Expectation effects in working memory training.

There is a growing body of research focused on developing and evaluating behavioral training paradigms meant to induce enhancements in cognitive function. It has recently been proposed that one mechanism through which such performance gains could be induced involves participants' expectations of improvement. However, no work to date has evaluated whether it is possible to cause changes in cognitive function in a long-term behavioral training study by manipulating expectations. In this study, positive or negative expectations about cognitive training were both explicitly and associatively induced before either a working memory training intervention or a control intervention. Consistent with previous work, a main effect of the training condition was found, with individuals trained on the working memory task showing larger gains in cognitive function than those trained on the control task. Interestingly, a main effect of expectation was also found, with individuals given positive expectations showing larger cognitive gains than those who were given negative expectations (regardless of training condition). No interaction effect between training and expectations was found. Exploratory analyses suggest that certain individual characteristics (e.g., personality, motivation) moderate the size of the expectation effect. These results highlight aspects of methodology that can inform future behavioral interventions and suggest that participant expectations could be capitalized on to maximize training outcomes.

Contrast response function estimation with nonparametric Bayesian active learning.

Multidimensional psychometric functions can typically be estimated nonparametrically for greater accuracy or parametrically for greater efficiency. By recasting the estimation problem from regression to classification, however, powerful machine learning tools can be leveraged to provide an adjustable balance between accuracy and efficiency. Contrast sensitivity functions (CSFs) are behaviorally estimated curves that provide insight into both peripheral and central visual function. Because estimation can be impractically long, current clinical workflows must make compromises such as limited sampling across spatial frequency or strong assumptions on CSF shape. This article describes the development of the machine learning contrast response function (MLCRF) estimator, which quantifies the expected probability of success in performing a contrast detection or discrimination task. A machine learning CSF can then be derived from the MLCRF. Using simulated eyes created from canonical CSF curves and actual human contrast response data, the accuracy and efficiency of the machine learning contrast sensitivity function (MLCSF) was evaluated to determine its potential utility for research and clinical applications. With stimuli selected randomly, the MLCSF estimator converged slowly toward ground truth. With optimal stimulus selection via Bayesian active learning, convergence was nearly an order of magnitude faster, requiring only tens of stimuli to achieve reasonable estimates. Inclusion of an informative prior provided no consistent advantage to the estimator as configured. MLCSF achieved efficiencies on par with quickCSF, a conventional parametric estimator, but with systematically higher accuracy. Because MLCSF design allows accuracy to be traded off against efficiency, it should be explored further to uncover its full potential.

Development and validation of a Spanish-language spatial release from masking task in a Mexican population.

This study validates a new Spanish-language version of the Coordinate Response Measure (CRM) corpus using a well-established measure of spatial release from masking (SRM). Participants were 96 Spanish-speaking young adults without hearing complaints in Mexico City. To present the Spanish-language SRM test, we created new recordings of the CRM with Spanish-language Translations and updated the freely available app (PART; https://ucrbraingamecenter.github.io/PART_Utilities/) to present materials in Spanish. In addition to SRM, we collected baseline data on a battery of non-speech auditory assessments, including detection of frequency modulations, temporal gaps, and modulated broadband noise in the temporal, spectral, and spectrotemporal domains. Data demonstrate that the newly developed speech and non-speech tasks show similar reliability to an earlier report in English-speaking populations. This study demonstrates an approach by which auditory assessment for clinical and basic research can be extended to Spanish-speaking populations for whom testing platforms are not currently available.

Remote auditory assessment using Portable Automated Rapid Testing (PART) and participant-owned devices.

Remote testing of auditory function can be transformative to both basic research and hearing healthcare; however, historically, many obstacles have limited remote collection of reliable and valid auditory psychometric data. Here, we report performance on a battery of auditory processing tests using a remotely administered system, Portable Automatic Rapid Testing. We compare a previously reported dataset collected in a laboratory setting with the same measures using uncalibrated, participant-owned devices in remote settings (experiment 1, n = 40) remote with and without calibrated hardware (experiment 2, n = 36) and laboratory with and without calibrated hardware (experiment 3, n = 58). Results were well-matched across datasets and had similar reliability, but overall performance was slightly worse than published norms. Analyses of potential nuisance factors such as environmental noise, distraction, or lack of calibration failed to provide reliable evidence that these factors contributed to the observed variance in performance. These data indicate feasibility of remote testing of suprathreshold auditory processing using participants' own devices. Although the current investigation was limited to young participants without hearing difficulties, its outcomes demonstrate the potential for large-scale, remote hearing testing of more hearing-diverse populations both to advance basic science and to establish the clinical viability of auditory remote testing.

UCancellation: A new mobile measure of selective attention and concentration.

Measuring selective attention in a speeded task can provide valuable insight into the concentration ability of an individual, and can inform neuropsychological assessment of attention in aging, traumatic brain injury, and in various psychiatric disorders. There are only a few tools to measure selective attention that are freely available, psychometrically validated, and can be used flexibly both for in-person and remote assessment. To address this gap, we developed a self-administrable, mobile-based test called "UCancellation" (University of California Cancellation), which was designed to assess selective attention and concentration and has two stimulus sets: Letters and Pictures. UCancellation takes less than 7 minutes to complete, is automatically scored, has multiple forms to allow repeated testing, and is compatible with a variety of iOS and Android devices. Here we report the results of a study that examined parallel-test reliability and convergent validity of UCancellation in a sample of 104 college students. UCancellation Letters and Pictures showed adequate parallel test reliability (r = .71-.83, p < 0.01) and internal consistency (ɑ = .73-.91). It also showed convergent validity with another widely used cancellation task, d2 Test of Attention (r = .43-.59, p < 0.01), and predicted performance on a cognitive control composite (r = .34-.41, p < 0.05). These results suggest that UCancellation is a valid test of selective attention and inhibitory control, which warrants further data collection to establish norms.

Antimicrobial Coating Prevents Ventilator-Associated Pneumonia in a 72 hour Large Animal Model.

BACKGROUND: The primary goal of this study was to demonstrate that endotracheal tubes coated with antimicrobial lipids plus mucolytic or antimicrobial lipids with antibiotics plus mucolytic would significantly reduce pneumonia in the lungs of pigs after 72 hours of continuous mechanical ventilation compared to uncoated controls. MATERIALS AND METHODS: Eighteen female pigs were mechanically ventilated for up to 72 hours through uncoated endotracheal tubes, endotracheal tubes coated with the antimicrobial lipid, octadecylamine, and the mucolytic, N-acetylcysteine, or tubes coated with octadecylamine, N-acetylcysteine, doxycycline, and levofloxacin (6 pigs per group). No exogenous bacteria were inoculated into the pigs, pneumonia resulted from the pigs' endogenous oral flora. Vital signs were recorded every 15 minutes and arterial blood gas measurements were obtained for the duration of the experiment. Pigs were sacrificed either after completion of 72 hours of mechanical ventilation or just prior to hypoxic arrest. Lungs, trachea, and endotracheal tubes were harvested for analysis to include bacterial counts of lung, trachea, and endotracheal tubes, lung wet and dry weights, and lung tissue for histology. RESULTS: Pigs ventilated with coated endotracheal tubes were less hypoxic, had less bacterial colonization of the lungs, and survived significantly longer than pigs ventilated with uncoated tubes. Octadecylamine-N-acetylcysteine-doxycycline-levofloxacin coated endotracheal tubes had less bacterial colonization than uncoated or octadecylamine-N-acetylcysteine coated tubes. CONCLUSION: Endotracheal tubes coated with antimicrobial lipids plus mucolytic and antimicrobial lipids with antibiotics plus mucolytic reduced bacterial colonization of pig lungs after prolonged mechanical ventilation and may be an effective strategy to reduce ventilator-associated pneumonia.

The Role of Acid Sphingomyelinase Inhibition in Repetitive Mild Traumatic Brain Injury.

BACKGROUND: Chronic traumatic encephalopathy is a consequence of repetitive mild traumatic brain injury (rmTBI). These injuries can result in psychiatric disorders that are treated with amitriptyline. Amitriptyline improves neuronal regeneration in major depression via inhibition of acid sphingomyelinase. We hypothesized that acid sphingomyelinase inhibition would preserve neuronal regeneration and decrease depressive symptoms following rmTBI in a murine model. METHODS: A murine model of rmTBI was established using a weight-drop method. Mice were subjected to mTBI every other day for 7 d. Mice received amitriptyline injection 2 h prior to each mTBI. After the final mTBI, mice underwent behavioral studies or biochemical analysis. Hippocampi were analyzed for markers of neurogenesis and phosphorylated tau aggregation. RESULTS: Mice that underwent rmTBI showed increased hippocampal phosphorylated tau aggregation 1 mo following rmTBI as well as decreased neuronal regeneration by bromodeoxyuridine uptake and doublecortin immunohistochemistry. Mice with either genetic deficiency or pharmacologic inhibition of acid sphingomyelinase demonstrated improved neuronal regeneration and decreased phosphorylated tau aggregation compared to untreated rmTBI mice. Behavioral testing showed rmTBI mice spent significantly more time in the dark and waiting to initiate feeding compared to sham mice. These behaviors were partially prevented by the inhibition of acid sphingomyelinase. CONCLUSIONS: We established a murine model of rmTBI that leads to tauopathy, depression, and impaired hippocampal neurogenesis. Inhibition of acid sphingomyelinase prevented the harmful neurologic and behavioral effects of rmTBI. These findings highlight an important opportunity to improve recovery or prevent neuropsychiatric decline in patients at risk for chronic traumatic encephalopathy.

Auditory-visual interactions in egocentric distance perception: Ventriloquism effect and aftereffect.

This study describes data on auditory-visual integration and visually-guided adaptation of auditory distance perception using the ventriloquism effect (VE) and ventriloquism aftereffect (VAE). In an experiment, participants judged egocentric distance of interleaved auditory or auditory-visual stimuli with the auditory component located from 0.7 to 2.04 m in front of listeners in a real reverberant environment. The visual component of auditory-visual stimuli was displaced 30% closer (V-closer), 30% farther (V-farther), or aligned (V-aligned) with respect to the auditory component. The VE and VAE were measured in auditory and auditory-visual trials, respectively. Both effects were approximately independent of target distance when expressed in logarithmic units. The VE strength, defined as a difference of V-misaligned and V-aligned response bias, was approximately 72% of the auditory-visual disparity regardless of the visual-displacement direction, while the VAE was stronger in the V-farther (44%) than the V-closer (31%) condition. The VAE persisted to post-adaptation auditory-only blocks of trials, although it was diminished. The rates of build-up/break-down of the VAE were asymmetrical, with slower adaptation in the V-closer condition. These results suggest that auditory-visual distance integration is independent of the direction of induced shift, while the re-calibration is stronger and faster when evoked by more distant visual stimuli.

Exponential spectro-temporal modulation generation.

Traditionally, real-time generation of spectro-temporally modulated noise has been performed on a linear amplitude scale, partially due to computational constraints. Experiments often require modulation that is sinusoidal on a logarithmic amplitude scale as a result of the many perceptual and physiological measures which scale linearly with exponential changes in the signal magnitude. A method is presented for computing exponential spectro-temporal modulation, showing that it can be expressed analytically as a sum over linearly offset sidebands with component amplitudes equal to the values of the modified Bessel function of the first kind. This approach greatly improves the efficiency and precision of stimulus generation over current methods, facilitating real-time generation for a broad range of carrier and envelope signals.

Temporal integration of monaural and dichotic frequency modulation.

Frequency modulation (FM) detection at low modulation frequencies is commonly used as an index of temporal fine-structure processing. The present study evaluated the rate of improvement in monaural and dichotic FM across a range of test parameters. In experiment I, dichotic and monaural FM detection was measured as a function of duration and modulator starting phase. Dichotic FM thresholds were lower than monaural FM thresholds and the modulator starting phase had no effect on detection. Experiment II measured monaural FM detection for signals that differed in modulation rate and duration such that the improvement with duration in seconds (carrier) or cycles (modulator) was compared. Monaural FM detection improved monotonically with the number of modulation cycles, suggesting that the modulator is extracted prior to detection. Experiment III measured dichotic FM detection for shorter signal durations to test the hypothesis that dichotic FM relies primarily on the signal onset. The rate of improvement decreased as duration increased, which is consistent with the use of primarily onset cues for the detection of dichotic FM. These results establish that improvement with duration occurs as a function of the modulation cycles at a rate consistent with the independent-samples model for monaural FM, but later cycles contribute less to detection in dichotic FM.

Acquisition of visual priors and induced hallucinations in chronic schizophrenia.

Prominent theories suggest that symptoms of schizophrenia stem from learning deficiencies resulting in distorted internal models of the world. To test these theories further, we used a visual statistical learning task known to induce rapid implicit learning of the stimulus statistics. In this task, participants are presented with a field of coherently moving dots and are asked to report the presented direction of the dots (estimation task), and whether they saw any dots or not (detection task). Two of the directions were more frequently presented than the others. In controls, the implicit acquisition of the stimuli statistics influences their perception in two ways: (i) motion directions are perceived as being more similar to the most frequently presented directions than they really are (estimation biases); and (ii) in the absence of stimuli, participants sometimes report perceiving the most frequently presented directions (a form of hallucinations). Such behaviour is consistent with probabilistic inference, i.e. combining learnt perceptual priors with sensory evidence. We investigated whether patients with chronic, stable, treated schizophrenia (n = 20) differ from controls (n = 23) in the acquisition of the perceptual priors and/or their influence on perception. We found that although patients were slower than controls, they showed comparable acquisition of perceptual priors, approximating the stimulus statistics. This suggests that patients have no statistical learning deficits in our task. This may reflect our patients' relative wellbeing on antipsychotic medication. Intriguingly, however, patients experienced significantly fewer (P = 0.016) hallucinations of the most frequently presented directions than controls when the stimulus was absent or when it was very weak (prior-based lapse estimations). This suggests that prior expectations had less influence on patients' perception than on controls when stimuli were absent or below perceptual threshold.

Dissociating electrophysiological correlates of contextual and perceptual learning in a visual search task.

Perceptual learning and contextual learning are two types of implicit visual learning that can co-occur in the same tasks. For example, to find an animal in the woods, you need to know where to look in the environment (contextual learning) and you must be able to discriminate its features (perceptual learning). However, contextual and perceptual learning are typically studied using distinct experimental paradigms, and little is known regarding their comparative neural mechanisms. In this study, we investigated contextual and perceptual learning in 12 healthy adult humans as they performed the same visual search task, and we examined psychophysical and electrophysiological (event-related potentials) measures of learning. Participants were trained to look for a visual stimulus, a small line with a specific orientation, presented among distractors. We found better performance for the trained target orientation as compared to an untrained control orientation, reflecting specificity of perceptual learning for the orientation of trained elements. This orientation specificity effect was associated with changes in the C1 component. We also found better performance for repeated spatial configurations as compared to novel ones, reflecting contextual learning. This context-specific effect was associated with the N2pc component. Taken together, these results suggest that contextual and perceptual learning are distinct visual learning phenomena that have different behavioral and electrophysiological characteristics.

We don't all look the same; detailed examination of peripheral looking strategies after simulated central vision loss.

Loss of central vision can be compensated for in part by increased use of peripheral vision. For example, patients with macular degeneration or those experiencing simulated central vision loss tend to develop eccentric viewing strategies for reading or other visual tasks. The factors driving this learning are still unclear and likely involve complex changes in oculomotor strategies that may differ among people and tasks. Although to date a number of studies have examined reliance on peripheral vision after simulated central vision loss, individual differences in developing peripheral viewing strategies and the extent to which they transfer to untrained tasks have received little attention. Here, we apply a recently published method of characterizing oculomotor strategies after central vision loss to understand the time course of changes in oculomotor strategies through training in 19 healthy individuals with a gaze-contingent display obstructing the central 10° of the visual field. After 10 days of training, we found mean improvements in saccadic re-referencing (the percentage of trials in which the first saccade placed the target outside the scotoma), latency of target acquisition (time interval between target presentation and a saccade putting the target outside the scotoma), and fixation stability. These results are consistent with participants developing compensatory oculomotor strategies as a result of training. However, we also observed substantial individual differences in the formation of eye movement strategies and the extent to which they transferred to an untrained task, likely reflecting both variations in learning rates and patterns of learning. This more complete characterization of peripheral looking strategies and how they change with training may help us understand individual differences in rehabilitation after central vision loss.

A method to characterize compensatory oculomotor strategies following simulated central vision loss.

Loss of central vision can be partially compensated by increased use of peripheral vision. For example, patients experiencing central vision loss due to disease (macular degeneration) or healthy participants trained with simulated central vision loss, tend to develop eccentric fixation spots for reading or other visual tasks. In both patients and in simulated conditions, there are substantial individual variations in the effective use of the periphery. The factors driving these individual differences are still unclear. Although early approaches have described some dimensions of these strategies, the field is still in its initial stages and important elements are often conflated when examining gaze patterns. Here, we propose a systematic approach to characterize oculomotor strategies in cases of central vision loss that distinguishes different components: saccadic re-referencing, saccadic precision, first saccade landing dispersion, fixation stability, latency of target acquisition, and percentage of trials that are useful. We tested this approach in healthy individuals trained with a gaze-contingent display obstructing the central 10 degrees of the visual field. The use of simulated scotoma helps overcome known challenges in clinical research, from recruitment and compliance to the diverse extent and nature of the visual loss. Importantly, this approach offers the ability to examine oculomotor strategies as they develop in controlled settings where viewing conditions are similar across participants. Results show substantial differences in characteristics of peripheral looking strategies, both across trials and individuals. This more complete characterization of peripheral looking strategies can help us understand individual differences in rehabilitation after central vision loss.

Portable Automated Rapid Testing (PART) for auditory assessment: Validation in a young adult normal-hearing population.

This study aims to determine the degree to which Portable Automated Rapid Testing (PART), a freely available program running on a tablet computer, is capable of reproducing standard laboratory results. Undergraduate students were assigned to one of three within-subject conditions that examined repeatability of performance on a battery of psychoacoustical tests of temporal fine structure processing, spectro-temporal amplitude modulation, and targets in competition. The repeatability condition examined test/retest with the same system, the headphones condition examined the effects of varying headphones (passive and active noise-attenuating), and the noise condition examined repeatability in the presence of recorded cafeteria noise. In general, performance on the test battery showed high repeatability, even across manipulated conditions, and was similar to that reported in the literature. These data serve as validation that suprathreshold psychoacoustical tests can be made accessible to run on consumer-grade hardware and perform in less controlled settings. This dataset also provides a distribution of thresholds that can be used as a normative baseline against which auditory dysfunction can be identified in future work.

Deep Neural Networks for Modeling Visual Perceptual Learning.

Understanding visual perceptual learning (VPL) has become increasingly more challenging as new phenomena are discovered with novel stimuli and training paradigms. Although existing models aid our knowledge of critical aspects of VPL, the connections shown by these models between behavioral learning and plasticity across different brain areas are typically superficial. Most models explain VPL as readout from simple perceptual representations to decision areas and are not easily adaptable to explain new findings. Here, we show that a well -known instance of deep neural network (DNN), whereas not designed specifically for VPL, provides a computational model of VPL with enough complexity to be studied at many levels of analyses. After learning a Gabor orientation discrimination task, the DNN model reproduced key behavioral results, including increasing specificity with higher task precision, and also suggested that learning precise discriminations could transfer asymmetrically to coarse discriminations when the stimulus conditions varied. Consistent with the behavioral findings, the distribution of plasticity moved toward lower layers when task precision increased and this distribution was also modulated by tasks with different stimulus types. Furthermore, learning in the network units demonstrated close resemblance to extant electrophysiological recordings in monkey visual areas. Altogether, the DNN fulfilled predictions of existing theories regarding specificity and plasticity and reproduced findings of tuning changes in neurons of the primate visual areas. Although the comparisons were mostly qualitative, the DNN provides a new method of studying VPL, can serve as a test bed for theories, and assists in generating predictions for physiological investigations.SIGNIFICANCE STATEMENT Visual perceptual learning (VPL) has been found to cause changes at multiple stages of the visual hierarchy. We found that training a deep neural network (DNN) on an orientation discrimination task produced behavioral and physiological patterns similar to those found in human and monkey experiments. Unlike existing VPL models, the DNN was pre-trained on natural images to reach high performance in object recognition, but was not designed specifically for VPL; however, it fulfilled predictions of existing theories regarding specificity and plasticity and reproduced findings of tuning changes in neurons of the primate visual areas. When used with care, this unbiased and deep-hierarchical model can provide new ways of studying VPL from behavior to physiology.

Validation of a matrix reasoning task for mobile devices.

Many cognitive tasks have been adapted for tablet-based testing, but tests to assess nonverbal reasoning ability, as measured by matrix-type problems that are suited to repeated testing, have yet to be adapted for and validated on mobile platforms. Drawing on previous research, we developed the University of California Matrix Reasoning Task (UCMRT)-a short, user-friendly measure of abstract problem solving with three alternate forms that works on tablets and other mobile devices and that is targeted at a high-ability population frequently used in the literature (i.e., college students). To test the psychometric properties of UCMRT, a large sample of healthy young adults completed parallel forms of the test, and a subsample also completed Raven's Advanced Progressive Matrices and a math test; furthermore, we collected college records of academic ability and achievement. These data show that UCMRT is reliable and has adequate convergent and external validity. UCMRT is self-administrable, freely available for researchers, facilitates repeated testing of fluid intelligence, and resolves numerous limitations of existing matrix tests.

Staphylococcus aureus Alpha-Toxin Disrupts Endothelial-Cell Tight Junctions via Acid Sphingomyelinase and Ceramide.

Staphylococcus aureus (S. aureus) infections are among the most common and severe infections, garnering notoriety in an era of increasing resistance to antibiotics. It is therefore important to define molecular mechanisms by which this pathogen attacks host cells. Here, we demonstrate that alpha-toxin, one of the major toxins of S. aureus, induces activation of acid sphingomyelinase and concomitant release of ceramide in endothelial cells treated with the toxin. Activation of acid sphingomyelinase by alpha-toxin is mediated via ADAM10. Infection experiments employing alpha-toxin-deficient S. aureus and the corresponding wild-type strain reveal that activation of acid sphingomyelinase in endothelial cells requires alpha-toxin expression by the pathogen. Activation of acid sphingomyelinase is linked to degradation of tight junctions in endothelial cells in vitro, which is blocked by pharmacological inhibition of acid sphingomyelinase. Most importantly, alpha-toxin induces severe degradation of tight junctions in the lung and causes lung edema in vivo, which is prevented by genetic deficiency of acid sphingomyelinase. These data indicate a novel and important role of the acid sphingomyelinase/ceramide system for the endothelial response to toxins and provide a molecular link between alpha-toxin and the degradation of tight junctions. The data also suggest that inhibition of acid sphingomyelinase may provide a novel treatment option to prevent lung edema caused by S. aureus alpha-toxin.

Neutrophils Kill Reactive Oxygen Species-Resistant Pseudomonas aeruginosa by Sphingosine.

BACKGROUND/AIMS: Cystic fibrosis (CF) is dominated by chronic inflammation and infection of the lung resulting in lung destruction and early death of patients. The lungs of CF patients are characterized by a massive accumulation of neutrophils. It requires definition why these massive numbers of neutrophils fail to eliminate typical CF pathogens like Staphylococcus aureus and Pseudomonas aeruginosa (P. aeruginosa) in CF lungs. METHODS: We determined ceramide, sphingosine and reactive oxygen species (ROS) in neutrophils from wildtype and CF mice and determined the effect of sphingosine and ROS alone or in combination on killing of different P. aeruginosa strains. RESULTS: We demonstrate that wildtype neutrophils are able to kill non-mucoid and mucoid clinical P. aeruginosa strains, while neutrophils from CF mice are insufficient to kill these P. aeruginosa strains, although both types of neutrophils infected with P. aeruginosa produce comparable levels of superoxide. All three analyzed P. aeruginosa strains are resistant to reactive oxygen species. The inability of CF neutrophils to kill P. aeruginosa is caused by a marked decrease of surface sphingosine levels in CF neutrophils. Wildtype neutrophils contain much higher concentrations of surface sphingosine than CF neutrophils. Further, wildtype neutrophils, but not CF neutrophils, release sphingosine, most likely as microparticles, upon infection. Sphingosine kills P. aeruginosa in vitro at low micromolar concentrations. Reconstitution of sphingosine in CF neutrophils restores their ability to kill these pathogens, demonstrating the significance of sphingosine for bacterial killing. CONCLUSION: The data provide evidence for a new paradigm explaining how neutrophils kill ROS-resistant P. aeruginosa, i.e. by sphingosine that kills P. aeruginosa at low concentrations. This mechanism is defective in CF neutrophils.

pH modulation ameliorates the red blood cell storage lesion in a murine model of transfusion.

BACKGROUND: Prolonged storage of packed red blood cells (pRBCs) induces a series of harmful biochemical and metabolic changes known as the RBC storage lesion. RBCs are currently stored in an acidic storage solution, but the effect of pH on the RBC storage lesion is unknown. We investigated the effect of modulation of storage pH on the RBC storage lesion and on erythrocyte survival after transfusion. METHODS: Murine pRBCs were stored in Additive Solution-3 (AS3) under standard conditions (pH, 5.8), acidic AS3 (pH, 4.5), or alkalinized AS3 (pH, 8.5). pRBC units were analyzed at the end of the storage period. Several components of the storage lesion were measured, including cell-free hemoglobin, microparticle production, phosphatidylserine externalization, lactate accumulation, and byproducts of lipid peroxidation. Carboxyfluorescein-labeled erythrocytes were transfused into healthy mice to determine cell survival. RESULTS: Compared with pRBCs stored in standard AS3, those stored in alkaline solution exhibited decreased hemolysis, phosphatidylserine externalization, microparticle production, and lipid peroxidation. Lactate levels were greater after storage in alkaline conditions, suggesting that these pRBCs remained more metabolically viable. Storage in acidic AS3 accelerated erythrocyte deterioration. Compared with standard AS3 storage, circulating half-life of cells was increased by alkaline storage but decreased in acidic conditions. CONCLUSIONS: Storage pH significantly affects the quality of stored RBCs and cell survival after transfusion. Current erythrocyte storage solutions may benefit from refinements in pH levels.