Study Time Points Research Articles

Maintaining glucose integrity ex-vivo: Impact of preanalytical specimen handling.

Adopting the WHO protocol for glucose analysis is arguably impractical in the routine clinical setting. Deviations may develop due to a lack of understanding regarding the impact of glycolysis on the accuracy of results. We sought to assess the stability of glucose in two different blood collection tubes (BCT), BD Vacutainer® FX 'Fl-Ox' and Greiner Vacuette® FC-Mix 'FC-Mix' stored at room temperature (RT:18-22°C) and 4°C over 8.5days. Each participant provided venous whole blood collected into 51 BCTs; 'Fl-Ox' (n = 26) and 'FC-Mix' (n = 25). One Fl-Ox sample from each participant was handled according to the WHO recommended method. The remaining BCTs were stored at 4°C/RT prior to analyses at designated study timepoints. Glucose was measured using the hexokinase assay on the Cobas® 8000 platform. Participants (n = 8, Male = 2) were aged 24-56years. Plasma glucose measured in FI-Ox BCTs according to the WHO sample-handling method had a median concentration of 5.73mmol/L (Range: 5.39-10.37mmol/L). Glucose decreased by greater than minimal difference (>0.26mmol/L) in blood collected into Fl-Ox and stored @4°C/RT within 24h of phlebotomy. FC-Mix BCT maintained glucose <0.26mmol/L @4°C over a period of 8.5days and up to 4days @RT when compared to the WHO recommended method. Glucose in FC-Mix BCT stored @4°C demonstrated the best agreement with results determined using the WHO specifications. When FC-Mix tubes were stored @RT, glucose was stable for 4days. These findings suggest that the FC-Mix BCT effectively inhibits glycolysis and should be introduced into routine clinical practice.

Telemedicine and HIV Care Quality Measures During the COVID-19 Pandemic.

During the COVID-19 pandemic, telemedicine was adopted to ensure continuity of HIV care. We examined how introducing televisits affected technical quality of care for people with HIV (PWH) during this time. PWH receiving HIV care at Howard Brown Health Centers and Northwestern University in Chicago, Illinois were included. HIV care quality indicators were calculated using data extracted from electronic medical records during 4 timepoints every 6 months from March, March 1, 2020 to September 1, 2021. Generalized linear mixed models estimated differences in indicators across timepoints within each site while controlling for multiple observations of individuals. Generalized linear mixed models were also used to compare differences in outcomes among PWH who attended all versus a combination of in-person and televisits versus no televisits across the study time periods. 6447 PWH were included in the analysis. Compared with prepandemic levels, there were significant declines in care utilization and processes of care measures. Measures of HIV virologic suppression, blood pressure control, and HbA1C <7% (in both people with and without diabetes) were stable with no significant differences noted across the study timepoints. Similar trends were observed across all age, race, and sex subgroups. In multivariable models, televisits were not associated with decreased HIV viral suppression. During the COVID-19 pandemic and rapid implementation of televisits, indicators of care utilization and processes of care decreased compared with prepandemic levels. Among PWH who remained in care, televisits were not associated with worse virologic, blood pressure, and glycemic control in PWH.

Low-Grade Inflammatory Mediators and Metalloproteinases Yield Synchronous and Delayed Responses to Mechanical Joint Loading.

Mechanical loading is an essential factor for the maintenance of joint inflammatory homeostasis and the sensitive catabolic-anabolic signaling cascade involved in maintaining cartilage tissue health. However, abnormal mechanical loading of the joint structural tissues can propagate joint metabolic dysfunction in the form of low-grade inflammation. To date, few studies have attempted to delineate the early cascade responsible for the initiation and perpetuation of stress-mediated inflammation and cartilage breakdown in human joints. Fifteen healthy human male participants performed a walking paradigm on a cross-tilting treadmill platform. Blood samples were collected before exercise, after 30 minutes of flat walking, after 30 minutes of tilted walking, and after an hour of rest. Serum concentrations of the following biomarkers were measured: interleukin (IL)-1β, IL-6, IL-10, tumor necrosis factor alpha (TNF)-α, matrix metalloproteinase (MMP)-1, MMP-3, MMP-9, MMP-13, transforming growth factor beta (TGF)-β, tissue inhibitor of matrix metalloproteinase 1 (TIMP)-1, and cartilage oligomeric protein (COMP). Luminex Multiplex analysis of serum showed increased concentrations of COMP, IL-1β, TNF-α, IL-10, and TGF-β from samples collected after flat and cross-tilted treadmill walking compared to baseline. Serum concentrations of MMP-1 and MMP-13 also increased, but primarily in samples collected after tilted walking. Pearson's correlation analysis showed positive correlations between the expression of COMP, TNF-α, IL-10, and MMP-13 at each study timepoint. Stress-mediated increases in serum COMP during exercise are associated with acute changes in pro and anti-inflammatory molecular activity and subsequent changes in molecules linked to joint tissue remodeling and repair.

Bidirectional associations between memory and depression moderated by sex and age: Findings from the CLSA

BackgroundResearch has struggled to understand the temporal relationship between cognition and depression. Some literature suggests that depression may be a risk factor for memory decline, while other work indicates that memory decline may precede depression symptoms. The purpose of this study was to clarify the temporal relationship between memory and depression, examining the moderating role of sex and age. MethodsData were drawn from two time points in the Canadian Longitudinal Study on Aging (CLSA). Memory was measured using a composite of immediate and delayed verbal recall scores, and depressive symptoms were measured using the Center for Epidemiologic Studies Short Depression Scale (CESD-10). Separate cross-lagged panel models (CLPMs) were run based on age (i.e., ages 45–64; ages 65+) and sex (n = 51,338). ResultsResults indicated bidirectional associations between depressive symptoms and memory such that depressive symptoms at baseline predicted memory at follow-up (β= 0.029–0.068, with all p-values <0.01) and memory at baseline predicted depressive symptoms at follow-up (β= 0.025–0.033, with all p-values <0.05). The only exception was in the older female group, where memory did not predict depressive symptoms (β= -0.006, p = 0.543). Depressive symptoms at baseline were a stronger predictor of memory at follow-up than memory at baseline was for depressive symptoms at follow-up in all groups except for older males. FindingsThe findings suggest small but consistent bidirectional associations between depression and memory in almost all sex/age groupings. Depressive symptoms tended to be a stronger predictor of memory than memory was for future depressive symptoms.

Prevention of contrast induced-acute kidney injury using coenzyme Q10 in patients with ST-segment elevation myocardial infarction undergoing primary percutaneous coronary intervention.

We assessed the potential effect of CoQ10 administration for the prevention of contrast induced-acute kidney injury (CI-AKI) in patients with ST-segment elevation myocardial infarction (STEMI) undergoing primary percutaneous coronary intervention (PCI). One hundred fifty STEMI patients who were candidates for primary PCI, along with intravenous saline hydration, randomly received a placebo or CoQ10. CoQ10 was administrated orally, 400 mg before the procedure and 200 mg twice daily after the procedure for three consecutive days. Serum creatinine concentration and corresponding creatinine clearance (estimated by the CKD Epidemiology Collaboration (CKD-EPI) creatinine equation) were measured at baseline and 24, 48, and 72 h after primary PCI. Furthermore, the serum level of superoxide dismutase (SOD), total antioxidant capacity (TAC), and malondialdehyde (MDA) was measured before and 72 h after primary PCI. The mean serum creatinine concentration before contrast administration was similar in the two groups (0.98 ± 0.08 versus 0.99 ± 0.09mg/dL). While in both study groups, compared to baseline, the mean serum creatinine concentration increased at 48 and 72 h after contrast exposure, the CoQ10 group showed a lower serum creatinine concentration than the placebo group (P-value = 0.017 and 0.004, respectively). However, comparing the mean values of creatinine clearance between the groups at the study time points did not demonstrate a statistically significant difference. CI-AKI, defined as a > 25% or 0.5 mg/dL increase in baseline serum creatinine concentration, occurred in 8.00% of the cases in the CoQ10 group versus 20.00% in the placebo group (P-value = 0.034). Furthermore, at 72 h, the CoQ10-treated group exhibited higher serum levels of SOD and TAC and a lower MDA level than the placebo-treated group. Our research's findings proposed CoQ10 supplementation as an adjuvant to saline hydration as a preventive approach against CI-AKI. The trial was registered at Iranian Registry of Clinical Trials ( https://www.irct.ir/trial/60435 , identifier code: IRCT20120215009014N414). Registration date: 2021-12-29.

Monitoring Cigarette Smoking and Relapse in Young Adults With and Without Remote Biochemical Verification: Randomized Brief Cessation Study.

Technological advancements to study young adult smoking, relapse, and to deliver interventions remotely offer conceptual appeal, but the incorporation of technological enhancement must demonstrate benefit over traditional methods without adversely affecting outcomes. Further, integrating remote biochemical verification of smoking and abstinence may yield value in the confirmation of self-reported smoking, in addition to ecologically valid, real-time assessments. The goal of this study was to evaluate the impact of remote biochemical verification on 24-hour self-reported smoking and biochemical verification agreement, retention, compliance with remote sessions, and abstinence during a brief, 5-week cessation attempt and relapse monitoring phase. Participants (N=39; aged 18-25 years; mean age 21.6, SD 2.1 years; n=22, 56% male; n=29, 74% White) who smoked cigarettes daily engaged in a 5-week cessation and monitoring study (including a 48-hour quit attempt and provision of tobacco treatment in the form of nicotine replacement therapy, brief cessation counseling, and financial incentives for abstinence during the 2-day quit attempt only). Smoking (cigarettes per day) was self-reported through ecological momentary assessment (EMA) procedures, and participants were randomized to either (1) the inclusion of remote biochemical verification (EMA + remote carbon monoxide [rCO]) 2× per day or (2) in-person, weekly CO (wCO). Groups were compared on the following outcomes: (1) agreement in self-reported smoking and breath carbon monoxide (CO) at common study time points, (2) EMA session compliance, (3) retention in study procedures, and (4) abstinence from smoking during the 2-day quit attempt and at the end of the 5-week study. No significant differences were demonstrated between the rCO group and the wCO (weekly in-person study visit) group on agreement between 24-hour self-reported smoking and breath CO (moderate to poor), compliance with remote sessions, or retention, though these outcomes numerically favored the wCO group. Abstinence was numerically higher in the wCO group after the 2-day quit attempt and significantly different at the end of treatment (day 35), favoring the wCO group. Though study results should be interpreted with caution given the small sample size, findings suggest that the inclusion of rCO breath added to EMA compared to EMA with weekly, in-person CO collection in young adults did not yield benefit and may have even adversely affected outcomes. Our results suggest that technological advancements may improve data accuracy through objective measurement but may also introduce barriers and burdens and could result in higher rates of missing data. The inclusion of technology to inform smoking cessation research and intervention delivery among young adults should consider (1) the research question and necessity of biochemical verification and then (2) how to seamlessly incorporate monitoring into personalized and dynamic systems to avoid the added burden and detrimental effects to compliance and honesty in self-report.

Plasma proteomic profiling identifies CD33 as a marker of HIV control in natural infection and after therapeutic vaccination

Biomarkers predicting the outcome of HIV-1 virus control in natural infection and after therapeutic interventions in HIV-1 cure trials remain poorly defined. The BCN02 trial (NCT02616874), combined a T-cell vaccine with romidepsin (RMD), a cancer-drug that was used to promote HIV-1 latency reversal and which has also been shown to have beneficial effects on neurofunction. We conducted longitudinal plasma proteomics analyses in trial participants to define biomarkers associated with virus control during monitored antiretroviral pause (MAP) and to identify novel therapeutic targets that can improve future cure strategies. BCN02 was a phase I, open-label, single-arm clinical trial in early-treated, HIV infected individuals. Longitudinal plasma proteomes were analyzed in 11 BCN02 participants, including 8 participants that showed a rapid HIV-1 plasma rebound during a monitored antiretroviral pause (MAP-NC, 'non-controllers') and 3 that remained off ART with sustained plasma viremia <2000 copies/ml (MAP-C, 'controllers'). Inflammatory and neurological proteomes in plasma were evaluated and integration data analysis (viral and neurocognitive parameters) was performed. Validation studies were conducted in a cohort of untreated HIV-1+ individuals (n=96) and invitro viral replication assays using an anti-CD33 antibody were used for functional validation. Inflammatory plasma proteomes in BCN02 participants showed marked longitudinal alterations. Strong proteome differences were also observed between MAP-C and MAP-NC, including in baseline timepoints. CD33/Siglec-3 was the unique plasma marker with the ability to discriminate between MAPC-C and MAP-NC at all study timepoints and showed positive correlations with viral parameters. Analyses in an untreated cohort of PLWH confirmed the positive correlation between viral parameters and CD33 plasma levels, as well as PBMC gene expression. Finally, adding an anti-CD33 antibody to invitro virus cultures significantly reduced HIV-1 replication and proviral levels in T cells and macrophages. This study indicates that CD33/Siglec-3 may serve as a predictor of HIV-1 control and as potential therapeutic tool to improve future cure strategies. Spanish Science and Innovation Ministry (SAF2017-89726-R and PID2020-119710RB-I00), NIH (P01-AI131568), European Commission (GA101057548) and a Grifols research agreement.

Di-(2-Ethylhexyl) Phthalate (DEHP) in House Dust in Canadian Homes: Behaviors and Associations with Housing Characteristics and Consumer Products

Background. Di-(2-ethylhexyl) phthalate (DEHP), which is ubiquitous in indoor environments, was the predominant phthalate measured in house dust in the Canadian CHILD Cohort and was found to be associated with a large increased risk of childhood asthma. Objective. To inform interventions by identifying sources of DEHP in dust and assessing behaviors related to DEHP concentrations in house dust. Methods. DEHP levels were measured in 726 dust samples collected at ~3 months of age in CHILD as well as in ~50 homes at two time points (June and November) in the CHILD pilot study. DEHP metabolites were measured in urine for a subset of the ~3-month-old infants. Housing characteristics were assessed at the time of dust and urine collection. Numerous factors from these surveys were investigated as potential sources of DEHP using univariate analyses and multivariable regressions. Correlations between DEHP in dust and urinary metabolites and between repeat dust samples were examined to study the relationship between dust measurement and DEHP exposure. Results. Overall, DEHP dust concentrations were higher for lower-income families. Homes with vinyl flooring in the kitchen and bathroom showed higher levels of DEHP than those without vinyl flooring. The quantity of vinyl furniture and the presence of mold were associated with higher DEHP concentrations, while the use of mattress covers reduced concentration. No other significant associations were found. DEHP concentrations in dust were consistent over 6 months, although the correlation between dust and DEHP metabolites in urine was low. Conclusion. DEHP in house dust persisted over multiple months, contributed to infant internal exposure, and was associated with specific housing characteristics. These findings may inform the public on their choice of building materials and products, as well as future policies, aimed at reducing the health risk associated with exposures in the indoor environment especially for children.

Enhanced podophyllotoxin production of endophyte Fusarium proliferatum TQN5T by host extract and phenylalanine.

Fermentation technology using endophytes is considered a potential alternative approach for producing pharmaceutical compounds like podophyllotoxin (PTOX). In this study, fungus TQN5T (VCCM 44284) was selected from endophytic fungi isolated from Dysosma versipellis in Vietnam for PTOX production through TLC. The presence of PTOX in TQN5T was further confirmed by HPLC. Molecular identification indicated TQN5T as Fusarium proliferatum with 99.43% identity. This result was asserted by morphological characteristics such as white cottony, filamentous colony, layer and branched mycelium, and clear hyphae septa. Cytotoxic assay indicated both biomass extract and culture filtrate of TQN5T presented strong cytotoxicity on LU-1 and HepG2 with IC50 of 0.11, 0.20, 0.041, and 0071, respectively, implying anti-cancer compounds were accumulated in the mycelium and secreted into the medium. Further, the production of PTOX in TQN5T was investigated in the fermentation condition supplemented with 10µg/ml of host plant extract or phenylalanine as elicitors. The results revealed a significantly higher amount of PTOX in the PDB + PE and PDB + PA at all studied time points in comparison with PDB (control). Especially, after 168h of culture, PTOX content in the PDB with plant extract reached the peak with 314µg/g DW which is 10% higher than the best yield of PTOX in previous studies, denoting F. proliferatum TQN5T as a promising PTOX producer. This is the first study on enhancing the PTOX production in endophytic fungi by supplementing phenylalanine-a precursor for PTOX biosynthesis in plants into fermented media, suggesting a common PTOX biosynthetic pathway between host plant and endophytes. KEY POINTS: • Fusarium proliferatum TQN5T was proven for PTOX production. • Both mycelia extract and spent broth extract of Fusarium proliferatum TQN5T presented strong cytotoxicity on cancer cell lines LU-1 and HepG2. • The supplementation of 10µg/ml host plant extract and phenylalanine into fermentation media of F. proliferatum TQN5T improved the yield of PTOX.

Blood collection tubes impact expression of activated CD4+ and CD8+ T cells in human whole blood assay

BackgroundT-lymphocyte subsets CD4 and CD8 play important role in host immune responses. However, little attention has been given to the impact of time lapse and the various anticoagulant blood collection tubes on the expression frequency and activation status of CD4+ and CD8+ T cells. To this end, we explore the impact of time (t<1 h and t=4 h) and collection tubes (EDTA and heparin) on the expression frequency and activation status of CD4+ and CD8+ T cells among healthy Ghanaian individuals. MethodsA cohort of healthy individuals (n=9) is recruited, and blood samples obtained in Ghana for the frequency of CD4+and CD8+ T cells at various time points (<1 h and 4 h). The proportions of activation of these immune markers were profiled using immunophenotyping. ResultsSignificant statistical differences in the activation frequency of CD69 expressing CD4+T cells (t < 1 h and t=4 h; p=0.02) and CD69 expressing CD8+ T cells from EDTA tubes at times (t < 1 h and t=4 h; p=0.05) was observed. No significant difference were observed with CD69 expressing cells in Heparin tubes. Notably, CD8+ T cell activation frequency was observed to be consistently higher than that of CD4+ T cell at the various study time points and in the collection tubes used. No marked alterations were observed witth the proportion of CD4+ and CD8+ T cells in the samples collected at the time points; <1 h and at 4 h. ConclusionThe study shows that activation of CD4+ and CD8+ T cells in EDTA tubes differed significantly between both time points (t <1 h and t=4 h) but not in the heparin collection tubes. Therefore, it is important to take into account the elapsed time and the type of blood collection tubes when performing phenotypic characterization of activated immune markers.

The relationship of fetal sex and maternal race and ethnicity with early and late pregnancy C-reactive protein and interleukin-8.

Promotion of a healthy pregnancy is dependent on a coordinated immune response that minimizes inflammation at the maternal-fetal interface. Few studies investigated the effect of fetal sex on proinflammatory biomarkers during pregnancy and whether maternal race could impact this association. We aimed to examine whether fetal sex could, independently of maternal race/ethnicity and the condition of pregnancy (normal vs. complicated), impact inflammatory markers (C-reactive protein [CRP] and interleukin-8 [IL-8] levels) in early and late pregnancy. This study was a cohort analysis using prospectively collected data from pregnant women who participated in the Vitamin Antenatal Asthma Reduction Trial (VDAART, N=816). Maternal serum CRP and IL-8 levels were measured in early and late pregnancy (10-18 and 32-38weeks of gestation, respectively). Five hundred and twenty-eight out of 816 pregnant women who participated in the trial had available CRP and IL-8 measurements at both study time points. We examined the association of fetal sex with early and late CRP and IL-8 levels and their paired sample difference. We further investigated whether maternal race/ethnicity, pregnancy complications (i.e., preeclampsia and gestational diabetes), and early pregnancy body mass index (BMI) could affect the association between these two biomarkers and fetal sex adjusting for potential confounders. For this purpose, we used generalized linear and logistic regression models on log-normalized early and late CRP and IL-8 levels as well as their split at median to form high and low groups. Women pregnant with male fetuses (266/528=56.5%) had higher CRP levels in early to mid-pregnancy (β=.18: 95% confidence interval [CI]: CI=0.03-0.32; p=.02). Twenty-seven percent (143/528) of the study subjects were Hispanic. Hispanic African American [AA] women and women of races other than White and AA had higher levels of CRP at early to mid-pregnancy compared with White women (β=.57; 95% CI: 0.17-0.97; p<.01 and β=.27; 95% CI: 0.05-0.48; p=.02, respectively). IL-8 levels were not associated with fetal sex in early and late pregnancy (p's>.05). Other factors such as gestational diabetes and early pregnancy BMI were associated with higher CRP levels and higher CRP and IL-8 levels, respectively. Dichotomizing log-normalized cytokine levels at the median in a sensitivity analysis, women with male fetuses had lower odds of high (above-median) IL-8 levels at early pregnancy. Also, women with races other than AA and White carrying male fetuses had higher odds of having high (above-median) late-pregnancy CRP and early-pregnancy IL-8 levels (adjusted odds ratio [aOR]=3.80, 95% CI: 0.24-1.23; p=.02 and aOR=3.57; 95% CI: 0.23-1.03; p=.02, respectively). Of the pregnancy complications, women with gestational diabetes mellitus had a higher paired difference of early and late pregnancy CRP levels (β=.38; 95% CI: 0.09-0.68; p=.01), but no difference in IL-8 levels (p's>.05). No associations between the inflammatory markers and preeclampsia were found. Fetal sex is associated with CRP in early pregnancy and an association with IL-8 in early pregnancy is implied. Our study further indicates that maternal race/ethnicity could be a contributing factor in the relationship between fetal sex and inflammatory responses during pregnancy. However, the specificity and level of the contribution might vary by type of cytokine, pregnancy stage, and other confounding factors such as BMI that may impact these associations.

Characteristics associated with differences in 24-hour device-measured and self-reported sleep, sedentary behaviour and physical activity in a sample of Australian primary school children

BackgroundHow much time children spend sleeping, being sedentary and participating in physical activity affects their health and well-being. To provide accurate guidelines for children’s time use, it is important to understand the differences between device-measured and self-reported use-of-time measures, and what may influence these differences. Among Australian primary school-aged children, this study aimed to describe the differences between device-measured and self-reported sleep, sedentary behaviour, light-intensity physical activity (LPA), and moderate-vigorous-intensity physical activity (MVPA), and to explore how sociodemographic and personal characteristics were associated with these differences.MethodsParticipants (n = 120, 67% female, age 9–11 years) were drawn from the Life on Holidays cohort study. Device measured use of time was from 7-day accelerometry worn over five timepoints in a 2-year period, and self-reported use of time was from 2-day Multimedia Activity Recall for Children and Adults (MARCA), conducted at the same timepoints. For each participant and measurement method, average daily time spent in sleep, sedentary time, LPA and MVPA was derived for any overlapping days (that had both types of measurement) across the study period. Participant characteristics were either obtained from baseline parental survey (age, sex, parental education, puberty) or derived from the average of direct measurements across the study timepoints (aerobic fitness from shuttle run, body mass index from anthropometric measurements, academic performance from national standardised tests). Differences between device-measured and self-reported use of time were described using Bland-Altmann plots. Compositional outcome linear-regression models were used to determine which participant characteristics were associated with differences by use-of-time measurement type.ResultsRelative to device-measured, self-reported daily LPA was underestimated by 83 min (35% difference), whilst sleep (+ 37 min; 6% difference), MVPA (+ 34 min; 33% difference) and sedentary time (+ 12 min; 3% difference) were overestimated. Characteristics underpinning the differences between measurement types were sex (χ2 = 11.9, p = 0.008), parental education (χ2 = 23.0, p = 0.001), aerobic fitness (χ2 = 10.7, p = 0.01) and academic performance (χ2 = 15.9, p = 0.001).ConclusionsAmong primary school-aged children, device-measured and self-reported use-of-time measurements should not be used interchangeably as there are systematic biases and differences relative to socio-demographic characteristics.

1-Year Prospective Study of the Relationship of Serial Exhaled Nitric Oxide Level and Asthma Control.

Previous studies found that the fractional nitric oxide concentration in exhaled breath (FeNO) levels in healthy Chinese adults was higher than in White adults. More understanding of serial changes of FeNO levels with asthma control in a real-life clinical setting would be important to explore the utility of this biomarker in routine asthma management. This study assessed the FeNO levels of Chinese asthma subjects with different levels of asthma control and the serial changes with respect to the changes in asthma control over 1 year. A 12-month prospective study (subjects recruited between November 2019 and January 2021) with serial measurement of FeNO levels at baseline, 4, 8 and 12 months. Asthma control was assessed by the Global Initiative for Asthma classification, Asthma Control Test (ACT) and Asthma Control Questionnaire (ACQ). Altogether, 136 subjects (mean age 51.51±15.09 years, 46[33.8%] male) had successful baseline FeNO measurements. At baseline, the FeNO levels did not show a statistically significant difference for controlled, partly controlled and uncontrolled asthma according to GINA classification, ACT and ACQ. FeNO levels decreased with improving asthma control and stayed at similar levels with unchanged or worsening asthma control for all subjects. For subjects with baseline blood eosinophil levels ≥300 cells/µL(n=59), FeNO levels decreased with improving asthma control, stayed similar without change for asthma control and increased with worsening asthma control. Receiver operating characteristic (ROC) analysis with the highest area under curve (AUC) for changes in FeNO levels for improving asthma control was between ≤ -10 to -25 ppb at various time points in the 12-month study. Changes in FeNO levels over time were associated with changes in clinical asthma control, particularly in those with higher blood eosinophil count and are likely more useful than a single time point measurement in managing asthma.

The Association Between Depression, Anxiety and COVID-19 Symptoms

Background: The variation of COVID-19 illness is not fully understood. There is a need for further identification of predictors for COVID-19-related health outcomes, which may improve the delivery of healthcare. The primary objective was to identify whether anxiety/depression symptoms are associated with the number of COVID-19 symptoms. The second objective was to examine differences in anxiety and depression symptoms between individuals with or without COVID-19 symptoms. Methods: 782 Virginians ages 18 to 87 years, enrolled from March to May 2021 and were followed-up for six months. Vibrent Health online platform was used to collect data. PHQ-9, GAD-7, and CDC's COVID-19 tracing form, were used to assess depression, anxiety, and COVID-19 symptoms, respectively. An MMRM test was used to examine whether anxiety and depression symptoms were associated with the number of COVID-19 symptoms. Age, race, sex, medical diagnoses, and COVID-19 related economic/social hardships were included as covariates. Mann-Whitney U tests were used to assess differences in anxiety/depression at all study time points. We conducted analyses using SAS 9.4, p-values &lt; .05 were considered significant. Results: Depression/anxiety symptoms, COVID-19 related economic/social hardships, and medical diagnoses, were significantly associated with the number of COVID-19 symptoms (p&lt;.05), whereas age, sex, and race were not (p&gt;.05). Overall, PHQ9 and GAD7 scores were consistently and significantly higher for individuals with COVID-19 symptoms than those without COVID-19 symptoms (p&lt;.05). Conclusions: The severity of depression and anxiety symptoms is linked to symptoms of COVID-19 over time. Physical and mental health integrated healthcare approaches may be necessary. Further investigation into causative mechanisms is needed.

Long-Term Chondrocyte Retention in Partially Decellularized Tracheal Grafts.

Decellularized tracheal grafts possess the biological cues necessary for tissue regeneration. However, conventional decellularization approaches to target the removal of all cell populations including chondrocytes lead to a loss of mechanical support. We have created a partially decellularized tracheal graft (PDTG) that preserves donor chondrocytes and the mechanical properties of the trachea. In this study, we measured PDTG chondrocyte retention with a murine microsurgical model. Murine in vivo time-point study. Research Institute affiliated with Tertiary Pediatric Hospital. PDTG was created using a sodium dodecyl sulfate protocol. Partially decellularized and syngeneic grafts were orthotopically implanted into female C57BL/6J mice. Grafts were recovered at 1, 3, and 6 months postimplant. Pre- and postimplant grafts were processed and analyzed via quantitative immunofluorescence. Chondrocytes (SOX9+, DAPI+) present in the host and graft cartilage was evaluated using ImageJ. Partial decellularization resulted in the maintenance of gross tracheal architecture with the removal of epithelial and submucosal structures on histology. All grafts demonstrated SOX9+ chondrocytes throughout the study time points. Chondrocytes in PDTG were lower at 6 months compared to preimplant and syngeneic controls. PDTG retained donor graft chondrocytes at all time points. However, PDTG exhibits a reduction in chondrocytes at 6 months. The impact of these histologic changes on cartilage extracellular matrix regeneration and repair remains unclear.

Evaluating a prototype digital mental health literacy intervention for children and young people aged 11–15 in Java, Indonesia: a mixed methods, multi-site case study evaluation

BackgroundThe Improving Mental Health Literacy Among Children and Young People in Indonesia (IMPeTUs) intervention is a co-produced, evidence-based digital intervention designed to improve anxiety and depression focused mental health literacy and self-management among people aged 11–15 in Java, Indonesia. This study aimed to evaluate the usability, feasibility and preliminary impact of our intervention.MethodsMixed methods, multi-site case studies based on a theory of change. Pre-and post-assessments of a range of outcomes and qualitative interviews/focus groups with children and young people (CYP), parents and facilitators. The intervention was implemented in 8 health, school and community sites across Java, Indonesia (Megelang, Jakarta and Bogor).Quantitative data designed to understand the impact of and feasibility of evaluating the intervention collected from 78 CYP who used the intervention were analysed descriptively. Qualitative data from interviews and focus groups collected from 56 CYP, 49 parents/caregivers and 18 facilitators were analysed using framework analysis.ResultsQualitative data analysis indicated high levels of usability and acceptability for the interface aesthetic, personalisation, message presentation and navigation. Participants reported minimal burden and no negative outcomes associated with the intervention. CYP, parents and facilitators identified a range of direct and spill over effects of interventions engagement, some of which were not anticipated at study outset. Quantitative data highlighted the feasibility of intervention evaluation, with high levels of recruitment and retention across study time points. Minimal changes were identified in outcomes pre-to-post intervention, which may in part be due to a lack of scale relevance and/or sensitivity to the intervention mechanisms indicated in the qualitative data.ConclusionsDigital mental health literacy applications are potentially an acceptable and feasible way to prevent burdens of common mental health problems amongst CYP in Indonesia. Our intervention and evaluative processes will be further refined prior to definitive evaluation.

Long-term effect of COVID-19 infection on kidney function among COVID-19 patients followed in post-COVID-19 recovery clinics in British Columbia, Canada.

We investigated the effect of Post-Acute COVID Syndrome or "long-COVID" on kidney function among patients followed in post-COVID recovery clinics (PCRC) in British Columbia, Canada. Long-COVID patients referred to PCRC between July 2020 to April 2022, aged ≥18years who had an estimated glomerular filtration rate (eGFR) value recorded at 3months from the coronavirus disease 2019 (COVID-19) diagnosis (index) date were included. Those requiring renal replacement therapy prior to index date were excluded. Primary outcome was change in eGFR and urine albumin-creatinine ratio (UACR) after COVID-19 infection. The proportion of patients in each of the six eGFR categories (<30, 30-44, 45-59, 60-89, 90-120 and >120mL/min/1.73m2) and three UACR categories (<3, 3-30 and >30mg/mmol) in all of the study time points were calculated. Linear mixed model was used to investigate change in eGFR over time. The study sample included 2212 long-COVID patients. Median age was 56years, 51% were male. Half (∼47%-50%) of the study sample had normal eGFR (≥90mL/min/1.73m2) from COVID-19 diagnosis to 12months post-COVID and <5% of patients had an eGFR <30mL/min/1.73m2. There was an estimated 2.96mL/min/1.73m2 decrease in eGFR within 1year after COVID-19 infection that was equivalent to 3.39% reduction from the baseline. Decline in eGFR was highest in patients hospitalized for COVID-19 (6.72%) followed by diabetic patients (6.15%). More than 40% of patients were at risk of CKD. People with long-COVID experienced a substantial decline in eGFR within 1year from the infection date. The prevalence of proteinuria appeared to be high. Close monitoring of kidney function is prudent among patients with persistent COVID-19 symptoms.

Protective effects of Centella asiatica extract on spatial memory and learning deficits in animal model of systemic inflammation induced by lipopolysaccharide

Background and aim Centella asiatica (L.) Urb. (Apiaceae) is a renowned medicinal plant being used in the Ayurvedic system for its pharmacological effects on the central nervous system such as rejuvenating, sedative, anxiolytic and memory-enhancing properties. The present study was designed to investigate the effect of Centella asiatica (CA) extract on inflammatory responses induced by lipopolysaccharide (LPS) and resulting changes in cognitive behavior. Materials and methods Adult male Sprague-Dawley rats were divided into 4 groups as control, LPS, CA and LPS + CA. The treatments with LPS (5 mg/kg) were intraperitoneally (i.p) injected on day 4 and CA ethanol extract (200 mg/kg) were given orally for 14 days. Morris Water Maze (MWM) test was performed to assess spatial learning and memory performance. Acute oral toxicity of the extract at the highest dose of 5000 mg/kg was also conducted. Results Single administration of LPS was able to significantly elicit learning and memory impairment (p < .05) when compared to the control groups. Treatment with CA significantly improved the impaired learning ability in which the LPS + CA rats took the shortest time and route to find the hidden platform (15.85 ± 2.68 s (p < .001); 352.43 ± 88.10 cm (p < .001) on day 5) and induced differential cytokine responses in the blood. No mortality and no significant variation in the body and organ weights between the control and the treated group was observed after 14 days of acute toxicity study. Hematological analysis and biochemical parameters revealed no toxic effects of the extract. Pathologically, neither gross abnormalities nor histopathological changes were observed. Discussion and conclusion Centella asiatica extract exhibited significant learning and memory enhancement potential in animal model. Hence, indicating its putative preventive therapeutic effects in neuroinflammation related diseases. KEY MESSAGE A single dose of lipopolysaccharide (LPS) (5 mg/kg) administered systemically to mimic the consequences of LPS-induced inflammatory responses was able to affect some behavioral modification of spatial memory at the time point of study. The study showed that the learning capability during the training trial was restored or ameliorated with the pre-emptive treatment of Centella asiatica extract (200 mg/kg). Centella asiatica extract improves spatial memory, learning deficits and regulates proinflammatory responses in systemic LPS-treated rats.

Usability of a novel lateral flow assay for the point-of-care detection of Neisseria gonorrhoeae: A qualitative time-series assessment among healthcare workers in South Africa.

Accurate and user-friendly rapid point-of-care diagnostic tests (POCT) are needed to optimize treatment of Neisseria gonorrhoeae, especially in low-resource settings where syndromic management is the standard of care for sexually transmitted infections. This study aimed to assess the acceptability and usability of a novel lateral flow assay and portable reader for the point-of-care detection of N. gonorrhoeae infection (NG-LFA). This mixed-methods study was conducted as part of a diagnostic performance and usability evaluation of a prototype NG-LFA for detection of N. gonorrhoeae in symptomatic men and women at primary healthcare facilities in the Buffalo City Metro, South Africa. The Standardized System Usability Scale (SUS) was administered, and in-depth interviews were conducted among healthcare professionals (HCPs) and fieldworkers (FWs) at pre-implementation, initial use and 3- and 6-month study implementation to assess user expectations, practical experience, and future implementation considerations for the NG-LFA. Data collection and analysis was guided by the Health Technology Adoption Framework, including new health technology attributes, learnability, satisfaction, and suitability. The framework was adapted to include perceived durability. A total of 21 HCPs and FWs were trained on the NG-LFA use. SUS scores showed good to excellent acceptability ranging from 78.8-90.6 mean scores between HCPs and FWs across study time points. All transcripts were coded using Dedoose and qualitative findings were organized by learnability, satisfaction, suitability, and durability domains. Usability themes are described for each time point. Initial insecurity dissipated and specimen processing dexterity with novel POCT technology was perfected over time especially amongst FWs through practical learning and easy-to-use instructions (learnability). Participants experienced both positive and negative test results, yielding perceived accuracy and minimal testing challenges overall (satisfaction). By 3- and 6-month use, both HCPs and FWs found the NG-LFA convenient to use in primary health care facilities often faced with space constraints and outlined perceived benefits for patients (suitability and durability). Findings show that the NG-LFA device is acceptable and usable even amongst paraprofessionals. High SUS scores and qualitative findings demonstrate high learnability, ease-of-use and suitability that provide valuable information for first-step scale-up requirements at primary healthcare level. Minor prototype adjustments would enhance robustness and durability aspects.

0203 Time-In-Bed Classroom Challenges: Incentivizing Undergraduate Students to Improve Sleep During Final Exams Week

Abstract Introduction Late bedtimes and all-nighters are ingrained into student culture, especially during final exam week. We previously found that extra credit incentives “to sleep 8 hours” encouraged longer sleep durations in small-sized classes. The current work investigated whether a modified approach—extending one’s time-in-bed—was scalable to larger-sized classes and dependent on the size of the incentive. Methods Across two studies, 129 undergraduate students completed baseline actigraphy and sleep diary monitoring early in Fall 2021 or Fall 2022. At the end of the semester, students could opt to take a time-in-bed challenge to earn large-sized (8 points; Study 1) or medium-sized (5 points; Study 2) extra credit on their final exam if they maintained an average time-in-bed of nine hours (or average total sleep time of eight hours). A smaller-sized extra credit (2 points) could be earned if the overall benchmarks were not met but one still improved on their baseline sleep duration by 20 minutes or sleep efficiency by 3%. Participants provided informed consent and the class instructor was kept blinded until after exams were graded. Results Most students attempted the time-in-bed sleep challenge (Study 1: 86.79%; Study 2: 92.11%). In Study 1, large-sized extra credit encouraged a 66 minute/night improvement in actigraphy-defined total sleep time from early-semester baseline (6.72 ± 1.18 hours) to final exams week (7.82 ± 0.89 hours; p&amp;lt;.001). In Study 2, a medium-sized extra credit opportunity improved actigraphy-defined total sleep time by 46 minutes/night from baseline (6.69 ± 0.80 hours) to finals week (7.46 ± 1.12 hours; p&amp;lt;.001). The interaction with incentive size was non-significant (p&amp;gt;.05). Sleep efficiency was similar across timepoints in Study 1 (83.6 ± 5.35 to 82.8 ± 5.18, p&amp;gt;.10) and Study 2 (82.9 ± 6.1 to 82.9 ± 6.47, p&amp;gt;.10). Students did not have to sacrifice total sleep time to perform well on their final exams (rs&amp;lt;.10); interestingly, students with higher sleep efficiencies performed better on their final exams (r=.28, p&amp;lt;.05). Conclusion The all-nighter culture of final exam week can be overcome by creating incentives for students to prioritize nighttime sleep. Support (if any) National Science Foundation (1920730 and 1943323).