Vitalant

Twenty-four-hour post-transfusion recovery (PTR24) is a key measure of red blood cell (RBC) quality. Traditionally assessed using radioactive chromium-51 (51Cr), supply and facility limitations have prompted the need for alternatives. Biotinylation of RBCs (BioRBCs) offers a non-radioactive method capable of distinguishing multiple RBC populations by flow cytometry. Researchers at three major blood centers developed a standardized BioRBC labeling protocol for pharmacokinetic studies. The multicenter development of the standardized protocol followed a two-phase design. First, inter- and intra-site flow cytometry variability was assessed using RBCs biotinylated at 6 and 18 μg/mL. Samples were stained with streptavidin-phycoerythrin (SA-PE), diluted, and analyzed in triplicates using shared flow cytometry templates. Second, inter-site variability of biotinylation was tested using paired RBC units split and processed simultaneously at each site. Separation index (SI) was calculated as: SI = (median BioRBC - median unlabeled)/(84th percentile unlabeled - median unlabeled)/0.995. Staining and analysis of BioRBCs demonstrated minimal inter- and intra-site variability when using identical protocols. Coefficients of variation for median fluorescence intensity were <7.9% within sites and <5.1% across sites. Biotinylation of paired RBC with 6 and 18 μg/mL of biotin resulted in comparable separation indices, suggesting reliable and reproducible biotinylation ratios. The standardized BioRBC protocol produces reliable, reproducible labeling with minimal inter-site variability. In vivo performance will be assessed in upcoming PTR24 studies comparing BioRBCs with dual radiolabeling using 51Cr and technetium-99m (99mTc).

Donidalorsen, a prekallikrein-directed antisense oligonucleotide indicated for prophylaxis of hereditary angioedema (HAE) attacks in patients aged ≥12 years, demonstrated efficacy and acceptable safety in the phase 3, placebo-controlled OASIS-HAE trial (NCT05139810). Here, we report 1-year results from the corresponding open-label extension (OLE) cohort of the OASISplus study (NCT05392114). OASISplus included patients who rolled over from OASIS-HAE. Patients who received donidalorsen 80 mg or placebo subcutaneously every 4weeks (Q4W) in OASIS-HAE received donidalorsen Q4W in OASISplus. Patients who received donidalorsen 80 mg or placebo every 8weeks (Q8W) in OASIS-HAE received donidalorsen Q8W or Q4W, if not attack-free in the final 8weeks of OASIS-HAE. The primary endpoint was safety (ie, incidence of treatment-emergent adverse events [TEAEs]). Secondary endpoints included the monthly rate of HAE attacks and Angioedema Quality of Life (AE-QoL). The OLE cohort included 83 patients (Q4W, n=69 [83%]; Q8W, n=14 [17%]). Of these, 75 (90.4%) completed Year 1, and 6 patients receiving donidalorsen Q8W in OASIS-HAE switched to the Q4W dosing group in the OLE. Median donidalorsen exposure was 392.3 days. From Weeks 0 to 52, reductions in mean HAE attack rate from OASIS-HAE baseline were 94% (Q4W) and 95% (Q8W), and patients reported clinically meaningful improvements in mean AE-QoL total score at Week 52 (Q4W, 28.1 points; Q8W, 26.7 points). Twenty-two (27%) patients reported treatment-related TEAEs; none were serious, and injection-site reactions were the most frequently reported. Donidalorsen demonstrated sustained reductions in HAE attack rate, improvements in QoL, and an acceptable safety profile after 1 year of treatment.

The role of aromatic gut-derived bacterial metabolites (GDBMs) in shaping immune cell metabolism and function remains poorly explored. Using ex vivo metabolomic profiling of paired plasma and CD4+ T-cells from people living with HIV-1 (PLWH), we identified a network of aromatic GDBMs whose cell-associated abundance, rather than systemic levels, was linked to broad alterations in CD4+ T-cell metabolic and functional states. Among these metabolites, p-cresol sulfate (PCS) emerged as a mechanistic prototype investigated in depth. Ex vivo flow cytometry and single-cell RNA sequencing of CD4+ T-cells stratified by cell-associated PCS levels revealed dose-dependent enrichment of transcriptional programs associated with impaired differentiation capacity, regulatory-like identity, and cellular senescence. Consistently, in vitro transcriptomic and proteomic analyses of PCS-exposed CD4+ T cells demonstrated induction of cell-cycle arrest, mitochondrial dysfunction, and senescence-associated programs, including upregulation of p16 and p21. Integration of these immunometabolic features with measurements of HIV-1 reservoir size in PLWH revealed that CD4+ T-cell states defined by cell-associated GDBMs track with intact proviral DNA levels in vivo. Together, these findings define a microbiome-derived axis that reshapes CD4+ T-cell metabolism and fate and promotes immune aging–associated states in PLWH. Our data suggest that cell-associated GDBMs may foster immunometabolic CD4+ T-cell states previously linked to long-term HIV-1 reservoir persistence in vivo.

Abstract Purpose Several clinical studies have shown correlations between certain physiological measurements and an autism spectrum disorder (ASD) diagnosis. Such findings, however, have generally not resulted in tangible progress towards practical translation. In fact, most studies have been retrospective in nature and compare biomarker profiles in children with an ASD diagnosis to those who are typically developing. A clinically meaningful test, however, requires an ASD diagnostic center to distinguish children from those with a developmental disorder pre-diagnosis. Methods This paper presents, for the first time, a double-blind case/control trial design in which metabolic profiles, collected at two developmental pediatric clinics, were collected from children on a diagnostic waitlist for the purpose of developing a blood-based test for ASD. Besides obtaining blood samples, the children underwent gold-standard clinical evaluations, including the Autism Diagnostic Observation Schedule (ADOS), Mullens Scale of Early Learning (MSEL), and Vineland Adaptive Behavior Scale (VABS). The analysis, together with a complete medical history and physical exam, allowed to confirm or rule-out suspected ASD using DSM-V criteria. The study was based on a cohort of 140 children between the ages 18-60 months, that were referred to a developmental pediatrician because of concerns in their development. Results 114 of these children received an ASD diagnosis, while 26 were diagnosed with non-ASD related developmental delays. Based on the measured metabolites, artificial intelligence-based classification algorithms allowed for an over 80% accuracy in predicting whether a sample came from a child diagnosed with ASD or not. Conclusion While these results need to be replicated in a larger study, especially involving more children with non-ASD related developmental delays, this is the first work using physiological measurements, coupled with AI, to support ASD diagnoses in a clinically relevant setting.

We present data from the TRuE-V1/TRuE-V2 rollover TRuE-V long-term extension study evaluating withdrawal and continuation of ruxolitinib cream treatment. Among patients with near-complete facial repigmentation in TRuE-V1/TRuE-V2, approximately 70% of those randomized to treatment continuation and approximately 40% of those randomized to treatment withdrawal maintained facial repigmentation indicative of treatment success (i.e. ≥ F-VASI 75). Lost repigmentation could be regained in a timely manner following reinitiation of open-label active treatment.

Objectives:Epigenetic clocks are biomarkers of aging. Epigenetic clocks are associated with early-life famine exposure. We investigated the impact of a cluster-randomized early-life nutrition intervention on epigenetic age.Methods:We analyzed follow-up data from participants in the INCAP Nutrition Supplementation Trial, conducted in 4 villages in eastern Guatemala. DNA methylation was measured in buffy coat samples using the Illumina Infinium™ MethylationEPICv2.0 array and standard quality control procedures.Epigenetic age was quantified using DunedinPACE, PhenoAge, and GrimAge. PhenoAge and GrimAge acceleration were calculated as residuals by regressing epigenetic age on chronological age. We used intent-to-treat difference-in-difference modeling to assess the impact of a protein-energy supplement provided during the first 1,000 days of life (conception to age 2y) on epigenetic age in middle adulthood. Covariates included sex, birth year, the trial supplement type (atole [intervention] vs. fresco [control]), exposure period of supplement (any of the first 1,000 days, other), and a random effect to account for sibships. The primary coefficient of interest was represented by the interaction between supplement type and exposure period.Results:The analysis included 1095 participants (mean age 45.0 y (SD 4.3); 60.3 % female, 40.3 % exposed to any atole during the first 1,000 days, mean DunedinPACE 1.2 (SD 0.1), Phenoage 46.7 y (SD 6.7), and GrimAge 56.3 y (SD 4.1). In difference-in-difference analyses, exposure to atole during any of the first 1,000-day period was associated with lower DunedinPACE (−0.03, 95% CI −0.06, −0.004), PhenoAge acceleration (− 1.91 y, 95% CI −3.43, −0.39), and GrimAge acceleration (−0.85 y, 95% CI −1.53, −0.11) compared to other exposures. Following additional adjustment for cell type proportions, the direction of the coefficients remained the same but were no longer statistically significant.Conclusions:Exposure to atole during the first 1,000 days was associated with modest reductions in epigenetic age as measured by DunedinPACE, PhenoAge, and GrimAge. These findings complement prior evidence of epigenetic age acceleration among individuals with early-life famine exposure.

Cryoprecipitated antihemophiliac factor (cryo) is primarily used to replenish fibrinogen in acquired coagulopathy. Little has been reported about its usage patterns within hospitals with respect to patient population, frequency of use, and dosing. Cryo production and usage data were collected over 5 years from July 1, 2016, to June 30, 2021, from nationally based blood collection centers in the United States (U.S. n = 2) and Canada (n = 2) and from eight large academic hospitals, respectively. Usage data for a similar product, purified fibrinogen concentrate were also collected from four hospitals. Two U.S. blood collectors reported increases in cryo production normalized to total whole blood collections from 17.4% in 2016 to 22.3% in 2021 and from 16.4% in 2016 to 20.2% in 2021. In contrast, in Canada cryo manufacturing increased slightly in one region (11.3%-12.9%) and decreased (8.0%-2.0%) in the other. Cryo utilization, defined as numbers of patients treated normalized to the inpatient census, and dose administered per patient did not consistently increase and differed significantly between hospitals participating in the study (p < .0001). Likewise, the departments that most frequently transfused cryo varied between hospitals. Similarly, variations in practice were observed for fibrinogen concentrate usage. While much of Canada and Europe have moved towards using fibrinogen concentrate, two large U.S. collectors have increased cryo distribution. The lack of standardization and variability in the clinical practice regarding the use of cryo and fibrinogen concentrate reported by study sites may be attributable to practitioner preference, availability, and/or cost rather than adherence to published evidence or guidelines.

The incidence of transfusion-transmitted babesiosis (TTB) has declined substantially in recent years following the introduction of geographically selective nucleic acid testing (NAT) by blood collectors. This approach has proven quite effective in interdicting parasite-infected units from US regions where Babesia is endemic and is complemented by donor screening questions about prior diagnosis (used in lieu of NAT in non-Babesia-endemic states). Despite these successes, however, breakthrough cases continue to occur due to limitations in the current geographically targeted testing strategy. Herein, we report a Minnesota donor reinstated almost 3.5 years after a Babesia NAT-positive donation whose first post-reinstatement donation tested NAT-negative. Three months later, a subsequent donation tested NAT-positive for Babesia microti. Notification of the hospital that transfused the NAT-negative red blood cell unit revealed a likely B. microti transmission. This reverse-notification TTB case was unlikely to have been the result of either a product from an asymptomatic untested donor from a non-endemic state infected during travel to an endemic area or a tick-borne infection of the transfusion recipient. More likely, TTB resulted from transfusion of a NAT-negative unit donated 3 months prior to the involved donor's subsequent NAT-positive collection. To our knowledge, this is the first reported potential Babesia NAT failure.