Single Donor Research Articles

Allogeneic chimeric antigen receptor cell therapies for cancer: progress made and remaining roadblocks.

Chimeric antigen receptor (CAR) T cells are revolutionizing cancer therapy, particularly for haematological malignancies, conferring durable and sometimes curative responses in patients with advanced-stage disease. The CAR T cell products currently approved for clinical use are all autologous and are often effective; however, in patients who are lymphopenic and/or heavily pretreated with chemotherapy, autologous T cells can be difficult to harvest in sufficient numbers or have functional impairments that might ultimately render them less efficacious. Moreover, autologous products take several weeks to produce, and each product can be used in only one patient. By contrast, allogeneic CAR T cells can be produced for many patients using T cells from a single healthy donor, can be optimized for safety and efficacy, can be instantly available for 'off-the-shelf' use and, therefore, might also be more cost-effective. Despite these potential advantages, the development of allogeneic CAR T cells has lagged behind that of autologous products, owing to the additional challenges such as avoiding graft-versus-host disease and host-mediated graft rejection. Over the past few years, the development of advanced genome-editing techniques has facilitated the generation of novel allogeneic CAR T cell products. Furthermore, CAR cell products derived from other cell types such as induced pluripotent stem cells and natural killer cells are being investigated for clinical use. In this Review, we discuss the potential of allogeneic CAR cell products to expand life-saving immunotherapy to a much broader population of patients in the coming years, the progress made to date and strategies to overcome remaining hurdles.

Cyclosporin A toxicity on endothelial cells differentiated from induced pluripotent stem cells: Assembling an adverse outcome pathway.

Cyclosporin A (CSA) is a potent immunosuppressive agent in pharmacologic studies. However, there is evidence for side effects, specifically in regard to vascular dysfunction. Its mode of action inducing endothelial cell toxicity is partially unclear, and a connection with an adverse outcome pathway (AOP) is not established yet. Therefore, we designed this study to get deeper insights into the mechanistic toxicology of CSA on angiogenesis. Stem cells, especially induced pluripotent stem cells (iPSCs) with the ability of differentiation to all organs of the body, are considered a promising in vitro model to reduce animal experimentation. In this study, we differentiated iPSCs to endothelial cells (ECs) as one cell type that in other studies would allow to generate cells or organoids from single donors. Flow cytometry and immunostaining confirmed our scalable differentiation protocol. Then dose and time course experiments assessing CSA cytotoxicity on iPS derived endothelial cells were performed. Transcriptomic data suggested CDA dependent induction of reactive oxygen species (ROS) and mitochondrial dysfunction, which was confirmed by in vitro experiments. Additionally, CSA impaired angiogenesis via ROS induction. Finally, we combined this information into an AOP, was developed based on here observed and literature based evidence for CSA-mediated endothelial cell toxicity. This AOP will help to design in vitro test batteries, model events observed in human toxicity studies, as well for predictive toxicology.

Stereoselective Radical Acylfluoroalkylation of Bicyclobutanes via N-Heterocyclic Carbene Catalysis.

Cyclobutanes are prominent structural components in natural products and drug molecules. With the advent of strain-release-driven synthesis, ring-opening reactions of bicyclo[1.1.0]butanes (BCBs) provide an attractive pathway to construct these three-dimensional structures. However, the stereoselective difunctionalization of the central C-C σ-bonds remains challenging. Reported herein is a covalent-based organocatalytic strategy that exploits radical NHC catalysis to achieve diastereoselective acylfluoroalkylation of BCBs under mild conditions. The Breslow enolate acts as a single electron donor and provides an NHC-bound ketyl radical with appropriate steric hindrance, which effectively distinguishes between the two faces of transient cyclobutyl radicals. This operationally simple method tolerates various fluoroalkyl reagents and common functional groups, providing a straightforward access to polysubstituted cyclobutanes (75 examples, up to >19:1 d.r.). The combined experimental and theoretical investigations of this organocatalytic system confirm the formation of the NHC-derived radical and provide an understanding of how stereoselective radical-radical coupling occurs.

Safety comparison of single-donor and pooled fecal microbiota transfer product preparation in ulcerative colitis: systematic review and meta-analysis.

Multiple studies have evaluated fecal microbiota transfer (FMT) in patients with ulcerative colitis (UC) using single-donor (SDN) and multidonor (MDN) products. Systematic review and meta-analysis were performed to compare the safety of SDN and MDN products. Systematic searches were performed in Web of Science, Scopus, PubMed, and Orbit Intelligence to identify studies that compared FMT products manufactured using SDN or MDN strategies against control treatment in patients with UC. Fifteen controlled studies were selected for meta-analysis (11 randomized controlled trials and 4 controlled cohort trials). Safety of each treatment type was assessed using the counts of adverse events and serious adverse events using fixed- and random-effects models. Significance of the indirect difference between FMT preparations was assessed using a network approach. Benefit-risk ratios were calculated by multiplicative utility model, incorporating geometric mean of risk ratios (RRs) of efficacy and safety. Safety data was collected for a total of 587 patients (193 exposed to SDN products, 114exposed to MDN products and 280 exposed to control treatment). The 12 studies showed similar overall safety event counts for MDN and SDN versus placebo (RRs: 0.90 and 1.09, respectively [P = 0.206 and P = 0.420, respectively]). Results indicated similar risk of safety events for MDN compared to SDN (RR: 0.83, P = 0.159). Positive benefit-risk ratios were demonstrated for MDN and SDN versus placebo (RRs: 1.70 and 1.16, respectively [P = 0.003 and P = 0.173, respectively]). MDN had a greater benefit-risk ratio compared to SDN (RR: 1.46, P = 0.072). Similar safety profiles were observed for MDN and SDN strategies. Alongside previously described superior efficacy, treatment with MDN has greater benefit-risk ratio than SDN in patients with UC. Further development of MDN FMT treatment for UC should be considered.

Exploring the potential of dermal grafting: a narrative review in plastic surgery

Dermal grafting (DG) has emerged as an innovative technique in plastic and reconstructive surgery, offering several advantages over traditional skin grafting methods. This review provides an in-depth exploration of DG, highlighting its applications, benefits, and future directions. The historical evolution of skin grafting is discussed, tracing the development of DG as a novel approach to address the limitations of conventional techniques. The review focuses on four key advantages of DG: (1) accelerated healing of donor sites; (2) improved aesthetic outcomes at recipient sites due to the elastic nature of dermal grafts; (3) increased graft availability by effectively at least doubling the amount of graft material obtained from a single donor site; and (4) utility in scar revision and reconstruction procedures, particularly in areas with restrictive scarring or contractures. Recent advancements, such as the development of a multiblade dermatome, have addressed the technical challenges associated with DG harvesting, potentially broadening the clinical adoption of this technique. Preliminary results from studies utilizing this new device have demonstrated its feasibility in producing dual grafts (split-thickness skin graft and dermal graft) concurrently, simplifying the surgical procedure. The review also explores future directions in DG, including further refinements to the multiblade dermatome, and clinical trials to validate long-term benefits. Overall, this review highlights the significant advantages of DG and its potential in advancements of plastic and reconstructive surgery, ultimately improving patient outcomes and quality of life.

Donor Parameters and Platelet Yield during Plateletpheresis: A Cross-sectional Prospective Study

Abstract Background: Platelet transfusion can be life-saving in bleeding patients with thrombocytopenia or reduced platelet function. Single donor platelets (SDP) unit use is preferred due to the various advantages like reduction in transfusion-transmitted diseases and transfusion reactions. The quality of SDP is determined by platelet yield. Therefore, we conducted this study to assess the donor parameters affecting platelet yield during plateletpheresis. Materials and Methods: This cross-sectional prospective study included 160 single-donor platelet apheresis procedures done for 1 year. The pre-donation platelet count of the donor was set to 250 × 109/L to get double-dose platelet. Plateletpheresis was performed using Trima Accel single-needle separator apheresis machine. The demographic and hematologic parameters of donors were correlated with their platelet yield. Data obtained were tabulated using version 20 of the Statistical Package for the Social Sciences (SPSS). A relationship between pre-donation donor variables and platelet yield was studied using the Pearson correlation coefficient (r). P < 0.05 was considered statistically significant. Results: There was a significant positive correlation between platelet yield and body mass index (BMI) (r = 1710 and P = 0.0304), total leukocyte count (TLC) (r = 0.2220 and P = 0.0048), and platelet count (r = 0.8880 and P < 0.0001). The mean platelet volume (MPV) showed a negative correlation with platelet yield (r = −0.1910 and P = 0.0156). Age, hemoglobin, hematocrit, and platelet distribution width showed no significant correlation with the platelet yield. Conclusion: Donor parameters such as pre-donation platelet count, BMI, TLC, and MPV need to be considered while recruiting the donors for the SDP procedure for better yield of platelets.

Echo Location: Distances to Galactic Supernovae From ASAS-SN Light Echoes and 3D Dust Maps

Light echoes occur when light from a luminous transient is scattered by dust back into our line of sight with a time delay due to the extra propagation distance. We introduce a novel approach to estimating the distance to a source by combining light echoes with recent three-dimensional dust maps. We identify light echoes from the historical supernovae Cassiopeia A and SN 1572 (Tycho) in nearly a decade of imaging from the All-Sky Automated Survey for Supernovae (ASAS-SN). Using these light echoes, we find distances of 3.6±0.1 kpc and 3.2−0.2+0.1 kpc to Cas A and Tycho, respectively, which are generally consistent with previous estimates but are more precise. These distance uncertainties are primarily dominated by the low distance resolution of the 3D dust maps, which will likely improve in the future. The candidate single degenerate explosion donor stars B and G in Tycho are clearly foreground stars. Finally, the inferred reddening towards each SN agrees well with the intervening column density estimates from X-ray analyses of the remnants.

Intramolecular Charge Transfer Inhibition Strategy toward a Desired Solvatochromic Fluorescent Platform: Visualization of Duple Organelles and Detection of Carbon Dioxide.

Solvatochromic fluorescent probes are crucial molecular tools to investigate and aggregate proteins' fold, visualize fine structures in biomembranes, and label different organelles in dual emission colors. However, solvatochromic fluorogens often displayed a weak emission at high polarity, hindering their bioimaging applications. To resolve this problem, herein, we propose an intramolecular charge transfer (ICT) inhibition strategy. The probe was designed with a single electronic donor and two acceptors in order to split and inhibit the ICT procedure. As a result, the probe displayed an intense emission at both low and high polarities and showed a large emission shift (84 nm) upon polarity change. Using the probe, we successfully imaged lipid droplets and the endoplasmic reticulum in different fluorescence colors. Moreover, the different degrees of lipid accumulation by oleic acid, stearic acid, and cholesterol (oleic acid > stearic acid > cholesterol) have been revealed. The lipid accumulation induced by the three lipids could be rapidly consumed under lipid-less conditions, and the lipids with stearic acid were the most difficult to be consumed. The biological results could facilitate the understanding and treatment of lipid accumulation and obesity. Furthermore, utilizing the polarity increase of diethylamine after the reaction with CO2, the ratiometric detection of CO2 has been achieved for the first time with the probe.

P1.06C.02 Description of A Single Center Thoracic Rapid Tissue Donation Program Experience

P1.06C.02 Description of A Single Center Thoracic Rapid Tissue Donation Program Experience

Measurement of Enhanced Spin-Orbit Coupling Strength for Donor-Bound Electron Spins in Silicon.

While traditionally considered a deleterious effect in quantum dot spin qubits, the spin-orbit interaction is recently being revisited as it allows for rapid coherent control by on-chip AC electric fields. For electrons in bulk silicon, spin-orbit coupling (SOC) is intrinsically weak, however, it can be enhanced at surfaces and interfaces, or through atomic placement. Here it is showed that the strength of the spin-orbit coupling can be locally enhanced by more than two orders of magnitude in the manybody wave functions of multi-donor quantum dots compared to a single donor, reaching strengths so far only reported for holes or two-donor system with certain symmetry. These findings may provide a pathway toward all-electrical control of donor-bound spins in silicon using electric dipole spin resonance (EDSR).

Insights into human norovirus cultivation in human intestinal enteroids.

Human noroviruses (HuNoVs) are a significant cause of epidemic and sporadic acute gastroenteritis worldwide. The lack of a reproducible culture system hindered the study of HuNoV replication and pathogenesis for almost a half-century. This barrier was overcome with our successful cultivation of multiple HuNoV strains in human intestinal enteroids (HIEs), which has significantly advanced HuNoV research. We optimized culture media conditions and generated genetically modified HIE cultures to enhance HuNoV replication in HIEs. Building upon these achievements, we now present new insights into this culture system, which involve testing different media, unique HIE lines, and additional virus strains. HuNoV infectivity was evaluated and compared in new HIE models, including HIEs generated from different intestinal segments of individual adult organ donors, HIEs from human intestinal organoids produced from directed differentiation of human embryonic stem cells that were then transplanted and matured in mice before making enteroids (H9tHIEs), genetically engineered (J4FUT2 knock-in [KI], J2STAT1 knockout [KO]) HIEs, as well as HIEs derived from a patient with common variable immunodeficiency (CVID) and from infants. Our findings reveal that small intestinal HIEs, but not colonoids, from adults, H9tHIEs, HIEs from a CVID patient, and HIEs from infants support HuNoV replication with segment and strain-specific differences in viral infection. J4FUT2-KI HIEs exhibit the highest susceptibility to HuNoV infection, allowing the cultivation of a broader range of genogroup I and II HuNoV strains than previously reported. Overall, these results contribute to a deeper understanding of HuNoVs and highlight the transformative potential of HIE cultures in HuNoV research.IMPORTANCEHuman noroviruses (HuNoVs) cause global diarrheal illness and chronic infections in immunocompromised patients. This paper reports approaches for cultivating HuNoVs in secretor positive human intestinal enteroids (HIEs). HuNoV infectivity was compared in new HIE models, including ones from (i) different intestinal segments of single donors, (ii) human embryonic stem cell-derived organoids transplanted into mice, (iii) genetically modified lines, and (iv) a patient with common variable immunodeficiency disease. HIEs from small intestine, but not colon, support HuNoV replication with donor, segment, and strain-specific variations. Unexpectedly, HIEs from one donor are resistant to GII.3 infection. The genetically modified J4FUT2 knock-in (KI) HIEs enable cultivation of a broad range of GI and GII genotypes. New insights into strain-specific differences in HuNoV replication in HIEs support this platform for advancing understanding of HuNoV biology and developing potential therapeutics.

Unveiling the Significance of tert-Butoxides in Transition Metal-Free Cross-Coupling Reactions.

The astounding reactivity of tert-butoxides in transition metal-free coupling reactions is driving the scientific community towards a new era of environmental friendly, as well as cost-effective, transformation strategies. Transition metal-catalyzed coupling reactions generate hazardous wastes and require harsh reaction conditions, mostly at elevated temperature, which increases not only costs but also environmental concerns regarding the methodology. Tert-butoxide-catalyzed/mediated coupling reactions have several advantages and potential applications. They can form carbon-carbon, carbon-heteroatom, and heteroatom-heteroatom bonds under mild reaction conditions. Mechanistic insights into these reactions include both ionic and radical pathways, with the fate of the intermediates depending on the reaction conditions and/or additives used in the reactions. Among all of the known tert-butoxides, potassium tert-butoxide has pronounced applications in transition metal-free coupling reactions as compared to other tert-butoxides, such as sodium and lithium tert-butoxides, because of the higher electropositivity of potassium compared to sodium and lithium. Moreover, potassium tert-butoxide can act as a source of base, nucleophile and single electron donors in various important transformations. In this review, we provide an extensive overview and complete compilation of transition metal-free cross-coupling reactions catalyzed/promoted by tert-butoxides during the past 10years.

THE POSSIBILITY OF DONATION OF ORGANS AND TISSUES

Organ and tissue donation is a very important subject in our society in general. I intend to discuss what organ donation is. The importance of organ and tissue donation. About the importance of awareness. Organ or tissue donation is an act by which the living individual decides to help in the treatment of other people. Donation may be from organs (kidney, liver, heart, pancreas and lung) or tissue (cornea, skin, bones, heart valves, cartilage, bone marrow and umbilical cord blood). The donation of organs such as the kidney, part of the liver and bone marrow can be done in life. For the donation of organs of deceased persons, only after the confirmation of the diagnosis of brain death can the procedure be performed. The most common is that it happens to people who suffer some kind of accident that causes head trauma, or who are victims of a stroke (stroke) and evolved to brain death. To understand the transplantation of the organs it is necessary to emphasize that the right to separate parts of the living or dead body integrate the personality, according to the understanding of Maria Helena Diniz (2006, p.249). Most of the time, organ transplantation may be the only life expectancy or the opportunity for a fresh start to about 40,000 patients waiting in the queue for Brazil. The gesture of family members who lose a loved one and decide to donate is an act of life, altruism and generosity, as a single donor can benefit at least ten people and make the difference between life and death. Brazil has the largest public transplant system in the world, responsible for financing about 95% of transplants. It is the second country in absolute number of transplants, behind only the United States.

6651 A Traceable Clinical Free Thyroxine Assay Based on Equilibrium Dialysis ID-LC/MS/MS

Abstract Disclosure: D. Spector: None. L. Zhang: None. A. Ribera: None. A. Doty: None. C. Tse: None. O. Sugahara: None. N. Vazquez: None. U. Danilenko: None. H.W. Vesper: None. Accurate diagnosis and effective treatment of thyroid diseases rely on reliable thyroid function tests. Serum free thyroxine (FT4) immunoassays (IAs) are most commonly used for FT4 measurements in clinical practice. However, FT4 IAs have not been standardized with large variations between different assay platforms. A reference measurement procedure (RMP) for FT4 based on well-defined equilibrium dialysis (ED) of serum has been established for CDC standardization program to support the standardization of clinical assays for patient care. In the current study, we aim to develop a routine clinical FT4 assay based on ED-LC/MS/MS that is traceable to the RMP. ED-LC/MS/MS analysis was performed using a commercially available micro-ED plate. FT4 in dialysate was extracted with a 96-well C18 SPE plate. FT4 in the samples was quantitated by using LC/MS/MS in positive ion mode. The assay was calibrated with the certified primary reference material. By using RMP as a reference, we compared the analysis of 40 single donor sera between the ED-LC/MS/MS assay and FT4 IA on Roche e401platform. ED-LC/MS/MS assay could resolve T4, T3, reverse triiodothyronine (rT3), and other interferences in the samples with C18 chromatography in 4 min. The linear range of the routine FT4 assay was from 0.5-100 pg/mL covering clinically relevant FT4 concentrations including hypo-, hyperthyroid patient samples. The intra and inter-run CV% of the assay in 20 days was 3.72 and 5.25%, respectively. The effect of minor changes to the method conditions was studied. It was determined that serum/buffer ratios from 1:1 to 1:2 in the ED inserts, speeds of shaker for ED, and ED incubation time, did not influence the FT4 measurement. However, elevated ED temperature at 38°C could result in a more than 10% increase in FT4 values. A microplate format for the procedures of ED and SPE is amiable to the application of an automation system which significantly improves the throughput of the procedure. Method comparison showed good agreement between ED-LC/MS/MS assay and RMP in Deming regression analysis with a slope close to 1, while there was significant bias of IA from RMP. ED-LC/MS/MS assay was also applied for pregnancy serum samples with minimal difference on the FT4 measurement from RMP. We have developed a routine clinical FT4 assay that is traceable to RMP. The accuracy, precision, and sensitivity of the assay are suitable for high-throughput FT4 measurements in clinical laboratories and large epidemiologic studies. We demonstrate that the developed method can provide accurate FT4 measurements in patients including pregnant women. Presentation: 6/3/2024

Isolation of Single Donors in ZnO.

The shallow donor in zinc oxide (ZnO) is a promising semiconductor spin qubit with optical access. Single indium donors are isolated in a commercial ZnO substrate using plasma focused ion beam (PFIB) milling. Quantum emitters are identified optically by spatial and frequency filtering. The indium donor assignment is based on the optical bound exciton transition energy and magnetic dependence. The emission stability of these single donors in terms of both intensity and frequency, alongside their transition linewidths less than twice the lifetime limit, highlight the promise of single In donors as optically accessible spin qubits. The optical stability of single donors after FIB fabrication is promising for optical device integration required for scalable quantum technologies based on single donors in direct band gap semiconductors.

Expedient measurement of total protein in human serum and plasma via the biuret method using fiber optic probe for patient samples and certified reference materials.

The biuret method is currently recognized as a reference measurement procedure for serum/plasma total protein by the Joint Committee for Traceability in Laboratory Medicine (JCTLM). However, as the reaction involved in this method is highly time-dependent, to ensure identical measurement conditions for calibrator and samples for high accuracy, a fast and simple measurement procedure is critical to ensure the precision and trueness of this method. We measured serum/plasma total protein using a Cary 60 spectrophotometer coupled with a fiber optic probe, which was faster and simpler than the conventional cuvette method. The biuret method utilizing alkaline solutions of copper sulfate and potassium sodium tartrate was added to the sample and calibrator (NIST SRM 927e) incubated for 1h before measurement. A panel of samples consisting of pooled human serum, single donor serum, and certified reference materials (CRMs) from three sources were measured for method validation. Sixteen native patient samples were measured using the newly developed biuret method and compared against clinical analyzers. Additionally, the results of three cycles of a local External Quality Assessment (EQA) Programme submitted by participating clinical laboratories were compared against the biuret method. Our biuret method using fiber optic probe demonstrated good precision with within-day relative standard deviation (RSD) of 0.04 to 0.23% and between-day RSD of 0.58%. The deviations between the obtained values and the certified values for all three CRMs ranged from -0.38 to 1.60%, indicating good method trueness. The routine methods using clinical analyzers were also found to agree well with the developed biuret method using fiber optic probe for EQA samples and native patient samples. The biuret method using a fiber optic probe represented a convenient and reliable way of measuring serum total protein. It also demonstrated excellent precision and trueness using CRMs and patient samples, which made the method a simpler candidate reference method for serum protein measurement.

Icelike water molecules with single hydrogen bond donor on the surface of nano anatase and rutile particles by IR spectroscopy

The hydration behavior of TiO2 nanoparticles was studied by molecular dynamics simulations and a novel IR ratio spectroscopy method. It was found that the hydration water at the titanium dioxide interface contains molecules with a large number of single hydrogen bond donors, and approximately five water layers were confined within the nano-grooves of the nanoparticles. The confinement effect was observed to enhance the strength of the hydrogen bonds and to slow down the movement of the water molecules around the surface of the nanoparticles.

A-038 Interlaboratory Comparison of Parathyroid Hormone Analytical Results Across Clinical Assays from 8 Manufacturers - Steps Towards Standardization

Abstract Background Parathyroid hormone (PTH) is an 84 amino acid peptide produced by the parathyroid gland and is a key biomarker used for the diagnosis and treatment of Chronic Kidney Disease-Mineral and Bone Disorder, as well as hypo- and hyperparathyroidism. PTH clinical assays were previously reported to exhibit significant inter-assay measurement variability, which limits the utility of PTH as a diagnostic biomarker and can result in the misclassification of patients. Thus, there is a need for PTH clinical assay standardization to improve clinical assay measurement agreement. The Centers for Disease Control and Prevention’s Clinical Standardization Program (CDC-CSP) will launch a PTH standardization program, which is in support of and coordinated with the IFCC Committee on Bone Metabolism. In preparation, the CDC-CSP conducted an Inter-Laboratory Comparison Study to evaluate the current status of PTH inter-assay variability and to investigate potential sources of variability. Methods Eight PTH assay manufacturer’s laboratories measured PTH in 42 deidentified, fresh-frozen single donor serum samples in duplicate over 2 independent runs. Sample PTH concentrations ranged from 1.3 - 324 pg/mL, based on initial screening values. Five manufacturers submitted data for 2 different assays, resulting in PTH measurements from 13 assays total. Data were submitted to the CDC CSP for analysis. Results Assay precision was <5% for 10/13 measurement procedures (MP) assessed and 3 MPs exhibited precisions between 5 - 18%. Assay-specific PTH measurements demonstrated good linear correlation compared to the all-assay mean concentration. Interestingly, inter-assay variability increased with increasing PTH concentrations, with individual sample standard deviations increasing from 2.9 - 123, with a mean SD of 24.1 and individual sample CVs increasing from 18.9 - 44.5%, with a mean CV of 24.2%. Conclusions This study provides new insights into PTH inter-assay variability and assay performance for assays commonly used in the clinical setting. The findings of this PTH Interlaboratory Comparison Study establish a baseline for PTH inter-assay variability and will guide future PTH standardization efforts.

Bilateral Lower Limb Injuries and the Need for Two Free Flaps: Simultaneous or Sequential

Abstract Objective To determine the choice of flap cover for patients presenting with bilateral lower limb trauma requiring free flap cover and to derive a step-wise guide to the planning of bilateral lower limb free flaps. Materials and Methods This was a retrospective study of patients over a 20 year period from 2000 to 2020 who presented with bilateral lower limb defects following trauma and were managed with two free flaps for wound cover, done either simultaneously or sequentially in the same admission. Results Of the 11 cases with 22 defects, there were 3 re-explorations with 1 flap loss managed with delayed fasciocutaneous flap cover. Donor morbidity was minimal requiring aspiration of seroma for 1 patient and secondary SSG for recipient area graft loss over muscle flap in 2 patients. Conclusion Use of single donor site ensures economy of time and tissue with the use of skin and muscle chimeric flaps. Harvesting of twin ALT flaps or the use of lattisimus dorsi as one of the flaps requires a 2 stage surgery to avoid prolonged operative time.

Dynamics and Evolution of Donor-derived Cytomegalovirus Infection in 3 Solid Organ Transplant Recipients With the Same Multiorgan Donor.

Cytomegalovirus (CMV) infection poses a significant risk to immunosuppressed transplant recipients, manifesting through primary infection, reinfection, or reactivation. We analyzed the emergence of drug resistance in CMV infection in 3 patients who were later found to have received an allograft from a shared, deceased donor. The seronegative transplant recipients developed symptomatic CMV infections after bowel/pancreas, kidney, or lung transplantation. Prospective Sanger sequencing was used to identify mutations in the viral DNA polymerase (DP) and protein kinase (PK). DP and PK variants were retrospectively quantified by targeted next-generation sequencing. The impact of the novel DP-A505G substitution on drug susceptibility was assessed using a recombinant virus. Whole-genome sequencing of clinical CMV samples was enabled through target DNA enrichment. The DP-A505G substitution was found in all patient samples and could be associated with a natural polymorphism. A subsequent review of the patients' clinical histories revealed that they had all received organs from a single donor. The CMV infection exhibited divergent evolution among the patients: patient 1 developed resistance to ganciclovir and foscarnet because of 2 DP mutations (V715M and V781I), patient 2 showed no genotypic resistance, and patient 3 developed ganciclovir (PK-L595S) and maribavir resistance (PK-T409M). Interpatient variation across the entire CMV genome was minimal, with viral samples clustering in phylogenetic analysis. All 3 transplant recipients were infected with the same donor-derived CMV strain and readily developed different drug susceptibility profiles. This underscores the importance of judicious antiviral drug use and surveillance in preventing antiviral resistance emergence.