Webcam-based eye tracking to detect mind wandering and comprehension errors.

Recent advances in computer vision have opened the door for scalable eye tracking using only a webcam. Such solutions are particularly useful for online educational technologies, in which a goal is to respond adaptively to students' ongoing experiences. We used WebGazer, a webcam-based eye-tracker, to automatically detect covert cognitive states during an online reading-comprehension task related to task-unrelated thought and comprehension. We present data from two studies using different populations: (1) a relatively homogenous sample of university students (N = 105), and (2) a more diverse sample from Prolific (N = 173, with < 20% White participants). Across both studies, the webcam-based eye-tracker provided sufficiently accurate and precise gaze measurements to predict both task-unrelated thought and reading comprehension from a single calibration. We also present initial evidence of predictive validity, including a positive correlation between predicted rates of task-unrelated thought and comprehension scores. Finally, we present slicing analyses to determine how performance changed under certain conditions (lighting, glasses, etc.) and generalizability of the results across the two datasets (e.g., training on the data Study 1 and testing on data from Study 2, and vice versa). We conclude by discussing results in the context of remote research and learning technologies.

Permanent urethral ligation after AUS cuff erosion: Is it ready for prime time?

AIMS: Although artificial urinary sphincter (AUS) has long been the gold standard treatment for severe stress urinary incontinence, poor tissue quality in patients with prior cuff erosions may preclude this option. Formal supravesical diversion and/or bladder neck closure comprise alternative salvage options but are associated with significant morbidity and mortality. We review our experience with permanent urethral ligation (PUL) among patients deemed not to be candidates for AUS replacement following cuff erosion. METHODS: From a single-center database of 396 patients undergoing AUS from 2014 to 2020, 20 men underwent PUL with suprapubic tube (SPT) diversion. Clinical characteristics and outcomes were evaluated. Quality of life (QOL) was assessed using chart review, Michigan Incontinence Symptom Index (M-ISI), and Patient Global Impression of Improvement (PGI-I). RESULTS: PUL resulted in continence in 18 (90%) men; 15 after the initial surgery and three after repeat ligation. Patients were elderly (average age 75) with significant comorbidities. A total of 11 (55.5%) patients experienced complications in the 90-day postoperative period (seven Clavien-Dindo Grade II, four Grade III). Over an average follow-up of 30.3 months (interquartile range: 15.75-48.75), four patients underwent cystectomy and one underwent perineal urethrostomy. In the remaining patients managed by PUL, 13 had satisfactory M-ISI scores and indicated overall improvement on PGI-I. CONCLUSIONS: For men with AUS cuff erosion who are poor candidates for replacement, PUL with chronic SPT drainage represents an acceptable alternative option to restore continence and improve QOL. Though complications are not uncommon, the morbidity profile still compares favorably to more invasive formal urinary diversion.

The Vibrio cholerae type VI secretion system can modulate host intestinal mechanics to displace gut bacterial symbionts.

Host-associated microbiota help defend against bacterial pathogens; however, the mechanisms by which pathogens overcome this defense remain largely unknown. We developed a zebrafish model and used live imaging to directly study how the human pathogen Vibrio cholerae invades the intestine. The gut microbiota of fish monocolonized by symbiotic strain Aeromonas veronii was displaced by V. cholerae expressing its type VI secretion system (T6SS), a syringe-like apparatus that deploys effector proteins into target cells. Surprisingly, displacement was independent of T6SS-mediated killing of A. veronii, driven instead by T6SS-induced enhancement of zebrafish intestinal movements that led to expulsion of the resident microbiota by the host. Deleting an actin cross-linking domain from the T6SS apparatus returned intestinal motility to normal and thwarted expulsion, without weakening V. cholerae's ability to kill A. veronii in vitro. Our finding that bacteria can manipulate host physiology to influence intermicrobial competition has implications for both pathogenesis and microbiome engineering.

The effect of habitats and fitness on species coexistence in systems with cyclic dominance.

Cyclic dominance between species may yield spiral waves that are known to provide a mechanism enabling persistent species coexistence. This observation holds true even in presence of spatial heterogeneity in the form of quenched disorder. In this work we study the effects on spatio-temporal patterns and species coexistence of structured spatial heterogeneity in the form of habitats that locally provide one of the species with an advantage. Performing extensive numerical simulations of systems with three and six species we show that these structured habitats destabilize spiral waves. Analyzing extinction events, we find that species extinction probabilities display a succession of maxima as function of time, that indicate a periodically enhanced probability for species extinction. Analysis of the mean extinction time reveals that as a function of the parameter governing the advantage of one of the species a transition between stable coexistence and unstable coexistence takes place. We also investigate how efficiency as a predator or a prey affects species coexistence.

Protein O-Mannosyltransferases Affect Sensory Axon Wiring and Dynamic Chirality of Body Posture in the Drosophila Embryo.

Genetic defects in protein O-mannosyltransferase 1 (POMT1) and POMT2 underlie severe muscular dystrophies. POMT genes are evolutionarily conserved in metazoan organisms. In Drosophila, both male and female POMT mutants show a clockwise rotation of adult abdominal segments, suggesting a chirality of underlying pathogenic mechanisms. Here we described and analyzed a similar phenotype in POMT mutant embryos that shows left-handed body torsion. Our experiments demonstrated that coordinated muscle contraction waves are associated with asymmetric embryo rolling, unveiling a new chirality marker in Drosophila development. Using genetic and live-imaging approaches, we revealed that the torsion phenotype results from differential rolling and aberrant patterning of peristaltic waves of muscle contractions. Our results demonstrated that peripheral sensory neurons are required for normal contractions that prevent the accumulation of torsion. We found that POMT mutants show abnormal axonal connections of sensory neurons. POMT transgenic expression limited to sensory neurons significantly rescued the torsion phenotype, axonal connectivity defects, and abnormal contractions in POMT mutant embryos. Together, our data suggested that protein O-mannosylation is required for normal sensory feedback to control coordinated muscle contractions and body posture. This mechanism may shed light on analogous functions of POMT genes in mammals and help to elucidate the etiology of neurological defects in muscular dystrophies.SIGNIFICANCE STATEMENT Protein O-mannosyltransferases (POMTs) are evolutionarily conserved in metazoans. Mutations in POMTs cause severe muscular dystrophies associated with pronounced neurological defects. However, neurological functions of POMTs remain poorly understood. We demonstrated that POMT mutations in Drosophila result in abnormal muscle contractions and cause embryo torsion. Our experiments uncovered a chirality of embryo movements and a unique POMT-dependent mechanism that maintains symmetry of a developing system affected by chiral forces. Furthermore, POMTs were found to be required for proper axon connectivity of sensory neurons, suggesting that O-mannosylation regulates the sensory feedback controlling muscle contractions. This novel POMT function in the peripheral nervous system may shed light on analogous functions in mammals and help to elucidate pathomechanisms of neurological abnormalities in muscular dystrophies.

Host Gut Motility Promotes Competitive Exclusion within a Model Intestinal Microbiota.

The gut microbiota is a complex consortium of microorganisms with the ability to influence important aspects of host health and development. Harnessing this "microbial organ" for biomedical applications requires clarifying the degree to which host and bacterial factors act alone or in combination to govern the stability of specific lineages. To address this issue, we combined bacteriological manipulation and light sheet fluorescence microscopy to monitor the dynamics of a defined two-species microbiota within a vertebrate gut. We observed that the interplay between each population and the gut environment produces distinct spatiotemporal patterns. As a consequence, one species dominates while the other experiences sudden drops in abundance that are well fit by a stochastic mathematical model. Modeling revealed that direct bacterial competition could only partially explain the observed phenomena, suggesting that a host factor is also important in shaping the community. We hypothesized the host determinant to be gut motility, and tested this mechanism by measuring colonization in hosts with enteric nervous system dysfunction due to a mutation in the ret locus, which in humans is associated with the intestinal motility disorder known as Hirschsprung disease. In mutant hosts we found reduced gut motility and, confirming our hypothesis, robust coexistence of both bacterial species. This study provides evidence that host-mediated spatial structuring and stochastic perturbation of communities can drive bacterial population dynamics within the gut, and it reveals a new facet of the intestinal host-microbe interface by demonstrating the capacity of the enteric nervous system to influence the microbiota. Ultimately, these findings suggest that therapeutic strategies targeting the intestinal ecosystem should consider the dynamic physical nature of the gut environment.

Designing and testing an educational innovation.

Technology provides an opportunity to develop innovations to improve the way we teach. Developing educational innovations must be approached in a methodical and thorough manner to identify the educational gap and validate the innovation's success in filling the gap. A successful educational innovation requires a stepwise approach that includes needs assessment, design of intervention, testing and analysis, evaluation of intervention, and determination of learning retention.

Activity monitoring in men's college soccer: a single season longitudinal study.

Performance in soccer has been characterized previously using time-motion analyses; however, it is unclear if men's college soccer shares performance characteristics with women's college or men's professional soccer. The purpose of this study was to compare proportions of matches spent walking, jogging, running, and sprinting in men's college soccer. Twenty-two male college soccer players wore global positioning system units during matches. Proportions of walking, jogging, running, high-speed running, and sprinting were calculated for each player based on time period (first half, second half, extra time) and outcome (win, loss, tie). Multivariate analyses of variance were run for each time period to compare positions. Means, 95% confidence intervals, and effect sizes were calculated for each position based on time period and match outcome. There were differences in low-speed and high-speed activities based on position, with forwards and midfielders demonstrating increased high-speed activities. Positional differences may require different physiological profiles and should be a consideration during training.

Comparative analysis of anticipated pain versus experienced pain in patients undergoing office vasectomy.

INTRODUCTION: Advances in vasectomy technique have minimized patient discomfort; however fear of pain remains a primary concern. The objective is to determine how the anticipation of pain associated with vasectomy compares with patient's actual intraoperative experienced pain levels. MATERIALS AND METHODS: A cohort of patients undergoing clinic vasectomy was analyzed. Using visual analog pain scale patients were asked to rate their anticipated pain score (APS) pre-procedure and then an experienced pain score (EPS) post-procedure. Patients were also stratified by APS scores (high versus low). Changes in pain score were compared across these groups. Pain scores stratified by age, race, narcotics use, psychiatric history, and prior surgical history were also compared. RESULTS: In the 172 patients included, the average pre-op APS was 5.2 (95% CI 4.3-5.6), while post-op EPS was 2.1 (95% CI 1.8-2.4). Patients were stratified into 'high' (6-10) and 'low' (0-5) pre-op APS groups. The average drop in pain scores was found to be significantly larger in the 'high' versus the 'low' APS groups (4.66 versus 1.65 p < 0.001). No statistical difference was noted in the change in pain scores based on age, race, prior surgical history, chronic narcotics use or psychiatric history. Both groups tolerated the procedure well, with the mean EPS of 2.56 for the 'high' and 1.73 for the 'low' APS group (p < 0.05). CONCLUSIONS: The actual pain experienced by a patient is significantly lower than their anticipation of vasectomy pain, which will aid clinicians in appropriately counseling patients and minimizing pre-procedural anxiety.

Alkaline polymer electrolyte membranes for fuel cell applications.

In this review, we examine the most recent progress and research trends in the area of alkaline polymer electrolyte membrane (PEM) development in terms of material selection, synthesis, characterization, and theoretical approach, as well as their fabrication into alkaline PEM-based membrane electrode assemblies (MEAs) and the corresponding performance/durability in alkaline polymer electrolyte membrane fuel cells (PEMFCs). Respective advantages and challenges are also reviewed. To overcome challenges hindering alkaline PEM technology advancement and commercialization, several research directions are then proposed.

Antibody responses in blood and saliva post COVID-19 bivalent booster do not reveal an Omicron BA.4/BA.5- specific response.

Introduction: Current SARS-CoV-2 strains continue to mutate and attempt to evade the antibody response elicited by previous exposures and vaccinations. In September of 2022, the first updated SARS-CoV-2 vaccines, designed to create immune responses specific for the variants circulating in 2022, were approved. These new vaccines, known commonly as the bivalent boost(er), include mRNA that encodes both the original Wuhan-Hu-1 spike protein as well as the spike protein specific to the Omicron BA.4 and BA.5 variants. Methods: We recruited volunteers from University of Massachusetts student, faculty and staff members to provide samples of blood and saliva at four different time points, including pre-boost and three times post boost and analyzed samples for antibody production as well as neutralization of virus. Results: Our data provide a comprehensive analysis of the antibody response following a single dose of the bivalent boost over a 6-month period and support previous findings that the response induced after the bivalent boost does not create a strong BA.4/BA.5-specific antibody response. Conclusion: We found no evidence of a specific anti-BA.4/BA.5 response developing over time, including in a sub-population of individuals who become infected after a single dose of the bivalent booster. Additionally, we present data that support the use of saliva samples as a reliable alternative to blood for antibody detection against specific SARS-CoV-2 antigens.

Investigation into the role of phosphatidylserine in modifying the susceptibility of human lymphocytes to secretory phospholipase A(2) using cells deficient in the expression of scramblase.

Normal human lymphocytes resisted the hydrolytic action of secretory phospholipase A(2) but became susceptible to the enzyme following treatment with a calcium ionophore, ionomycin. To test the hypothesis that this susceptibility requires exposure of the anionic lipid phosphatidylserine on the external face of the cell membrane, experiments were repeated with a human Burkitt's lymphoma cell line (Raji cells). In contrast to normal lymphocytes or S49 mouse lymphoma cells, most of the Raji cells (83%) did not translocate phosphatidylserine to the cell surface upon treatment with ionomycin. Those few that did display exposed phosphatidylserine were hydrolyzed immediately upon addition of phospholipase A(2). Interestingly, the remaining cells were also completely susceptible to the enzyme but were hydrolyzed at a slower rate and after a latency of about 100s. In contradistinction to the defect in phosphatidylserine translocation, Raji cells did display other physical membrane changes upon ionomycin treatment that may be relevant to hydrolysis by phospholipase A(2). These changes were detected by merocyanine 540 and trimethylammonium diphenylhexatriene fluorescence and were common among normal lymphocytes, S49 cells, and Raji cells. The levels of these latter effects corresponded well with the relative rates of hydrolysis among the three cell lines. These results suggested that while phosphatidylserine enhances the rate of cell membrane hydrolysis by secretory phospholipase A(2), it is not an absolute requirement. Other physical properties such as membrane order contribute to the level of membrane susceptibility to the enzyme independent of phosphatidylserine.

Supporting student success on the practical nurse (PN) licensure exam: The Health Education Systems Incorporated (HESI) PN Exit Exam Study.

BACKGROUND: Practical Nurses or Vocational Nurses (PNs/VNs) complete a 12-month certificate program and must successfully complete the National Council Licensure Exam for Practical Nurses (NCLEX-PN) prior to practice. While the scope of their practice is more circumscribed than the Registered Nurse (RN), they contribute significantly to healthcare throughout the U.S.A. Current research to support their learning and success is needed. Elsevier's Health Education Systems Incorporated (HESI) PN Exit Exam (E2) has been used to determine students' NCLEX-PN readiness for over 20 years. Given regular updates to the NCLEX test plan, ongoing research is needed to assess the E2's continued predictive validity and examine E2 program policies to enhance PN/VN student success. OBJECTIVES: To examine the predictive validity of the E2 on NCLEX-PN first-time pass rates (NCLEX-PN FTPR) and determine which E2-related program policies are related to higher E2 scores and NCLEX-PN FTPR. METHOD: Sixteen PN/VN program directors participated in the study, providing E2 program policy information and NCLEX-FTPR outcomes for 1371 students who took the E2 between 2018 and 2021. RESULTS: Students passed the NCLEX-PN 96-98 % of the time when they scored 900 or higher on the E2 and passed 94 %-96 % of the time when they scored 850 or higher. Out of 16 programs, most required E2 test preparation (n = 13, 81.25 %) and allowed students more than one E2 attempt (n = 11; 68.75 %). Requiring minimum E2 scores and specific E2 test preparation were associated with higher NCLEX-PN pass rates, in each case mediated by higher E2 scores. Requiring remediation for students with low initial E2 scores led to higher E2 scores but not higher NCLEX-PN pass rates. CONCLUSIONS: This study provides evidence for program policies and educational resources to support students' NCLEX-PN success. Setting an expected performance benchmark and requiring PN students to engage in E2 preparation prior to their first attempt are recommended strategies emerging from this study.

Modulation of colonic immunometabolic responses during Clostridioides difficile infection ameliorates disease severity and inflammation.

Clostridioides difficile infection (CDI) is the leading cause of antibiotic-associated diarrhea, and its clinical symptoms can span from asymptomatic colonization to pseudomembranous colitis and even death. The current standard of care for CDI is antibiotic treatment to achieve bacterial clearance; however, 15 to 35% of patients experience recurrence after initial response to antibiotics. We have conducted a comprehensive, global colonic transcriptomics analysis of a 10-day study in mice to provide new insights on the local host response during CDI and identify novel host metabolic mechanisms with therapeutic potential. The analysis indicates major alterations of colonic gene expression kinetics at the acute infection stage, that are restored during the recovery phase. At the metabolic level, we observe a biphasic response pattern characterized by upregulated glycolytic metabolism during the peak of inflammation, while mitochondrial metabolism predominates during the recovery/healing stage. Inhibition of glycolysis via 2-Deoxy-D-glucose (2-DG) administration during CDI decreases disease severity, protects from mortality, and ameliorates colitis in vivo. Additionally, 2-DG also protects intestinal epithelial cells from C. difficile toxin damage, preventing loss of barrier integrity and secretion of proinflammatory mediators. These data postulate the pharmacological targeting of host immunometabolic pathways as novel treatment modalities for CDI.

The impacts of learning analytics and A/B testing research: a case study in differential scientometrics.

BACKGROUND: In recent years, research on online learning platforms has exploded in quantity. More and more researchers are using these platforms to conduct A/B tests on the impact of different designs, and multiple scientific communities have emerged around studying the big data becoming available from these platforms. However, it is not yet fully understood how each type of research influences future scientific discourse within the broader field. To address this gap, this paper presents the first scientometric study on how researchers build on the contributions of these two types of online learning platform research (particularly in STEM education). We selected a pair of papers (one using A/B testing, the other conducting learning analytics (LA), on platform data of an online STEM education platform), published in the same year, by the same research group, at the same conference. We then analyzed each of the papers that cited these two papers, coding from the paper text (with inter-rater reliability checks) the reason for each citation made. RESULTS: After statistically comparing the frequency of each category of citation between papers, we found that the A/B test paper was self-cited more and that citing papers built on its work directly more frequently, whereas the LA paper was more often cited without discussion. CONCLUSIONS: Hence, the A/B test paper appeared to have had a larger impact on future work than the learning analytics (LA) paper, even though the LA paper had a higher count of total citations with a lower degree of self-citation. This paper also established a novel method for understanding how different types of research make different contributions in learning analytics, and the broader online learning research space of STEM education.

Splenogonadal fusion presenting as a testicular mass, a case report and literature review.

Splenogonadal fusion is a rare, frequently misdiagnosed, congenital anomaly in which the splenic tissue is abnormally attached to the gonadal or mesonephric remnants. It is commonly found as an incidental finding at autopsy, during orchiopexy or hernia repair. However, it can present as a testicular mass or as an acute scrotal pathology such as testicular torsion or epididymoorchitis. It poses as a diagnostic challenge preoperatively and often leads to unnecessary orchiectomy. We present a case of a 15-year-old male who presented with a long-standing left testicular mass thought to be a testicular tumor. Resection of the lesion along with partial left orchiectomy was done and histopathologic evaluation revealed splenogonadal fusion.

Computational modeling of complex bioenergetic mechanisms that modulate CD4+ T cell effector and regulatory functions.

We built a computational model of complex mechanisms at the intersection of immunity and metabolism that regulate CD4+ T cell effector and regulatory functions by using coupled ordinary differential equations. The model provides an improved understanding of how CD4+ T cells are shaping the immune response during Clostridioides difficile infection (CDI), and how they may be targeted pharmacologically to produce a more robust regulatory (Treg) response, which is associated with improved disease outcomes during CDI and other diseases. LANCL2 activation during CDI decreased the effector response, increased regulatory response, and elicited metabolic changes that favored Treg. Interestingly, LANCL2 activation provided greater immune and metabolic modulation compared to the addition of exogenous IL-2. Additionally, we identified gluconeogenesis via PEPCK-M as potentially responsible for increased immunosuppressive behavior in Treg cells. The model can perturb immune signaling and metabolism within a CD4+ T cell and obtain clinically relevant outcomes that help identify novel drug targets for infectious, autoimmune, metabolic, and neurodegenerative diseases.

NCLEX-RN readiness: HESI Exit Exam validity and nursing program policies.

BACKGROUND: The HESI® Exit Exam (E2) has been used to assess student readiness for the NCLEX-RN® examination for over two decades. PURPOSE: In this study, we examined the relationship between students' average E2 scores and their first time NCLEX-RN pass or fail outcomes to understand the validity of the E2 as a predictor of NCLEX-RN readiness. In addition, we examined the relationship between policies governing E2 use in programs and success on the NCLEX-RN. METHODS: Over 40 programs across the United States participated in the study, providing NCLEX-RN outcome information for 3686 students that took the E2 in 2018-2019 and information about policies. RESULTS: Students who achieved an average E2 score of 850 and above had a NCLEX-RN first-time pass rate (FTPR) of 96.33%. The NCLEX-RN FTPR was 97.29% for students whose average E2 score was 900 and above. Policies such as test preparation requirements and remediation for the E2 were associated with statistically significantly higher student NCLEX-RN FTPR. CONCLUSION: This study builds upon past evidence about the validity of the E2 and extends research in new directions by demonstrating the role of specific E2 policies in supporting students' success on the NCLEX-RN.

Neutralizing Antibodies and Cytokines in Breast Milk After Coronavirus Disease 2019 (COVID-19) mRNA Vaccination.

OBJECTIVE: To evaluate immune responses to coronavirus disease 2019 (COVID-19) mRNA-based vaccines present in breast milk and transfer of the immune responses to breastfeeding infants. METHODS: We enrolled 30 lactating women who received mRNA-based COVID-19 vaccines from January through April 2021 in this cohort study. Women provided serial milk samples, including milk expressed before vaccination, across 2-3 weeks after the first dose, and across 3 weeks after the second dose. Women provided their blood, spotted on cards (dried blood spots), 19 days after the first dose and 21 days after the second dose. Stool samples from the breastfed infants were collected 21 days after mothers' second vaccination. Prepandemic samples of milk, dried blood spots, and infant stool were used as controls. Milk, dried blood spots, and infant stool were tested by enzyme-linked immunosorbent assay for receptor-binding domain (RBD)-specific immunoglobulin (Ig)A and IgG. Milk samples were tested for the presence of neutralizing antibodies against the spike and four variants of concern: D614G, Alpha (B.1.1.7), Beta (B.1.351), and Gamma (P.1). Levels of 10 cytokines were measured in milk samples. RESULTS: Milk from COVID-19-immunized women neutralized the spike and four variants of concern, primarily driven by anti-RBD IgG. The immune response in milk also included significant elevation of interferon-γ. The immune response to maternal vaccination was reflected in breastfed infants: anti-RBD IgG and anti-RBD IgA were detected in 33% and 30% of infant stool samples, respectively. Levels of anti-RBD antibodies in infant stool correlated with maternal vaccine side effects. Median antibody levels against RBD were below the positive cutoffs in prepandemic milk and infant stool samples. CONCLUSION: Humoral and cellular immune responses to mRNA-based COVID-19 vaccination are present in most women's breast milk. The milk anti-RBD antibodies can neutralize severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike and variants of concern. Anti-RBD antibodies are transferred to breastfed infants, with the potential to confer passive immunity against SARS-CoV-2.

Durable antibody and effector memory T cell responses in breastmilk from women with SARS-CoV-2.

Background: Given that only 25% of pregnant women elect to receive a COVID-19 vaccine, maternal SARS-CoV-2 infection remains an important route of conferring protective passive immunity to breastfed infants of mothers who are not vaccinated. Methods: We enrolled 30 lactating participants between December 2020 and March 2021 who had a positive PCR-test and their first COVID-19 symptoms within the previous 21 days. Participants were asked to provide serial bilateral milk samples at 12 timepoints (~ every 3 days) over a period of 35 days. A second set of samples was collected at least four months after the beginning of the first set. Participants also were asked to provide their dried blood spots and infant stool samples. All samples were tested for receptor-binding domain (RBD)-specific immunoglobulin (Ig)A, IgG, and IgM. Milk samples were assessed for neutralizing ability against the spike protein and four SARS-CoV-2 variants: D614G, Alpha (B.1.1.7), Beta (B.1.351), and Gamma (P.1). Permeability of the breast epithelium was assessed by measuring the sodium to potassium ions (Na:K) in milk. Using flow cytometry, memory CD4 and CD8 T cells (CD45RO+ and CCR7+/-) and mucosal-homing CD4 and CD8 T cells (CD103+) were determined in cells from milk expressed at 35 days and at least 4 months after their first milk donation. Results: Milk antibodies from SARS-CoV-2 positive participants neutralized the spike complex. Milk from 73, 90, and 53% of participants had binding reactivities to RBD-specific IgA, IgG, and IgM, respectively. In contrast to blood spots, which showed increased levels of IgG, but not IgA or IgM, the COVID-19 response in milk was associated with a robust IgA response. Twenty-seven percent of participants had increased breast-epithelium permeability, as indicated by Na:K ≥ 0.6. The percentage of CD45RO+CCR7- effector-memory T cells in the day ≥120 milk samples was significantly higher than day 35 samples (P< 0.05). Conclusions: Antibodies in milk from participants with recent SARS-CoV-2 infection and those who recovered can neutralize the spike complex. For the first time we show that breastmilk T cells are enriched for mucosal memory T cells, further emphasizing the passive protection against SARS-CoV-2 conferred to infants via breastmilk.