Using mHealth to reduce disparities in Black maternal health: Perspectives from Black rural postpartum mothers.

BACKGROUND: Racial disparities are evident in maternal morbidity and mortality rates globally. Black women are more likely to die from pregnancy and childbirth than any other race or ethnicity. This leaves one of the largest gaps in women's health to date. OBJECTIVES: mHealth interventions that connect with women soon after discharge may assist in individualizing and formalizing support for mothers in the early postpartum period. To aid in developing an mHealth application, Black postpartum mothers' perspectives were examined. DESIGN: Utilizing the Sojourner Syndrome Framework and Maternal Mortality & Morbidity Measurement Framework, group interview discussion guides were developed to examine the facilitators and barriers of postpartum transitional care for rural Black women living in the United States to inform the development of a mobile health application. METHODS: In this study, seven group interviews were held with Black mothers, their support persons, and healthcare providers in rural Georgia to aid in the development of the Prevent Maternal Mortality Using Mobile Technology (PM3) mobile health (mHealth) application. Group interviews included questions about (1) post-birth experiences; (2) specific needs (e.g. clinical, social support, social services, etc.) in the postpartum period; (3) perspectives on current hospital discharge processes and information; (4) lived experiences with racism, classism, and/or gender discrimination; and (5) desired features and characteristics for the mobile app development. RESULTS: Fourteen out of the 78 screened participants were eligible and completed the group interview. Major discussion themes included: accessibility to healthcare and resources due to rurality, issues surrounding race and perceived racism, mental and emotional well-being in the postpartum period, and perspectives on the PM3 mobile application. CONCLUSION: Participants emphasized the challenges that postpartum Black women face in relation to accessibility, racism and discrimination, and mental health. The women favored a culturally relevant mHealth tool and highlighted the need to tailor the application to address disparities.

An in vitro CRISPR screen of cell-free DNA identifies apoptosis as the primary mediator of cell-free DNA release.

ABTRACT: Clinical circulating cell-free DNA (cfDNA) testing is now routine, however test accuracy remains limited. By understanding the life-cycle of cfDNA, we might identify opportunities to increase test performance. Here, we profile cfDNA release across a 24-cell line panel and utilize a cell-free CRISPR screen (cfCRISPR) to identify mediators of cfDNA release. Our panel outlines two distinct groups of cell lines: one which releases cfDNA fragmented similarly to clinical samples and purported as characteristic of apoptosis, and another which releases larger fragments associated with vesicular or necrotic DNA. Our cfCRISPR screens reveal that genes mediating cfDNA release are primarily involved with apoptosis, but also identify other subsets of genes such as RNA binding proteins as potential regulators of cfDNA release. We observe that both groups of cells lines identified primarily produce cfDNA through apoptosis. These results establish the utility of cfCRISPR, genetically validate apoptosis as a major mediator of DNA release in vitro, and implicate ways to improve cfDNA assays.

Examining the Perceptions of mHealth on Racial and Ethnic Disparities in Postpartum Health for Black Women: A Scoping Review.

Background. Several disparities exist for Black mothers during the postpartum period, including but not limited to increased maternal mortality and morbidity rates, decreased access to care, and limited access to resources. Given the racial discrepancies in attention to postpartum care, coupled with the critical importance of the postpartum period for preventing adverse maternal health outcomes, research is warranted to explore how mobile health (mHealth) applications may help to alleviate maternal health disparities by optimizing postpartum care and addressing barriers to care for postpartum Black women. Thus, this review examines the perceptions of mHealth applications and their utility in health outcomes among postpartum Black women. Methods. We undertook a comprehensive literature search using PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) guidelines. We included peer-reviewed articles published between 2010 and 2022 that were written in English, utilized mHealth as a primary intervention, and focused on postpartum health and access to resources, primarily among Black women in the United States. Results. A total of eight articles were included in our synthesis, encompassing mobile phone-based interventions for Black women. Cultural tailoring was included in five studies. Interventions that incorporated tailored content and fostered interactions reported high rates of follow-up. Conclusions. Tailored mHealth interventions can effectively promote behavior change and improve health care outcomes for Black women. However, there is a critical need for more research to assess user engagement and retention and whether these improvements indicate long-term sustainability.

Draft genome sequence of Exiguobacterium sp. strain MMG028 isolated from a salt marsh.

Here, we report the draft genome sequence of Exiguobacterium sp. strain MMG028, isolated from Rose Creek, San Diego, CA, USA, assembled and analyzed by undergraduate students participating in a marine microbial genomics course. A genomic comparison suggests that MMG028 is a novel species, providing a resource for future microbiology and biotechnology investigations.

The agency domain and behavioral interactions: assessing positive animal welfare using the Five Domains Model.

Animal welfare denotes how an animal experiences their life. It represents the overall mental experiences of an animal and is a subjective concept that cannot be directly measured. Instead, welfare indicators are used to cautiously infer mental experiences from resource provisions, management factors, and animal-based measures. The Five Domains Model is a holistic and structured framework for collating these indicators and assessing animal welfare. Contemporary approaches to animal welfare management consider how animals can be given opportunities to have positive experiences. However, the uncertainty surrounding positive mental experiences that can be inferred has resulted in risk-averse animal welfare scientists returning to the relative safety of positivism. This has meant that aspects of positive welfare are often referred to as animal 'wants'. Agency is a concept that straddles the positivist-affective divide and represents a way forward for discussions about positive welfare. Agency is the capacity of individual animals to engage in voluntary, self-generated, and goal-directed behavior that they are motivated to perform. Discrete positive emotions are cautiously inferred from these agentic experiences based on available knowledge about the animal's motivation for engaging in the behavior. Competence-building agency can be used to evaluate the potential for positive welfare and is represented by the Behavioral Interactions domain of the Five Domains Model. In 2020, The Model was updated to, amongst other things, include consideration of human-animal interactions. The most important aspect of this update was the renaming of Domain 4 from "Behavior" to "Behavioral Interactions" and the additional detail added to allow this domain's purpose to be clearly understood to represent an animal's opportunities to exercise agency. We illustrate how the Behavioral Interactions domain of The Model can be used to assess animals' competence-building agency and positive welfare. In this article, we use the examples of sugar gliders housed in captivity and greyhounds that race to illustrate how the agentic qualities of choice, control, and challenge can be used to assess opportunities for animals to exercise agency and experience positive affective engagement.

Linking bacterial tetrabromopyrrole biosynthesis to coral metamorphosis.

An important factor dictating coral fitness is the quality of bacteria associated with corals and coral reefs. One way that bacteria benefit corals is by stimulating the larval to juvenile life cycle transition of settlement and metamorphosis. Tetrabromopyrrole (TBP) is a small molecule produced by bacteria that stimulates metamorphosis with and without attachment in a range of coral species. A standing debate remains, however, about whether TBP biosynthesis from live Pseudoalteromonas bacteria is the primary stimulant of coral metamorphosis. In this study, we create a Pseudoalteromonas sp. PS5 mutant lacking the TBP brominase gene, bmp2. Using this mutant, we confirm that the bmp2 gene is critical for TBP biosynthesis in Pseudoalteromonas sp. PS5. Mutation of this gene ablates the bacterium's ability in live cultures to stimulate the metamorphosis of the stony coral Porites astreoides. We further demonstrate that expression of TBP biosynthesis genes is strongest in stationary and biofilm modes of growth, where Pseudoalteromonas sp. PS5 might exist within surface-attached biofilms on the sea floor. Finally, we create a modular transposon plasmid for genomic integration and fluorescent labeling of Pseudoalteromonas sp. PS5 cells. Our results functionally link a TBP biosynthesis gene from live bacteria to a morphogenic effect in corals. The genetic techniques established here provide new tools to explore coral-bacteria interactions and could help to inform future decisions about utilizing marine bacteria or their products for coral restoration.

A modular plasmid toolkit applied in marine bacteria reveals functional insights during bacteria-stimulated metamorphosis.

A conspicuous roadblock to studying marine bacteria for fundamental research and biotechnology is a lack of modular synthetic biology tools for their genetic manipulation. Here, we applied, and generated new parts for, a modular plasmid toolkit to study marine bacteria in the context of symbioses and host-microbe interactions. To demonstrate the utility of this plasmid system, we genetically manipulated the marine bacterium Pseudoalteromonas luteoviolacea, which stimulates the metamorphosis of the model tubeworm, Hydroides elegans. Using these tools, we quantified constitutive and native promoter expression, developed reporter strains that enable the imaging of host-bacteria interactions, and used CRISPR interference (CRISPRi) to knock down a secondary metabolite and a host-associated gene. We demonstrate the broader utility of this modular system for testing the genetic tractability of marine bacteria that are known to be associated with diverse host-microbe symbioses. These efforts resulted in the successful conjugation of 12 marine strains from the Alphaproteobacteria and Gammaproteobacteria classes. Altogether, the present study demonstrates how synthetic biology strategies enable the investigation of marine microbes and marine host-microbe symbioses with potential implications for environmental restoration and biotechnology. IMPORTANCE Marine Proteobacteria are attractive targets for genetic engineering due to their ability to produce a diversity of bioactive metabolites and their involvement in host-microbe symbioses. Modular cloning toolkits have become a standard for engineering model microbes, such as Escherichia coli, because they enable innumerable mix-and-match DNA assembly and engineering options. However, such modular tools have not yet been applied to most marine bacterial species. In this work, we adapt a modular plasmid toolkit for use in a set of 12 marine bacteria from the Gammaproteobacteria and Alphaproteobacteria classes. We demonstrate the utility of this genetic toolkit by engineering a marine Pseudoalteromonas bacterium to study their association with its host animal Hydroides elegans. This work provides a proof of concept that modular genetic tools can be applied to diverse marine bacteria to address basic science questions and for biotechnology innovations.

Deep Learning Segmentation of the Nucleus Basalis of Meynert on 3T MRI.

BACKGROUND AND PURPOSE: The nucleus basalis of Meynert is a key subcortical structure that is important in arousal and cognition and has been explored as a deep brain stimulation target but is difficult to study due to its small size, variability among patients, and lack of contrast on 3T MR imaging. Thus, our goal was to establish and evaluate a deep learning network for automatic, accurate, and patient-specific segmentations with 3T MR imaging. MATERIALS AND METHODS: Patient-specific segmentations can be produced manually; however, the nucleus basalis of Meynert is difficult to accurately segment on 3T MR imaging, with 7T being preferred. Thus, paired 3T and 7T MR imaging data sets of 21 healthy subjects were obtained. A test data set of 6 subjects was completely withheld. The nucleus was expertly segmented on 7T, providing accurate labels for the paired 3T MR imaging. An external data set of 14 patients with temporal lobe epilepsy was used to test the model on brains with neurologic disorders. A 3D-Unet convolutional neural network was constructed, and a 5-fold cross-validation was performed. RESULTS: The novel segmentation model demonstrated significantly improved Dice coefficients over the standard probabilistic atlas for both healthy subjects (mean, 0.68 [SD, 0.10] versus 0.45 [SD, 0.11], P = .002, t test) and patients (0.64 [SD, 0.10] versus 0.37 [SD, 0.22], P < .001). Additionally, the model demonstrated significantly decreased centroid distance in patients (1.18 [SD, 0.43] mm, 3.09 [SD, 2.56] mm, P = .007). CONCLUSIONS: We developed the first model, to our knowledge, for automatic and accurate patient-specific segmentation of the nucleus basalis of Meynert. This model may enable further study into the nucleus, impacting new treatments such as deep brain stimulation.

Linking bacterial tetrabromopyrrole biosynthesis to coral metamorphosis.

An important factor dictating coral fitness is the quality of bacteria associated with corals and coral reefs. One way that bacteria benefit corals is by stimulating the larval to juvenile life cycle transition of settlement and metamorphosis. Tetrabromopyrrole (TBP) is a small molecule produced by bacteria that stimulates metamorphosis in a range of coral species. A standing debate remains, however, about whether TBP biosynthesis from live Pseudoalteromonas bacteria is the primary stimulant of coral metamorphosis. In this study, we create a Pseudoalteromonas sp. PS5 mutant lacking the TBP brominase gene, bmp2 . Using this mutant, we confirm that the bmp2 gene is critical for TBP biosynthesis in Pseudoalteromonas sp. PS5. Mutation of this gene ablates the bacterium's ability in live cultures to stimulate the metamorphosis of the stony coral Porites astreoides . We further demonstrate that expression of TBP biosynthesis genes is strongest in stationary and biofilm modes of growth, where Pseudoalteromonas sp. PS5 might exist within surface-attached biofilms on the sea floor. Finally, we create a modular transposon plasmid for genomic integration and fluorescent labeling of Pseudoalteromonas sp. PS5 cells. Our results functionally link a TBP biosynthesis gene from live bacteria to a morphogenic effect in corals. The genetic techniques established here provide new tools to explore coral-bacteria interactions and could help to inform future decisions about utilizing marine bacteria or their products for restoring degraded coral reefs.

A modular plasmid toolkit applied in marine Proteobacteria reveals functional insights during bacteria-stimulated metamorphosis.

A conspicuous roadblock to studying marine bacteria for fundamental research and biotechnology is a lack of modular synthetic biology tools for their genetic manipulation. Here, we applied, and generated new parts for, a modular plasmid toolkit to study marine bacteria in the context of symbioses and host-microbe interactions. To demonstrate the utility of this plasmid system, we genetically manipulated the marine bacterium Pseudoalteromonas luteoviolacea , which stimulates the metamorphosis of the model tubeworm, Hydroides elegans . Using these tools, we quantified constitutive and native promoter expression, developed reporter strains that enable the imaging of host-bacteria interactions, and used CRISPR interference (CRISPRi) to knock down a secondary metabolite and a host-associated gene. We demonstrate the broader utility of this modular system for rapidly creating and iteratively testing genetic tractability by modifying marine bacteria that are known to be associated with diverse host-microbe symbioses. These efforts enabled the successful transformation of twelve marine strains across two Proteobacteria classes, four orders and ten genera. Altogether, the present study demonstrates how synthetic biology strategies enable the investigation of marine microbes and marine host-microbe symbioses with broader implications for environmental restoration and biotechnology.

The association between platelet indices and presence and severity of psoriasis: a systematic review and meta-analysis.

The role of platelet function indices-platelet count (PLT), mean platelet volume (MPV), platelet distribution width (PDW), plateletcrit (PCT), immature platelet fraction (IPF), and platelet mass index (PMI)-in psoriasis is uncertain. This systematic review and meta-analysis aimed to evaluate the association of these platelet biomarkers with both presence and severity of psoriasis. We searched MEDLINE (Ovid), Embase (Ovid), and the Cochrane Library from inception to November 2021. To evaluate the association of platelet function indices and psoriasis, we recorded mean differences (MD) and 95% confidence intervals (CI) as well as correlation coefficients (r) for each included study, and generated summary estimates using random-effects inverse-variance modelling. We screened 1,079 unique studies, and included 33 studies with 6724 patients in the quantitative analyses. Compared with controls, patients with psoriasis had higher PLT (MD 12.86 × 109/L, 95% CI 6.34-19.39, p < 0.001), MPV (MD 0.61fL, 95% CI 0.31-0.92, p < 0.001), and PCT (MD 0.05%, 95% CI 0.01-0.09, p = 0.010), but similar PDW (MD 0.16%, 95% CI -0.46-0.79, p = 0.610). Psoriasis Area and Severity Index (PASI) was weakly correlated with PLT (r 0.17, 95% CI 0.06-0.28, p = 0.003), MPV (r 0.36, 95% CI 0.22-0.49, p < 0.001), and PDW (r 0.17, 95% CI 0.08-0.26, p < 0.001). Study numbers were insufficient to judge the relationship of IPF and PMI with psoriasis presence, or PCT, IPF, and PMI with psoriasis severity. In summary, PLT, MPV, and PCT are significantly elevated in patients with psoriasis, and PLT, MPV, and PDW are weakly correlated with PASI. Future studies are needed to evaluate the independent diagnostic and prognostic potentials of these biomarkers in patients with psoriasis.

Adaptive Language Mapping Paradigms for Presurgical Language Mapping.

BACKGROUND AND PURPOSE: Functional MR imaging is widely used for preoperative language assessment in candidates for resective neurosurgery. Language mapping paradigms that are adaptive to participant performance have the potential to engage the language network more robustly and consistently, resulting in more accurate functional maps. The aim of the current study was to compare two adaptive paradigms with the recommended language mapping paradigms that constitute the current standard of care. MATERIALS AND METHODS: Seventy-three patients undergoing fMRI for language lateralization and/or localization completed an adaptive semantic matching paradigm, an adaptive phonological judgment paradigm, and two standard paradigms: sentence completion and word generation. The paradigms were compared in terms of the degree to which they yielded lateralized language maps and the extent of activation in frontal, temporal, and parietal language regions. RESULTS: The adaptive semantic paradigm resulted in the most strongly lateralized activation maps, the greatest extent of frontal and temporal activations, and the greatest proportion of overall satisfactory language maps. The adaptive phonological paradigm identified anterior inferior parietal phonological encoding regions in most patients, unlike any of the other paradigms. CONCLUSIONS: The adaptive language mapping paradigms investigated have several psychometric advantages compared with currently recommended paradigms. Adoption of these paradigms could increase the likelihood of obtaining satisfactory language maps in each individual patient.

Ocular biodistribution of cysteamine delivered by a sustained release microsphere/thermoresponsive gel eyedrop.

The objective of the investigation was to determine the ocular biodistribution of cysteamine, a reducing agent used for treatment of cystine crystals in cystinosis, following topical administration of a sustained release formulation and traditional eyedrop formulation. To the right eye only, rabbits received a 50 µL drop of 0.44% cysteamine eyedrops at one drop per waking hour for 2, 6, 12, and 24 h. A second group received one 100 µL drop of a sustained release formulation containing encapsulated cysteamine microspheres suspended in a thermoresponsive gel. Upon serial sacrifice, ocular tissues from both eyes and plasma were obtained and quantified for cysteamine using LC-MS/MS. Cysteamine was detected in the cornea, aqueous humor and vitreous humor. Systemic plasma concentrations of cysteamine from treatment groups were below the limit of detection. As expected, 0.44% cysteamine eyedrops when administered hourly maintained drug concentrations within the cornea at a magnitude 5 times higher than a single dose of the sustained release formulation over 12 h. The sustained release formulation maintained cysteamine presentation across 12 h from a single drop. These studies demonstrate distribution of cysteamine to the eye following topical administration, including high drug uptake to the cornea and low systemic distribution.

Novel Combined Lidocaine/Povidone Iodine Delivery System for Preintravitreal Injection.

Purpose: Intravitreal injection has become a popular treatment for various retina disorders and dramatically increased over the past few years. In traditional preintravitreal injection, the preparation steps are time consuming for practitioners who perform a significant number of injections per day. Besides, lidocaine gel (L-Gel) shows a potential absorption barrier on the antibacterial effect of povidone iodine (PI). Methods: In this study, we describe a L/PI gel system as an alternative approach to address these issues for traditional preinjection drug administration. Lidocaine and PI are loaded in a thermoresponsive gel instilled as a liquid to the lower fornix that transitions to a stable, solid gel depot. Results and Conclusion: The gel demonstrated decrease in conjunctival touch sensitivity and sufficient bacteria killing with a single step, suggesting a significant decrease in the time required and less potential for drug inhibition due to sequential administration.

NOTCH1 PEST domain variants are responsive to standard of care treatments despite distinct transformative properties in a breast cancer model.

Activating variants in the PEST region of NOTCH1 have been associated with aggressive phenotypes in human cancers, including triple-negative breast cancer (TNBC). Previous studies suggested that PEST domain variants in TNBC patients resulted in increased cell proliferation, invasiveness, and decreased overall survival. In this study, we assess the phenotypic transformation of activating NOTCH1 variants and their response to standard of care therapies. AAV-mediated gene targeting was used to isogenically incorporate 3 NOTCH1 variants, including a novel TNBC frameshift variant, in two non-tumorigenic breast epithelial cell lines, MCF10A and hTERT-IMEC. Two different variants at the NOTCH1 A2241 site (A2441fs and A2441T) both demonstrated increased transformative properties when compared to a non-transformative PEST domain variant (S2523L). These phenotypic changes include proliferation, migration, anchorage-independent growth, and MAPK pathway activation. In contrast to previous studies, activating NOTCH1 variants did not display sensitivity to a gamma secretase inhibitor (GSI) or resistance to chemotherapies. This study demonstrates distinct transformative phenotypes are specific to a given variant within NOTCH1 and these phenotypes do not correlate with sensitivities or resistance to chemotherapies or GSIs. Although previous studies have suggested NOTCH1 variants may be prognostic for TNBC, our study does not demonstrate prognostic ability of these variants and suggests further characterization would be required for clinical applications.

Gut microbiota and age shape susceptibility to clostridial enteritis in lorikeets under human care.

BACKGROUND: Enteritis is a common cause of morbidity and mortality in lorikeets that can be challenging to diagnose and treat. In this study, we examine gut microbiota in two lorikeet flocks with enteritis (Columbus Zoo and Aquarium-CZA; Denver Zoo-DZ). Since 2012, the CZA flock has experienced repeated outbreaks of enteritis despite extensive diet, husbandry, and clinical modifications. In 2018, both CZA and DZ observed a spike in enteritis. Recent research has revealed that the gut microbiota can influence susceptibility to enteropathogens. We hypothesized that a dysbiosis, or alteration in the gut microbial community, was making some lorikeets more susceptible to enteritis, and our goal was to characterize this dysbiosis and determine the features that predicted susceptibility. RESULTS: We employed 16S rRNA sequencing to characterize the cloacal microbiota in lorikeets (CZA n = 67, DZ n = 24) over time. We compared the microbiota of healthy lorikeets, to lorikeets with enteritis, and lorikeets susceptible to enteritis, with "susceptible" being defined as healthy birds that subsequently developed enteritis. Based on sequencing data, culture, and toxin gene detection in intestinal contents, we identified Clostridium perfringens type A (CZA and DZ) and C. colinum (CZA only) at increased relative abundances in birds with enteritis. Histopathology and immunohistochemistry further identified the presence of gram-positive bacilli and C. perfringens, respectively, in the necrotizing intestinal lesions. Finally, using Random Forests and LASSO models, we identified several features (young age and the presence of Rhodococcus fascians and Pseudomonas umsongensis) associated with susceptibility to clostridial enteritis. CONCLUSIONS: We identified C. perfringens type A and C. colinum associated with lorikeet necrohemorrhagic enteritis at CZA and DZ. Susceptibility testing of isolates lead to an updated clinical treatment plan which ultimately resolved the outbreaks at both institutions. This work provides a foundation for understanding gut microbiota features that are permissive to clostridial colonization and host factors (e.g. age, prior infection) that shape responses to infection.

CCL2 loaded microparticles promote acute patency in silk-based vascular grafts implanted in rat aortae.

Cardiovascular disease is the leading cause of death worldwide, often associated with coronary artery occlusion. A common intervention for arterial blockage utilizes a vascular graft to bypass the diseased artery and restore downstream blood flow; however, current clinical options exhibit high long-term failure rates. Our goal was to develop an off-the-shelf tissue-engineered vascular graft capable of delivering a biological payload based on the monocyte recruitment factor C-C motif chemokine ligand 2 (CCL2) to induce remodeling. Bi-layered silk scaffolds consisting of an inner porous and outer electrospun layer were fabricated using a custom blend of Antherea Assama and Bombyx Mori silk (lyogel). Lyogel silk scaffolds alone (LG), and lyogel silk scaffolds containing microparticles (LGMP) were tested. The microparticles (MPs) were loaded with either CCL2 (LGMP+) or water (LGMP-). Scaffolds were implanted as abdominal aortic interposition grafts in Lewis rats for 1 and 8 weeks. 1-week implants exhibited patency rates of 50% (7/14), 100% (10/10), and 100% (5/5) in the LGMP-, LGMP+, and LG groups, respectively. The significantly higher patency rate for the LGMP+ group compared to the LGMP- group (p = 0.0188) suggests that CCL2 can prevent acute occlusion. Immunostaining of the explants revealed a significantly higher density of macrophages (CD68+ cells) within the outer vs. inner layer of LGMP- and LGMP+ constructs but not in LG constructs. After 8 weeks, there were no significant differences in patency rates between groups. All patent scaffolds at 8 weeks showed signs of remodeling; however, stenosis was observed within the majority of explants. This study demonstrated the successful fabrication of a custom blended silk scaffold functionalized with cell-mimicking microparticles to facilitate controlled delivery of a biological payload improving their in vivo performance. STATEMENT OF SIGNIFICANCE: This study outlines the development of a custom blended silk-based tissue-engineered vascular graft (TEVG) for use in arterial bypass or replacement surgery. A custom mixture of silk was formulated to improve biocompatibility and cellular binding to the tubular scaffold. Many current approaches to TEVGs include cells that encourage graft cellularization and remodeling; however, our technology incorporates a microparticle based delivery platform capable of delivering bioactive molecules that can mimic the function of seeded cells. In this study, we load the TEVGs with microparticles containing a monocyte attractant and demonstrate improved performance in terms of unobstructed blood flow versus blank microparticles. The acellular nature of this technology potentially reduces risk, increases reproducibility, and results in a more cost-effective graft when compared to cell-based options.

Evaluating extraction methods to study canine urine microbiota.

The urinary microbiota is the collection of microbes present in urine that may play a role in host health. Studies of urine microbiota have traditionally relied upon culturing methods aimed at identifying pathogens. However, recent culture-free sequencing studies of the urine microbiota have determined that a diverse array of microbes is present in health and disease. To study these microbes and their potential role in diseases like bladder cancer or interstitial cystitis, consistent extraction and detection of bacterial DNA from urine is critical. However, urine is a low biomass substrate, requiring sensitive methods to capture DNA and making the risk of contamination high. To address this challenge, we collected urine samples from ten healthy dogs and extracted DNA from each sample using five different commercially available extraction methods. Extraction methods were compared based on total and bacterial DNA concentrations and bacterial community composition and diversity assessed through 16S rRNA gene sequencing. Significant differences in the urinary microbiota were observed by dog and sex but not extraction method. The Bacteremia Kit yielded the highest total DNA concentrations (Kruskal-Wallis, p = 0.165, not significant) and the highest bacterial DNA concentrations (Kruskal-Wallis, p = 0.044). Bacteremia also extracted bacterial DNA from the greatest number of samples. Taken together, these results suggest that the Bacteremia kit is an effective option for studying the urine microbiota. This work lays the foundation to study the urine microbiome in a wide range of urogenital diseases in dogs and other species.

Alterations in gut microbiota linked to provenance, sex, and chronic wasting disease in white-tailed deer (Odocoileus virginianus).

Chronic wasting disease (CWD) is a fatal, contagious, neurodegenerative prion disease affecting both free-ranging and captive cervid species. CWD is spread via direct or indirect contact or oral ingestion of prions. In the gastrointestinal tract, prions enter the body through microfold cells (M-cells), and the abundance of these cells can be influenced by the gut microbiota. To explore potential links between the gut microbiota and CWD, we collected fecal samples from farmed and free-ranging white-tailed deer (Odocoileus virginianus) around the Midwest, USA. Farmed deer originated from farms that were depopulated due to CWD. Free-ranging deer were sampled during annual deer harvests. All farmed deer were tested for CWD via ELISA and IHC, and we used 16S rRNA gene sequencing to characterize the gut microbiota. We report significant differences in gut microbiota by provenance (Farm 1, Farm 2, Free-ranging), sex, and CWD status. CWD-positive deer from Farm 1 and 2 had increased abundances of Akkermansia, Lachnospireacea UCG-010, and RF39 taxa. Overall, differences by provenance and sex appear to be driven by diet, while differences by CWD status may be linked to CWD pathogenesis.