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Nicotinic receptors (nAChRs) have long been investigated as targets for modulation of pain and inflammation. The α7 subtype of nAChR has more recently been demonstrated to have an important role in pain and inflammation. These receptors are expressed widely in neuronal and immune cells, including myeloid lineage cells, which mediate the endogenous cholinergic anti-inflammatory pathway in cultured immune cells. Complete knock-out of α7-nAChRs in mice results in increased inflammation and hyperalgesia, while α7-nAChR gain of function mice experience attenuation of hyperalgesia and inflammation. However, the mechanism of α7-nAChR attenuation of inflammation is not fully understood. We hypothesize that α7-nAChRs expressed on myeloid lineage cells play an important role in nociception and inflammation in vivo. Conditional knock-out mice deficient in α7-nAChR expression in myeloid cells (α7 cKO) and functionally wild-type (WT) mice were generated using Cre-loxP methodology. Acute inflammation was modeled in the mice using a surgical model in which the hind paw was incised and a chemical model in which mice were injected with LPS. Chronic inflammation was modeled using CFA injected in the dorsal hind paw. Evoked thermal hypersensitivity, mechanical hypersensitivity, and paw edema were measured. Pro-inflammatory cytokine expression in the dorsal root ganglia (DRG) and spinal cord after CFA injection was measured using qRT-PCR. In the surgical and chemical acute inflammatory pain model, all mice in the treatment group experienced increased paw withdrawal latency and increased paw edema, but there was no significant difference in thermal or mechanical hyperalgesia or paw diameter between α7 cKO and WT groups. In the chronic inflammatory pain model, however, a greater degree of hyperalgesia and paw edema was seen in α7 cKO mice compared to WT mice. Additionally, expression of pro-inflammatory cytokines IL-1β and TNF-α was increased in the DRG. In the acute inflammatory pain model, the absence of a significant difference between α7 cKO and WT mice indicates that α7-nAChRs in myeloid lineage cells may not play an important role in the modulation of acute inflammation. In contrast, in the chronic inflammatory model, significantly increased hyperalgesia and paw edema of α7 cKO mice suggests that deficiency of α7-nAChRs in myeloid cells contributes to increased inflammatory response. This is further demonstrated by the elevation of pro-inflammatory cytokine expression in the DRG after CFA, indicating that α7-nAChRs in myeloid lineage cells mediate attenuation of chronic inflammation, making them useful targets for the treatment of chronic pain.

AMA Research Challenge 2024 

The Role of Alpha-7 Nicotinic Acetylcholine Receptors in Myeloid Lineage Cells in Acute and Chronic Inflammatory Pain

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Background:
Psychedelic compounds have long been known for their ability to profoundly alter human consciousness, even at microgram doses. Psilocybin, in particular, has recently garnered significant attention due to its potential therapeutic benefits in treating many mental health disorders. Despite promising clinical results, the neural mechanisms underlying psilocybin's effects remain poorly understood. Psilocybin acts on the brain via multiple neurotransmitters, including serotonin and dopamine. On a whole-brain level, psilocybin decreases activity in the default mode network (DMN) and medial prefrontal cortex (mPFC), while increasing connectivity between the DMN, frontoparietal network, and salience network. These changes are thought to underlie the "ego dissolution" and synesthesia-like effects often reported during psilocybin-induced mystical experiences. However, a major knowledge gap remains in linking these global network effects to the initial receptor-level actions of the drug.
Methods:
To better understand how psilocybin affects the brain, we are combining molecular biology, in vivo imaging, and novel behavioral assays. First, we treated mice with either psilocybin or saline and isolated nuclei from the medial prefrontal cortex (mPFC) 24 hours later for single-nucleus RNA sequencing. This allowed us to identify distinct neuronal cell types (e.g., excitatory vs. inhibitory, different cortical layers) based on their unique gene expression profiles. Next, we performed live 1- and 2-photon microscopy in freely-moving and head-fixed mice with implanted lenses over the mPFC to visualize excitatory neural activity during and after psilocybin administration. Finally, we developed a novel behavioral assay using a high-fidelity camera to record mouse movement on a 2-dimensional linear track, enabling a detailed, AI-driven analysis of kinematic parameters.
Results:
Our molecular profiling experiment revealed that psilocybin treatment upregulated genes involved in excitatory synapse assembly. Excitatory neurons in layers V and VI were particularly affected, including a specific subtype enriched for the serotonin 2C receptor. Our 2-photon imaging data showed that psilocybin decreased mPFC activity in response to learned, positively salient cues. Ongoing analysis of recently acquired 1-photon imaging data will uncover the acute effects of psilocybin in real-time. Behaviorally, acute psilocybin administration led to decreased rearing and grooming, changes in gait and stance, and preserved overall mobility.
Conclusions:
We hope that this work will contribute to the understanding of how psychedelics affect the brain on a molecular, behavioral and neural network level both acutely and longer-term. In doing so, we may begin to elucidate how the natural function of the brain utilizes these pathways for neuroplasticity and consciousness.

Uncovering the molecular, neural circuit, and behavioral mechanisms of psilocybin action

Background : Every year there are more than 15,000 new spinal cord injuries in the United States.1 Of those, a complete spinal cord transection, characterized by a lesion through both the motor and sensory tracts, is one of the most devastating injuries to patients. During the subacute and chronic phases of injury, glial cells in vertebrae species accumulate at the lesion site to provide a protective barrier through scar formation. Although the glial scar is imperative in protecting against additional tissue loss, it blocks axon regeneration, causing additional loss of function. A powerful model that lacks this glial scar formation is the zebrafish (Danio rerio). It’s genetic commonalities to humans and larval translucency make it one of the more resourceful models when studying new pharmacological therapeutics for spinal cord injury.2 4-Aminopyridine (4-AP) is a therapeutic that is inhibitory to voltage-gated potassium channels (Kv) allowing for increasing action potentials across demyelinated axons.3 It is essential to understand 4-AP’s therapeutic effect on glial cells spinal cord injury patients in the future and the zebrafish model allows a window to discover its role in glial cell proliferation, migration, and remodeling.

Methods: A complete spinal cord injury was performed on an Tg(gfap:EGFP) x Tg(elavl3:mcherry) transgenic zebrafish line five days post fertilization. On one day post injury, continuous dosing of E2 recovery buffer supplemented with either DMSO or 4-AP was given to an equal number of transected zebrafish and uninjured zebrafish. Utilizing the Leica Thunder System, Z stack images at 20x were taken from day of injury until five days post injury to analyze pixel intensity of the glial cell bridging to ensure a complete transection and progress of bridging. Zebrafish were fed one day during the recovery period and multi-well plates were replenished with new recovery buffer each day after imaging.

Results: Continuous dosing of 4-AP one-day post injury increased glial cell proliferation at both the rostral and caudal ends of the lesion site. The 4-AP zebrafish also had narrower injury sites by the fifth day post injury. The DMSO zebrafish showed higher proliferation at the rostral lesion site.

Conclusion: In closing, it was found that continuous dosing of 4-AP one day post injury enhances glial cell bridging, promoting regeneration of the spinal cord in the zebrafish model. Varying the dosing schedules and monitoring locomotion behaviors are needed to advance the knowledge of 4-AP's effects on regeneration after spinal cord injury.

References:

(Spinal Cord Injury Prevalence In The U.S. | Reeve Foundation (christopherreeve.org)).

Effectiveness of zebrafish models in understanding human diseases—A review of models - ScienceDirect)

Neuroprotective Properties of 4-Aminopyridine - PMC (nih.gov) Neuroprotective Properties of 4-Aminopyridine - PMC (nih.gov)

Building bridges, not walls: spinal cord regeneration in zebrafish - PMC (nih.gov)


Understanding 4-AP's Capabilites to Enhance Axonal & Glial Spinal Cord Regeneration after Injury Utilizing a Zebrafish Model

As the field of genomics continues to grow, the identification of substantial genes as it relates to rare diseases remains a significant challenge. Traditionally, researchers work backwards starting from the disease to determine associated proteins. Then use expensive and time-consuming methods, such as mass spectroscopy or immunofluorescence staining, to determine genetic codes. While this method may work with single gene disorders, it is lacking when dealing with complex, polygenic diseases. To address this, we aimed to develop a novel method of discovering genes that displayed similar phenotypes when knocked out in mice using data from the International Mouse Phenotyping Consortium (IMPC), streamlining gene identification for complex genetic conditions.
We used the National Organization for Rare Diseases (NORD) to select the Map2K2 gene, associated with cardiofaciocutaneous syndrome, as our starting gene. We then used the IMPC database to identify phenotypes associated with Map2k2 knockout mice. These phenotypes were organized into an Excel spreadsheet. Next, we searched the IMPC database for genes that exhibited the same phenotypes when knocked out in mice. This information was compiled into separate Excel tabs, one for each phenotype. We then applied variations of a conditional format to visualize the frequencies of each gene, assigning specific colors based on the number of hits. 
The IMPC database revealed five phenotypes associated with Map2k2 knockout mice: increased mean corpuscular volume, decreased total body fat, decreased body weight, abnormal gait, and decreased body length. Each phenotype had varying amounts of contributing genes, the most being decreased body weight with 403 associated genes, and the least being abnormal gait with 241 associated genes. Of the 947 unique genes found, 129 genes contributed to two phenotypes, 24 contributed to three phenotypes, and four contributed to four phenotypes. Notably, the gene Slc38a10 was the only gene associated with all five Map2k2 knockout phenotypes.
While the Map2k2 gene’s involvement in the MAPK signaling pathway has been extensively studied, literature regarding interacting genes has been sparse. Our analysis identified Slc38a10 as a potential interacting gene, suggesting interplay between the pathways regulated by Map2k2 and Slc38a10. The Slc38a10 gene encodes an amino acid transporter crucial for metabolism and neurotransmission. Given the role of Map2k2 in the MAPK signaling pathway, it is plausible that these two genes have downstream interactions.
Our gene detection method proved to be inexpensive and quick, offering a valuable tool for future genetic research. We hope that novel approaches like ours will contribute to enhancing the understanding of complex genetic disorders and advancing personalized medicine.


Unlocking Gene Interactions: Novel Insights from the IMPC Database

Introduction
Normal weight type-2 diabetes (NWD), characterized by body mass index (BMI) below 25 kg/m2, is more prevalent amongst Asian Americans (AA). While recent studies have shown strength training (ST) was superior to aerobic (AER) or combination training (COMB) in lowering HbA1c levels in individuals with NWD, it is unclear if these exercises have the same effects across AA subgroups, which differentiate due to cultural norms.

Methods
We used data from a 9-month randomized control trial for NWD (conducted in San Francisco Bay Area in 2019-2020) to evaluate the effectiveness of weekly planned ST, AER, and COMB training sessions for Chinese and Asian Indian populations. Our primary outcome was change in diabetic control (HbA1c). Secondary outcomes included changes in body composition (e.g. fat and lean mass) and muscle strength (e.g. leg extension).We performed intention-to-treat (ITT) and per-protocol analysis (PP, restricted to those with 50% minimum adherence) to evaluate changes to HbA1c and body composition using repeated measures ANOVA and paired t-tests.

Results
Of 129 participants, 59 were Chinese (19 ST, 20 AER, 20 COMB), and 70 were Asian Indian (26 ST, 20 AER, 24 COMB). Average baseline HbA1c was 7.5% for both groups (p=0.96). Chinese participants (77.3%) showed higher adherence than Asian Indian (56.6%) participants. Chinese participants showed reduced HbA1c for each intervention group (ITT: ST -0.42% [-0.742, -0.11], AER -0.6% [-0.92 -0.28], and COMB -0.13% [-0.43, 0.17], all p < 0.05), with AER intervention yielding the largest decrease (p<0.01). Results from PP provided a similar conclusion. Asian Indians had no meaningful reductions in HbA1c levels ( ITT: ST -0.36% [-0.83, -0.11], AER 0.26% [-0.27, 0.78], and COMB -0.42% [-0.97, 0.13]. For body composition, Chinese participants showed significantly reduced FMI-Z (ST: -1.03 [-1.94, -0.12], AER: -0.17 [-0.31, -0.04], COMB: -0.17 [-0.23, -0.10]) in all intervention groups. Asian Indian participants only showed significant reduction in FMI-Z (fat mass adjusted for height, age, sex) in the ST group -0.22 [-0.40, -0.03]. Body composition changes were associated with improvement in HbA1c.

Conclusion
Overall, AA persons in over 50% of prescribed exercise improved their diabetic control. In Chinese participants, all interventions lowered HbA1c with higher reductions observed in ST and AER interventions. They also experienced significant reductions in fat mass in all exercise interventions. Asian Indian participants showed no significant reductions in HbA1c in any exercise intervention, but showed significant reductions in fat mass in ST. By recognizing unique cultural and social factors that influence AA behaviors and choices, healthcare professionals can develop tailored interventions that foster cultural understanding, thus empowering individuals to take control of their health and reduce the burden of diabetes.

Is Strength or Cardio Training More Effective at Affecting Diabetes Control & Body Composition for Asians? Subgroup Analysis of a Randomized Control Trial.

Introduction: Blood lead screening has become a critical preventive health element of pediatric well-child appointments, as exposure to lead can lead to developmental delays and many other serious health consequences. However, research shows that clinics face difficulty carrying out lead screening recommendations. 
Purpose: This project evaluated rates of lead screening at the UT St. Francis Family Medicine Residency clinic according to the Tennessee Department of Health (TN DOH) guidelines and proposed quality improvement strategies based on several factors that potentially prevent routine screenings. 
Methods: A retrospective chart review was conducted using a random sample of 353 pediatric patients aged 12 to 72 months who had established UT-St. Francis as their primary care clinic. We utilized a retrospective chart review to determine if patients had 12- and 24-month well-child appointments, if lead screenings were completed at these times, and the blood lead levels reported at each screening. We also investigated whether any acute medical issues were coded in patient charts for these well-child encounters and the rates of “catch-up” screenings among patients who had never been screened for lead. To assess physician awareness of lead screening recommendations, an anonymous questionnaire was distributed to the UT St. Francis Family Medicine residents, attendings, and fellows.​
Results: 47% of the sampled patients were screened for lead at 12-month well-child appointments, and 26% were screened at 24 months. This study identified several likely clinical barriers to the completion of the TN DOH recommended lead screenings, which included clinical workflow and few systems-based reminders but excluded physician awareness. Based on these results, we developed several suggestions for future quality improvement projects. 
Conclusions: Our recommendations directly address the identified causes of missed screenings from this study’s analyses and hold potential to increase future lead screening rates based on the clinic’s specific needs. Future study should investigate the short- and long-term effects of these recommendations on subsequent lead screening rates.

Lead Screening in Pediatric Patients at the UT St. Francis Family Medicine Clinic​

Background
Since the COVID-19 pandemic’s onset, utilizing virtual visits has increased significantly. Although a beneficial aspect of implementing virtual visits is to increase access, there have been few studies with a focus on the opinions of African Americans or uninsured patients, populations where distrust in physicians and healthcare is more prevalent. It has been reported that patient-physician trust leads to better self-reported health outcomes, patient satisfaction, health behaviors, quality of life, and decreased symptom severity. Therefore, identifying ways to
increase trust is essential to improving healthcare, especially among these groups.
Our study aims to evaluate the knowledge, attitudes, and experiences of the uninsured community living in Detroit regarding virtual care and trust in physicians and the healthcare system. We hypothesize that factors such as virtual care and free clinic use will be correlated with increased trust.
Methods
This project involves three student-run free clinics in a large Midwestern metropolitan city. We are conducting a survey-based research study evaluating the knowledge, attitudes, and experiences of uninsured people with virtual healthcare services, physician trust, and trust in the overall healthcare system using combined, previously published, survey instruments. A report of the demographics and average responses will analyze the data. Then, using t-tests and ANOVA via the SPSS platform, the effect of virtual care and free clinic use on trust will be evaluated.
Results
Data collection is currently in progress. At this point in time, we have a sample size of 48 uninsured participants above the age of 18 years and approximately an 89% response rate. We aim to have over 50 participants by the conference deadline in order to extrapolate more meaningful interpretations from data analysis. Currently, 71.8% of the participants are African American and 75% earn less than $40,000 per year. Our initial findings show that 84.4% of participants have utilized free clinics before, with 96.4% of those reporting an extremely or mostly positive experience. Furthermore, preliminary analysis finds that 90.6% of participants trust their physician’s judgment about their medical care. In regard to telemedicine, 43.8% have previously accessed health care via phone or video chat, although only 23.8% of users report obtaining “better access to healthcare services by use of telemedicine”.
Conclusion
Further research is necessary to illuminate perceptions of access and trust in free clinics and telehealth services among underserved populations and to determine mechanisms to implement to improve these views.

Perception of Trust, Virtual Care, and Access Among the Uninsured Population: A Multi-Site Study at Student-Run Free Clinics

Background: Trauma, characterized by persistent emotional distress following exposure to a distressing event, is linked to adverse physical and mental health outcomes. Both childhood and adulthood trauma have been associated with increased risk of chronic diseases and psychiatric disorders in adulthood. According to literature, African Americans, compared to their White counterparts, experience higher levels of trauma and greater prevalence of chronic diseases. This study investigates whether African American adults with a history of trauma, regardless of
timing, exhibit a higher chronic disease burden. Secondarily, the study explores the relationship between traumatic experiences and emotional/psychiatric issues within this population.

Methods: This secondary data analysis utilized data from the Health and Retirement Study to examine the association between chronic disease burden and lifetime trauma, childhood trauma, and adulthood trauma. Multiple linear regression analyses were conducted to assess the relationship between each trauma category and chronic disease burden. Multiple logistic regression models were also implemented to examine associations between trauma categories and emotional/psychiatric problems, adjusting for sociodemographic and health-related variables.

Results: Trauma, regardless of timing, was significantly associated with increased chronic disease burden (childhood: p = .008, adulthood: p < .001, lifetime: p < .001) and higher likelihood of emotional/psychiatric problems (childhood: OR = 1.368, 95% CI = 1.220, 1.534; adulthood: OR = 1.375, 95% CI = 1.185, 1.596; lifetime: OR = 1.284, 95% CI = 1.192, 1.382) in adulthood among African American adults. These associations remained robust after adjusting for age, sex, education, BMI, and smoking status.

Conclusion: This study highlights the profound impact of trauma on both physical and mental health outcomes in African American adults. Beyond the well-established biological mechanisms linking trauma to adult health, these findings demonstrate the critical need for healthcare providers to address trauma-related concerns in clinical practice. Encouraging patients to adopt healthy coping strategies and facilitating access to professional support would be essential steps in mitigating the adverse health effects of trauma in this population.

The Association Between Trauma and Chronic Disease Burden in African American Adults

Background: Hairstyles common in Black communities, braids, locs, twists, (BLTs) can present as artifacts, posing unique diagnostic challenges in interpreting radiological imaging, though limited research has been conducted on the current climate of their interpretation.

Purpose: To present a review of hair artifacts in literature and investigate the comfortability and familiarity of physicians and radiology technicians in assessing braids, twists, and locs (BTLs) as hair artifacts on radiological imaging.

Methods: A web-based survey on the experiences and awareness of BTLs on imaging was distributed to physicians in multiple departments within a nationally respected academic medical center. Survey responses were collected between April 2023 through August 2023 and included demographic information, career length, how often they came across hair artifacts in imaging and a quiz to distinguish pathology from hair artifact on chest x-ray images. Respondents also provided reports of their comfortability and familiarity in identifying BTL’s in plain film images.

Results: 46 surveys were collected from various departments, including Temple Radiology (41.3%), Temple Emergency Medicine (21.7%), Temple Jeanes Campus (26.1%), and St. Lukes Bethlehem Radiology Group (2.2%). Demographic data in Table 1 reveal the Radiologist cohort had an average experience of 9.40 years (SD=12.41), Emergency Medicine cohort had 7.11 years (SD=7.08), and Internist cohort had 0.5 years (SD=0.71). Career length (p < 0.0001) and patient load (p = 0.029) significantly affected self-reported scores (p < 0.0001), with longer careers and larger loads associated with higher score of comfort and familiarity among study participants.

Conclusions: Our findings demonstrated that the number of years in practice and the number of patients per day improved physicians’ and technicians' awareness of how Black hairstyles can affect medical imaging. This study presented a radiological phenomenon lacking in literature and provided valuable insights into the impact of experience on physicians' ability to identify and interpret radiographic images affected by styles that can mimic pathology. The conclusions conveyed a need for early education of BLTs in medical training. Further applications of this research may be directed towards comparing pathology localized in lung apices to hairstyles popular in the Black community, assessing the validity of an educational module in increasing awareness, and expanding on patient populations to explore further associations.

The Imperative for Early Education of Common Black Hairstyles in Radiological Imaging: A Pilot Study


Background
This study investigates potential cost reduction by replacing erythromycin with azithromycin for managing PPROM. Effective strategies are vital due to the high risks associated with PPROM. Targeting Hispanic PPROM patients at Centro Medico Episcopal San Lucas in Ponce, Puerto Rico, this research focuses on comparing the cost-effectiveness of azithromycin and erythromycin in extending pregnancy latency.

Methods
Using a retrospective study design with predictive model elements, patient data from 2020 to 2021 at Centro Medico Episcopal San Lucas was analyzed, projecting the expected number of PPROM diagnoses in 2022. Detailed cost data for azithromycin and erythromycin were obtained from hospital’s pharmacy. Costs for erythromycin encompassed the 250mg IV bag and vial, 100 mL saline bag, primary line for initial dosing, and erythromycin 500 mg tablets for treatment continuation. Azithromycin costs were evaluated for each dosing tablet (1g, 250mg, and 500mg). Average total cost of treatment per year was computed based on the projected number of PPROM patients in 2022. Statistical analysis, including a 95% confidence interval, guided the estimation of the sample size.

Results
The expected number of patients with PPROM in 2022 at Centro Médico Episcopal San Lucas is 136 patients. The combined IV and PO erythromycin regimen, incurs a per- patient cost of $449.30, resulting in a total cost of $60,880 for 136 expected PPROM cases in 2022. In contrast, azithromycin costs per patient varies from $1.16 (1g = 4 tablets of 250mg) and $0.98 (1g = 2 tablets of 500mg) with an estimated total cost of $163.50 and $132.80, respectively, for 136 average expected patients in 2022. A population size of 114 measurements are needed for the confidence level to be at 95%. A 99% cost reduction is projected if azithromycin is employed as standard of care for PPROM compared to erythromycin. The economic advantage of azithromycin lies in its lower cost and single-dose regimen, which is effective for managing PPROM. This regimen benefits from improved pharmacokinetics, slower elimination, and rapid intracellular uptake and concentration.

Conclusion
In conclusion, our study provides insights into the potential cost- reduction with the substitution of azithromycin for erythromycin as the standard treatment for PPROM, without compromising clinical efficacy. This study lays the groundwork for conducting a larger clinical trial to further evaluate the safety, efficacy and economic benefits of this treatment approach. The findings of such a trial could have significant implications for optimizing PPROM management protocols and improving healthcare resource utilization.


The Journey for Optimizing Care: Cost-Informed Perspectives on Erythromycin Vs. Azithromycin for PPROM Management in Southern Puerto Rico

There is currently a gap in knowledge regarding the knowledge, trust, and barriers of the Hispanic community regarding the influenza vaccine. A descriptive, mixed quantitative survey was administered to 100 participants at flu clinics as well as various locations throughout Metro Detroit between September 2023 and September 2024 to identify their levels of knowledge, trust, and barriers regarding influenza vaccination. There were a significant proportion of individuals with misconceptions regarding the vaccine, with 59% of respondents unware that it was safe for pregnant women, and 67% under the belief that it could interact with medications. Reported barriers to vaccination included lack of time (31%) and other/unspecified (47%), while only 12% reported a lack of insurance. Contrary to our hypothesis, our study shows that Hispanic patients of Metro Detroit may have sufficient levels of knowledge and trust regarding the influenza vaccine. Barriers may primarily be time-related, as opposed to access-related.

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The Role of Alpha-7 Nicotinic Acetylcholine Receptors in Myeloid Lineage Cells in Acute and Chronic Inflammatory Pain

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Uncovering the molecular, neural circuit, and behavioral mechanisms of psilocybin action

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