Ebp1 p48 promotes oncogenic activities in human colon cancer cells through regulation of TIF-90-mediated ribosomal RNA synthesis.

The ErbB3-binding protein 1 (Ebp1) has been reported as either an oncogenic regulator or a tumor suppressor in a variety of cancers. Here, we show that Ebp1 p48, a predominant expression isoform, is highly expressed in the majority of human colon tumor cells compared with normal adjacent tissues and its expression is required for the oncogenic activities of these cells. Depletion of Ebp1 expression in primary colon cancer cells inhibits cell proliferation, colony forming, and invasion in vitro as well as tumor formation in vivo and enhances cell sensitivity to irradiation. We further demonstrated that Ebp1 interacts with TIF-90, a splice variant of transcription initiation factor IA (TIF-IA) of the RNA polymerase I complex, allowing for regulation of ribosomal RNA (rRNA) synthesis and oncogenesis in human colon cancer cells. Moreover, Ebp1 expression is essential for Akt protected TIF-90 stability by preventing TIF-90's ubiquitination by Mdm2 and hence, its proteasomal degradation. The results of the present study support a mechanism of underlying oncogenic activities by means of Ebp1 through regulation of TIF-90-mediated rRNA synthesis and suggest the potential therapeutic treatment of colon cancer by targeting Ebp1 and its signaling.

The role of ErbB3 binding protein 1 in cancer: Friend or foe?

ErbB3, a member of the epidermal growth factor receptor family, reportedly plays an essential role in the regulation of cancer progression and therapeutic resistance. Numerous studies have indicated that ErbB3 binding protein 1 (Ebp1), a binding partner for ErbB3, plays an important regulatory role in the expression and function of ErbB3, but there is no agreement as to whether Ebp1 also has an ErbB3-independent function in cancer and how it might contribute to tumorigenesis. In this review, we will discuss the different functions of the two Ebp1 isoforms, p48 and p42, that may be responsible for the potentially dual role of Ebp1 in cancer growth.

A targeted immunomic approach identifies diagnostic antigens in the human pathogen Babesia microti.

BACKGROUND: Babesia microti is a protozoan parasite responsible for the majority of reported cases of human babesiosis and a major risk to the blood supply. Laboratory screening of blood donors may help prevent transfusion-transmitted babesiosis but there is no Food and Drug Administration-approved screening method yet available. Development of a sensitive, specific, and highly automated B. microti antibody assay for diagnosis of acute babesiosis and blood screening could have an important impact on decreasing the health burden of B. microti infection. STUDY DESIGN AND METHODS: Herein, we take advantage of recent advances in B. microti genomic analyses, field surveys of the reservoir host, and human studies in endemic areas to apply a targeted immunomic approach to the discovery of B. microti antigens that serve as signatures of active or past babesiosis infections. Of 19 glycosylphosphatidylinositol (GPI)-anchored protein candidates (BmGPI1-19) identified in the B. microti proteome, 17 were successfully expressed, printed on a microarray chip, and used to screen sera from uninfected and B. microti-infected mice and humans to determine immune responses that are associated with active and past infection. RESULTS: Antibody responses to various B. microti BmGPI antigens were detected and BmGPI12 was identified as the best biomarker of infection that provided high sensitivity and specificity when used in a microarray antibody assay. CONCLUSION: BmGPI12 alone or in combination with other BmGPI proteins is a promising candidate biomarker for detection of B. microti antibodies that might be useful in blood screening to prevent transfusion-transmitted babesiosis.

Prevalence and Genetic Characterization of Histomonas meleagridis in Chickens in Vietnam.

This study was performed to investigate the prevalence and to characterize the genetic diversity of Histomonas meleagridis isolates in chickens in southern Vietnam. A total of 194 chickens, randomly selected from 18 backyard and 18 commercial flocks, were screened for H. meleagridis infection using both macroscopic diagnosis and an 18S rRNA gene-based PCR method. Overall, 12.9% of birds, representing 19 flocks, showed gross lesions typical for histomonosis whereas 25.3% of the birds from 29 flocks were positive by PCR assay. Following initial diagnostic approaches, H. meleagridis-positive samples were further analyzed by sequencing three different genomic loci; the 18S rRNA, alpha-actinin1, and rpb1. Thirteen samples from 12 flocks were genetically identified as H. meleagridis, demonstrating a flock and sample prevalence of 33.3% and 6.7%, respectively. There was no significant difference in prevalence between different farm types, age groups, and seasonality. Genetic analysis demonstrated minor heterogeneity of Vietnamese isolates with 99% homology to H. meleagridis sequences from the database. This is the first survey of the prevalence and genetic characterization of H. meleagridis in chickens in Vietnam.

A graphene oxide based composite granule for methylene blue separation from aqueous solution: Adsorption, kinetics and thermodynamic studies.

Graphene oxide and chitosan composite material using as a high-efficiency and low-cost granular adsorbent for methylene blue removal was fabricated via self-assembling method. The effects of pH value, contact time, initial concentration, adsorbent dose, temperature, and recyclic stability on the adsorption performance of methylene blue in aqueous solution were investigated in detail. Desorption process with the effects of solvents, contact time, and temperature were also conducted carefully in this study. The adsorption kinetics and adsorption isotherm of dye adsorption process showed that dye adsorption process was fitted to the pseudo-second-order kinetic model and the Freundlich adsorption isotherm, indicating a physical adsorption process with multilayer adsorption. The intra-particle diffusion model indicated that the dye adsorption by the granular adsorbent was strongly happened during the first 4 h. The thermodynamic study showed that the adsorption was a spontaneous and exothermic process and dye ions were condensed onto the surface of adsorbent. The maximum adsorption capacity of dye on the granular adsorbent was calculated as 951.35 mg/g and the adsorbent could maintain its adsorption performance after six cycles. In general, this study provided an efficient, cost-effective, and recyclable the granular adsorbent for dye separation from aqueous solution.

Formulations of poly(vinyl alcohol) functionalized silk fibroin nanoparticles for the oral delivery of zwitterionic ciprofloxacin.

Fibroin nanoparticles (FNP) have been employed in numerous biomedical applications. However, limited research has focused on the oral delivery of FNP and in-depth molecular interactions between the encapsulated drug and FNP. Therefore, this work developed the FNP, functionalized with poly(vinyl alcohol) (PVA), to orally deliver the zwitterionic ciprofloxacin, focused on the molecular interactions. The particles were formulated using both desolvation (the drug precipitated during the particles formulation) and adsorption (the drug adsorbed on the particles surfaces) methods. The optimal formula possessed a size of ~630 nm with narrow size distribution (measured by DLS method), spherical shape (determined by SEM), and moderate drug loading (confirmed by FT-IR, XRD, and DSC techniques) of ~50% for the desolvation method and ~43% for the adsorption method. More than 80% of the drug molecules resided on the particle surfaces, mainly via electrostatic forces with fibroin. The drug was physically adsorbed onto FNP, which followed Langmuir model and pseudo second-order kinetics. In the in-vitro simulated gastric condition at pH 1.2, the ciprofloxacin bound strongly with FNP via electrostatic forces, thus hindering the drug release (< 40%). Contrastingly, in the simulated intestinal condition at pH 6.8, the particles could control the drug release rates dependent on the PVA amount, with up to ~100% drug release. Lastly, the particles possessed adequate antibacterial activities on Bacillus subtilis, Escherichia coli, and Salmonella enterica, with MIC of 128, 8, and 32 µg/mL, respectively. In summary, the FNP and PVA functionalized FNP could be a potential oral delivery system for zwitterionic drugs.

Nanocellulose-alginate composite beads for improving Ciprofloxacin bioavailability.

Nanocellulose is a potential material utilized in numerous biomedical applications. However, its hydrophilic characteristic and uncontrolled encapsulated drug release hinders nanocellulose uses in oral drug administration. Thus, this work developed novel nanocellulose/alginate composite (CNC/Alg) beads for oral delivery and bioavailability enhancement of a model drug, Ciprofloxacin (CIP). CNC was green synthesized employing electrolysis process from sugarcane bagasse. CNC/Alg beads were formulated by dropwise adding CNC-Alg mixture in CaCl2 solution at room temperature. CIP was incorporated into CNC/Alg beads by adsorption technique. X-ray diffractometry and Fourier-transform infrared spectra images showed that the beads were effectively produced with high crystallinity of 75.5 %, and the typical bond of cellulose and alginate. Within 4 h of adsorption, CIP loading efficiency reached 45.27 %, with 87.2 % molecules in the zwitterionic state. The adsorption followed Elovich and pseudo-second-order models, indicating a multi-mechanism including both physical and chemical adsorptions. Importantly, in gastrointestinal tract, the beads could protect CIP from acidic stomach environment while releasing it sustainably in simulated intestinal condition (75.05 %). The beads also showed strong antibacterial activity against both Gram(-) and Gram(+) bacteria, as evidenced by low IC50 and minimum inhibitory concentration values. Finally, CNC/Alg beads could improve CIP bioavailability for effective oral drug delivery route.

Prevalence and molecular characterization of Cryptosporidium in ostriches (Struthio camelus) on a farm in central Vietnam.

This study was performed to determine the prevalence and molecular characterization of Cryptosporidium in ostriches on a farm in Khanh Hoa province, central Vietnam. A total of 464 ostrich fecal samples were examined Cryptosporidium oocysts using the modified Ziehl-Neelsen staining method, and 110 (overall prevalence 23.7%) were identified as positive by microscopy. Prevalence of Cryptosporidium in animals of <45 days, 45-60 days, 61-90 days, 91 days-12 months and >12 months was 23.5% (16/68), 33.3% (22/66), 35.2% (68/193), 0 and 5.8% (4/69), respectively (p<0.05). The majority of positive samples scored as the 3+ level of intensity of infection were from 61 to 90 days ostriches. Molecular analysis in the 18S ribosomal RNA, 70 kDa heat shock protein and actin genes demonstrated the presence of only Cryptosporidium avian genotype II in ostriches in central Vietnam.

Dynamic antibody responses to the Mycobacterium tuberculosis proteome.

Considerable effort has been directed toward controlling tuberculosis, which kills almost two million people yearly. High on the research agenda is the discovery of biomarkers of active tuberculosis (TB) for diagnosis and for monitoring treatment outcome. Rational biomarker discovery requires understanding host-pathogen interactions leading to biomarker expression. Here we report a systems immunology approach integrating clinical data and bacterial metabolic and regulatory information with high-throughput detection in human serum of antibodies to the entire Mycobacterium tuberculosis proteome. Sera from worldwide TB suspects recognized approximately 10% of the bacterial proteome. This result defines the M. tuberculosis immunoproteome, which is rich in membrane-associated and extracellular proteins. Additional analyses revealed that during active tuberculosis (i) antibody responses focused on an approximately 0.5% of the proteome enriched for extracellular proteins, (ii) relative target preference varied among patients, and (iii) responses correlated with bacillary burden. These results indicate that the B cell response tracks the evolution of infection and the pathogen burden and replicative state and suggest functions associated with B cell-rich foci seen in tuberculous lung granulomas. Our integrated proteome-scale approach is applicable to other chronic infections characterized by diverse antibody target recognition.

Molecular characterization of Cryptosporidium in pigs in central Vietnam.

Little information is available on the epidemiology of Cryptosporidium in pigs in central Vietnam. The aims of this study were to investigate the prevalence and to characterize the genotype distribution of Cryptosporidium isolates in pigs in this region. A total of 193 pig fecal samples were screened for the presence of Cryptosporidium oocysts using the modified Ziehl-Neelsen staining method, and 28 (overall prevalence 14.5 %) were identified as positive by microscopic observation. Positive samples were further analyzed by polymerase chain reaction amplification and sequencing. Genetic identification based on the 18S ribosomal RNA and 70 kDa heat shock protein genes revealed that pigs in Vietnam are infected with two species/genotypes (Cryptosporidium suis and Cryptosporidium pig genotype II). This study is the first molecular characterization of Cryptosporidium in pigs in Vietnam. The presence of these host-adapted species/genotypes suggests that pigs may not pose a significant public health risk in this area. More extensive studies are necessary to ascertain the zoonotic potential of Cryptosporidium in porcine hosts in Vietnam.

Proteome-scale antibody responses and outcome of Mycobacterium tuberculosis infection in nonhuman primates and in tuberculosis patients.

BACKGROUND: Biomarkers of progression from latent Mycobacterium tuberculosis infection to active tuberculosis are needed. We assessed correlations between infection outcome and antibody responses in macaques and humans by high-throughput, proteome-scale serological studies. METHODS: Mycobacterium tuberculosis proteome microarrays were probed with serial sera from macaques representing various infection outcomes and with single-point human sera from tuberculosis suspects. Fluorescence intensity data were analyzed by calculating Z scores and associated P values. Temporal changes in macaque antibody responses were analyzed by polynomial regression. Correlations between human responses and sputum bacillary burden were assessed by quantile and hurdle regression. RESULTS: Macaque outcome groups exhibited distinct antibody profiles: early, transient responses in latent infection and stable antibody increase in active and reactivation disease. In humans, antibody levels and reactive protein numbers increased with bacillary burden. Responses to a subset of 10 proteins were more tightly associated with disease state than reactivity to the broader reactive proteome. CONCLUSIONS: Integration of macaque and human data reveals dynamic properties of antibody responses in relation to outcome and leads to actionable findings for translational research. These include the potential of antibody responses to detect acute infection and preclinical tuberculosis and to identify serodiagnostic proteins for the spectrum of bacillary burden in tuberculosis.

Ultrasensitive and amplification-free detection of SARS-CoV-2 RNA using an electrochemical biosensor powered by CRISPR/Cas13a.

This study proposed a CRISPR/Cas13a-powered electrochemical multiplexed biosensor for detecting SARS-CoV-2 RNA strands. Current SARS-CoV-2 diagnostic methods, such as reverse transcription PCR (RT-PCR), are primarily based on nucleic acid amplification (NAA) and reverse transcription (RT) processes, which have been linked to significant issues such as cross-contamination and long turnaround times. Using a CRISPR/Cas13a system integrated onto an electrochemical biosensor, we present a multiplexed and NAA-free strategy for detecting SARS-CoV-2 RNA fragments. SARS-CoV-2 S and Orf1ab genes were detected in both synthetic and clinical samples. The CRISPR/Cas13a-powered biosensor achieved low detection limits of 2.5 and 4.5 ag/µL for the S and Orf1ab genes, respectively, successfully meeting the sensitivity requirement. Furthermore, the biosensor's specificity, simplicity, and universality may position it as a potential rival to RT-PCR.

Onto the differences in formulating micro-/nanoparticulate drug delivery system from Thai silk and Vietnamese silk: A critical comparison.

Silk fibroin is a natural polymer with physicochemical properties heavily dependent on its silkworm sources and cultivation conditions. Hence, this study critically compared the characteristics and capacity to generate micro-/nanoparticles of fibroin extracted from the Thai silk and Vietnamese silk. Both Thai fibroin (SFT) and Vietnamese fibroin (SFV) were extracted and fabricated into micro-/nanoparticles using the same methods of desalination and condensation, respectively. Firstly, the amino acid compositions of SFT and SFV were determined and found to be similar, suggesting that the different cultivation conditions did not alter the fibroin chemical contents. Secondly, utilizing various analytical techniques, the SFT structure revealed less heavy chains, more light chains and P-25 glycoproteins, and lower crystallinity than those of SFV. Accordingly, compared to the particles formed by SFT, the SFV-based particles were significantly bigger (∼1700 nm vs. ∼150 nm), and possessed less drug (Amphotericin B) entrapment efficiency (64.3 ± 4.4% vs. 79.3 ± 5.1%), higher hemototoxicity, and less biostability in the blood. Conclusively, these differences add more insights for the appropriate applications of each fibroin kind to best promote its qualities and effectiveness.

Prevalence of Fasciola in cattle and of its intermediate host Lymnaea snails in central Vietnam.

The aims of this study were to investigate the prevalence of natural Fasciola infections in both the definitive hosts (cattle) and the intermediate hosts (Lymnaea snails) in central Vietnam. A total of 1,075 fecal samples, randomly collected from cattle in Binh Dinh, Khanh Hoa, and Phu Yen provinces, were examined for Fasciola eggs by a sedimentation method. The overall prevalence of Fasciola was 45.3 %. A subset of the animals (235) was also screened for antibodies against Fasciola by an enzyme-linked immunosorbent assay. Overall, 46.3 % of these animals were shedding Fasciola eggs while 87.2 % were Fasciola seropositive. A lower prevalence of Fasciola was observed in calves ≤ 2 years of age (37.6 %) compared to that in cattle >2 years of age (53.7 %) (p < 0.05). The prevalence in the rainy season (50.8 %) was significantly different to that in the dry season (38.1 %) (p < 0.05). Of the 3.269 Lymnaea viridis and 1.128 Lymnaea swinhoei examined, 31 (0.95 %) and seven (0.62 %), respectively, were found to be infected with Fasciola. This appears to be the first epidemiological survey of the prevalence of Fasciola in cattle and snails in these three provinces in central Vietnam.

Rapid, multiplexed, and nucleic acid amplification-free detection of SARS-CoV-2 RNA using an electrochemical biosensor.

Considering the worldwide health crisis associated with highly contagious severe respiratory disease of COVID-19 outbreak, the development of multiplexed, simple and rapid diagnostic platforms to detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is in high demand. Here, a nucleic acid amplification-free electrochemical biosensor based on four-way junction (4-WJ) hybridization is presented for the detection of SARS-CoV-2. To form a 4-WJ structure, a Universal DNA-Hairpin (UDH) probe is hybridized with two adaptor strands and a SARS-CoV-2 RNA target. One of the adaptor strands is functionalized with a redox mediator that can be detected using an electrochemical biosensor. The biosensor could simultaneously detect 5.0 and 6.8 ag/µL of S and Orf1ab genes, respectively, within 1 h. The biosensor was evaluated with 21 clinical samples (16 positive and 5 negative). The results revealed a satisfactory agreement with qRT-PCR. In conclusion, this biosensor has the potential to be used as an on-site, real-time diagnostic test for COVID-19.

Mapping SARS-CoV-2 Antibody Epitopes in COVID-19 Patients with a Multi-Coronavirus Protein Microarray.

The rapid worldwide spread of SARS-CoV-2 has accelerated research and development for controlling the COVID-19 pandemic. A multi-coronavirus protein microarray was created containing full-length proteins, overlapping protein fragments of various lengths, and peptide libraries from SARS-CoV-2 and four other human coronaviruses. Sera from confirmed COVID-19 patients as well as unexposed individuals were applied to multicoronavirus arrays to identify specific antibody reactivity. High-level IgG, IgM, and IgA reactivity to structural proteins S, M, and N of SARS-CoV-2, as well as accessory proteins such as ORF3a and ORF7a, were observed that were specific to COVID-19 patients. Antibody reactivity against overlapping 100-, 50-, and 30-amino acid fragments of SARS-CoV-2 proteins was used to identify antigenic regions. Numerous proteins of SARS-CoV, Middle East respiratory syndrome coronavirus (MERS-CoV), and the endemic human coronaviruses HCoV-NL63 and HCoV-OC43 were also more reactive with IgG, IgM, and IgA in COVID-19 patient sera than in unexposed control sera, providing further evidence of immunologic cross-reactivity between these viruses. Whereas unexposed individuals had minimal reactivity against SARS-CoV-2 proteins that poorly correlated with reactivity against HCoV-NL63 and HCoV-OC43 S2 and N proteins, COVID-19 patient sera had higher correlation between SARS-CoV-2 and HCoV responses, suggesting that de novo antibodies against SARS-CoV-2 cross-react with HCoV epitopes. Array responses were compared with validated spike protein-specific IgG enzyme-linked immunosorbent assays (ELISAs), showing agreement between orthologous methods. SARS-CoV-2 microneutralization titers were low in the COVID-19 patient sera but correlated with array responses against S and N proteins. The multi-coronavirus protein microarray is a useful tool for mapping antibody reactivity in COVID-19 patients. IMPORTANCE With novel mutant SARS-CoV-2 variants of concern on the rise, knowledge of immune specificities against SARS-CoV-2 proteins is increasingly important for understanding the impact of structural changes in antibody-reactive protein epitopes on naturally acquired and vaccine-induced immunity, as well as broader topics of cross-reactivity and viral evolution. A multi-coronavirus protein microarray used to map the binding of COVID-19 patient antibodies to SARS-CoV-2 proteins and protein fragments as well as to the proteins of four other coronaviruses that infect humans has shown specific regions of SARS-CoV-2 proteins that are highly reactive with patient antibodies and revealed cross-reactivity of these antibodies with other human coronaviruses. These data and the multi-coronavirus protein microarray tool will help guide further studies of the antibody response to COVID-19 and to vaccination against this worldwide pandemic.

Quantitative Determination of Triiodothyronine by Electrochemical Impedance Spectroscopic Biosensor Using Gold Nanoparticle-Modified Electrode.

A label-free electrochemical impedimetric immunosensor for the detection of Triiodothyronine-a thyroid hormone that functions as the biomarker for monitoring for thyroid dysfunction was developed. The gold nanoparticle-modified electrode was employed to achieve the sensitive determination of Triiodothyronine at a low concentration level. The gold nanoparticle layer on the gold electrode was generated by chronoamperometry method and its resulting characteristics were investigated by scanning electron microscopy. Redox probe [Fe(CN)6]3-/4- and electrochemical impedance spec-troscopy was used for both evaluation of the immobilization of anti-Triiodothyronine antibody on the electrode surface and quantitative determination of target Triiodothyronine in different concentrations. The electrode with absorbed antibodies showed significant changes in charge transfer resistance upon binding the antigen, which resulted in an increase in normalized impedance change as the addition of antigen concentrations over a dynamic linear range of 0.01-100 ng/ml. These results indicated that the proposed immunosensor could be a potential alternative method for determination of Triiodothyronine in clinics with the advantage of low cost and less time-consuming.

2D Materials in Development of Electrochemical Point-of-Care Cancer Screening Devices.

Effective cancer treatment requires early detection and monitoring the development progress in a simple and affordable manner. Point-of care (POC) screening can provide a portable and inexpensive tool for the end-users to conveniently operate test and screen their health conditions without the necessity of special skills. Electrochemical methods hold great potential for clinical analysis of variety of chemicals and substances as well as cancer biomarkers due to their low cost, high sensitivity, multiplex detection ability, and miniaturization aptitude. Advances in two-dimensional (2D) material-based electrochemical biosensors/sensors are accelerating the performance of conventional devices toward more practical approaches. Here, recent trends in the development of 2D material-based electrochemical biosensors/sensors, as the next generation of POC cancer screening tools, are summarized. Three cancer biomarker categories, including proteins, nucleic acids, and some small molecules, will be considered. Various 2D materials will be introduced and their biomedical applications and electrochemical properties will be given. The role of 2D materials in improving the performance of electrochemical sensing mechanisms as well as the pros and cons of current sensors as the prospective devices for POC screening will be emphasized. Finally, the future scopes of implementing 2D materials in electrochemical POC cancer diagnostics for the clinical translation will be discussed.

Silk Fibroin-Based Biomaterials for Biomedical Applications: A Review.

Since it was first discovered, thousands of years ago, silkworm silk has been known to be an abundant biopolymer with a vast range of attractive properties. The utilization of silk fibroin (SF), the main protein of silkworm silk, has not been limited to the textile industry but has been further extended to various high-tech application areas, including biomaterials for drug delivery systems and tissue engineering. The outstanding mechanical properties of SF, including its facile processability, superior biocompatibility, controllable biodegradation, and versatile functionalization have allowed its use for innovative applications. In this review, we describe the structure, composition, general properties, and structure-properties relationship of SF. In addition, the methods used for the fabrication and modification of various materials are briefly addressed. Lastly, recent applications of SF-based materials for small molecule drug delivery, biological drug delivery, gene therapy, wound healing, and bone regeneration are reviewed and our perspectives on future development of these favorable materials are also shared.

Age of transfused blood: an independent predictor of mortality despite universal leukoreduction.

BACKGROUND: The transfusion of relatively older stored blood has been associated with an increased risk of multiple organ failure, infection, and death. It remains unknown whether this phenomenon is mitigated by transfusion of leukoreduced red cell units. The purpose of this study was to evaluate the influence of stored blood age on mortality in injured patients who universally received leukoreduced blood. METHODS: Trauma patients who received > or = 1 unit of blood during the first 24 hours after hospital arrival were selected for inclusion. Patients were stratified both according to total units and "old" units (> or = 14 days) versus "young" units (< 14 days) received in the initial 24 hours. Odds ratios and 95% confidence intervals (CIs) were calculated for the association between mortality and the age and amount of blood transfused, adjusted for age, sex, injury severity, injury mechanism, number of units transfused, and length of stay. RESULTS: Over 7.5 years, 1,813 patients met study criteria. Among patients who received a total of 1 to 2 or 3 to 5 units in the first 24 hours, there was no association between the amount and age of transfused blood and mortality. For patients who received a total of > or = 6 units, the presence of > or = 3 units of young blood was associated with a 3.8-fold increased odds of death (CI: 1.1-12.7), compared with a 7.8-fold (CI: 2.3-26.3) increased odds of death associated with the presence of > or = 3 units of old blood (p = 0.0024). CONCLUSION: Although larger volumes of blood, irrespective of age, are associated with increased odds of mortality, the transfusion of blood stored beyond 2 weeks appears to potentiate this association despite a practice of universal leukoreduction. For patients who receive relatively smaller transfusion volumes, blood age appears to have no effect on mortality.