Background: Urine testing is common among hospitalized patients with indwelling urinary catheters, yet variation in collection practices may compromise diagnostic accuracy and contribute to unnecessary downstream interventions. Guidelines recommend avoiding sampling from indwelling catheters when feasible to reduce contamination and detection of asymptomatic colonization. However, real-world compliance and its relationship to diagnostic urine test results, subsequent UTI diagnosis, and antimicrobial prescribing remain incompletely characterized. Methods: We conducted a retrospective medical record review of adult patients who underwent urine testing between March and November 2024 and had an indwelling urinary catheter or catheter removal within the prior 24 hours. Urine samples were classified as compliant if obtained after catheter removal and noncompliant if collected with a catheter in place. Analyses included all urine samples, with urinalysis-specific analyses limited to samples with available results. Urine collection practices, urinalysis parameters, urine culture outcomes, provider-documented UTI diagnoses, concordance with symptom-based criteria and IDSA/NHSN definitions, and antimicrobial prescribing following urine testing (receipt, timing, route, and duration) were summarized descriptively and stratified by compliance status. Results: A total of 921 urine samples were included; 663 had an initial urinalysis with reflex urine culture, while the remainder underwent urine culture alone. Samples compliant with recommended collection practices accounted for 59% of cases. Among samples with urinalysis, overall positivity was similar in compliant and noncompliant samples (57% vs 55%), with comparable urinalysis component positivity across compliance groups. Urine culture positivity was also similar across compliance groups in both reflex-culture and culture-only samples (29% vs 27% and 27% vs 29%). Gram-negative organisms predominated (13-15%), followed by fungi (7-12%) and Gram-positive organisms (2-4%), with fungal isolates more common among noncompliant samples. Antimicrobial therapy was prescribed in 19% of compliant samples compared with 15% of noncompliant samples and was frequently initiated before culture results in both groups. Median treatment duration was similar (7 vs 6 days). Provider-documented UTI diagnoses were common (80% vs 84%); however, only a portion met standardized UTI definitions (IDSA criteria: 41% compliant vs 32% noncompliant), with greater concordance observed among compliant samples. Conclusion: Recommended urine collection practices were followed in just over half of samples, with similar urine test positivity across compliance groups. Differences in organism distribution, antimicrobial prescribing, and concordance between provider-documented and standardized UTI diagnoses suggest that urine collection practices may influence diagnostic classification and antimicrobial decision-making in hospitalized adults.

Background: Nasal colonization with Staphylococcus aureus is a well-established risk factor for central line–associated bloodstream infections (CLABSIs). Many institutions have adopted screening and targeted decolonization strategies using intranasal mupirocin; however, real-world implementation remain incompletely characterized. Methods: We conducted a retrospective cohort study at University Hospitals Cleveland Medical Center to evaluate implementation of S. aureus screening and decolonization protocol among hospitalized adults undergoing central venous catheter (CVC) or peripherally inserted central catheter (PICC) placement. All patients aged ≥18 years admitted between October 1, 2023, and May 5, 2025, who had a catheter placed were included. Patients with documented bloodstream infection at the time of catheter placement were excluded. Primary implementation outcomes were: (1) proportion of patients screened for S. aureus colonization, and (2) proportion of colonized patients who received prophylactic intranasal mupirocin. Secondary outcomes included overall mupirocin use, bloodstream infection (BSI), and CLABSI rates. Mixed effects logistic regression was performed to identify factors associated with screening uptake. Results: Among 11,107 central lines placed, 63.4% underwent S. aureus screening. Of those screened, 22.6% tested positive for Staph aureus. Only 21.2% of colonized patients received intranasal mupirocin, corresponding to an overall mupirocin use rate of 5.3%. Across all central lines placed during the study period, the overall BSI and CLABSI rates were 0.91% and 0.67%, respectively. In adjusted analyses, screening was more likely among patients with arm insertion sites (OR 1.68, 95% CI 1.34–2.12), left-sided catheter (OR 1.18, 95% CI 1.07–1.29), increasing catheter lumens (three lumens: OR 1.27, 95% CI 1.11–1.47; four lumens: OR 2.22, 95% CI 1.58–3.12), ICU location (OR 1.66, 95% CI 1.49–1.84), and hematology–oncology services (OR 1.51, 95% CI 1.12–2.03). Although catheter type itself was not independently associated with screening, patients receiving PICCs demonstrated a positive, though non-significant, trend toward higher screening uptake compared with CVCs (OR 1.15, 95% CI 0.91–1.47). Tunneled catheters (OR 0.87, 95% CI 0.79–0.97) and a higher number of prior inpatient admissions were associated with lower odds of screening, while demographic characteristics and most comorbidities were not. Conclusions: S. aureus screening uptake among patients with central lines was low, and gaps at subsequent steps in the process resulted in fewer than one quarter of colonized patients receiving mupirocin. This stepwise breakdown in implementation highlights opportunities to improve protocol reliability by reducing decision points through more standardized approaches.

Bees are key species for pollination and apiculture. Within the multiple biotic threats, parasites are one of the main players involved in bee health. Among them, trypanosomatid parasites have been the focus of recent studies that have placed them as one of the most prevalent microorganisms in the digestive tract of bees around the world. Here, we review what is known in epidemiology, cell biology and genetics of these parasites in bees with a particular focus on the work made on honeybees. We also discuss the possible implications for honeybee health and describe research gaps to be explored both from the honeybee host and trypanosomatid parasites sides.

Alcohol use is influenced by genetic and environmental factors. We examined the interactive effects between genome-wide polygenic risk scores for alcohol use (alc-PRS) and social support in relation to alcohol use among European American (EA) and African American (AA) adults across sex and developmental stages (emerging adulthood, young adulthood, and middle adulthood). Data were drawn from 4,011 EA and 1,274 AA adults from the Collaborative Study on the Genetics of Alcoholism who were between ages 18–65 and had ever used alcohol. Participants completed the Semi-Structured Assessment for the Genetics of Alcoholism and provided saliva or blood samples for genotyping. Results indicated that social support from friends, but not family, moderated the association between alc-PRS and alcohol use among EAs and AAs (only in middle adulthood for AAs); alc-PRS was associated with higher levels of alcohol use when friend support was low, but not when friend support was high. Associations were similar across sex but differed across developmental stages. Findings support the important role of social support from friends in buffering genetic risk for alcohol use among EA and AA adults and highlight the need to consider developmental changes in the role of social support in relation to alcohol use.

Back pain is one of the largest drivers of workplace injury and lost productivity in industries around the world. Back injuries were one of the leading reasons in resulting in days away from work at 38.5% across all occupations, increasing for manual laborers to 43%. While the cause of the back pain can vary across occupations, for materiel movers it is often caused from repetitive poor lifting. To reduce the issues, the Aerial Porter Exoskeleton (APEx) was created. The APEx uses a hip-mounted, powered exoskeleton attached to an adjustable vest. An onboard computer calculates the configuration of the user to determine when to activate. Lift form is assisted by using a novel lumbar brace mounted on the sides of the hips. Properly worn, the APEx holds the user upright while providing additional hip torque through a lift. This was tested by having participants complete a lifting test with the exoskeleton worn in the “on” configuration compared with the exoskeleton not worn. The APEx has been shown to deliver 30 Nm of torque in lab testing. The activity recognition algorithm has also been shown to be accurate in 95% of tested conditions. When worn by subjects, testing has shown average peak reductions of 14.9% BPM, 8% in VO2 consumption, and an 8% change in perceived effort favoring the APEx.

The sustainability concept seeks to balance how present and future generations of humans meet their needs. But because nature is viewed only as a resource, sustainability fails to recognize that humans and other living beings depend on each other for their well-being. We therefore argue that true sustainability can only be achieved if the interdependent needs of all species of current and future generations are met, and propose calling this ‘multispecies sustainability’. We explore the concept through visualizations and scenarios, then consider how it might be applied through case studies involving bees and healthy green spaces.

Frailty is increasingly used in clinical settings to describe a physiological state resulting from a combination of age-related co-morbidities. Frailty also has a strong ‘lay’ meaning that conjures a particular way of being. Recent studies have reported how frail older people perceive the term frailty, showing that frailty is often an unwanted and resisted label. While there are many scores and measures that clinicians can use to determine frailty, little has been published regarding how health-care professionals use and make sense of the term. This paper reports the findings of a qualitative study that explored how health professionals perceive frailty. Forty situated interviews were conducted with health-care professionals working in an emergency department in the English Midlands. The interview talk was analysed using discourse analysis. The findings show that the health professionals negotiate an ‘ideological dilemma’ – a tension between contradictory sets of meanings and consequences for action – based on their ‘lay’ and clinical experience of the term frailty. It is concluded that this dilemma could have a negative impact on the assessment of frailty depending on the system of assessment used.

OBJECTIVES/SPECIFIC AIMS: Focal cartilage injuries of the knee joint are common and present a treatment challenge due to minimal intrinsic repair. Cartilage tissue engineering techniques currently used in clinical practice are expensive, cumbersome, and often ineffective in patients with mechanical or medical comorbidities. To address these issues, we developed an acellular nanofibrous scaffold with encapsulated growth factors designed to enhanced articular cartilage repair. Our goal is to evaluate this technology in vitro and pilot a large animal model for eventual translation into human subjects. METHODS/STUDY POPULATION: Hyaluronic acid (HA, 65 kDa) will be methacrylated (~40% modification, MeHA) and conjugated with cell-adhesive (RGD) groups. A solution of 4% wt/vol MeHA, 2% wt/vol polyethylene oxide (900 kDa), 0.05% wt/vol Irgacure 2959, and 0.005% wt/vol stromal cell-derived factor-1α (SDF-1α) and/or transforming growth factor-β3 (TGF-β3) will be prepared in ddH2O. The solution will be electrospun onto a rotating mandrel to achieve a dry scaffold thickness of 0.5 mm. The scaffold matt will be UV cross-linked and 5 mm-diameter samples will be cut out. Four groups of scaffolds will be prepared: MeHA, MeHA+SDF, MeHA+TGF, MeHA+SDF+TGF. All groups will be evaluated for fiber diameter, swell thickness, equilibrium compressive modulus, degradation rate, and growth factor release rate over 4 weeks (n=10). Scaffolds will also be seeded with juvenile porcine MSCs (5×104) in 200 μL of medium incubated for 24 hours. Seeded scaffolds will be evaluated for equilibrium compressive modulus, cell infiltration, and chondrogenesis at 4 and 8 weeks (n=10). Scaffolds will then be evaluated in a juvenile Yucatan minipig cartilage defect model. In total, 6 animals will undergo bilateral knee surgery to create four 4 mm-diameter full-thickness cartilage defects in each trochlear grove. All defects will receive microfracture to release marrow elements. Each knee will receive 2 scaffolds of the same group (replicates) with paired microfracture controls, resulting in a sample size of 3. Animals will be sacrificed at 12 weeks and defects will be evaluated via non-destructive indentation testing for mechanical properties, microCT for defect fill and subchondral bone morphology, and histology for ICRS II Visual Histological Assessment Scoring. RESULTS/ANTICIPATED RESULTS: Our preliminary studies have shown reliable replication of electrospun MeHA scaffolds. We anticipate cross-linking density to correlate positively with compressive modulus, and negatively with swell thickness, degradation rate, and growth factor release rate. We anticipate the addition of SDF-1α and TGF-β3 to increase cell infiltration and chondrogenesis, respectively, within seeded scaffolds. Similarly, we expect minipig defects treated with growth factor-releasing scaffolds to show greater mechanical properties, defect fill, and ICRS II score compared with MeHA scaffolds without growth factor. DISCUSSION/SIGNIFICANCE OF IMPACT: This study has the potential to show how an HA-based cell-free scaffold can be augmented with 2 growth factors that act synergistically to improve cartilage repair in a large animal model. This technology would improve upon the cell-free scaffolds already used clinically for autologous matrix-induced chondrogenesis and is directly translatable.

OBJECTIVES/SPECIFIC AIMS: A hallmark of progressive HIV-1 infection is the massive activation and depletion of the gut barrier protective CD4 T helper subsets (Th17 and Th22) in the intestinal mucosa. The loss of these cells is thought to contribute to microbial translocation and systemic immune activation that occurs during chronic infection. In addition to the loss of protective Th subsets, we previously showed that chronically HIV-1 infected individuals have an altered colonic mucosal microbiome, which is in part characterized by a lower relative abundance of bacteria that produce the short-chain fatty acid butyrate in conjunction with increased relative abundance of gram-negative pathobionts. This dysbiosis was linked to markers of mucosal and systemic immune activation in these individuals. Following up on these clinical observations, we sought to understand how a loss of butyrate might contribute to HIV-associated inflammation. Initial studies showed that the addition of butyrate to cultured lamina propria mononuclear cells (LPMC) resulted in decreased pathobiont-driven gut T cell activation, HIV-1 infection levels and production of IL-17 and IFNy. Since the gut barrier protective Th17 and Th22 subsets are preferentially infected and depleted, which is critical to HIV-1 pathogenesis, we wanted to determine the mechanism by which butyrate modulates activation of these important Th subsets in the gut. METHODS/STUDY POPULATION: Total LPMCs or purified LP CD4 T cells were isolated from human jejunal tissue (n=3–6), labeled with CFSE and cultured with TCR/CD28 beads to mimic APC driven T cell activation, with the addition of butyrate at physiologic doses(0–2 mM). Four days after culture, secreted cytokine(IL-17 and IFNy) levels were measured by ELISA. Cells were then short-term (4 hr) mitogenically stimulated (PMA/Ionomycin) in the presence of a golgi transport inhibitor. Total CD4 T cell activation (CD38+/HLA-DR+, CD25+), proliferation (CFSElow), and frequencies of intracellular cytokines were measured by multi-color flow cytometry. Paired t-tests were performed to determine statistical significance. RESULTS/ANTICIPATED RESULTS: Butyrate inhibited LP CD4 T cell activation (p=0.013) and proliferation (p=0.015) within total LPMCs stimulated with TCR/CD28 beads in a dose-dependent manner, with significant activity starting at 0.125 mM. Quantification of total secreted cytokines revealed that butyrate significantly decreased both IL-17 and IFNy production after 4 days of culture at 0.0625 mM and 0.25 mM of butyrate, respectively. Assays using purified LP CD4 T cells demonstrated that butyrate directly decreased LP CD4 T cell activation, proliferation and cytokine production in response to TCR/CD28 stimulation. Studies on specific T helper subsets revealed that butyrate inhibited proliferation of Th17 cells at lower concentrations (IC50:0.147 mM) compared with Th1 (IC50:0.229 mM) and Th22 (IC50:0.258 mM) and Th non-IL-22/IL-17/IFNy producing (IC50:2.14 mM) subsets. In addition, it appeared there was a paradoxical increase of HIV-1 infection levels at lower concentrations of butyrate (0.125 mM). DISCUSSION/SIGNIFICANCE OF IMPACT: The addition of butyrate to activated LP CD4 T cells decreases TCR-mediated activation in a dose-dependent manner, and butyrate acts directly on purified LP CD4 T cell populations independent of other cell populations. Butyrate differentially inhibited the proliferation of Th17, Th1, and Th22 subsets, with Th17 cells being the most sensitive to butyrate but increased the infection levels of all T helper subsets at low concentrations. Further studies are needed to determine the mechanism of butyrate’s actions on LP Th cells and the sensitivity of Th17 cells to the inhibitory effects of butyrate. These results could help direct targeted manipulation of the colonic microbiome of HIV-1 infected individuals to help resolve inflammation and limit the impact of the infection in the gut mucosa and systemically.

Coronary artery disease after bone marrow transplantation is rare in children and young adults. We report the case of a 21-year-old who developed coronary artery disease and acute myocardial infarction secondary to graft versus host disease following bone marrow transplantation. Physicians caring for young patients after bone marrow transplantation should be aware of the potential for coronary artery disease and evaluate appropriately.

Healthy adults (n 30) participated in a placebo-controlled, randomised, double-blinded, cross-over study consisting of two 28 d treatments (β2-1 fructan or maltodextrin; 3×5 g/d) separated by a 14-d washout. Subjects provided 1 d faecal collections at days 0 and 28 of each treatment. The ability of faecal bacteria to metabolise β2-1 fructan was common; eighty-seven species (thirty genera, and four phyla) were isolated using anaerobic medium containing β2-1 fructan as the sole carbohydrate source. β2-1 fructan altered the faecal community as determined through analysis of terminal restriction fragment length polymorphisms and 16S rRNA genes. Supplementation with β2-1 fructan reduced faecal community richness, and two patterns of community change were observed. In most subjects, β2-1 fructan reduced the content of phylotypes aligning within the Bacteroides, whereas increasing those aligning within bifidobacteria, Faecalibacterium and the family Lachnospiraceae. In the remaining subjects, supplementation increased the abundance of Bacteroidetes and to a lesser extent bifidobacteria, accompanied by decreases within the Faecalibacterium and family Lachnospiraceae. β2-1 Fructan had no impact on the metagenome or glycoside hydrolase profiles in faeces from four subjects. Few relationships were found between the faecal bacterial community and various host parameters; Bacteroidetes content correlated with faecal propionate, subjects whose faecal community contained higher Bacteroidetes produced more caproic acid independent of treatment, and subjects having lower faecal Bacteroidetes exhibited increased concentrations of serum lipopolysaccharide and lipopolysaccharide binding protein independent of treatment. We found no evidence to support a defined health benefit for the use of β2-1 fructans in healthy subjects.