venous catheter (CVC) sepsis, venous thrombosis, and intestinal failure-associated liver disease. Chyme reinfusion therapy restores distal gut function but has only been evaluated in non-randomised cohort studies. What is new: This first randomised controlled trial of device-assisted chyme reinfusion demonstrates that 43 per cent of participants can completely cease parenteral nutrition by 60 days, with a 70 per cent reduction in intestinal losses, high participant satisfaction and an acceptable risk profile. Potential impact on future practice: Early initiation of device-assisted chyme reinfusion in suitable patients with double enterostomy or enterocutaneous fistula reduces parenteral nutrition dependence, avoids associated complications and costs, and facilitates rehabilitation before reconstructive surgery.

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Associations between Exposure to Perfluoroalkyl Substances with Subsequent Body Composition and Glycemic Responses to Bariatric Surgery

Sankara, S.; Smith, M. R.; Eick, S. M.; Valvi, D.; Burley, T. M.; Walker, D. I.; Lin, E.; Hechenbleikner, E. M.; Gonzalez Ramirez, L. A.; Nesbeth, P.-D. C.; Vellanki, P.; Gower, B. A.; McConnell, R.; Jones, D. P.; Alvarez, J. A.; Chatzi, V. L.; Ziegler, T. R.

2026-04-01 surgery 10.64898/2026.03.30.26349786 medRxiv
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Per- and polyfluoroalkyl substances (PFAS) are chemicals linked to obesity and metabolic dysfunction, but their role in bariatric surgery remains poorly understood. This prospective pilot study examined correlations between plasma PFAS concentrations, body composition, and glycemic measures in adults undergoing bariatric surgery. Thirty-two patients (91% female; 66% Black; mean age 43 years) were enrolled preoperatively; twenty-two completed follow-up at a mean 8.6 months post-surgery. Three PFAS (PFHxS, PFNA, and PFOS) were quantified by plasma liquid chromatography-mass spectrometry; body composition and insulin sensitivity were assessed by dual-energy X-ray absorptiometry and intravenous glucose tolerance testing. At baseline, higher plasma PFNA and PFOS concentrations tracked with lower total lean mass ({rho}s = -0.46 and -0.48, respectively) and lean mass index ({rho}s = -0.46 and -0.42), and PFNA was inversely correlated with body weight ({rho}s = -0.40). No baseline associations were observed with adiposity or glycemic indices. Postoperatively, PFHxS concentrations decreased (median = -1.103 ng/mL, p < 0.001), whereas PFNA and PFOS did not change. Average PFNA was positively correlated with postoperative changes in HOMA-IR ({rho}s = 0.51) and total lean mass ({rho}s = 0.49). No significant associations were observed for average PFHxS or PFOS. These findings suggest that PFNA and PFOS may be linked to reduced lean tissue at baseline, and that PFNA burden modestly tracks with attenuated metabolic and body composition recovery. In an ANCOVA, baseline PFNA was not significantly associated with postoperative HOMA-IR or total lean mass. Larger, longitudinal studies are needed to clarify how PFAS influence these associations.

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Effects of Glucagon-Like Peptide-1 (GLP-1) Agonists on Surgical Wound Healing: A Single Institution Pilot Study

Adams, J. C.; Pullmann, D.; Belostotsky, H.; Mestvirishvili, T.; Chiu, E.; Oh, C.; Rabbani, P. S.

2026-04-22 surgery 10.64898/2026.04.21.26351321 medRxiv
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ObjectiveThis study evaluates the impact of systemic GLP-1 receptor agonist (GLP-1RA) use on surgical wound healing in high-risk surgical populations, including patients with diabetes, and implications for perioperative planning and healing outcomes. ApproachThis pilot retrospective cohort study compared adult surgery patients with non-healing postoperative wounds by their GLP-1RA use. Outcomes included healing status, time to wound closure, and number of surgical interventions. ResultsThe cohort included 35 non-GLP-1RA users and 16 GLP-1RA users with comparable baseline characteristics, except for significant higher prevalence of venous insufficiency among users. Though median time to closure was similar for all patients, users required fewer surgical interventions and their wounds reached closure in significant difference from non-users. Among patients with diabetes, all GLP-1RA users healed significantly compared to non-users. InnovationThe impact of GLP-1RA therapy on wound healing in high-risk reconstructive and soft-tissue surgery remains poorly defined. This pilot cohort addresses that gap, offering an early signal that GLP-1RA use is associated with improved wound healing and fewer postoperative interventions. These findings may inform perioperative practice by identifying a systemic pharmacologic factor that optimizes surgical outcomes in high-risk populations. ConclusionGLP-1RA use was associated with higher healing rates and fewer interventions, particularly among patients with diabetes. These findings support a beneficial role in surgical wound healing and warrant larger multi-site studies.

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Seroprevalence and Trends of Transfusion-Transmissible Infections Among Blood Donors in the Volta Region, Ghana: A Four-Year Retrospective Study

Hanu, E. K.; Ayanku, S.; Akuba, L.; Tetteh, C.; Akweh, T. Y.; Kwasie, D. A.; Bawu, S. B.; Fianko, K.; Dongdem, A. Z.

2026-05-06 epidemiology 10.64898/2026.05.04.26352407 medRxiv
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BackgroundBlood transfusion is a life-saving intervention; however, transfusion-transmissible infections (TTIs) such as human immunodeficiency virus (HIV), hepatitis B virus (HBV), hepatitis C virus (HCV), and syphilis remain major public health concerns, particularly in low-and middle-income countries. This study assessed the seroprevalence and temporal trends of TTIs among blood donors in the Volta Region of Ghana and to identify the demographic factors associated with seropositivity MethodsA retrospective cross-sectional study was conducted using secondary data from blood donors at Ho Teaching Hospital and Hohoe Regional Hospital between January 2020 and December 2023. Data from 6,147 eligible donors were extracted and analyzed using STATA version 17. Descriptive statistics summarized prevalence, while chi-square or Fishers exact tests assessed associations. Multivariable logistic regression was used to identify predictors of TTI seropositivity at a 5% significance level. ResultsThe overall prevalence of TTIs was 8.1%, with syphilis (3.6%) being the most prevalent infection, followed by HBV (1.8%), HCV (1.8%), and HIV (1.0%). All infections peaked in 2022 before declining in 2023. Older age ([&ge;]50 years) and year of donation were significant predictors of TTI positivity. In Hohoe, male donors had lower odds of HCV infection compared to females (aOR = 0.13; 95% CI: 0.06-0.28). ConclusionsAlthough TTI prevalence was relatively low, temporal increases and age-related disparities highlight the need for strengthened donor screening, targeted recruitment of voluntary donors, and enhanced surveillance strategies to ensure blood safety.

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Cost of Goods Sold Analysis for Manufacturing mRNA-Based Cell and Gene Therapies

Lieberthal, R. D.; Buontempo, P.; Harmon, B.; Omosule, A.; Washabaugh, M.; Whittaker, A.

2026-05-06 health economics 10.64898/2026.05.04.26352406 medRxiv
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BackgroundCell and gene therapies (CGT) represent a transformative class of medical interventions, yet their high production costs limit patient access. Understanding the structure of manufacturing costs is essential for informing policies that can expand access to these therapies. ObjectiveThis study develops and applies a cost-of-goods-sold (COGS) model to analyze the contributors to manufacturing costs for mRNA-based CGT, with application to a wide range of current and future therapies. MethodsAn Excel-based COGS model was constructed based on cost categories for CGT. Two mRNA-based products at commercial scale were used to populate the model: an mRNA vaccine and a therapeutic mRNA gene therapy. Cost inputs were drawn from vendor pricing, peer-reviewed and grey literature, and expert consultation with CGT manufacturing specialists. Three scenarios (worst, base, and best case) were modeled across six cost categories: materials, consumables, capital, labor, licenses, and royalties. A tornado diagram sensitivity analysis was conducted to identify key cost drivers. The mRNA vaccine was used to build and validate the model strucutre using publicly available data sources. The therapeutic mRNA therapy was used as the main use case for illustration and sensitivity analysis. ResultsUnder base-case assumptions, the estimated cost per dose for the therapeutic mRNA product is $56.09, ranging from $3.68 (best case) to $383.22 (worst case). Licensing and royalty fees together account for approximately 83% of total base-case COGS ($6,996,000 and $6,960,000 per production run, respectively, out of $16,825,597 total). Excluding these fees, material costs represent the largest remaining share (61%), followed by consumables (34%), capital (4%), and labor (1%). Sensitivity analysis confirms that licensing and royalty assumptions are the dominant source of uncertainty in the model. ConclusionsLicensing and royalty fees are the primary driver of mRNA-based CGT production costs and represent the greatest opportunity for cost reduction through policy intervention. Strategic priorities for cost reduction should focus on optimizing reagent utilization, increasing platform potency, and expanding use of contract development and manufacturing organizations (CDMOs) to reduce capital and labor costs. Key PointsProducing an example mRNA gene therapy costs about $56 per dose to manufacture, driven almost entirely driven by fees paid to patent holders for the underlying technology. Licensing and royalty fees cost roughly 83 cents of every dollar spent on these new biopharmaceutical products. Until that changes, the gap between what therapies cost to make and what patients and payers are charged will remain very wide.

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Second multistate outbreak of tuberculosis caused by a bone allograft product

Schildknecht, K. R.; Williams, P. M.; Schwartz, N. G.; Haddad, M. B.; Stewart, R. J.; Annambhotla, P.; Basavaraju, S. V.; Nabity, S. A.; Keh, C. E.; Calvet, H. M.; Zahn, M. M.; Beltran, R.; Cortez, A.; Lomeli, A.; Percak, J. M.; Gooze, L. L.; Coloma, M.; Shaw, T.; Davidson, P. J.; Smith, S. R.; Dickson, R. P.; Kaul, D. R.; Gonzalez, A. R.; Rodriguez, G.; Decimo, A.; Sanchez, A.; Armitige, L. Y.; Stapleton, J.; Lacassagne, M.; Brown, C.; Zheng, C.; Ali, J.; Wolfe, A. W.; Young, L. R.; Ariail, K.; Behm, H.; Jordan, H. T.; Spencer, M.; Nilsen, D. M.; Goradia, R.; Montoya Denison, B.; Burgos, M.;

2026-05-06 transplantation 10.64898/2026.04.29.26351868 medRxiv
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Tuberculosis screening is not mandatory for prospective tissue donors. In 2021 and 2023, two different bone allograft products caused nationwide tuberculosis outbreaks. We assessed the morbidity and mortality of the second outbreak and reviewed donor and tissue screening to identify deficiencies. Thirty-six people residing in nine states received the product during spinal and dental procedures. Twenty-seven recipients had tuberculosis infection, 11 had microbiologic or imaging evidence of tuberculosis disease, and two died from tuberculosis within 12 months of outbreak detection. Another recipient died from tuberculosis nearly 3 years after product implantation. The bone donor died of pneumonia and septic shock. Polymerase chain reaction testing of the product before and after distribution did not detect Mycobacterium tuberculosis. Mycobacterial culture was not performed until after outbreak detection, when M. tuberculosis was isolated from 2 of 6 unused product units. This outbreak demonstrates persistent gaps in tissue transplant safety. Appropriate selection of donors and mycobacterial culture of donated tissues could reduce but not eliminate the risk of M. tuberculosis transmission. Therefore, it is important that clinicians monitor tissue recipients and promptly report adverse events to tissue establishments and health authorities.

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Predicting Patient-Reported Appearance Satisfaction After Facial Skin Cancer Reconstruction: Development and Internal Validation of a Multivariable Prediction Model

Ottenhof, M. M. J.

2026-04-03 surgery 10.64898/2026.04.01.26349978 medRxiv
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Patient-reported outcomes have become standard in facial skin cancer surgery, yet clinicians currently lack validated tools to predict postoperative appearance satisfaction from preoperative patient characteristics. We developed and internally validated a prediction model for appearance satisfaction three months after facial skin cancer reconstruction. A prospective cohort study enrolled 287 patients at a tertiary referral center (2017-2018); 111 patients with complete data were included in the primary analysis. Patients completed the FACE-Q Skin Cancer Module preoperatively and at three months postoperatively. Our multivariable linear regression model incorporated age, sex, comorbidities, smoking status, and baseline appearance satisfaction. The model explained 23.0% of variance in postoperative appearance satisfaction (R2 = 0.23; adjusted R2 = 0.19; p < 0.001). Baseline appearance satisfaction (B = 0.48; 95% CI 0.28-0.68; p < 0.001) and female sex (B = -7.16; 95% CI -12.52 to -1.81; p = 0.009) emerged as independent predictors. Bootstrap resampling (500 iterations) yielded an optimism-corrected R2 of 0.17, supporting acceptable internal validity. Mean appearance satisfaction remained stable from baseline (54.8 +/- 13.8) to three months (57.0 +/- 16.4; p = 0.27). Baseline appearance satisfaction and female sex independently predict postoperative appearance satisfaction following facial skin cancer reconstruction. External validation in independent cohorts is warranted before clinical implementation.

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Metabolic profiling of cultured erythroblast for the production of transfusion-ready cultured red blood cells

Gallego-Murillo, J. S.; van Lakwijk, I.; Yagci, N.; Reisz, J. A.; Pozo Garcia, V.; D'Alessandro, A.; van der Wielen, L. A. M.; von Lindern, M.; Wahl, S. A.; Van den akker, E.

2026-06-02 cell biology 10.64898/2026.06.02.729469 medRxiv
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Transfusion-ready red blood cells can be cultured ex vivo from hematopoietic progenitors. Despite its promising outlook, a cultured transfusion unit cannot be produced at competitive costs. Large media volumes are required to maintain a maximum erythroblast cell density of 1-2.106 cells/mL during the erythroblast proliferation stage. To identify the origin of the cell density limitation, we investigated the cellular support and metabolomic phenotype using different media formulations and feeding regimens. Media that were exposed to an increasing density of erythroblasts (termed spent media) displayed a proportional decrease in erythroblast proliferation support. A 1:1 combination of spent media with fresh media (not previously exposed to the cells) restored growth for all tested conditions. Filtering both fresh and spent media with a 3 kDa cut-off filter, and subsequent recombination of the two fractions, indicated that exhaustion of the small molecular weight fraction (<3 kDa) was primarily responsible for growth limitation. We performed targeted and untargeted metabolomics analysis, for both the intra- and extracellular compartments, following seeding in fresh medium (12, 24, 36 h). We observed degradation of nucleosides, depletion of amino acids, and a decrease in intermediates of the glutathione-ascorbate, {gamma}-glutamyl and cysteine-methionine cycles. The latter compounds suggested an increase in oxidative stress in high density erythroblast cultures. Elimination of nucleosides from the medium led to a lower accumulation of purine salvage intermediates, and a 30% increase in cell productivity. In conclusion, we demonstrate that high-density erythroid cultures are subject to metabolic stress, defining critical constraints for scalable culture expansion.

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Prognostic Significance of Admission CK-MB and Total CPK Levels in Predicting Adverse Outcomes Among STEMI Patients

Rehman, M. U.

2026-04-15 cardiovascular medicine 10.64898/2026.04.14.26350841 medRxiv
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BackgroundST-elevation myocardial infarction (STEMI) is reported to be a leading cause of mortality worldwide. While cardiac troponins are the gold standard for myocardial injury detection but creatine kinase-MB (CK-MB) and total creatine phosphokinase (CPK) retain prognostic use in resource-limited settings. ObjectiveTo evaluate the prognostic significance of admission CK-MB and CPK levels in STEMI patients and to assess their association with hematological parameters for integrated risk stratification. MethodsThis cross-sectional study enrolled 15 consecutive STEMI patients from the Punjab Institute of Cardiology, Lahore, during January 2024. Comprehensive laboratory analysis including cardiac biomarkers (CK-MB, CPK, troponin-I, LDH), complete blood count, renal function, serum electrolytes, and metabolic parameters, was performed on admission. Pearson correlation and comparative statistical analyses were also conducted to assess the relationships between cardiac biomarkers and hematological indices. ResultsThe cohort includes 15 patients (mean age 50.1 {+/-} 12.2 years; 73.3% male). Cardiac biomarker elevation was prevalent: CK-MB was elevated in 12/15 (80%), CPK was elevated in 12/15 (80%), with concordant elevation in 11/15 (73.3%), which indicates extensive myocardial necrosis. Troponin-I showed the highest elevation rate at 13/15 (86.7%). Hematological abnormalities included anemia (60%), WBC elevation (53.3%), and RBC reduction (40%). Random glucose averaged 150.80 {+/-} 63.55 mg/dL, with 66.7% highlighted the hyperglycemia. Remarkably, electrolyte balance was preserved in all of the patients (0% sodium, potassium, and bicarbonate abnormalities), indicating maintained homeostasis. Pearson correlation analysis revealed a significant correlation between CK-MB and CPK (r = 0.615, p = 0.0126), while correlations between cardiac biomarkers and hematological parameters were weak (p > 0.05). Risk stratification identified 53.3% of patients as high-risk who required intensive management. ConclusionsCK-MB and CPK demonstrate significant concordance and retain prognostic value in STEMI patients, particularly in resource-limited settings where troponin access may be constrained. While troponin-I remains the most sensitive biomarker, combined assessment of conventional cardiac enzymes supports reliable evaluation of myocardial injury. Hematological parameters reflect systemic response but show limited correlation with cardiac biomarkers.

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HbF/F-cell and the Phenotype of Sickle Cell Disease

Wilks, A.; Lofters, J.; Lee, J.; Milton-Hicks, J.; Klings, E.; Steinberg, M.

2026-06-04 hematology 10.64898/2026.06.02.26354737 medRxiv
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Fetal hemoglobin (HbF) prevents the polymerization of sickle hemoglobin (HbS). HbF, measured usually as a percent of total hemoglobin (%HbF), is inversely associated with the severity of sickle cell disease (SCD) but fails to capture the distribution of HbF concentrations within red blood cells (RBCs). The relative proportion of HbF and HbS within a RBC is reflected by the HbF:HbS ratio whereas HbF/F-cell quantifies the absolute amount of HbF/RBC. While correlated, HbF:HbS ratio and HbF/F-cell are not interchangeable. In the context of mean corpuscular hemoglobin (MCH), HbF/F-cell approximates whether sufficient HbF is present to inhibit HbS polymerization. We examined the association of mean HbF/F-cell with sub-phenotypes of sickle cell disease in three independent cohorts. Both %HbF and HbF/F-cell were significantly associated with multiple clinical and laboratory features of SCD; however, HbF/F-cell demonstrated stronger associations with clinical severity measures across cohorts. Higher HbF/F-cell was associated with fewer clinical events, reduced hemolysis, and mortality. Changes in HbF/F-cell after hydroxyurea treatment were associated with ~11-13% reduction in acute events in patients with <1 pg increase and >60% reduction with a >5 pg increase in HbF/F-cell. For each pg increase in HbF/F-cell there was ~6% reduction in the rate of acute events. As a surrogate for the distribution of HbF concentrations among F-cells, HbF/F-cell adds physiologically relevant insights that could guide prognosis and treatment

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Prenatal diagnosis of sickle cell disease by amniocentesis using FTA technology in a context of precariousness in sub-Saharan Africa: Challenges and perspectives

KAMUANYA, N. C.; LOKOMBA, V. B.; MIKOBI, E. K. B.; MIKOBI, H. T. M.; LUKUSA, P. T.; Mikobi, T. M.

2026-03-24 genetic and genomic medicine 10.64898/2026.03.20.26348948 medRxiv
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Sickle cell disease (SCD) is the most common inherited hemoglobinopathy worldwide. Improving the quality of life of people with SCD requires prenatal and neonatal screening. Our primary objective was to demonstrate that prenatal diagnosis of SCD is possible even in situations of poverty. Secondarily, we described the socioeconomic profile of couples seeking molecular diagnosis of SCD in Kinshasa, Democratic Republic of Congo. Methods This was a cross-sectional study conducted in Kinshasa between January 2020 and December 2025. During this study period, 107 couples underwent prenatal diagnosis. Prenatal diagnosis was performed using amniocentesis with FTA Elute technology. This diagnosis was confirmed at birth using cord blood DNA extracted via the conventional salting-out technique. Results The mean age of the pregnant women was 28 {+/-} 4 years. Eighty-one couples (75.7%) were Christian, nine couples (8.4%) were Muslim, and seventeen couples (15.8%) were animist. Eighty-two couples (76.6%) were known heterozygous AS couples, eleven (10.2%) were heterozygous couples, and fourteen (13.0%) were couples composed of one homozygous SS and one heterozygous AS partner. All pregnancies were singleton. Socioeconomic status was upper middle class (39.2%). The AS genotype was found in 79% of the fetuses. One intrauterine fetal death was observed after amniocentesis. In terms of handling, the FTA Elute technology reduces DNA extraction time to 30 minutes. It is easy to use. Results are available in less than 24 hours. Conclusion The FTA Elute technology is a reliable, less expensive, and easy-to-use prenatal screening technique for sickle cell disease. Sample transport and storage conditions are better suited to resource-limited settings.

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An IgG-Optimized Enzyme-Linked Lectin Assay (ELLA) for Quantitative Analysis of Immunoglobulin Glycosylation

Wiggins, C. D.; Lauffenburger, D. A.

2026-06-01 bioengineering 10.64898/2026.05.28.728458 medRxiv
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Antibody Fc glycosylation is modulated in a variety of disease and immune response contexts, altering downstream functional responses including antibody-dependent cellular cytotoxicity through modified immune cell Fc receptor binding. Accessible, high-throughput glycosylation assays such as enzyme-linked lectin assays (ELLAs) are essential to advance understanding of glycosylation regulation and function. However, current ELLA protocols lack standardization and optimization, and results are reported out in arbitrary absorbance units, limiting reproducibility and cross-study comparability. We developed an optimized multi-lectin parallel ELLA with three specific improvements: systematic optimization of incubation times and reagent concentrations; incorporation of Protein A for IgG specificity; and use of commercially available bovine fetuin B as a quantitative surrogate standard for cross-study reproducibility. Our panel of 8 lectins, SNA, RCA, LCA, PHA-E, PHA-L, MAL-I, WGA, and DSL, cover the major IgG glycoforms. We demonstrate that our ELLA panel can reveal biologically relevant cytokine-induced plasticity of IgG glycosylation profiles in immortalized B cells.

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Modeling competitive transplantation using HLA-mismatched human hematopoietic stem cells

Idowu, A. M.; Ropa, J.; Hurwitz, S. N.

2026-03-20 cell biology 10.64898/2026.03.18.712629 medRxiv
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BackgroundCompetitive transplantation is essential for defining intrinsic repopulating capacity of murine hematopoietic stem and progenitor cells (HSPCs), yet comparable assays for human cells have been limited by the lack of a robust in vivo platform. MethodsHere, we describe a novel competitive transplantation method in humanized NOD.Cg-KitW-41J Tyr + Prkdcscid Il2rgtm1Wjl/ThomJ (NBSGW) mice that enables simultaneous engraftment and longitudinal tracking of distinct human grafts within a shared microenvironment. ResultsUsing human leukocyte antigen-mismatched donor CD34+ cells, this method facilitates standard flow cytometry panels to track multiple donor cell chimerism, lineage output, and HSPC composition. The experimental framework may be adapted to different mouse models, conditioning strategies, donor sources, and treatments. ConclusionsOverall, this humanized competitive repopulation assay fills a critical translational gap and offers a flexible foundation for advancing mechanistic discovery in human hematopoietic biology and improving clinical strategies for stem cell transplantation.

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A portable molecular laboratory for rapid genotyping in the field: application to sickle cell disease

Grunder, F.; Haemmerli, A.-F.; Bokembya, C. I. N.; Hennart, S.; Helmers, M.; Porret, N. A.; Graz, B.; Choudja Ouabo, C.; Abriel, H.

2026-05-12 genetic and genomic medicine 10.64898/2026.05.05.26352080 medRxiv
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BackgroundSickle cell disease (SCD) is the most common recessive genetic disorder, caused by pathogenic variants of the HBB gene. SCD is associated with a range of clinical manifestations, including vaso-occlusive crises, infections, and severe anaemia, which contribute to increased morbidity and mortality. The frequency of pathogenic alleles is high in Sub-Saharan African countries, with heterozygous carriers reaching up to 25% of the population. Several methods can be employed for molecular diagnostics, with HBB gene sequencing being the most precise. However, access to DNA analyses and sequencing in Low- and Middle-Income Countries (LMICs), where SCD prevalence is high, is limited. Understanding genetic profiles is crucial at both individual and population levels, as it can guide public health strategies and facilitate accurate genetic counselling. AimThis feasibility study aimed to demonstrate that a portable medical genetic laboratory (in suitcases) can be used to genotype individuals for the HBB A, S, and C alleles and their combinations within a few hours outside of a laboratory setting. Methods and resultsWe established a portable medical genetics laboratory capable of DNA extraction and isothermal DNA amplification using a commercially available kit for the A, S, and C alleles of the HBB gene. During one single study day, this portable lab was set up in a room where the Swiss Association of Patients with SCD was holding its annual meeting. We analysed the samples of 27 participants who were aware of their A, S, or C status. We collected buccal swabs and dried blood samples for genotyping. Genotype results for all participants were obtained within five hours after sample collection. In four cases, we observed discrepancies between the buccal swab and blood genotypes; three were resolved upon repeat testing, and one reflected donor chimerism following hematopoietic stem-cell transplantation. ConclusionsThis study demonstrates the feasibility and efficiency of using a portable medical genetics laboratory for rapid genotyping of HBB SCD alleles in community settings.This approach can improve access to molecular diagnostics in resource-limited environments. Such tools have the potential to significantly enhance local capabilities for genetic screening, counselling, and public health planning in regions heavily affected by SCD.

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An AI-Powered Smartphone Application for Universal and Standardized Reading and Interpretation of Lateral Flow Assays

Bermejo-Pelaez, D.; Darias, O.; Pastor, L.; Valles, R.; Diez, N.; Lin, L.; Garcia-Villena, J.; Cuadrado, D.; Vladimirov, A.; Alamo, E.; Postigo, M.; Rodriguez-Dominguez, M.; Canton, R.; Rodriguez-Tudela, J. L.; Alastruey Izquierdo, A.; Bohorquez, L. C.; Rubio, J. M.; Dacal, E.; Luengo-Oroz, M.

2026-05-18 public and global health 10.64898/2026.05.14.26352875 medRxiv
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Introduction. Lateral flow assays (LFAs) are indispensable rapid diagnostic tools in healthcare, enabling point-of-care diagnosis critical for patient management and support disease burden assessment and surveillance when results are properly recorded. However, misinterpretation errors and unreported cases remain a concern. A quality-assured, affordable Ai-powered tool, supporting the decision-making during result interpretation could promote proper disease monitoring and epidemiological surveillance. Here, we describe the performance of a universal AI model to digitize and interpret results from multiple LFA types through a smartphone application, a step that could ultimately enable standardized and digitally reportable test outcomes. Methods. The AI algorithm was evaluated in 17 LFA types, including both 2-band and 3-band tests for different diseases and manufacturers. The model was trained on a dataset of 22,576 images captured under diverse lighting conditions with different smartphone models and using a custom mobile application, TiraSpot (Spotlab, Madrid, Spain). To assess generalizability, a leave-one-out cross-validation was applied, where in each LFA type was iteratively excluded from training and used for testing. Model performance was evaluated using bootstrapping on the inference dataset. Results. In the assessment of the model's ability to generalize to new LFA types not previously analyzed (not included during development), the model achieved an overall AUC of 94.3% for second band detection. This overall performance was enhanced to 99.3% (Sensitivity=98,6%; Specificity=98%) after training with 50 images of each LFA type, highlighting the benefit of additional data for specific LFA types. For the third band detection, where less training data was available, the system achieved an overall AUC of 83.9% for unseen LFAs, improving to 94.2% (Sensitivity=92.9%; Specificity=87,9%) after training with 50 images of each LFA type. Conclusion. This system demonstrates the feasibility of an AI-powered universal digital reader for interpreting LFA results from diverse test types using smartphone-captured images. Its compatibility with standard smartphones makes it a universal tool, enabling reliable LFA interpretation across devices and settings. By standardizing test interpretation and digitizing results, this tool could support decision making in result interpretation, enhancing epidemiological surveillance, particularly in resource-limited settings. Its adaptability across various infections highlights its potential to improve diagnostic consistency and support disease management in diverse healthcare settings.

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Efficacy of the PragmaVAC Manual Negative Pressure Wound Therapy Device to Treat Acute Traumatic Wounds in a Conflict Setting: A Retrospective Cohort Study from Gaza

Ramadan, I.; Hariri, M.; Shalakhti, O.; Alawa, J.; Godier-Furnemont, A.; Traboulsi, A. A.-R.; MOWAFI, H.

2026-06-10 surgery 10.64898/2026.06.04.26354740 medRxiv
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Abstract: Background: Acute war-related traumatic wounds present significant challenges due to significant soft-tissue damage/loss, risk of contamination, limited access to antimicrobial therapy, need for delayed closure, and limited access to surgical and wound care. Negative Pressure Wound Therapy (NPWT) has been used effectively to reduce the volume of soft-tissue defects, edema, and infection in traumatic wounds, and to promote growth of healthy granulation tissue. However, conventional NPWT devices are costly and electricity-dependent, limiting their utility in conflict settings. Methods: This retrospective cohort study evaluated the use of PragmaVAC, a manually operated, electricity-independent NPWT device, in patients across three hospitals in Gaza with conflict-related wounds that were deemed by the treating surgeon to be unsuitable for primary closure. Secondary analysis was performed of clinical records of patients treated with the PragmaVac NPWT device to assess ability to achieve a primary outcome of wound bed with healthy granulation tissue, time to primary outcome, and rates of adverse effects. Secondary outcome of wound closure and closure method was also assessed. Results: Treatment with PragmaVAC manual NPWT was prescribed to 88 patients. Of those, 27 (31%) had incomplete documentation of their wound healing or were lost to follow up. The remaining 61 (69%) had complete documentation of their wound healing, complications, and final outcome with 59 (67%) successful closure and 2(2%) failure. Conclusion: The use of the PragmaVAC NPWT device provided a safe, effective wound care option to achieve wound closure for large conflict-related traumatic wounds in resource-limited settings. Future studies may further evaluate such use through prospective trials, evalutions of patients' experiences with manual NPWT, and evaluating outcomes beyond primary wound closure to include medium- and long-term complications, cosmesis, and cost of therapy.