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Infection from SARS-CoV-2 elicits an immune response to the nucleocapsid (N) and spike proteins (subunits S1 and S2). In this study, we set out to understand the utility of the multiplexed Quidel Sofia 2 SARS-CoV-2 IgG Antibody Fluorescent Immuno-Assay (FIA) that measures IgG antibodies against these three primary SARS-CoV-2 antigens from a single sample in 15 minutes. Using this assay with samples that were collected prior to the COVID-19 pandemic (n=816) and diseased state samples (n=99), the specificities for the three antigens were 98.4-99.9% and 98.0-100.0%, respectively. A longitudinal study was designed to collect weekly fingerstick, venous whole blood, serum and plasma samples from subjects vaccinated with the Moderna or Pfizer/BioNtech mRNA vaccines. The majority of these enrolled subjects had no known prior infection while a subset was known to have had prior COVID-19 infection. We found that the fingerstick whole blood samples performed as effectively as serum, plasma, and venous whole blood samples with a 95.8-99.5% agreement allowing physicians in a near-patient setting to rapidly provide results to their patients. Additionally, as this assay measures an IgG response against three viral proteins, S1, S2 and N, we were able to characterize immune response between i) naturally infected subjects, ii) vaccinated subjects with no prior infection, iii) vaccinated subjects with known prior infection, and iv) vaccinated subjects with prior asymptomatic exposure/infection. The Quidel Sofia 2 SARS-CoV-2 IgG FIA will aid in providing insights to the protective humoral responses as an increasing number of the world population is vaccinated against SARS-CoV-2.

Background Leukoreduction (LR) of platelet concentrate (PC) has evolved as the standard to mitigate risks of alloimmunization, clinical refractoriness, acute transfusion reactions (ATRs), and cytomegalovirus infection, but does not prevent transfusion-associated graft-versus-host disease (TA-GVHD). Amotosalen-ultraviolet A pathogen reduction (A-PR) of PC reduces risk of transfusion-transmitted infection and TA-GVHD. In vitro data indicate that A-PR effectively inactivates WBCs and infectious pathogens. Study design and methods A sequential cohort study evaluated A-PR without LR, gamma irradiation, and bacterial screening in hematopoietic stem cell transplant (HSCT) recipients. The first cohort received conventional PC (control) processed without LR, but with gamma irradiation and bacterial screening. The second cohort received A-PR PC (test) processed without: LR, bacterial screening, or gamma irradiation. The primary efficacy outcome was the 1-hour corrected count increment. The primary safety outcome was treatment-emergent ATR. Secondary outcomes included clinical refractoriness, and 100-day status for engraftment, TA-GVHD, HSCT-GVHD, infections, and mortality. Results Mean corrected count increment (× 103 ) of 33 test PC recipients was similar (18.9 ± 8.8 vs. 16.6 ± 8.4; p = 0.296) to that of 31 control PC recipients. Test recipients had a reduced, but nonsignificant, incidence of ATR (test = 9.1%, Control = 19.4%; p = 0.296). The frequencies of clinical refractoriness (0 of 33 vs. 4 of 31 patients) and refractory transfusions (6.6% vs. 19.3%) were lower in the test cohort (p = 0.05 and 0.02), respectively. No patient in either cohort had TA-GVHD. Day 100 engraftment, HSCT-GVHD, mortality, and infectious disease complications were similar between cohorts. Conclusions This study indicated that A-PR PC without LR, gamma irradiation, or bacterial screening is feasible for support of HSCT.

Background Transfusion-associated graft-versus-host disease (TA-GVHD) is a rare, often fatal complication of blood transfusion that can occur in immunocompromised or immunocompetent recipients and is the result of viable T cells present in the blood components transfused. Study design and methods We examined the TA-GVHD clinical case literature including numerous original clinical case reports and several comprehensive case series. We also evaluated recent in vitro experimental data on the inhibition of T cell proliferation, comparing the effect of a specific pathogen inactivation (PI) technology to that of the Food and Drug Administration-recommended gamma irradiation dose of 2500 cGy. Results We identified 12 published TA-GVHD cases with atypical/milder or delayed symptom presentations and/or an atypical clinical course; these included cases attributed to leukoreduced or suboptimally irradiated units. We summarize recent in vitro data using a sensitive limiting dilution assay that establish that, compared to irradiation at the recommended 2500 cGy dose, PI using amotosalen/ultraviolet A, or amustaline/glutathione achieves a greater degree of inhibition of T cell proliferation. Conclusion We propose that TA-GVHD has a spectrum of disease severity indicating that additional mild cases may still occur but be undiagnosed and/or underreported, opening up the possibility, supported by in vitro experimental data, that irradiation at the currently established dose may not be fully protective. Furthermore, since many US institutions use component irradiation selectively only for immunocompromised patients, immunocompetent recipients are not fully protected. PI technologies appear to be at least equal to, if not better than, gamma irradiation in abrogating the ability of T cells to proliferate, and if applied to all blood components, protection against TA-GVHD would be an additional benefit that would allow for the elimination of component irradiation.

This paper reports on in situ measurements of the Linear-Energy-Transfer (LET) spectra of galactic cosmic rays and their progeny and of trapped Van Allen belt protons as recorded by a Pulse Height Analyzer (PHA) radiation spectrometer which flew on the STS-95 DISCOVERY mission on the Hubble Orbital Systems Test (HOST) cradle. The Shuttle was launched on 29 October 1998 and had a mission duration of 8.5 days during the minimum phase of the solar activity cycle. The orbit of the STS-95 was about 550 km altitude and 28.5° inclination. Close agreement was seen between radiation environment model predictions and the measurements of the PHA. Agreement is obtained by considering the directionality of the radiation interacting with the Shuttle structure.

Background The INTERCEPT Blood System pathogen reduction technology (PRT), which uses amotosalen and ultraviolet A light treatment (amotosalen/UV-PRT), inactivates pathogens in plasma and platelet components (PCs). This review summarizes data describing the inactivation efficacy of amotosalen/UVA-PRT for a broad spectrum of viruses and parasites. Methods Twenty-five enveloped viruses, six nonenveloped viruses (NEVs), and four parasites species were evaluated for sensitivity to amotosalen/UVA-PRT. Pathogens were spiked into plasma and PC at high titers. Samples were collected before and after PRT and assessed for infectivity with cell cultures or animal models. Log reduction factors (LRFs) were defined as the difference in infectious titers before and after amotosalen/UV-PRT. Results LRFs of ≥4.0 log were reported for 19 pathogens in plasma (range, ≥4.0 to ≥7.6), 28 pathogens in PC in platelet additive solution (PC-PAS; ≥4.1-≥7.8), and 14 pathogens in PC in 100% plasma (PC-100%; (≥4.3->8.4). Twenty-five enveloped viruses and two NEVs were sensitive to amotosalen/UV-PRT; LRF ranged from >2.9 to ≥7.6 in plasma, 2.4 or greater to greater than 6.9 in PC-PAS and >3.5 to >6.5 in PC-100%. Infectious titers for four parasites were reduced by >4.0 log in all PC and plasma (≥4.9 to >8.4). Conclusion Amotosalen/UVA-PRT demonstrated effective infectious titer reduction for a broad spectrum of viruses and parasites. This confirms the capacity of this system to reduce the risk of viral and parasitic transfusion-transmitted infections by plasma and PCs in various geographies.

Background Risk of transfusion-transmitted (TT) malaria is mainly associated with whole blood (WB) or red blood cell (RBC) transfusion. Risk mitigation relies mostly on donor deferral while a limited number of countries perform blood testing, both negatively impacting blood availability. This study investigated the efficacy of the pathogen reduction system using amustaline and glutathione (GSH) to inactivate Plasmodium falciparum in WB. Study design and methods WB units were spiked with ring stage P. falciparum infected RBCs. Parasite loads were measured in samples at time of infection, after 24 hours at room temperature (RT), and after a 24-hour incubation at RT post-treatment with 0.2 mM amustaline and 2 mM GSH. Serial 10-fold dilutions of the samples were inoculated to RBC cultures and maintained up to 4 weeks. Parasitemia was quantified by cytometry. Results The P. falciparum viability assay has a limit of detection of a single live parasite per sample. Input parasite titer was >5.7 log10 TCID50 per mL. A 24-hour incubation at RT paused parasite development in controls, but they retained viability and infectivity when tested in culture. In contrast, no infectious parasites were detected in the amustaline/GSH-treated sample after 4 weeks of culture. Conclusion A robust level of P. falciparum inactivation was achieved in WB using amustaline/GSH treatment. Parasite log reduction was >5.7 log10 TCID50 per mL. Development of such a pathogen reduction system may provide an opportunity to reduce the risk of TT malaria and improve blood availability.

The talk was given in the International Conference " El Greco in terre d’Umbria. Per una nuova geografia dei soggiorni di Domínikos Theotokópoulos tra Creta e l’Italia.", Bettona, 28-30 September 2018. The paper is included in the proceedings of the Conference, Printed version: ISBN 978-88-943497-6-4 (Sistema Museo, August 2021); e-book: ISBN: 978-84-9927-566-6 (Cordova University Press, 2020). Abstract: The transition of Theotokópoulos from Candia to Venice represents for many scholars a reference point for the beginning of a new era in his painting, which gradually, yet definitely, strays from the art of icon painting. However, a closer examination and study of two of his works ‒ the Triptych of Modena in the Galleria Estense of Modena and the Baptism of Christ in the Historical Museum of Crete (HMC) – focusing on the materials he used and on his painting technique in this period, reveals essential constant and identifiable traits in his work: they form the connective tissue of a transition from the post-Byzantine (Cretan) tradition to the western style of the great painter., Part of the Data interpreted in this paper has been acquired in the framework of the project Pro.Di.G.I.2017 – Progetto Diagnostica Greco Italia (2017), Studio multidisciplinare sul periodo di passaggio dall'iconografia al manierismo di El Greco (Domínikos Theotokópoulos. Candia 1541 – Toledo 1614), conceived and lead by Prof. Lionello Puppi as Access project to the E-RIHS.it (IPERION CH.it) Research Infrastructure.

Background Chikungunya virus, a mosquito-borne arbovirus, often co-circulates with the Zika, dengue, and yellow fever viruses in Aedes mosquito-infested areas where cases of arbovirus transfusion-transmitted infections have been reported. Building on past experience to help maintain the availability of safe components during major outbreaks of chikungunya virus in La Reunion, Italy, and Thailand and of Zika virus in the Pacific, the Caribbean, and the Americas, pathogen inactivation is a mitigation strategy to reduce the risk of transfusion-transmitted infection. Inactivation of chikungunya virus was investigated for platelets in 100% plasma using amotosalen/ultraviolet A light, and in red blood cells using amustaline/glutathione. Study design and methods Platelets in 100% plasma and red blood cells (RBCs) were spiked with chikungunya virus. Infectious chikungunya virus titers were measured in contaminated blood products before and after treatment with amotosalen/ultraviolet A light for platelets in 100% plasma and after treatment with amustaline/glutathione for RBCs. Viral infectivity was quantified by plaque assay. Results The mean chikungunya virus infectivity titers before inactivation were 6.50 log10 plaque-forming units/mL for platelets in 100% plasma and 7.60 log10 plaque-forming units/mL for RBCs. No infectivity was detected after amotosalen/ultraviolet A light or amustaline/glutathione treatment, corresponding to greater than 6.5 log10 plaque-forming units/mL and greater than 7.1 log10 plaque-forming units/mL of inactivation, respectively. Conclusion Robust levels of chikungunya virus inactivation were achieved for platelets in 100% plasma and for RBC components. The licensed amotosalen/ultraviolet A light technology and the amustaline/glutathione pathogen-reduction system under development may provide an opportunity for comprehensive mitigation of the risk of chikungunya virus transfusion-transmitted infection by plasma, platelets, and RBCs.

Physicochemical properties of nanoparticles, such as size, shape, surface charge, density, and porosity play a central role in biological interactions and hence accurate determination of these characteristics is of utmost importance. Here we propose tunable resistive pulse sensing for simultaneous size and surface charge measurements on a particle-by-particle basis, enabling the analysis of a wide spectrum of nanoparticles and their mixtures. Existing methodologies for measuring zeta potential of nanoparticles using resistive pulse sensing are significantly improved by including convection into the theoretical model. The efficacy of this methodology is demonstrated for a range of biological case studies, including measurements of mixed anionic, cationic liposomes, extracellular vesicles in plasma, and in situ time study of DNA immobilisation on the surface of magnetic nanoparticles. The high-resolution single particle size and zeta potential characterisation will provide a better understanding of nano-bio interactions, positively impacting nanomedicine development and their regulatory approval.

BACKGROUND Dengue virus (DENV) is an arbovirus primarily transmitted through mosquito bite; however, DENV transfusion-transmitted infections (TTIs) have been reported and asymptomatic DENV RNA–positive blood donors have been identified in endemic countries. DENV is considered a high-risk pathogen for blood safety. One of the mitigation strategies to prevent arbovirus TTIs is pathogen inactivation. In this study we demonstrate that the amustaline and glutathione (S-303/GSH) treatment previously found effective against Zika virus in red blood cells (RBCs) is also effective in inactivating DENV. STUDY DESIGN AND METHODS Red blood cells were spiked with high levels of DENV. Viral RNA loads and infectious titers were measured in the untreated control and before and after pathogen inactivation treatment of RBC samples. DENV infectivity was also assessed over five successive cell culture passages to detect any potential residual replicative virus. RESULTS The mean ± SD DENV titer in RBCs before inactivation was 6.61 ± 0.19 log 50% tissue culture infectious dose (TCID50)/mL and the mean viral RNA load was 8.42 log genome equivalents/mL. No replicative DENV was detected either immediately after completion of treatment using S-303/GSH or after cell culture passages. CONCLUSION Treatment using S-303/GSH inactivated high levels of DENV in RBCs to the limit of detection. In combination with previous studies showing the effective inactivation of DENV in plasma and platelets using the licensed amotosalen/UVA system, this study demonstrates that high levels of DENV can be inactivated in all blood components.

BACKGROUND Concerned over the risk of Zika virus (ZIKV) transfusion transmission, public health agencies recommended the implementation of mitigation strategies for its prevention. Those strategies included the use of pathogen inactivation for the treatment of plasma and platelets. The efficacy of amotosalen/ultraviolet A to inactivate ZIKV in plasma had been previously demonstrated, and the efficacy of inactivation in platelets with the same technology was assumed. These studies quantify ZIKV inactivation in platelet components using amotosalen/ultraviolet A. STUDY DESIGN AND METHODS Platelet components were spiked with ZIKV, and ZIKV infectious titers and RNA loads were measured by cell culture-based assays and real-time polymerase chain reaction in spiked platelet components before and after photochemical treatment using amotosalen/ultraviolet A. RESULTS The mean ZIKV infectivity titers and RNA loads in platelet components before inactivation were either 4.9 log10 plaque forming units per milliliter, or 4.4 log10 50% tissue culture infective dose per milliliter and 7.5 log10 genome equivalents per milliliter, respectively. No infectivity was detected immediately after amotosalen/ultraviolet A treatment. No replicative virus remained after treatment, as demonstrated by multiple passages on Vero cell cultures; and ZIKV RNA was not detected from the first passage after inactivation. Additional experiments in this study demonstrated efficient inactivation to the limit of detection in platelets manufactured in 65% platelet additive solution, 35% plasma, or 100% plasma. CONCLUSION As previously demonstrated for plasma, robust levels of ZIKV inactivation were achieved in platelet components. With inactivation of higher levels of ZIKV than those reported in asymptomatic, RNA-reactive blood donors, the pathogen-inactivation system using amotosalen/ultraviolet A offers the potential to mitigate the risk of ZIKV transmission by plasma and platelet transfusion.

BACKGROUND The potential for Zika virus (ZIKV) transfusion-transmission (TT) has been demonstrated in French Polynesia and Brazil. Pathogen inactivation (PI) of blood products is a proactive strategy to inactivate TT pathogens including arboviruses. Inactivation of West Nile, dengue, Zika, and chikungunya viruses was previously demonstrated by photochemical treatment with amotosalen and ultraviolet A (UVA) illumination. In this study, we evaluated ZIKV inactivation in red blood cell (RBC) components by a chemical approach that uses amustaline (S-303) and glutathione (GSH). STUDY DESIGN AND METHODS RBC components were spiked with a high titer of ZIKV. Viral titers (infectivity) and ZIKV RNA loads (reverse transcription–polymerase chain reaction) were measured in spiked RBCs before and after S-303 and GSH treatment and confirmed using repetitive passages in cell culture. A mock-treated arm validated the approach by demonstrating stability of the virus (infectivity and RNA load) during the process. RESULTS The mean ZIKV infectivity titer and RNA load in RBCs were 5.99 ± 0.2 log 50% tissue culture infectious dose (TCID50)/mL and 7.75 ± 0.16 log genomic equivalents/mL before inactivation. No infectivity was detected immediately after S-303 and GSH treatment and after five serial passages in cell culture. CONCLUSION Complete ZIKV inactivation of more than 5.99 log TCID50/mL in RBCs was achieved using S-303 and GSH at levels higher than those found in asymptomatic ZIKV-infected blood donors. Therefore, the S-303 and GSH PI system is promising for mitigating the risk of ZIKV TT.

Background and Objectives Blood Centre logistics, staffing and donor scheduling may be optimized if pathogen inactivation (PI) of platelets can be delayed until Day 1, but bacteria may rapidly grow during this time. This study evaluates bacterial PI performed 24 and 30 h after collection. Materials and Methods PAS-3 platelet units were collected on the Amicus and subsequently inoculated (3–53 CFU/unit) with 1of 5 transfusion relevant bacterial species (n = 3/organism). Units were then stored for either 24 ± 0·3 or 30 ± 0·3 h at 20–24°C with agitation, subsequently treated with amotosalen and UVA, and stored for 7 days. Samples were taken before and after inactivation, on Days 2, 5 and 7 for BacT/ALERT testing, and on Days 5 and 7 for plate counts. Results All samples from units taken prior to inactivation either demonstrated positive plate culture counts, or, in untreated positive controls, were culture-positive during storage. All contaminated units treated with amotosalen and UVA 24 after inoculation were culture-negative on all days tested. With inactivation performed 30 h following inoculation, one of 15 units (1-of-3 replicates) was culture-positive with Klebsiella pneumonia (1 × 109 CFU/ml) by Day 5. Conclusion Photochemical treatment did not inactivate 1 of 15 units to sterility in apheresis platelets stored in PAS with a 30-h delay between contamination and treatment, but did inactivate 15 of 15 units with a 24-h delay.

BACKGROUND Pathogen inactivation methods are increasingly used to reduce the risk of infections after transfusion of blood products. Photochemical treatment (PCT) of platelets (PLTs) and plasma with amotosalen and ultraviolet A (UVA) light inactivates pathogens and white blood cells through formation of adducts between amotosalen and nucleic acid that block replication, transcription, and translation. The same adducts block the amplification of nucleic acids using polymerase chain reaction (PCR) in a manner that correlates with the number of adducts formed, providing a direct quality control (QC). Current QC measures for PCT rely on indirect methods that measure the delivered UVA dose or percent residual amotosalen after illumination, rather than directly measuring nucleic acid modification. STUDY DESIGN AND METHODS Endogenous mitochondrial DNA (mtDNA), which is detectable in PLT and plasma units, was chosen as a target for the quantification of photochemically induced modifications. DNA was extracted from untreated or amotosalen and UVA–treated PLTs or plasma, and mtDNA fragments of variable lengths were quantified using a real-time PCR inhibition assay. RESULTS PCT induced increasing real-time PCR inhibition of mtDNA amplification for larger amplicon sizes. Amplification was unaffected by treatment with amotosalen or UVA alone, whereas up to 3 log inhibition was observed after PCT. Blinded PCR testing of a panel of 110 samples each, from PLT or plasma components prepared for routine use within a blood center, allowed 100% discrimination between untreated and treated units. CONCLUSION Our initial findings indicate that an adequately sensitive, quantitative real-time PCR inhibition assay targeting mtDNA could provide a valuable tool to confirm and monitor PCT.

BACKGROUND Red blood cell (RBC) transfusion risks could be reduced if a robust technology for pathogen inactivation of RBC (PI-RBCs) were to be approved. MATERIALS AND METHODS Estimates of per-unit and per-patient aggregate infectious risks for conventional RBCs were calculated; the latter used patient diagnosis as a determinant of estimated lifetime exposure to RBC units. Existing in vitro data for the two technologies under development for producing PI-RBCs and the status of current clinical trials are reviewed. RESULTS Minimum and maximum per-unit risk were calculated as 0.0003% (1 in 323,000) and 0.12% (1 in 831), respectively. The minimum estimate is for known lower-risk pathogens while the maximal estimate also includes an emerging infectious agent (EIA) and endemic area Babesia risk. Minimum and maximum per-patient lifetime risks by diagnosis grouping were estimated as 1.5 and 3.3%, respectively, for stem cell transplantation (which includes additional risk for cytomegalovirus transmission); 1.2 and 3.7%, respectively, for myelodysplastic syndrome; and 0.2 and 44%, respectively, for hemoglobinopathy. DISCUSSION There is potential for PI technologies to reduce infectious RBC risk and to provide additional benefits (e.g., prevention of transfusion-associated graft-versus-host disease and possible reduction of alloimmunization) due to white blood cell inactivation. PI-RBCs should be viewed in the context of having a fully PI-treated blood supply, enabling a blood safety paradigm shift from reactive to proactive. Providing insurance against new EIAs. Further, when approved, the use of PI for all components may catalyze operational changes in blood donor screening, laboratory testing, and component manufacturing.

The total medical-biological impact of the Earth's space radiation environment on humans is a function of combined EVA and non-EVA exposure. In either case, the correct assessment of the eventual health risk to crew members is crucial to the success and viability of a project or mission. Aside from the medical-biological aspect itself, the validity of any assessment depends entirely on the existence of good and reliable models providing the high quality data that is needed for such evaluations, which should contain time histories of storm and substorm events, their intensities, their frequency of occurence, and their duration. Prediction requirements, advantageous and desirable model developments and improvements, and systems that need to be designed and tested, which would alert space crews and maintenance personnel about impending radiation danger are outlined.

Background and objectives Chikungunya virus (CHIKV) infections have been reported in all continents, and the potential risk for CHIKV transfusion-transmitted infections (TTIs) was demonstrated by the detection of CHIKV RNA-positive donations in several countries. TTIs can be reduced by pathogen inactivation (PI) of blood products. In this study, we evaluated the efficacy of amustaline and glutathione (S-303/GSH) to inactivate CHIKV in red-blood-cell concentrates (RBCs). Material and methods Red-blood-cells were spiked with high level of CHIKV. Infectious titres and RNA loads were measured before and after PI treatment. Residual CHIKV infectivity was also assessed after five successive cell culture passages. Results The mean CHIKV titres in RBCs before inactivation was 5·81 ± 0·18 log10 50% tissue culture infectious dose (TCID50 )/mL, and the mean viral RNA load was 10·49 ± 0·15 log10 genome equivalent (GEq)/mL. No CHIKV TCID was detected after S-303 treatment nor was replicative CHIKV particles and viral RNA present after five cell culture passages of samples obtained immediately after S-303 treatment. Conclusion Chikungunya virus was previously shown to be inactivated by the PI technology using amotosalen and ultraviolet A light for the treatment of plasma and platelets. This new study demonstrates that S-303/GSH can inactivate high titres of CHIKV in RBCs.

INTRODUCTION Transfusion-associated graft-versus-host disease (TA-GVHD) is a rare complication after transfusion of components containing viable donor T cells. Gamma irradiation with doses that stop T-cell proliferation is the predominant method to prevent TA-GVHD. Treatment with pathogen inactivation methodologies has been found to also be effective against proliferating white blood cells, including T cells. In this study, T-cell inactivation was compared, between amotosalen/ultraviolet A (UVA) treatment and gamma-irradiation (2500 cGy), using a sensitive limiting dilution assay (LDA) with an enhanced dynamic range. METHODS AND MATERIALS Matched plasma units (N = 8), contaminated with 1 × 106 peripheral blood mononuclear cells (PBMCs) per mL, were either treated with amotosalen/UVA or gamma irradiation, or retained as untreated control. Posttreatment, cells were cultured under standardized conditions. T-cell proliferation was determined by the incorporation of 3 H-thymidine and correlated with microscopic detection. RESULTS Range-finding experiments showed that after gamma irradiation (2500 cGy), significant T-cell proliferation could be observed at a 1 × 107 cell culture density, some proliferation at 1 × 106 , and none at 1 × 105 cells/well. Based on these facts, a quantitative comparison was carried out between amotosalen/UVA at the highest challenge of 1 × 107 PBMCs/well, and gamma irradiation at 1 × 106 and 1 × 105 PBMCs/well. Complete inactivation of the T cells after amotosalen/UVA treatment was observed, equivalent to greater than 6.2 log inactivation. Complete inactivation of the T cells was also observed after gamma irradiation when 1 × 105 PBMCs/well were cultured (>4.2 log inactivation). Proliferation was observed when 1 × 106 PBMCs/well were cultured (≤5.2 log inactivation) after gamma irradiation. CONCLUSION Amotosalen/UVA treatment more effectively inactivates T cells than the current standard of gamma irradiation (2500 cGy) for the prevention of TA-GVHD.

Background and Objectives A photochemical treatment process (PCT) utilizing amotosalen and UVA light (INTERCEPT™ Blood System) has been developed for inactivation of viruses, bacteria, parasites and leucocytes that can contaminate blood components intended for transfusion. The objective of this study was to further characterize the safety profile of INTERCEPT-treated platelet components (PCT-PLT) administered across a broad patient population. Materials and Methods This open-label, observational haemovigilance programme of PCT-PLT transfusions was conducted in 21 centres in 11 countries. All transfusions were monitored for adverse events within 24 h post-transfusion and for serious adverse events (SAEs) up to 7 days post-transfusion. All adverse events were assessed for severity (Grade 0–4), and causal relationship to PCT-PLT transfusion. Results Over the course of 7 years in the study centres, 4067 patients received 19 175 PCT-PLT transfusions. Adverse events were infrequent, and most were of Grade 1 severity. On a per-transfusion basis, 123 (0·6%) were classified an acute transfusion reaction (ATR) defined as an adverse event related to the transfusion. Among these ATRs, the most common were chills (77, 0·4%) and urticaria (41, 0·2%). Fourteen SAEs were reported, of which 2 were attributed to platelet transfusion (

This paper proves that voice over Internet protocol (IP) technology can be combined with satellite transmission in air traffic control (ATC) applications provided that the systems used to implement this approach have been meticulously designed to meet the end-to-end delay requirements and to certify the quality of service they offer under strenuous conditions.We propose a modified E-model and prove it can be used as a radio network planning tool, because once the requirements that were drawn from it were fulfilled, the users of the resulting service accepted it as sufficient in terms of call quality. The approach has been applied on the SATWAYS system, cofunded by the European Space Agency, and designed and implemented to extend the coverage of radio frequencies within airspaces when the commonly used landline communication infrastructure is either too costly or impossible to install and maintain. The paper provides evidence that the way to an all-IP ATC network is wide open.

BACKGROUND With an increasing number of recognized transfusion-transmitted (TT) babesiosis cases, Babesia microti is the most frequently TT parasite in the United States. We evaluated the inactivation of B. microti in red blood cells (RBCs) prepared in Optisol (AS-5) using amustaline and glutathione (GSH) and in platelet components (PCs) in 100% plasma using amotosalen and low-energy ultraviolet A (UVA) light. STUDY DESIGN AND METHODS Individual RBCs and apheresis PCs were spiked with B. microti–infected hamster RBCs (iRBCs) to a final concentration of 106 iRBCs/mL and treated with the respective inactivation systems according to the manufacturer's instruction. Samples were collected before (control) and after (test) each treatment. Dilutions of the control samples to 10−6 were inoculated into hamsters, while the test samples were inoculated neat or at 10−1 dilution. At 3 and 5 weeks postinoculation, hamsters were evaluated for B. microti infection by microscopic observation of blood smears and 50% infectivity titers (ID50) were determined. Log reduction was calculated as control log ID50 minus test log ID50. RESULTS Parasitemia was detected in hamsters injected with as low as 100,000-fold diluted control samples, while no parasites were detectable in the blood smears of any hamsters receiving neat test samples. Mean log reduction was more than 5 log/mL by amustaline/GSH for RBCs and more than 4.5 log/mL by amotosalen/UVA for PCs. CONCLUSION B. microti was inactivated to the limit of detection in RBCs and PCs after the respective inactivation treatment. Complete inactivation of B. microti was achieved in this animal infectivity model, and pathogen reduction treatment inhibited transmission of infection.

IntroductionRib fractures in elderly patients are associated with increased morbidity and mortality. Predicting which patients are at risk for complications is an area of debate. Current models use anatomic, physiologic or laboratory parameters in isolation to answer this question. The ‘RibScore’ is an anatomic model that assesses fracture severity. Given that frailty is a major driver of adverse outcomes in the elderly, we hypothesize that the combined analysis of fracture severity, physiologic reserve and current pulmonary function are better predictors of respiratory compromise in this population.MethodsThis is a retrospective chart review of 263 trauma patients age ≥55 from January 2014 to June 2017. Criteria included blunt mechanism and ≥ 1 rib fracture identified by CT. Variables indicating adverse pulmonary outcomes were defined by: pneumonia, respiratory failure and tracheostomy. Three models were assessed: (1) RibScore, (2) Modified Frailty Index (mFI) and (3) initial partial pressure of carbondioxide (PaCO2).ResultsA total of 263 patients met inclusion criteria. 13% developed pulmonary complications. Increased RibScore, mFI and PaCO2 were each statistically associated with risk of complications. Receiver operating characteristics area under the curve analysis of individual models predicted complications with the following concordance statistic (CS): anatomic (RibScore) yielded a CS of 0.79 (95% CI 0.69 to 0.89); physiologic (mFI) yielded a CS of 0.83 (95% CI 0.75 to 0.91) and laboratory (PaCO2) yielded a CS of 0.88 (95% CI 0.80 to 0.95). The PaCO2 had the highest discriminative ability of the three individual models. Combining all three models yielded the best performance with a CS of 0.90 (95% CI 0.81 to 0.97).DiscussionThe RibScore maintains discriminative ability in the elderly. However, models based on mFI and PaCO2 individually outperform the RibScore. A combination of all three models yields the highest discriminative ability. This combined approach is best for assessing the severity of rib fractures and prediction of complications in the elderly.Level of evidencePrognostic Study, Level III.

Introduction: HSCT patients require platelet components (PC) for support of thrombocytopenia. Donors are tested for selected viruses to prevent transfusion-transmission infection (TTI), and PC are screened for bacteria and irradiated to prevent transfusion associated graft versus host disease (TA-GVHD). Leukocyte reduction (LR) is commonly, but not universally, used to mitigate the risks of acute transfusion reactions (ATR), allo-immunization, clinical refractoriness, and CMV TTI. However, donor testing, PC bacterial screening, and LR do not provide complete protection from ATR, transfusion related sepsis (TRS), refractoriness, CMV TTI, or residual risks of window period and emerging pathogen TTI. Amotosalen-UVA pathogen reduction treatment (A-PRT) inactivates a broad spectrum of pathogens (e.g. Zika), inhibits leukocyte antigen presentation and cytokine synthesis, and inhibits T-cell proliferation. A-PRT with LR has replaced bacterial screening, CMV serology, and gamma irradiation. Objective: To control cost, the Hong Kong Red Cross Blood Transfusion Service (HKRCBTS) has used whole blood platelet-rich-plasma (PRP) PC without LR. Based on data showing effective leukocyte inactivation by A-PRT, the HKRCBTS evaluated use of A-PRT buffy coat PC (BC PC) in platelet additive solution (PAS) without LR, bacterial screening, and gamma irradiation to manage costs, salvage more donor plasma, reduce TTI risk, and simplify production. Study Design and Methods : An open-label, prospective, sequential, two cohort study at Queen Mary Hospital, Hong Kong enrolled HSCT patients for up to 30 days of PC support with 100-day follow up. Cohorts were comparable for primary disease and type of HSCT. The first cohort received PRP PC prepared from CPD whole blood, a current HKRCBTS standard PC (Control-C). Five ABO-matched PRP PCs in 100% plasma were pooled without LR to yield ≥ 3 x 1011 platelets. C-PCs were screened for bacteria, gamma irradiated (2,500 cGy), stored up to 5 days at 22- 24 °C; and contained ~ 5 x 109 leukocytes. For the 2nd cohort, buffy coat PCs (BC PC) were prepared from CPD whole blood in PAS (SSP+, Macopharma, France). Five ABO-matched BC PC in 35% plasma-65% PAS were pooled, and prepared with A-UVA PRT to yield ≥ 3 x 1011 platelets (Test-T). T-PCs were not LR, not screened for bacteria, not gamma irradiated, stored for up to 5 days at 22- 24 °C; and contained ~ 5 x 108 leukocytes. HSCT patients ≥ 18 years expected to require PC were included. Exclusion criteria were: history of clinical refractoriness to platelet transfusion (2 successive 1-HR corrected count increments (CCI) < 5.0 x 103), immune thrombocytopenia, or disorders confounding 1-HR CCI determination. For both cohorts, a transfusion threshold of 10 x 109/L was specified for stable patients unless adjusted to 20 x 109/L for sepsis. Patients were not screened for HLA antibody at study entry. The efficacy outcome was the 1-hour (HR) count increment (CI), and the safety outcome was ATR incidence. Secondary (post-hoc) outcomes were the 1-HR CCI and the incidence of clinical refractoriness (two successive CCI < 5x103 per patient and transfusions with CCI < 5x103). Results: 64 patients were transfused (31 Control and 33 Test). Demographics were similar between cohorts (Table). The majority of patients had allogeneic HSCT from related donors. Mean pre-transfusion platelet counts were < 16 x 109/L. The mean PC dose and storage duration were similar between cohorts (Table). The 1-HR count increments (CI) and 1-HR CCI responses were within therapeutic ranges for both cohorts (Table). The proportion of transfusions with CCI responses < 5.0 x 103 and the proportion of patients with clinical refractoriness were less in the Test cohort (Table). ATRs trended lower with A-PRT (Test = 9.1%, Control = 19.4%, p=0.296).No patients had TRS or TA-GVHD reported during the active transfusion period. Day-100 engraftment (Table), HSCT-GVHD, mortality, and infectious disease complications were similar between cohorts. Conclusions: This exploratory study of A-PRT PC without LR demonstrated therapeutic 1-HR CCI responses, safety, and conserved HSCT engraftment. The proportions of transfusions with CCI < 5.0x103 and the proportion of patients with clinical refractoriness were less for A-PRT PC than conventional PC. A-PRT of pooled whole blood BC PC without LR offers the potential to improve PC supply, and to reduce the net cost of PRT PC by replacing other interventions. Table Table. Disclosures Corash: Cerus Corporation: Employment, Equity Ownership. Liu:Cerus Corporation: Employment, Equity Ownership. Huang:Cerus Corporation: Employment, Equity Ownership. Vermeij:Cerus Corporation: Employment, Equity Ownership. Stassinopoulos:Cerus Corporation: Employment, Equity Ownership. Benjamin:Cerus Corporation: Employment, Equity Ownership.

In the last few years, the transfusion medicine community has been paying special attention to emerging vector-borne diseases transmitted by arboviruses. Zika virus is the latest of these pathogens and is responsible for major outbreaks in Africa, Asia and, more recently, in previously infection-naïve territories of the Pacific area. Many issues regarding this emerging pathogen remain unclear and require further investigation. National health authorities have adopted different prevention strategies. The aim of this review article is to discuss the currently available, though limited, information and the potential impact of this virus on transfusion medicine.

Background Herpes zoster (HZ) develops in up to 50% of unvaccinated individuals, accounting for >1 million cases annually in the United States. A live attenuated HZ vaccine (LAV) is Food and Drug Administration approved for those age 50 years or older, though Advisory Committee on Immunization Practices recommendations are only for those age 60 years or older. LAV efficacy is ~70% for persons 50–59 years of age, with lower efficacy in older adults. A new 2-dose adjuvanted subunit vaccine (SUV) has >95% efficacy in persons 50–69 years of age and remains ~90% efficacious in persons vaccinated at age 70 years. Methods To estimate the relative cost-effectiveness of SUV, LAV, and no vaccination (NoV) strategies, a Markov model was developed based on published data on vaccine efficacy, durability of protection, quality of life, resource utilization, costs, and disease epidemiology. The perspective was US societal, and the cycle length was 1 year with a lifelong time horizon. SUV efficacy was estimated to wane at the same rate as LAV. Outcomes evaluated included lifetime costs, discounted life expectancy, and incremental cost-effectiveness ratios (ICERs). Results For individuals vaccinated at age 50 years, the ICER for LAV vs NoV was $118 535 per quality-adjusted life-year (QALY); at age 60 years, the ICER dropped to $42 712/QALY. SUV was more expensive but had better ICERs than LAV. At age 50, the ICER was $91 156/QALY, and it dropped to $19 300/QALY at age 60. Conclusions Vaccination with SUV was more cost-effective than LAV in all age groups studied. Vaccination with SUV at age 50 years appears cost-effective, with an ICER, This manuscript describes a cost-effectiveness analysis utilizing a Markov decision model that demonstrates vaccination with the two-dose adjuvanted subunit herpes zoster vaccine, given current pricing, offers very good value in comparison to the live attenuated herpes zoster vaccine or no vaccination strategy. Even at higher prices, faster waning rates, and poorer adherence to the second dose, our model suggests the subunit vaccine remains cost-effective at a willingness-to-pay threshold of $100,000 per quality-adjusted life-year.

Pathogen inactivation (PI) is an alternative approach to blood safety that is gaining widespread use.1 PI for platelets (PLTs) has been in use in Europe for more than 10 years and one of these systems, the INTERCEPT Blood System, was recently approved by the Food and Drug Administration (FDA).2 There are costs for implementing PI, but it is also likely that there will be offsetting savings. The availability of PI PLTs may have substantial economic impact both in terms of operating costs for the facilities which collect and prepare them and in the acquisition costs to hospitals. The purpose of this study was to carry out a detailed cost analysis of the operational activities that may be affected by the adoption of PI PLTs and to project those that will apply to either blood centers or hospitals. The costs will affect hospitals in different ways depending on whether they purchase all their PLTs from an outside supplier or if they produce some fraction of the PLTs. While PI PLTs may reduce the costs associated with transfusion-transmitted disease treatment those costs were not considered in this project. This study focuses on PLTs collected by apheresis because in the most recent survey in the United States, 91% of the total of 2,516,000 PLT units was collected by apheresis.3 In the United States, blood centers collected approximately 89% of the apheresis PLT supply while hospitals collected 11%.3 Collection can be done with different devices and the PLTs can be stored for up to 5 days in either donor plasma or PLT additive solution and plasma mix at room temperature. When PLTs are collected at a blood center, testing is done along with standardized whole blood testing; however, when PLTs are collected at a hospital, samples are often sent to a different facility for testing. Bacterial screening of each unit of PLTs is performed at the place of collection but when performed, point-of-release (PoR) testing is usually done at the hospital.

Introduction : Depending on the conditioning intensity and graft source, mosthematopoietic stem cell transplant(HSCT) patients invariably experience a cytopenic phase of at least 2-3 weeks and require transfusion support until engraftment. All transfused blood components post-transplant need to be gamma irradiated to prevent transfusion associated graft versus host disease (TA-GVHD). Platelet component (PC) manufacturing methodologies differ around the globe. Currently, Hong Kong HSCT patients are supported with individual platelet rich plasma (PRP) PCs without leukocyte filtration (NLF), tested by short-term aerobic cultures for bacterial contamination (STABC), and gamma irradiated. When new PC manufacturing methodologies are considered for adoption, it is important to assess the immediate and longer term effects on transfused patients. Amotosalen+UVA light (A-UVA) pathogen reduction technology (INTERCEPT™ Blood System, Cerus Corporation, Concord, CA) inactivates bacteria, viruses, parasites and leukocytes by efficient covalent adduct modification of DNA and RNA. Treatment with A-UVA prevents TA- GVHD in an animal model, inhibits clonal T cell proliferation, prevents allogeneic antigen stimulation in mixed lymphocyte reactions, and inhibits transcription mediated cytokine production and early activation antigen expression (Corash et al, BMT 2004). The common strategy of prescribing gamma irradiated PC only for suspected "high risk" patients is sub-optimal, as failure to identify recipients at risk for TA-GVHD was responsible for 50% of reported cases (Kopolovic et al. Blood 2016). Treatment of PC with A-UVA addresses two key issues for HSCT patients: bacterial contamination, and universal prevention of TA-GVHD. Here, we compared the clinical support of HSCT patients with conventional PCs (C-PC) vs. an equivalent HSCT group supported with A-UVA PCs (I-PC) using a sequential patient cohort design within a single HSCT clinical center. Methods : I-PCs were prepared from 5 pooled ABO-matched, NLF buffy coat PCs in platelet additive solution and treated with A-UVA replacing gamma irradiation and bacterial detection. C-PCs were prepared from 5 ABO-matched, STABC-negative, NLF PRP PCs, and treated with gamma irradiation. Each patient could receive up to 5 PC transfusions. The primary efficacy endpoint was the 1-hour corrected count increment (CCI) and the primary safety endpoint was the proportion of patients (P) with acute transfusion reactions (ATR). Follow-up data on mortality, hematopoiesis engraftment, and immune status were collected up to 100 days post HSCT. Results: Patient demographics and type of HSCT were similar between patient cohorts. 33 patients received 76 A-UVA PCs and 31 patients received 89 C-PCs. The mean days of platelet support (p=0.618) and mean 1-hour CCIs per patient averaged for all transfusions were comparable (p=0.296) between the I- PC and conventional C-PC cohorts (Table). The proportion of patients with AEs was lower (p=0.021) for the I-PC group (Table), and no related SAEs were observed during the entirety of trial. Survival at 100 days post HSCT and rates of remission were similar between the cohorts (Table). The ATR rate trended lower, although not significantly different (p=0.296), in the I- PC group (Table). Follow-up data showed that the patients had comparable neutrophil and platelet engraftment (Table) with comparable immune system reconstitution by 100 days post HSCT. No cases of TA-GVHD were observed in either cohort. Conclusions: A-UVA-treated PCs prepared without LF, gamma irradiation, and bacterial detection can replace C-PCs for support of HSCT patients resulting in comparable post transfusion CCI responses and short and intermediate term clinical outcomes, while offering additional protection against transfusion transmitted bacteria and emerging or untested pathogens. Table Table. Disclosures Sim Pui Yin: Cerus Corporation: Other: Investigator sponsored trial. Leung Yuk Yan:Cerus Corporation: Other: Investigator sponsored trial. Lam Chun Kit:Cerus Corporation: Other: Investigator sponsored trial. Tsoi Wai:Cerus Corporation: Other: Investigator sponsored trial. Lee Cheuk:Cerus Corporation: Other: Investigator sponsored trial. Lie Kwok Wai:Cerus Corporation: Other: Investigator sponsored trial. Huang:Cerus Corporation: Employment. Rico:Cerus Corporation: Employment. Lin:Cerus corp: Employment. Corash:Cerus Corporation: Employment. Stassinopoulos:Cerus Corporation: Employment.

COX-2 is an important mediator of inflammation that significantly influences tumor angiogenesis, invasion and metastasis. Here, we have investigated the role of COX-2 in modifying the number of activated cancer associated fibroblasts (CAFs) and in altering the extracellular matrix (ECM) in a breast cancer model. To investigate the role of COX-2 in modulating the ECM, we used an MDA-MB-231 cell clone (Clone 13) expressing a short hairpin RNA (shRNA) to downregulate COX-2 [1]. Clone 13 cells were characterized for significantly lower basal and TPA-induced COX-2 and PGE2 expression compared to parental MDA-MB-231 cells using ELISA (PGE2), western blot (COX-2 protein) and q-RT-PCR (COX-2 mRNA). Tumors were derived from parental (n = 5) and Clone 13 (n = 6) MDA-MB-231 cells following inoculation in the mammary fat pad in SCID mice. Tumors were excised at ∼ 500 mm3 and immunohistochemically stained to quantify vessel density (CD31) and activated CAFs (α-smooth muscle actin (SMA)) in 5 μm thick formalin fixed sections. Stained sections were digitally scanned and positive staining quantified using manufacturer supplied software (Aperio Technologies, CA). Clone 13 tumors showed delayed tumor growth compared to parental MDA-MB-231 tumors. We have previously observed that collagen 1 (Col1) fiber density and fiber volume were significantly lower in COX-2 reduced Clone 13 tumors compared to parental tumors [2]. While cancer cells shape Col1 fiber patterns through the secretion of various enzymes, Col1 fiber is laid down by activated CAFs within or around the tumor. Quantification of activated CAFs by immunohistochemistry for α-SMA in the tumors, and immunoblotting for α-SMA of crude protein extracted from the tumors, revealed significantly fewer CAFs and significantly reduced levels of α-SMA protein in Clone 13 tumors compared to parental MDA-MB-231 tumors. We previously observed a significant decrease in permeability as well as reduced influx and efflux of macromolecular transport in Clone 13 tumors compared to parental tumors, but no difference in vascular volume [2]. Immunohistochemistry for CD31 staining of endothelial cells did not detect a significant difference in CD31 density between Clone 13 and parental tumors further confirming our previous observations about vascular volume. These data reveal the multi-faceted effects of COX-2 in modifying the structure and function of the ECM, and identify the ability to attract and activate fibroblasts as one mechanism by which COX-2 modifies the ECM. Acknowledgements: We thank Mr. Gary Cromwell for technical assistance. This work was supported by NIH R01CA82337 and P50 CA103175. References: [1] Stasinopoulos, I., et al., Mol Cancer Res, 2007; [2], Stasinopoulos, I., et al., AACR, 2013 Chicago. Citation Format: Balaji Krishnamachary, Ioannis Stassinopoulos, Samata M. Kakkad, Marie-France Penet, Desmond Jacob, Flonne Wildes, Yelena Mironchik, Arvind Pathak, Meiyappan Solaiyappan, Zaver M. Bhujwalla. Cyclooxygenase-2 downregulation reduces activated fibroblasts and modifies the extracellular matrix in MDA-MB-231 breast cancer xenograft. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4021. doi:10.1158/1538-7445.AM2015-4021