Understanding the Role of Schools in Community Prevention of Childhood Obesity

Childhood obesity is considered an epidemic with the percentage of obesity among children in the United States more than tripling in the past 30 years (Institute of Medicine, 2012). Many factors in the environment contribute to this problem including increased availability of unhealthy foods and limited access to healthy foods, increased screen time and lack of accessible and safe places for physical activity (Center for Disease Control and Prevention, 2013). Since many of these factors are often shared by a community due to common experience, successful interventions should target the community. When planning community interventions, it is important to link community resources such as city parks with neighborhoods where children go to school and live. Community partners that include city parks, neighborhoods, schools, local government, and health care professionals are essential to developing successful childhood obesity prevention practices. Past reviews of studies found that nutrition education, increased opportunities for physical activity and use of the school as a resource are crucial in preventing and curbing childhood obesity (Bleich, Segal, Wu, Wilson & Wang, 2013; Waters et al., 2011). / / This honors project was conducted in rural eastern North Carolina at a school-based health center located in a public school in collaboration with a school nurse. The school has a student population of 460 in 5th through 8th grade. In eastern North Carolina 17.4% of children aged 2-17 years are obese (North Carolina State Center for Health Statistics, 2011). The purpose of this project is integrate community resources related to healthy food choices and physical activity with school-age children in the school setting. The first objective of this project was to interview 4-5 key informants, about what community resources, such as parks and community gardens are available and how the school uses these resources to improve food choices and physical activity. The second objective was to provide written recommendations on the integration of community resources with school-age children to increase healthy food choices and physical activity levels. Public health interventions used in this project were collaboration and coalition building with community partners.

β-Amyloid Deposition is Shifted to the Vasculature and Memory Impairment is Exacerbated When Hyperhomocysteinemia is Induced in APP/PS1 Transgenic Mice

INTRODUCTION: Vascular dementia is the second most common cause of dementia after Alzheimer\u27s disease (AD). In addition, it is estimated that almost half of all AD patients have significant cerebrovascular disease comorbid with their AD pathology. We hypothesized that cerebrovascular disease significantly impacts AD pathological progression. METHODS: We used a dietary model of cerebrovascular disease that relies on the induction of hyperhomocysteinemia (HHcy). HHcy is a significant clinical risk factor for stroke, cardiovascular disease and type 2 diabetes. In the present study, we induced HHcy in APP/PS1 transgenic mice. RESULTS: While total β-amyloid (Aβ) load is unchanged across groups, Congophilic amyloid deposition was decreased in the parenchyma and significantly increased in the vasculature as cerebral amyloid angiopathy (CAA; vascular amyloid deposition) in HHcy APP/PS1 mice. We also found that HHcy induced more microhemorrhages in the APP/PS1 mice than in the wild-type mice and that it switched the neuroinflammatory phenotype from an M2a biased state to an M1 biased state. Associated with these changes was an induction of the matrix metalloproteinase protein 2 (MMP2) and MMP9 systems. Interestingly, after 6 months of HHcy, the APP/PS1 mice were cognitively worse than wild-type HHcy mice or APP/PS1 mice, indicative of an additive effect of the cerebrovascular pathology and amyloid deposition. CONCLUSIONS: These data show that cerebrovascular disease can significantly impact Aβ distribution in the brain, favoring vascular deposition. We predict that the presence of cerebrovascular disease with AD will have a significant impact on AD progression and the efficacy of therapeutics

Recovery Strategies in Endurance Athletes

In order to achieve optimal performance, endurance athletes, whether at the professional or recreational level, need to implement a variety of recovery strategies that are specific to their individual training and competition. Recovery is a multidimensional process involving physiological, psychological, emotional, social, and behavioral aspects. PURPOSE: The purpose of the current study is to examine current implementation, beliefs, and sources of information associated with recovery strategies in endurance athletes. METHODS: Participants included 240 self-identified endurance athletes (m=112, f=126, non-binary=1, and preferred not to answer=1) across different sports (cycling=57, running=100, triathlon = 61, other=22; pro=13, current/former collegiate athlete=67, recreational=160). The participants ranged in age from 18 to 79 years old (40.7±13.73) and were primarily white (n=213). Participants completed an online survey through Qualtrics on demographics, recovery strategies used in practice and competition, perceived benefit of the strategy, and their sources of information regarding recovery strategies. RESULTS: Hydration was the most frequently reported recovery strategy in both training (91.3%, n = 219) and competition r (80.4%, n = 193). Nutrition followed in training (78.8%, n = 189) and competition (78.3%, n = 188). Sleep was the third most utilized strategy in training (77.9%, n = 187) and competition (76.7%, n = 184). The least used recovery strategy was ultrasound in training (1.3%, n = 3) and competition (1.7% n = 4). Chi-squared analyses showed no significant differences between training and competition for the use of the recovery strategies. Hydration was reported as a beneficial recovery strategy after practice (96.5%, n = 223) and competition (94.7%, n = 200) by the greatest percentage of participants, with sleep the second most frequently considered beneficial (91.4%, n = 213; 92.5%, n = 199), followed by nutrition (91.3%, n = 211; 92.5%, n = 198). The participants were most likely to get their information about recovery strategies from a fellow athlete (46.3%, n = 111), a coach (42.5%, n = 102), or a website (30.4%, n = 74). CONCLUSION: Hydration is the most common recovery strategy and found to be beneficial in both training and competition settings. These athletes are relying on the people around them and websites for information and recommendations. Endurance athletes should be educated on other strategies to address the multidimensionality of recovery. These findings will be useful for healthcare professionals, practitioners, and coaches in understanding recovery strategies in endurance athletes

Toll-Like Receptors and Dectin-1, a C-Type Lectin Receptor, Trigger Divergent Functions in CNS Macrophages

Spinal cord injury (SCI) activates macrophages, endowing them with both reparative and pathological functions. The mechanisms responsible for these divergent functions are unknown but are likely controlled through stochastic activation of different macrophage receptor subtypes. Various danger-associated molecular patterns released from dying cells in the injured spinal cord likely activate distinct subtypes of macrophage pattern recognition receptors, including bacterial toll-like receptors (TLRs) and fungal C-type lectin receptors (e.g., dectin-1). To determine the in vivo consequences of activating these receptors, ligands specific for TLR2 or dectin-1 were microinjected, alone or in combination, into intact spinal cord. Both ligands elicit a florid macrophage reaction; however, only dectin-1 activation causes macrophage-mediated demyelination and axonal injury. Coactivating TLR2 reduced the injurious effects of dectin-1 activation. When injected into traumatically injured spinal cord, TLR2 agonists enhance the endogenous macrophage reaction while conferring neuroprotection. Indeed, dieback of axons was reduced, leading to smaller lesion volumes at the peak of the macrophage response. Moreover, the density of NG2+ cells expressing vimentin increased in and near lesions that were enriched with TLR2-activated macrophages. In dectin-1-null mutant (knock-out) mice, dieback of corticospinal tract axons also is reduced after SCI. Collectively, these data support the hypothesis that the ability of macrophages to create an axon growth-permissive microenvironment or cause neurotoxicity is receptor dependent and it may be possible to exploit this functional dichotomy to enhance CNS repair. SIGNIFICANCE STATEMENT: There is a growing appreciation that macrophages exert diverse functions in the injured and diseased CNS. Indeed, both macrophage-mediated repair and macrophage-mediated injury occur, and often these effector functions are elicited simultaneously. Understanding the mechanisms governing the reparative and pathological properties of activated macrophages is at the forefront of neuroscience research. In this report, using in vitro and in vivo models of relevance to traumatic spinal cord injury (SCI), new data indicate that stochastic activation of toll-like and c-type lectin receptors on macrophages causes neuroprotection or neurotoxicity, respectively. Although this manuscript focuses on SCI, these two innate immune receptor subtypes are also involved in developmental processes and become activated in macrophages that respond to various neurological diseases

Transition from an M1 to a Mixed Neuroinflammatory Phenotype Increases Amyloid Deposition in APP/PS1 Transgenic Mice

BACKGROUND: The polarization to different neuroinflammatory phenotypes has been described in early Alzheimer\u27s disease, yet the impact of these phenotypes on amyloid-beta (Aβ) pathology remains unknown. Short-term studies show that induction of an M1 neuroinflammatory phenotype reduces Aβ, but long-term studies have not been performed that track the neuroinflammatory phenotype. METHODS: Wild-type and APP/PS1 transgenic mice aged 3 to 4 months received a bilateral intracranial injection of adeno-associated viral (AAV) vectors expressing IFNγ or green fluorescent protein in the frontal cortex and hippocampus. Mice were sacrificed 4 or 6 months post-injection. ELISA measurements were used for IFNγ protein levels and biochemical levels of Aβ. The neuroinflammatory phenotype was determined through quantitative PCR. Microglia, astrocytes, and Aβ levels were assessed with immunohistochemistry. RESULTS: AAV expressing IFNγ induced an M1 neuroinflammatory phenotype at 4 months and a mixed phenotype along with an increase in Aβ at 6 months. Microglial staining was increased at 6 months and astrocyte staining was decreased at 4 and 6 months in mice receiving AAV expressing IFNγ. CONCLUSIONS: Expression of IFNγ through AAV successfully induced an M1 phenotype at 4 months that transitioned to a mixed phenotype by 6 months. This transition also appeared with an increase in amyloid burden suggesting that a mixed phenotype, or enhanced expression of M2a and M2c markers, could contribute to increasing amyloid burden and disease progression

Determining the Role of IL-4 Induced Neuroinflammation in Microglial Activity and Amyloid-ß Using BV2 Microglial Cells and APP/PS1 Transgenic Mice

Background Microglia are considered the resident immune cells of the central nervous system (CNS). In response to harmful stimuli, an inflammatory reaction ensues in which microglia are activated in a sequenced spectrum of pro- and antiinflammatory phenotypes that are akin to the well-characterized polarization states of peripheral macrophages. A “classically” activated M1 phenotype is known to eradicate toxicity. The transition to an “alternatively” activated M2 phenotype encompasses neuroprotection and repair. In recent years, inflammation has been considered an accompanying pathology in response to the accumulation of extracellular amyloid-β (Aβ) in Alzheimer’s disease (AD). This study aimed to drive an M2a-biased immune phenotype with IL-4 in vitro and in vivo and to determine the subsequent effects on microglial activation and Aβ pathology. Methods In vitro, exogenous IL-4 was applied to BV2 microglial cell cultures to evaluate the temporal progression of microglial responses. In vivo, intracranial injections of an adeno-associate-virus (AAV) viral vector were performed to assess long-term expression of IL-4 in the frontal cortex and hippocampus of Aβ-depositing, APP/PS1 transgenic mice. Quantitative real-time PCR was used to assess the fold change in expression of biomarkers representing each of the microglial phenotypes in both the animal tissue and the BV2 cells. ELISAs quantified IL-4 expression and Aβ levels. Histological staining permitted quantification of microglial and astrocytic activity. Results Both in vitro and in vivo models showed an enhanced M2a phenotype, and the in vivo model revealed a trend toward a decreased trend in Aβ deposition. Conclusions In summary, this study offers insight into the therapeutic potential of microglial immune response in AD

Determining the role of IL-4 induced neuroinflammation in microglial activity and amyloid-β using BV2 microglial cells and APP/PS1 transgenic mice

Background Microglia are considered the resident immune cells of the central nervous system (CNS). In response to harmful stimuli, an inflammatory reaction ensues in which microglia are activated in a sequenced spectrum of pro- and antiinflammatory phenotypes that are akin to the well-characterized polarization states of peripheral macrophages. A “classically” activated M1 phenotype is known to eradicate toxicity. The transition to an “alternatively” activated M2 phenotype encompasses neuroprotection and repair. In recent years, inflammation has been considered an accompanying pathology in response to the accumulation of extracellular amyloid-β (Aβ) in Alzheimer’s disease (AD). This study aimed to drive an M2a-biased immune phenotype with IL-4 in vitro and in vivo and to determine the subsequent effects on microglial activation and Aβ pathology. Methods In vitro, exogenous IL-4 was applied to BV2 microglial cell cultures to evaluate the temporal progression of microglial responses. In vivo, intracranial injections of an adeno-associate-virus (AAV) viral vector were performed to assess long-term expression of IL-4 in the frontal cortex and hippocampus of Aβ-depositing, APP/PS1 transgenic mice. Quantitative real-time PCR was used to assess the fold change in expression of biomarkers representing each of the microglial phenotypes in both the animal tissue and the BV2 cells. ELISAs quantified IL-4 expression and Aβ levels. Histological staining permitted quantification of microglial and astrocytic activity. Results Both in vitro and in vivo models showed an enhanced M2a phenotype, and the in vivo model revealed a trend toward a decreased trend in Aβ deposition. Conclusions In summary, this study offers insight into the therapeutic potential of microglial immune response in AD

Acute multi-sgRNA knockdown of KEOPS complex genes reproduces the microcephaly phenotype of the stable knockout zebrafish model

Until recently, morpholino oligonucleotides have been widely employed in zebrafish as an acute and efficient loss-of-function assay. However, off-target effects and reproducibility issues when compared to stable knockout lines have compromised their further use. Here we employed an acute CRISPR/Cas approach using multiple single guide RNAs targeting simultaneously different positions in two exemplar genes (osgep or tprkb) to increase the likelihood of generating mutations on both alleles in the injected F0 generation and to achieve a similar effect as morpholinos but with the reproducibility of stable lines. This multi single guide RNA approach resulted in median likelihoods for at least one mutation on each allele of >99% and sgRNA specific insertion/deletion profiles as revealed by deep-sequencing. Immunoblot showed a significant reduction for Osgep and Tprkb proteins. For both genes, the acute multi-sgRNA knockout recapitulated the microcephaly phenotype and reduction in survival that we observed previously in stable knockout lines, though milder in the acute multi-sgRNA knockout. Finally, we quantify the degree of mutagenesis by deep sequencing, and provide a mathematical model to quantitate the chance for a biallelic loss-of-function mutation. Our findings can be generalized to acute and stable CRISPR/Cas targeting for any zebrafish gene of interest

Understanding the Role of Schools in Community Prevention of Childhood Obesity

Childhood obesity is considered an epidemic with the percentage of obesity among children in the United States more than tripling in the past 30 years (Institute of Medicine, 2012). Many factors in the environment contribute to this problem including increased availability of unhealthy foods and limited access to healthy foods, increased screen time and lack of accessible and safe places for physical activity (Center for Disease Control and Prevention, 2013). Since many of these factors are often shared by a community due to common experience, successful interventions should target the community. When planning community interventions, it is important to link community resources such as city parks with neighborhoods where children go to school and live. Community partners that include city parks, neighborhoods, schools, local government, and health care professionals are essential to developing successful childhood obesity prevention practices. Past reviews of studies found that nutrition education, increased opportunities for physical activity and use of the school as a resource are crucial in preventing and curbing childhood obesity (Bleich, Segal, Wu, Wilson & Wang, 2013; Waters et al., 2011). / / This honors project was conducted in rural eastern North Carolina at a school-based health center located in a public school in collaboration with a school nurse. The school has a student population of 460 in 5th through 8th grade. In eastern North Carolina 17.4% of children aged 2-17 years are obese (North Carolina State Center for Health Statistics, 2011). The purpose of this project is integrate community resources related to healthy food choices and physical activity with school-age children in the school setting. The first objective of this project was to interview 4-5 key informants, about what community resources, such as parks and community gardens are available and how the school uses these resources to improve food choices and physical activity. The second objective was to provide written recommendations on the integration of community resources with school-age children to increase healthy food choices and physical activity levels. Public health interventions used in this project were collaboration and coalition building with community partners.

The Effects of Face Coverings on Perceived Exertion and Attention Allocation during a Stepping Task

The COVID-19 pandemic has impacted the entire world from lockdowns to various recommended restrictions including social distancing and wearing face coverings. In a safe environment, cardiovascular exercise is important for both physical health and mental health. The current study examined the effects of face coverings on rating of perceived exertion and attention allocation during an exertive stepping task. Participants completed a stepping task with a weighted vest at 20% of their bodyweight until volitional fatigue with a face covering (n = 23) or without a face covering (n = 31). Results revealed a non-significant difference (p = 0.25) in the duration of the stepping task (in seconds) between the no face covering (M = 455.81, SD = 289.77) and face covering (M = 547.83, SD = 285.93) conditions. Results indicated increases in perceived exertion (p < 0.001) and heart rate (p < 0.001) as time progressed across the four time points (i.e., 30 s, 1/3 time to exhaustion, 2/3 time to exhaustion, and exhaustion) in both conditions. No significant differences were found between the conditions for RPE (p = 0.09) and heart rate (p = 0.50). Participants wearing a face covering were more internally focused across the duration of the stepping task (p = 0.05). This study has relevance for applied practitioners implementing physical activity interventions that require face coverings