Clickable Granular Hydrogel Scaffolds for Delivery of Neural Progenitor Cells to Sites of Spinal Cord Injury.

Spinal cord injury (SCI) is a serious condition with limited treatment options. Neural progenitor cell (NPC) transplantation is a promising treatment option, and the identification of novel biomaterial scaffolds that support NPC engraftment and therapeutic activity is a top research priority. The objective of this study is to evaluate in situ assembled poly (ethylene glycol) (PEG)-based granular hydrogels for NPC delivery in a murine model of SCI. Microgel precursors are synthesized by using thiol-norbornene click chemistry to react four-armed PEG-amide-norbornene with enzymatically degradable and cell adhesive peptides. Unreacted norbornene groups are utilized for in situ assembly into scaffolds using a PEG-di-tetrazine linker. The granular hydrogel scaffolds exhibit good biocompatibility and do not adversely affect the inflammatory response after SCI. Moreover, when used to deliver NPCs, the granular hydrogel scaffolds supported NPC engraftment, do not adversely affect the immune response to the NPC grafts, and successfully support graft differentiation toward neuronal or astrocytic lineages as well as axonal extension into the host tissue. Collectively, these data establish PEG-based granular hydrogel scaffolds as a suitable biomaterial platform for NPC delivery and justify further testing, particularly in the context of more severe SCI.

Exposure to Memory-Relevant versus Memory-Irrelevant Aging Stereotypes Differentially Affects Memory Self-Perceptions and Memory Test Scores of Young, Middle, and Older Age Adults.

This study examined the combined influence of assimilation and contrast effects on the memory self-efficacy and objective memory of young (ages 18-25, n = 114), middle-age (ages 26-59, n = 48), and older (ages 60-98, n = 59) adults. We reminded participants that they matched positive, not negative, memory-relevant or memory-irrelevant stereotypes of aging either before (experimental conditions) or after (control condition) they completed a memory self-efficacy questionnaire and took a memory test. Participants exposed to memory-relevant aging stereotypes prior to other measures reported higher memory self-efficacy than those exposed to memory-irrelevant stereotypes; this effect did not depend on age group. In contrast, the effect of stereotype exposure on memory performance differed with age. Young and older, but not middle-aged, adults showed differences in their memory scores depending on whether they were exposed to memory-relevant, memory-irrelevant or no aging stereotypes. In general, exposure to stereotypes (particularly those relevant to memory) had a negative influence on memory that contrasted with its positive effect on memory self-efficacy. Together, these results indicate that exposure to aging stereotypes can have opposing effects on the memory self-efficacy and objective test performance of adults of various ages and that the relevance of the stereotypes to the cognitive domain being assessed matters.

Severity of Schistosoma haematobium co-infection with malaria in school-children is potentially modulated by host CD14 gene variants.

OBJECTIVE: Schistosomiasis remains a chronic disease of global importance, especially in many rural areas of the world where co-infection with Plasmodium falciparum is common. It is critical to decipher the role of single or co-infected disease scenarios on immune system regulation in such individuals and how such co-infections can either ameliorate or complicate immune response and the consequent disease outcome. First, 10 ml of urine samples, collected between 10:00 am and 2:00 pm, was filtered for diagnosis of schistosomiasis, while egg count, indicative of disease severity, was determined by microscopy. Furthermore, genomic DNA samples extracted from dried blood spots collected on filter paper from one hundred and forty-four Schistosoma haematobium-infected school-children was tested for P. falciparum parasite positivity by an allele-specific nested-PCR analysis of merozoite surface protein (msp)-1 and -2 genes and a real-time PCR assay. In addition, among P. falciparum parasite-positive individuals, we carried out a Taqman SNP genotyping assay to extrapolate the effect of host CD14 (-159 C/T; rs2569190) genetic variants on schistosome egg count. RESULTS: Of the 144 individuals recruited, P. falciparum parasite positivity with msp-1 gene were 34%, 43% and 55% for MAD20, RO33 and K1 alleles respectively. Of the co-infected individuals, CD14 genetic variants ranged from 18.8% vs 21.5%, 33.3% vs 44.4%, 9.7% vs 11.8% for single versus schistosome co-infection for the wild type (CC), heterozygous (CT) and mutant (TT) variants respectively. Though the mean egg count for co-infected individuals with CD14 wild type (33.7 eggs per 10 ml of urine) and heterozygote variants (37.5 eggs per 10 ml of urine) were lower than that of schistosome infection alone (52.48 and 48.08 eggs/10 ml of urine respectively), it lacked statistical significance (p-value 0.12 and 0.29), possibly reflecting the benefit of the CD14 activation in schistosome plus malaria co-infection and not schistosome infection alone. In addition, the lower mean egg count in co-infected individuals reveal the benefit of downstream Th1 immune response mitigated by CD14 innate activation that is absent in schistosome infection alone.

Differences in Anatomical Outcomes Between Early Chronic and Far Chronic Time-Points After Transplantation of Spinal Cord Neural Progenitor Cells in Mice.

Spinal cord injury (SCI) affects millions of people worldwide. Neural progenitor cell (NPC) transplantation is a promising treatment for regenerating lost spinal cord tissue and restoring neurological function after SCI. We conducted a literature search and found that less than a quarter of experimental rodent cell and tissue transplantation studies have investigated anatomical outcomes at longer than 4 months post-transplantation. This is a critical topic to investigate, given that stem and progenitor cell therapies would need to remain in place throughout the lifetime of an individual. We sought to determine how commonly assessed anatomical outcomes evolve between early and far chronic time-points post-NPC transplantation. At either 8 weeks or 26 weeks following transplantation of NPCs into sites of cervical SCI, we evaluated graft neuronal density, astroglial cell density, graft axon outgrowth, and regeneration of host axon populations into grafts in male and female mice. We found that graft neuronal density does not change over time, but the numbers of graft-associated astrocytes and glial fibrillary acidic protein intensity is significantly increased in the far chronic phase compared with the early chronic time-point. In addition, graft axon outgrowth was significantly decreased at 26 weeks post-transplantation compared with 8 weeks post-transplantation. In contrast, corticospinal axon regeneration into grafts was not diminished over time, but rather increased significantly from early to far chronic periods. Interestingly, we found that graft neuronal density is significantly influenced by sex of the host animal, suggesting that sex-dependent processes may shape graft composition over time. Collectively, these results demonstrate that NPC transplants are dynamic and that commonly assessed outcome measures associated with graft efficacy evolve over the weeks to months post-transplantation into the spinal cord.

Developmental stage of transplanted neural progenitor cells influences anatomical and functional outcomes after spinal cord injury in mice.

Neural progenitor cell (NPC) transplantation is a promising therapeutic strategy for replacing lost neurons following spinal cord injury (SCI). However, how graft cellular composition influences regeneration and synaptogenesis of host axon populations, or recovery of motor and sensory functions after SCI, is poorly understood. We transplanted developmentally-restricted spinal cord NPCs, isolated from E11.5-E13.5 mouse embryos, into sites of adult mouse SCI and analyzed graft axon outgrowth, cellular composition, host axon regeneration, and behavior. Earlier-stage grafts exhibited greater axon outgrowth, enrichment for ventral spinal cord interneurons and Group-Z spinal interneurons, and enhanced host 5-HT+ axon regeneration. Later-stage grafts were enriched for late-born dorsal horn interneuronal subtypes and Group-N spinal interneurons, supported more extensive host CGRP+ axon ingrowth, and exacerbated thermal hypersensitivity. Locomotor function was not affected by any type of NPC graft. These findings showcase the role of spinal cord graft cellular composition in determining anatomical and functional outcomes following SCI.

The Emerging Role of Biological Sex in Cell Therapy for Spinal Cord Injury.

Neural progenitor cell (NPC) transplantation is a promising potential therapy for replacing spinal cord neurons and glial cells following spinal cord injury (SCI). Despite the rapid advancement of NPC transplantation to SCI clinical trials, we still lack understanding of fundamental biology underlying how NPC grafts interact with the injured host nervous system. Our recent study demonstrated a potent effect of biological sex mismatch between donor and host on graft immune rejection. Here we discuss the implications of this study in the context of clinical trials for SCI, and important topics for future research in SCI cell transplantation.

Preparedness of Pediatric Subspecialty Fellows to Address Emotional and Mental Health Needs Among Children With Chronic Medical Conditions.

This survey study investigates pediatric subspecialty fellows' attitudes about and perceived competence in addressing emotional and mental health needs of pediatric patients with chronic medical conditions.

Transcription Factor Hb9 Is Expressed in Glial Cell Lineages in the Developing Mouse Spinal Cord.

Hb9 (Mnx1) is a transcription factor described as a spinal cord motor neuron (MN)-specific marker and critical factor for the postmitotic specification of these cells. To date, expression of Hb9 in other cell types has not been reported. We performed a fate-mapping approach to examine distributions of Hb9-expressing cells and their progeny ("Hb9-lineage cells") within the embryonic and adult spinal cord of Hb9cre;Ai14 mice. We found that Hb9-lineage cells are distributed in a gradient of increasing abundance throughout the rostrocaudal spinal cord axis during embryonic and postnatal stages. Furthermore, although the majority of Hb9-lineage cells at cervical spinal cord levels are MNs, at more caudal levels, Hb9-lineage cells include small-diameter dorsal horn neurons, astrocytes, and oligodendrocytes. In the peripheral nervous system, we observed a similar phenomenon with more abundant Hb9-lineage Schwann cells in muscles of the lower body versus upper body muscles. We cultured spinal cord progenitors in vitro and found that gliogenesis was increased by treatment with the caudalizing factor FGF-8B, while glial tdTomato expression was increased by treatment with both FGF-8B and GDF-11. Together, these observations suggest that early and transient expression of Hb9 in spinal cord neural progenitors may be induced by caudalizing factors such as FGF and GDF signaling. Furthermore, our work raises the possibility that early Hb9 expression may influence the development of spinal cord macroglia and Schwann cells, especially at caudal regions. Together, these findings highlight the importance of using caution when designing experiments using Hb9cre mice to perform spinal cord MN-specific manipulations.

Effects of biological sex mismatch on neural progenitor cell transplantation for spinal cord injury in mice.

Despite advancement of neural progenitor cell transplantation to spinal cord injury clinical trials, there remains a lack of understanding of how biological sex of transplanted cells influences outcomes after transplantation. To address this, we transplanted GFP-expressing sex-matched, sex-mismatched, or mixed donor cells into sites of spinal cord injury in adult male and female mice. Biological sex of the donor cells does not influence graft neuron density, glial differentiation, formation of the reactive glial cell border, or graft axon outgrowth. However, male grafts in female hosts feature extensive hypervascularization accompanied by increased vascular diameter and perivascular cell density. We show greater T-cell infiltration within male-to-female grafts than other graft types. Together, these findings indicate a biological sex-specific immune response of female mice to male donor cells. Our work suggests that biological sex should be considered in the design of future clinical trials for cell transplantation in human injury.

Regulatory network of miRNA, lncRNA, transcription factor and target immune response genes in bovine mastitis.

Pre- and post-transcriptional modifications of gene expression are emerging as foci of disease studies, with some studies revealing the importance of non-coding transcripts, like long non-coding RNAs (lncRNAs) and microRNAs (miRNAs). We hypothesize that transcription factors (TFs), lncRNAs and miRNAs modulate immune response in bovine mastitis and could potentially serve as disease biomarkers and/or drug targets. With computational analyses, we identified candidate genes potentially regulated by miRNAs and lncRNAs base pair complementation and thermodynamic stability of binding regions. Remarkably, we found six miRNAs, two being bta-miR-223 and bta-miR-24-3p, to bind to several targets. LncRNAs NONBTAT027932.1 and XR_003029725.1, were identified to target several genes. Functional and pathway analyses revealed lipopolysaccharide-mediated signaling pathway, regulation of chemokine (C-X-C motif) ligand 2 production and regulation of IL-23 production among others. The overarching interactome deserves further in vitro/in vivo explication for specific molecular regulatory mechanisms during bovine mastitis immune response and could lay the foundation for development of disease markers and therapeutic intervention.

Integrating Datasets on Public Health and Clinical Aspects of Sickle Cell Disease for Effective Community-Based Research and Practice.

Sickle cell disease (SCD) is a genetic disease that has multiple aspects including public health and clinical aspects. The goals of the research study were to (1) understand the public health aspects of sickle cell disease, and (2) understand the overlap between public health aspects and clinical aspects that can inform research and practice beneficial to stakeholders in sickle cell disease management. The approach involved the construction of datasets from textual data sources produced by experts on sickle cell disease including from landmark publications published in 2020 on sickle cell disease in the United States. The interactive analytics of the integrated datasets that we produced identified that community-based approaches are common to both public health and clinical aspects of sickle cell disease. An interactive visualization that we produced can aid the understanding of the alignment of governmental organizations to recommendations for addressing sickle cell disease in the United States. From a global perspective, the interactive analytics of the integrated datasets can support the knowledge transfer stage of the SICKLE recommendations (Skills transfer, Increasing self-efficacy, Coordination, Knowledge transfer, Linking to adult services, and Evaluating readiness) for effective pediatric to adult transition care for patients with sickle cell disease. Considering the increased digital transformations resulting from the COVID-19 pandemic, the constructed datasets from expert recommendations can be integrated within remote digital platforms that expand access to care for individuals living with sickle cell disease. Finally, the interactive analytics of integrated expert recommendations on sickle cell disease management can support individual and team expertise for effective community-based research and practice.

Preparing Future Pediatricians to Meet the Behavioral and Mental Health Needs of Children.

The increasing prevalence of behavioral and/or mental health (B/MH) problems among children, adolescents, and young adults is rapidly forcing the pediatric community to examine its professional responsibility in response to this epidemic. Stakeholders involved in pediatric workforce training were brought together in April 2018, invited by the American Board of Pediatrics and the National Academies of Sciences, Engineering, and Medicine, to consider facilitators and barriers for pediatrician training to enhance care for B/MH problems and to catalyze commitment to improvement efforts. During the interactive meeting, parents, young adult patients, and trainees, together with leaders of pediatric training programs and health care organizations, acknowledged the growing B/MH epidemic and discussed past and current efforts to improve training and care, including integrated delivery models. Attendees committed in writing to making a change within their department or organization to improve training. There also was agreement that organizations that set the standards for training and certification bear some responsibility to ensure that future pediatricians are prepared to meet the needs of children and adolescents. Reports on commitments to change 12 months after the meeting indicated that although attendees had encountered a variety of barriers, many had creatively moved forward to improve training at the program or organizational level. This article describes the context for the April 2018 meeting, themes arising from the meeting, results from the commitments to change, and 3 case studies. Taken together, they suggest we, as a pediatric community, can and must collaborate to improve training and, by extension, care.

RecET-Mediated Recombineering in Acinetobacter baumannii.

Acinetobacter baumannii rapidly acquires antibiotic resistance, and its genome encodes mechanisms to tolerate biocides and desiccation, enhancing its persistence in hospital settings. Tools to rapidly dissect the A. baumannii genome are needed to understand cellular factors that contribute to its resiliency at a genetic and mechanistic level. While a substantial amount of clinical data has documented the global rise of A. baumannii as an antibiotic-resistant pathogen, genetic tools to dissect its molecular details have been limited. This procedure describes a recombination-mediated genetic engineering (recombineering) system for targeted genome editing of A. baumannii. This system can perform directed mutagenesis on wide-ranging genes and operons and has broad application in various strains of A. baumannii.

Isolation of Lipid Cell Envelope Components from Acinetobacter baumannii.

With the increasing occurrence of antibiotic resistance among Acinetobacter sp., the race is on for researchers to not only isolate resistant isolates but also utilize basic and applied microbiological techniques to study mechanisms of resistance. For many antibiotics, the limit of efficacy against Gram-negative bacteria is dependent on its ability to permeate the outer membrane and access its target. As such, it is critical that researchers be able to isolate and analyze the lipid components of the cell envelope from any number of Acinetobacter sp. that are either resistant or sensitive to antibiotics of interest. The following chapter provides in-depth protocols to confirm the presence or absence of lipooligosaccharide (LOS) in Acinetobacter sp., isolate lipid A, and glycerophospholipids and analyze them using qualitative (mass spectrometry) and semiquantitative (thin-layer chromatography) methods.

Discovery of Next-Generation Antimicrobials through Bacterial Self-Screening of Surface-Displayed Peptide Libraries.

Peptides have great potential to combat antibiotic resistance. While many platforms can screen peptides for their ability to bind to target cells, there are virtually no platforms that directly assess the functionality of peptides. This limitation is exacerbated when identifying antimicrobial peptides because the phenotype, death, selects against itself and has caused a scientific bottleneck that confines research to a few naturally occurring classes of antimicrobial peptides. We have used this seeming dissonance to develop Surface Localized Antimicrobial Display (SLAY), a platform that allows screening of unlimited numbers of peptides of any length, composition, and structure in a single tube for antimicrobial activity. Using SLAY, we screened ∼800,000 random peptide sequences for antimicrobial function and identified thousands of active sequences, dramatically increasing the number of known antimicrobial sequences. SLAY hits present with different potential mechanisms of peptide action and access to areas of antimicrobial physicochemical space beyond what nature has evolved. VIDEO ABSTRACT.

Exploring Costing Methods for Environmental Health Services in North Carolina.

All local health departments in North Carolina are mandated to provide a defined set of environmental health services, yet few have the tools to understand the costs incurred in delivering these services. The objectives of this study were to (1) derive cost estimates for 2 commonly provided environmental health services-food and lodging inspections and on-site water services-and (2) explore factors that drive variations in costs, focusing on the roles of economies of scale and scope. Using data from 15 local health departments in North Carolina, we found that costs varied substantially. A bivariate analysis found evidence of economies of scale: higher volumes of services were associated with lower costs per service. Providing a greater scope of services, however, was not consistently associated with reduced costs. In-depth cost data provide public health officials with key information when deciding how to best serve their communities.

Use and Perceived Impact of the County Health Rankings Report in Florida and North Carolina.

OBJECTIVE: Examine overall level of and variation in local health department (LHD) use and perceived impact of the County Health Rankings report (Rankings) in Florida (2010, 2011) and North Carolina (2010-2012, 2013). DESIGN: Two cross-sectional surveys among LHDs. PARTICIPANTS: Local health directors and relevant staff. MAIN OUTCOME MEASURES: Use of the Rankings was measured by asking respondents if their LHD had used the Rankings in any of 10 ways and through assessment of community engagement. Perceived impact was measured by amount of attention the Rankings received from various stakeholders and whether they had already produced or would likely produce any of 7 possible results. RESULTS: Overall, LHDs used the Rankings most often to educate staff in Florida (78%) and North Carolina (56%). Engagement with community groups around the Rankings was variable. Media engagement, through press releases (41%; 40%) or interviews (51%; 36%) in Florida and North Carolina, was moderate. Florida LHDs used the Rankings in more ways and significantly more frequently than North Carolina LHDs. There were few significant differences in perceived impact by state. At least a moderate amount of attention was received from media in Florida (52%) and North Carolina (46%). Twenty-percent of LHDs reported the Rankings received at least moderate attention from the general public in both states and 38% (Florida) and 33% (North Carolina) from policy makers. Tangible benefits to communities from the Rankings, such as having already influenced adoption of new policies, were modest in Florida (3%) and North Carolina (11%). CONCLUSIONS: Results suggest that tangible benefits to communities from use of the Rankings have yet to be fully realized but are encouraging. More effective media engagement could produce the community awareness necessary to maximize the Rankings' potential to mobilize communities for health improvement. State variation in Rankings use suggests that more support to LHDs may be helpful.

Factors Driving Local Health Departments' Partnerships With Other Organizations in Maternal and Child Health, Communicable Disease Prevention, and Chronic Disease Control.

OBJECTIVES: To describe levels of partnership between local health departments (LHDs) and other community organizations in maternal and child health (MCH), communicable disease prevention, and chronic disease control and to assess LHD organizational characteristics and community factors that contribute to partnerships. DATA SOURCES: Data were drawn from the National Association of County & City Health Officials' 2013 National Profile Study (Profile Study) and the Area Health Resources File. LHDs that received module 1 of the Profile Study were asked to describe the level of partnership in MCH, communicable disease prevention, and chronic disease control. Levels of partnership included "not involved," "networking," "coordinating," "cooperating," and "collaborating," with "collaborating" as the highest level of partnership. Covariates included both LHD organizational and community factors. Data analyses were conducted using Stata 13 SVY procedures to account for the Profile Study's survey design. RESULTS: About 82%, 92%, and 80% of LHDs partnered with other organizations in MCH, communicable disease prevention, and chronic disease control programs, respectively. LHDs having a public health physician on staff were more likely to partner in chronic disease control programs (adjusted odds ratio [AOR] = 2.33; 95% confidence interval [CI], 1.03-5.25). Larger per capita expenditure was also associated with partnerships in MCH (AOR = 2.43; 95% CI, 1.22-4.86) and chronic disease prevention programs (AOR = 1.76; 95% CI, 1.09-2.86). Completion of a community health assessment was associated with partnership in MCH (AOR = 7.26; 95% CI, 2.90-18.18), and chronic disease prevention (AOR = 5.10; 95% CI, 2.28-11.39). CONCLUSION: About 1 in 5 LHDs did not have any partnerships in chronic disease control. LHD partnerships should be promoted to improve care coordination and utilization of limited health care resources. Factors that might promote LHDs' partnerships include having a public health physician on staff, higher per capita expenditure, and completion of a community health assessment. Community context likely influences types and levels of partnerships. A better understanding of these contextual factors may lead to more complete and effective LHD partnerships.

Adoption of Evidence-Based Interventions in Local Health Departments: "1-2-3 Pap NC".

Descriptions of barriers and facilitators to adoption of evidence-based interventions in local health departments (LHDs) are limited. This study was conducted by the North Carolina Public Health Practice-Based Research Network to identify factors associated with adoption of an evidence-based human papillomavirus video intervention, "1-2-3 Pap NC," in North Carolina LHDs. A sequential mixed-method study design was used. Data from the 2013 National Profile of Local Health Departments were used to test associations between LHD characteristics and adoption of the intervention. Qualitative, key stakeholder interviews with LHD directors provided the context for quantitative data. Data collection and analysis continued from March 3, 2014, to September 15, 2014. Overall, 28% of North Carolina health jurisdictions (33 of 100 counties) implemented the intervention. Of the three channels used to deliver the intervention to clients, most LHDs opted to show the video in the exam room (42%), followed by website/other social media (36%) and video loop in the lobby/waiting room (22%). In logistic regression, gender of the director (female) was significantly and positively associated with adoption of the intervention (AOR=4.44, p<0.05). Being a first-time director was marginally significant (AOR=0.28, p=0.074), suggesting first-time directors were less likely to adopt. Qualitative results suggested that aspects of communication (awareness and positive attitudes) and agency directors' evaluation of resources, balanced against intervention complexity and flexibility, competing priorities, and mandates, influenced adoption. Adoption of evidence-based interventions by LHDs is critical to improve population health. Practice-based research can contribute to understanding facilitators and modifying barriers to this process.