Institute Grant Research Articles

Adapting Legal Research for a First-Generation Audience

One segment of our diverse law school population are students who are the first in their family to attend college, and by extension, law school. What can we as librarians do to advance a more inclusive classroom experience for our first-generation students? Washburn University, as part of a Title III Strengthening Institutions Grant, provides a summer program for faculty to enhance inclusive pedagogy and curriculum related to firstgeneration students. The author was a part of the program in 2022, and this paper will address the redesign of a legal research course based on the training from the program. The discussion will include an overview of the University provided training for the redesign, the concepts utilized in the class reboot geared toward fostering inclusion for first-generation students, and an extensive analysis of the final product.

Combination drug screen targeting glioblastoma core vulnerabilities reveals pharmacological synergisms

Pharmacological synergisms are an attractive anticancer strategy. However, with more than 5000 approved-drugs and compounds in clinical development, identifying synergistic treatments represents a major challenge. High-throughput screening was combined with target deconvolution and functional genomics to reveal targetable vulnerabilities in glioblastoma. The role of the top gene hit was investigated by RNA interference, transcriptomics and immunohistochemistry in glioblastoma patient samples. Drug combination screen using a custom-made library of 88 compounds in association with six inhibitors of the identified glioblastoma vulnerabilities was performed to unveil pharmacological synergisms. Glioblastoma 3D spheroid, organotypic exvivo and syngeneic orthotopic mouse models were used to validate synergistic treatments. Nine targetable vulnerabilities were identified in glioblastoma and the top gene hit RRM1 was validated as an independent prognostic factor. The associations of CHK1/MEK and AURKA/BET inhibitors were identified as the most potent amongst 528 tested pairwise drug combinations and their efficacy was validated in 3D spheroid models. The high synergism of AURKA/BET dual inhibition was confirmed in exvivo and invivo glioblastoma models, without detectable toxicity. Our work provides strong pre-clinical evidence of the efficacy of AURKA/BET inhibitor combination in glioblastoma and opens new therapeutic avenues for this unmet medical need. Besides, we established the proof-of-concept of a stepwise approach aiming at exploiting drug poly-pharmacology to unveil druggable cancer vulnerabilities and to fast-track the identification of synergistic combinations against refractory cancers. This study was funded by institutional grants and charities.

The utility of transcriptomics in the conservation of sensitive and economically important species

The utility of transcriptomics in the conservation of sensitive and economically important species

P-642 Estrogen deprivation and related risks in primary ovarian insufficiency with various hormone therapy initiation time points - evaluating 122,785 Asian women

Abstract Study question Does the initiation time point and/or duration of hormone therapy affect the long-term health consequences in primary ovarian insufficiency (POI)? Summary answer POI women with immediate hormone therapy following their diagnosis showed significantly reduced risk of metabolic syndrome and dyslipidemia, and such effect magnified with aging. What is known already POI women are exposed to extended duration of estrogen deficiency and premature aging, consequently associated with higher risks of cardiovascular, metabolic, osteoporotic and neurodegenerative diseases. The first line of treatment is hormone therapy (HT), preferably until the age of natural menopause. “Timing hypothesis” in terms of hormone replacement therapy (HRT) in menopause has globally changed the treatment protocol, but its evidence in POI is scarce. Study design, size, duration This population-based retrospective cohort study included 122,785 women registered to the National Health Insurance in South Korea from 2009 to 2015. The study group with POI diagnosis contained 24,557 women, determined by the International Classification of Disease, tenth revision (ICD-10) code E283 in the database. Women aged more than 40 years with the E283 code were considered being previously-diagnosed POI. The 1:4 age-matching control group with the systematic random-sampling method included 98,228 women. Participants/materials, setting, methods The participants’ demographic data, diagnosis codes, use of inpatient and outpatient services, pharmacy dispensing claims and mortality data were retrieved and statistically analyzed. According to their use of HT, the study group was further divided into two groups: with (n = 13,240) or without HT (n = 11,317); their routine health check-up data collected in the database were then compared. In all analysis, a p-value <0.05 was considered statistically significant. Main results and the role of chance Compared to the control group, POI women had significantly lower body mass index (BMI), blood pressure, serum total cholesterol and triglyceride (TG), but the incidence rate ratio (IRR) was significantly higher in POI regarding thyroid diseases, type 2 diabetes, dyslipidemia, hypertension, osteoporosis and cardiovascular diseases (IRR with 95% confidence interval (CI) of 1.731[1.691; 1.772], 1.300[1.260; 1.342], 1.499[1.470; 1.529], 1.100[1.071; 1.129], 2.449[2.388;2.513] and 1.468[1.392; 1.547], respectively). Among POI women overall, the HT users had significantly lower BMI, lower systolic blood pressure but similar diastolic blood pressure, and lower serum total cholesterol and TG, compared to the non-users. In terms of IRRs for chronic diseases, the HT users under the age of 40 years did not show any statistically significant difference in comparison to the non-users; however, the HT users aged more than 40 years had significantly lower IRRs for type 2 diabetes, dyslipidemia and hypertension compared to the non-users (0.876[0.822; 0.933], 0.873[0.840; 0.907], 0.849[0.805; 0.896], respectively). Such effect became more statistically vivid when the participants grew older (IRR[95% CI] of 0.849[0.749; 0.963] for thyroid diseases; 0.806[0.708; 0.917] for type 2 diabetes; 0.652[0.592; 0.718] for dyslipidemia; 0.739[0.662; 0.825] for hypertension; 0.835[0.752; 0.928] for osteoporosis; and 0.890[0.728; 1.089] for cardiovascular disease, respectively). Limitations, reasons for caution The current study relied on the health insurance claim data which did not include detailed patient medical record including menstruation pattern and obstetrical/gynecological histories. Also, critical biochemical assay results such as anti-mullerian hormone or pituitary hormones were not available because they were not covered by the national health insurance program. Wider implications of the findings Agreeing with the previous literature, the immediate use of HT is associated with the reduced incidence of chronic diseases in POI women, and its extended use exerts more significant results with aging. If acknowledged early and initiated with HT, previously-inevitable health risks with POI could be conceivably compensated. Trial registration number The current study was supported by Biomedical Research Institute Grant (#202201970001), Pusan National University Hospital.

Association between left ventricular strain and fibrosis in severe aortic regurgitation with preserved ejection fraction: a magnetic resonance and histology study

Abstract Funding Acknowledgements Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Institutional Grant Na Homolce Hospital Background The presence and extent of myocardial fibrosis are associated with poor outcomes in patients with severe aortic regurgitation. In the present study, we tested a correlation between left ventricular (LV) strain derived by using feature-tracking cardiac magnetic resonance (MRI) and diffuse myocardial fibrosis percentage (DMF%) at quantitative histology. Methods A study population included 23 patients (age 51 ± 12 years, 78% males) who underwent perioperative myocardial biopsy during aortic valve surgery for severe aortic regurgitation and who had a satisfactory pre-surgery MRI. Myocardial specimens obtained with a deep needle technique from the basal interventricular septum were stained with picrosirius red, digitalized, and analyzed using Image J software by a pathologist blinded to imaging findings. The DMF% was calculated as a ratio of the total collagen content to the total area. MRI-derived global longitudinal-(GLS), circumferential- (GCS), and radial (GRS) strain, as well as their ratio to heart rate and systolic blood pressure, were assessed using a feature-tracking technique by a certified imager blinded to histological analysis. Results Compared to normal values, all patients showed increased DMF% (19 ± 9%) and decreased GLS (15 ± 2%) despite preserved LV ejection fraction (64 ± 7%). Out of all the imaging parameters, only GLS/heart rate ratio, GCS/heart rate ratio and extracellular volume fraction showed a significant correlation with DMF% (Table 1, Figure 1). In contrast, GLS and GCS only tended to correlate with DMF%. The remaining parameters including GRS did not show a significant association. Patients with focal midwall scar in late gadolinium enhancement imaging (n=8, 35%), which was mainly localized in the basal septum, had similar GLS as than patients without focal scar (14.5±2.3 vs 15.3±2.3, p = 0.768). Conclusions Feature-tracking-derived ratios of LV GLS and GCS over heart rate are the most promising indices for non-invasive assessment of diffuse myocardial fibrosis in severe aortic regurgitation with preserved ejection fraction. Figure 1 Legend Correlation of a GLS/HR ratio 1A) and GLS 1B) with diffuse myocardial fibrosis %. Myocardial histology stained with Picrosirius red (collagen fibers are in red).

Hypersensitivity Reactions to Native E. coli L-asparaginase in Children With Acute Lymphoblastic Leukemia Treated in Trial ALL-BFM 2000: Impact of Treatment Schedule and Type of Glucocorticoid in Induction.

Hypersensitivity Reactions to Native E. coli L-asparaginase in Children With Acute Lymphoblastic Leukemia Treated in Trial ALL-BFM 2000: Impact of Treatment Schedule and Type of Glucocorticoid in Induction.

0157 Physiological responses to acute psychosocial stress in adolescents with insomnia

Abstract Introduction Insomnia often develops during adolescence, with girls being at greater risk than boys. Alterations in response to psychosocial stress have been linked to the etiology of the disorder, but it remains unclear whether the reactivity of the major stress systems (hypothalamic-pituitary-adrenal (HPA) axis and autonomic nervous system (ANS)) in response to stress is altered in adolescents with insomnia. Methods Forty-seven post-pubertal adolescents (age range: 16-20 years, 28 female) with (N=16; insomnia group) and without (N=31; control group) insomnia symptomatology underwent two randomized nights of polysomnographic (PSG) laboratory recordings. Participants underwent an experimental pre-bedtime stress anticipation protocol (stress night) and a control night during which they engaged in neutral activities before bedtime. The morning after the stress night, participants underwent the Trier Social Stress Test (TSST), a standardized protocol that requires participants to complete verbal and arithmetic tasks in front of observers. On both nights, heart rate (HR) was measured across the night and cortisol was collected upon awakening and 30 minutes later to calculate cortisol awakening responses (CAR). Stress perception, salivary cortisol (HPA activity), and HR were measured during the TSST. Results There were no differences in nocturnal HR recovery or in the CAR on the stress night compared with the control night in either group. Morning pre-TSST stress levels did not differ between groups. Both groups exhibited significant increases in HR in response to the TSST (p< 0.05). HR was higher in girls compared to boys in the control group at baseline (p< 0.05). Potential group differences in HPA responses to the TSST were inconclusive due to the apparent confounding effect of the CAR on cortisol responses to stress: many participants did not show a cortisol response to the task and larger CARs were associated with an attenuated cortisol response. Conclusion There were no significant differences in HR responses to anticipation of experimental stress during the night or in response to the morning TSST between adolescents with and without insomnia. Further work is needed to examine whether altered responses to stress may emerge if insomnia becomes chronic. Support (if any) This study was supported by the National Heart, Lung and Blood Institute (NHLBI) grant R01HL139652(MdZ).

0306 Circadian Disrupting Light Exposure within Pediatric Hospital Rooms

Abstract Introduction Pediatric patients undergoing stem cell transplant (SCT) often experience extended hospitalizations. Inappropriate lighting during admissions is known to disrupt sleep and circadian health, which can result in worse health outcomes. This study aimed to describe patterns of light across 24-hour periods in two inpatient rooms of children undergoing SCT. Methods Wall-mounted light meters (Extech SDL400) continuously tracked noise and light levels at one-minute intervals in two patient rooms over six months. We determined the percent of time that patients were exposed to non-optimal light levels (< 448 lux during the daytime, >18 lux in the evening, >2 lux at night); thresholds were based on converting recent consensus-based melanopic equivalent daylight illuminance guidelines to vertically measured illuminance thresholds. We assessed the frequency that light exceeded thresholds associated with night wakings (>150 lux). Wilcoxon rank-sum tests compared light levels between the two rooms. Room A had a single northwest-facing window overlooking a narrow courtyard. Room B had an unobstructed window facing north. Results On average, hospitalized pediatric SCT patients spent 50% of their time in inappropriate light conditions. They were exposed to light spikes >150 lux on 2% of nights, with overnight light peaking at 513 lux. When comparing average daytime and nighttime light levels between the two rooms, Room A was significantly dimmer than Room B, p <.001. In the dimmer room, light levels never reached the recommended threshold during the daytime, and the light was too bright for 19% of the evening and 17% of the nighttime. In the brighter room, light levels were too dim during 97% of the daytime and too bright during 59% of the evening and 30% of the nighttime. Conclusion During the day, pediatric SCT patients are rarely exposed to light bright enough to preserve a healthy circadian rhythm. Hospitals have an opportunity to significantly improve the sleep and circadian health for children already at elevated risk for health morbidities during transplant. Support (if any) This research was generously supported by grant funding from Pedals for Pediatrics (PI: Zhou) and an institutional research training grant (T32DK063929).

Medication use by insurance coverage in subjects with coronary heart disease in the Southern Cone of Latin America

Abstract Funding Acknowledgements Type of funding sources: Other. Main funding source(s): This work was supported by the National Heart, Lung, and Blood Institute (NHLBI) grant number HHSN268200900029C. Background Clinical guidelines recommend cardioprotective medication use in coronary heart disease (CHD) patients. Underuse of these drugs is common among patients of low resources, which may be explained by insufficient access in the public sector. Aim We aim to assess medication use and barriers to care by insurance coverage in subjects with CHD in the Southern Cone of Latin America. Methods We analysed cross-sectional data from 593 participants with CHD residing in Argentina, Chile and Uruguay, within the CESCAS community-based cohort study. Participants were categorized as covered by public insurance only or having additional coverage (social security or private insurance). We calculated the prevalence of recommended medications use, mean number of medications, use of ≥ 1 and ≥ 2 drugs, and reported barriers to needed care in insured and uninsured participants. Differences between coverage groups were assessed with univariable and multivariable analysis (logistic and Poisson regression) adjusted by age, sex, previous revascularization, educational level and barriers. Results Medication use ranged from 39.8% (lipid-lowering) to 84.1% (antihypertensives). Mean number of medications was 1.8 (SE 0.1), 95.4% used ≥ 1 drugs, and 59.8% used ≥ 2 drugs. There were no significant differences by coverage in medication use, except for higher ACE-inhibitor and nitrate use among participants in the public sector (35.8% vs. 53.2%, p=0.001; and 1.5% vs. 5.2%, p=0.047 respectively). In the multivariable analysis, having public coverage only was not significantly associated with medication use, but it was with reporting barriers to care (OR 2.06, 95% CI 1.21-3.52). Conclusion Insurance coverage was not associated with the use of most medications in CHD patients in the Southern Cone of Latin America, possibly due to public health programs providing them free of charge. Uninsured participants more often face barriers to access medical care, and future studies should address these healthcare inequalities.

Risk factor clustering in men and women with CHD in the Southern Cone of Latin America

Abstract Funding Acknowledgements Type of funding sources: Other. Main funding source(s): This work was supported by the National Heart, Lung, and Blood Institute (NHLBI) grant number HHSN268200900029C. Background Presence of multiple risk factors (RF) increases the risk for cardiovascular morbidity and mortality, and this is especially important in patients with coronary heart disease (CHD). Purpose The current study investigates sex differences in the presence of multiple cardiovascular RF in subjects with established CHD in the southern Cone of Latin America. Methods We analyzed cross-sectional data from the 634 participants aged 35-74 with CHD from the community-based CESCAS Study. We calculated the prevalence for counts of cardiometabolic (hypertension, dyslipidemia, obesity, diabetes) and lifestyle (current smoking, unhealthy diet, low physical activity, excessive alcohol consumption) RF. Differences in RF number between men and women were tested with age-adjusted Poisson regression. We identified the most common RF combinations among participants with ≥ 4 RF. We performed a subgroup analysis by educational level. Results The prevalence of cardiometabolic RF ranged from 76.3% (hypertension) to 26.8% (diabetes), and the prevalence of lifestyle RF from 81.9% (unhealthy diet) to 4.3% (excessive alcohol consumption). Obesity, central obesity, diabetes and low physical activity were more common in women, while excessive alcohol consumption and unhealthy diet were more common in men. Close to 85% of women and 81.5% of men presented with ≥ 4 RF. Women presented with a higher number of overall (relative risk (RR) 1.05, 95% CI 1.02-1.08) and cardiometabolic RF (1.17, 1.09-1.25). These sex differences were found in participants with primary education (RR women overall RF 1.08, 1.00-1.15, cardiometabolic RF 1.23, 1.09-1.39), but were diluted in those with higher educational attainment. The most common RF combination was hypertension/dyslipidemia/obesity/unhealthy diet. Conclusion Women in the CESCAS study showed a higher burden of multiple cardiovascular RF. Sex differences persisted in participants with low educational attainment, and women with low educational level had the highest RF burden.

Next Steps Toward Improving the Quality, Relevance, and Implementation of Chronic Limb-Threatening Ischemia Research Through Stakeholder Engagement

Stakeholder-engaged research (SER), in which patients and other stakeholders (including academics) engage as research, has been shown to improve the quality, feasibility, relevance, and implementation of research. There is scant SER in vascular surgery. Attitudes toward SER have not been assessed among academic vascular surgery stakeholders (clinicians and clinical researchers). Therefore, it is unclear how to increase SER to improve the care and outcomes of patients with vascular disease. As part of a Patient-Centered Outcomes Research Institute grant to increase stakeholder-centricity and sustainable engagement in chronic limb-threatening ischemia (CLTI) research, we developed an anonymous online survey based off qualitative interviews with 10 academic stakeholders. Survey responses are presented descriptively, grouped by prior SER experience. There were 42 responses to the survey for analysis. Of these, 19 had prior SER experience and 23 did not. A total of 89% of experienced respondents (ER) were moderately or extremely familiar with the term SER vs 35% of inexperienced respondents (IR). In general, ER and IR agreed about the benefits of SER, including improved relevance of research questions (80% ER, 79% IR) and selection of more relevant outcome measures (67% and 63%, respectively; Table I). However, ER more commonly reported that SER was the ethically/morally correct thing to do (60% vs 32%). There were noticeable differences in experienced/perceived drawbacks to SER, particularly in relation to the research design process (reported to be slower by 77% of ER vs 36% of IR) and the scientific interest of SER (reported to be less interesting by 8% of ER vs 29% of IR; Table I). Respondents agreed that lack of awareness was the largest barrier to increasing CLTI-related SER. However, IR identified some additional significant barriers (Table II). ER thought stakeholder training materials would be the most helpful resource to increase CLTI-related SER (73% vs 50%), whereas IR preferred assistance identifying and coordinating stakeholders (55% vs 71%; Table II). ER were more likely to feel that it was very or extremely important to partner with stakeholders in future research design/conduct (72% vs 22%). Researchers experienced with engaging stakeholders balanced a belief in its benefits with a realistic understanding of its associated difficulties. Some inexperienced researchers were strongly interested in future SER but highlighted several barriers to entry. Future efforts to increase SER in CLTI should focus on raising awareness of its benefits and the process, while lowering barriers to entry and assisting experienced researchers with well-defined pain points.Table IExperienced and perceived benefits and drawbacks to stakeholder-engaged research (SER)SER-experienced respondentsSER-inexperienced respondentsBenefits associated with SERN = 15N = 19 More relevant questions8079 More relevant outcomes6763 More ethical6032 Researcher-patient relationships5347 More relevant intervention5358 Follow-up feasibility4737 Recruitment and retention4047 Researcher satisfaction3316 Easier implementation2026 Easier funding1321 Easier publication75Drawbacks associated with SERN = 13N = 14 Slower design7736 Slower conduct4636 More expensive2336 Harder to get funding1521 Less interesting829 Harder to publish814 Less meaningful80 Less satisfying00Data are presented as percentage. Open table in a new tab Table IIBarriers and facilitators to stakeholder-engaged research (SER)SER-experienced respondentsSER-inexperienced respondentsBarriers to SER Awareness6967 Methodologic expertise3840 Interest3833 Access to stakeholders3153 Stakeholder knowledge3153 Stakeholder selection criteria2340 Stakeholder health status2327 Lack of dissemination opportunities150 Lack of regulatory mandate1520 Lack of funding820 Logistical difficulties820Potential facilitators for SER Stakeholder training materials7350 Stakeholder identification and coordination5571 Methodology collaborations4536 Funding aggregation2764 Methodologic training materials2736 Aggregation of regulatory guidance2714 Aggregation of dissemination opportunities914Data are presented as percentage. Open table in a new tab

No time for complacency on COVID-19 in Europe

No time for complacency on COVID-19 in Europe

In Response.

Department of Anesthesiology, Center of Anesthesiology and Intensive Care Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany, [email protected] Department of Anesthesiology, Center of Anesthesiology and Intensive Care Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany, Outcomes Research Consortium, Cleveland, Ohio Department of Anesthesiology, Center of Anesthesiology and Intensive Care Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany Accepted for publication July 26, 2022. Conflicts of Interest: B.S. has received honoraria for consulting, honoraria for giving lectures, and refunds of travel expenses from Edwards Lifesciences (Irvine, CA). B.S. has received honoraria for consulting, institutional restricted research grants, honoraria for giving lectures, and refunds of travel expenses from Pulsion Medical Systems (Feldkirchen, Germany). B.S. has received institutional restricted research grants, honoraria for giving lectures, and refunds of travel expenses from CNSystems Medizintechnik (Graz, Austria). B.S. has received institutional restricted research grants from Retia Medical (Valhalla, NY). B.S. has received honoraria for giving lectures from Philips Medizin Systeme Böblingen (Böblingen, Germany). B.S. has received honoraria for consulting, institutional restricted research grants, and refunds of travel expenses from Tensys Medical (San Diego, CA).

The perinatal outcomes of women treated for Asherman syndrome: a propensity score-matched cohort study.

Do the perinatal outcomes of patients following hysteroscopic treatment for Asherman syndrome (AS) differ from that of a control population? Perinatal complications including placental issues, high blood loss, and prematurity in women after treatment for AS should be considered as moderate to high risk, especially in patients who have undergone more than one hysteroscopy (HS) or repeated postpartum instrumental revisions of the uterine cavity (Dilation and Curettage; D&C). The detrimental impact of AS on obstetrics outcomes is commonly recognized. However, prospective studies evaluating perinatal/neonatal outcomes in women with AS history are sparse, and the characteristics accounting for the respective morbidity of AS patients remain to be elucidated. We conducted a prospective cohort study utilizing data from patients who underwent HS treatment for moderate to severe AS in a single tertiary University-affiliated hospital (enrolled between 01 January 2009 and March 2021), and who consequently conceived and progressed to at least 22nd gestational week of pregnancy. Perinatal outcomes were compared to a control population without an AS history, retrospectively enrolled concomitantly at the time of delivery for each patient with AS. Maternal and neonatal morbidity was assessed as well as the characteristics-related risk factors of AS patients. Our analytic cohort included a total of 198 patients, 66 prospectively enrolled patients with moderate to severe AS and 132 controls. We used multivariable logistic regression to calculate a propensity score to match 1-1 women with and without AS history based on demographic and clinical factors. After matching, 60 pairs of patients were analysed. Chi-square test was used to compare perinatal outcomes between the pairs. Spearman's correlation analysis was utilized to investigate the correlation between perinatal/neonatal morbidity and the characteristics-related factors of AS patients. The odds ratio (OR) for the associations was calculated by logistic regression. Among the 60 propensity matched pairs, the AS group more frequently experienced overall perinatal morbidity, including abnormally invasive placenta (41.7% vs 0%; P < 0.001), retained placenta requiring manual or surgical removal (46.7% vs 6.7%; P < 0.001), and peripartum haemorrhage occurrence (31.7% vs 3.3%; P < 0.001). Premature delivery (<37 gestational weeks) was reported more frequently also for patients with AS (28.3% vs 5.0%; P < 0.001). However, no increased frequency of intra-uterine growth restriction or worsened neonatal outcomes were observed in AS group. Univariable analysis of risk factors for AS group morbidity outcomes revealed that the main factor related to abnormally invasive placenta was two or more HS procedures (OR 11.0; 95% CI: 1.33-91.23), followed by two or more D&Cs preceding AS treatment (OR 5.11; 95% CI: 1.69-15.45), and D&C performed postpartum as compared to post abortion (OR 3.0; 95% CI: 1.03-8.71). Similarly, two or more HS procedures were observed as the most important factor for retained placenta (OR 13.75; 95% CI: 1.66-114.14), followed by two or more preceding D&Cs (OR 5.16; 95% CI: 1.67-15.9). Premature birth was significantly associated with the number of preceding D&Cs (OR for two or more, 4.29; 95% CI: 1.12-14.91). Although the cohort of patients with AS was enrolled prospectively, a baseline imbalance was intrinsically involved in the retrospective enrolment of the control group. However, to reduce the risk of bias, confounding factors were adjusted for using propensity score matching. The limitation to the generalization of our reported results is the single institution design in which all patients were treated for AS in one tertiary medical centre. Within our search scope, our study represents one of the first and largest prospective studies of perinatal and neonatal outcomes in moderate to severe AS patients with a prospectively analysis of the risks factors of characteristics significantly influencing reported morbidities among patients with AS. The study was supported by the Charles University in Prague [UNCE 204065] and by the institutional grant of The General Faculty Hospital in Prague [00064165]. No competing interests were declared. N/A.

Robust T cell responses to the Pfizer/BioNTech vaccine compared to infection and evidence of attenuated CD8+ T cell responses due to COVID-19

Abstract The COVID-19 pandemic has resulted in rapid development of several novel vaccine platforms, including the mRNA-based Pfizer/BioNTech vaccine. As of October 2022, more than 3.8 billion Pfizer/BioNTech vaccines were administrated to people in 180 countries. Meanwhile, the total number of COVID-19 cases has reached 627 million globally. We recently developed a new antigen-specific T cell staining tool— ”spheromer”that is able to label T cells more efficiently and capture a broader swath of the repertoire than any other available platform. Using spheromers loaded with SARS-CoV-2 peptides, we applied a panel covers 49 epitopesspanning the entire genome to characterize the magnitude and durability of antigen-specific T cell responses at single-epitope resolution. Using very sensitive “spheromer” peptide-MHC multimer reagents, we analyzed healthy subjects receiving two doses of the Pfizer/BioNTech BNT162b2 vaccine. Vaccination resulted in very robust peripheral blood responses to predicted SARS-CoV-2 spike peptides bound to either class-I or class-II MHC alleles. These peaked at 9.8% and 7.5% for the dominant CD4+ (HLA-DRB1*15:01/S191) and CD8+ (HLA-A*02/S691) T cell epitopes. Interestingly, the antigen-specific CD4+ and CD8+ T cell responses were asynchronous, with the peak CD4+ T cell responses occurring one week post the second vaccination (boost), whereas CD8+ T cells didn’t peak until two weeks later. These responses were much higher than in COVID-19 patients. We also found that prior SARS-CoV-2 infectiondecreased the CD8+ T cell response post vaccination by 2.4 to 41.4-fold at peak, and that the remaining cells were less responsive, suggesting that the virus can profoundly damage an individuals’ CD8+ T cell response. 1. Howard Hughes Medical Institute and NIAID grant AI057229 to Mark.M.Davis; 2. Bill and Melinda Gates Foundation grant OPP1113682, Center for Human Systems Immunology, to Mark.M.Davis; 3. U01 AI140498 to to Mark.M.Davis.

Impactos de las políticas gubernamentales e institucionales - de garantía de derechos - en las trayectorias formativas de los estudiantes de Turismo de la FHyCS – UnaM Impactos das políticas governamentais e institucionais - garantia de direitos - sobre as trajectórias educacionais dos estudantes de Turismo na FHyCS - UNaM

In the analysis of the multicausalities that have an impact on the high levels of university drop-out rates in the Tourism courses at the Faculty of Humanities and Social Sciences, it is considered important to address the impact of national and institutional programmes and grant projects that seek to mitigate this situation. In this way, the set of scholarships provided by the institution has been analysed, based on a quantitative study of data collection by means of surveys to admissions of the cohorts 2017 and 2018, which determined which have been the most demanded scholarships. Finally, we have described a programme - PATFEs - its scope and limitations, as a policy designed to address the complex process of transition between secondary and higher education, which has not been formally recognised by the teaching staff of Tourism degree courses.

Understanding NKT cells through sulfatide analogues

Abstract The goal of this research is to further investigate NKT cell activation using sulfatide analogues. NKT cells have been shown to play a significant role in mediating inflammatory immune responses and because of this, there has been strong interest in pharmacologically regulating this compartment to fight cancer. NKT cells recognize lipids that are loaded on CD1d molecules on the surface of cells, and it has been shown that structural modifications of these lipids directly affect NKT cell activation and polarization. Our work is focused on the impact of modifications of sulfatide, which is a glycolipid that can be loaded onto CD1d which is known to activate sulfatide-reactive (generally type II) NKT cells when presented by CD1d monomers on plastic. We have numerous sulfatide analogues with modifications (based on previous work with alpha-galactosylceramide analogues) on the acyl chain, sphingosine chain, and changes of beta-anomeric to alpha-anomeric. These analogues have been screened with NKT cell hybridomas, DN32 and XV19, to assess their effect on activation of type I and type II NKT cells, respectively. We observed that alterations in the sphingosine or acyl chain in sulfatides do not always mimic the effects on activity of similar changes in alpha-galactosylceramide analogues and that numerous sulfatide analogues can activate DN32 (not sulfatide-reactive) when presented on CD1d monomer or bone marrow-derived dendritic cells. Our results provide further insight into NKT cell activation and have identified sulfatide analogues that may be applicable to cancer therapy by activating type I NKT cells. NCI, NIH intramural research program (NIA-C-004020), and institutional grants of NIH (R01 GM111849)

Dependence of cardiomyocyte force, velocity, and power generation on ATP concentration

Theoretical analysis and computer modeling of myocardial energy metabolism under physiological and heart failure conditions predicts that cytoplasmic ATP and ADP levels both fall roughly in proportion to reductions in total adenine nucleotide pool observed in heart failure. Assessing sub-maximal force and velocity in loaded myocardial muscle at MgATP levels in the range of 2 to 8 mM, Beard et al. ( Biophys. J. 2022. 121: 3213–3223) observed effects of MgATP on myocardial power generation that suggest that although maximal force and velocity are not substantially sensitive to differences in [MgATP] that occur in the normal versus failing myocardium, maximal power generation, which occurs at submaximal force and velocity, is sensitive to [MgATP]. Previously developed computational models are not able to reproduce the observed dependence on MgATP. The goals of the present study are to construct and analyze a computational model of muscle dynamics to analyze data on the dependence of maximal force, velocity, and power generation on [MgATP] in order to determine a potential mechanism to explain the MgATP-dependent phenomena. Model development and identification was guided by dynamic measurements on permeabilized mouse trabecular fibers. The data include force-velocity relation and time constant of tension development. The mathematical model used to simulate cardiomyocyte crossbridge mechanics was based on the previous crossbridge and calcium models from Tewari et al. (J Mol Cell Cardiol, 2016. 96: 11-25) and Campbell et al. (Biophys J, 2018. 115: 543-553). The developed model includes strain- and metabolite-dependent transition between attachment states, and ATP-dependent destabilization of the super-relaxed (SRX) myosin state. Model-based analysis of force and velocity dyanmics is consistent with the hypothesis that ATP assocition with the myosin protein influence the kinetics and thermodynamics of transitions between the SRX and disorded (DRX) states. Funding: Department of Veterans Affairs Merit Review Award I01BX000740 (A.J.B.) and National Heart, Lung, and Blood Institute grant R01 HL154624 (A.J.B., D.A.B.) This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Assessment of donor compatibility in human solid organ transplantation and evaluation of allogeneic responses using a novel humanized mouse model.

Abstract The end result of solid organ transplantation is dictated by selection of a well-matched donor, leading to allograft survival. An allogeneic immune response is the main immunological barrier for successful organ transplantation. Donor and recipient human leukocyte antigen (HLA) mismatching results in poor graft acceptance, inflammation and rejection. The current evaluations for donor-recipient HLA incompatibility does not provide adequate information on the immunogenicity of individual HLA mismatches and impact of non-HLA-related alloantigens, especially in vivo. Here we demonstrate a novel method for analysis of alloimmune responsiveness between donor and recipient in vivoby introducing a humanized mouse model. Using molecular, cellular, and genomic analyses, we demonstrated that a recipient’s personalized humanized mouse provided the most sensitive assessment of allogeneic responsiveness to potential donors. In our study, HLA typing and mixed lymphocyte reaction (MLR) for assessment of allogeneic response was indistinguishable between 2 related donors of the recipient. Contrastingly in recipient’s humanized mouse model, the donor selected by HLA typing induced the strongest allogeneic response with increased infiltration of cytotoxic GzmB +CD8 +T cells. Moreover, the same donor induced upregulation of genes involved in the allograft rejection identified by transcriptomic analysis of graft infiltrating CD45 +cells. Humanized mouse model determined the lowest degree of recipient-donor alloimmune response, allowing for better selection of donor and minimized immunological risk of allograft rejection in transplantation. This approach can be used to evaluate alloresponses in cell based therapies as well. This study was supported by research funding from the Carlos and Marguerite Mason Trust, and by U.S. National Institutes of Health, National Cancer Institute Grant CA 172230

The clock gene Bmal1 inhibits kidney expression of SGLT2 and glucose reabsorption via the NR1d1/NRF1 pathway

Background &amp; Aims: The nuclear receptor subfamily 1, group D, member 1 (Nr1d1, also called Rev-erbα) is highly expressed in the kidney, and its transcriptional activity is increased by the clock gene Bmal1 (ARNTL or MOP3). Here we studied the roles and functional interactions of Bmal1 and Nr1d1 in mouse kidney, particularly in regulating the expression of the glucose transporter SGLT2 (SLC5A2) and glucose reabsorption. Methods: We performed histologic and biochemical analyses of kidney tissues from male C57BL/6 and Bmal1-knockout mice and performed gene expression analyses to identify genes regulated by Bmal1. The effects of fasting and refeeding on renal expression of Sglt2 was examined in male C57BL/6, Bmal1-knockout, and Nr1d1-knockout mice. Some mice were given hemin to stimulate Nr1d1 gene expression (daily for 1 week) at 12:00 with 100 μg/kg/day; renal glucose reabsorption, and glucose and fatty acid levels were measured. Renal proximal tubule epithelial cells (RPTECs) were generated from C57BL/6 and Bmal1-knockout mice and glucose uptake determined. Luciferase reporter assays were performed using HK2 cell. Results: We found that kidney SGLT2 mRNA and protein expression and glucose absorption were increased in Bmal1-knockout mice compared with C57BL/6 mice. Expression of SGLT2 was affected by fasting and refeeding in C57BL/6 mice, but not in Bmal1-knockout or Nr1d1-knockout mice. Bmal1-knockout mice also presented with higher levels of renal fatty acids compared with C57BL/6 mice. Hemin injection reduced expression of SGLT2, decreased glucose re-absorption in RPTECs, and increased levels of fatty acid in kidney; these changes were not observed in Bmal1-knockout mice. Nuclear respiratory factor 1 (NRF1), which regulates glucose, activated transcription of Sglt2. Fasting caused MAPK signing pathway to phosphorylate Bmal1, which activated NR1d1 and inhibited NRF1 activity and thereby reduced Sglt2 expression. Conclusions: The data indicate that fasting activates Bmal1 and Nr1d1, which inhibits NRF1 and leads to repression of renal SGLT2 expression and glucose reabsorption. This is the first study that mechanistically links food intake to the regulation of kidney glucose transport through a circadian clock gene. NIH National Heart, Lung, and Blood Institute grant R56 HL137912-01, and American Heart Association grant-in-aid 16GRNT30960027 to PX; NIH R01DK112042 to VV This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.