bluuebunny/crossref_metadata_2025 huggingface dataset:
Single Cell Sequencing of Myeloid Cells in Human Hypertension
Authors: Smart Duncan , Madhur Meena S. , Van Beusecum Justin P.
Published: May 2022

Hypertension is a major modifiable risk factor for death and cardiovascular disease, yet its causes are largely unknown. Monocytes and monocyte‐derived dendritic cells (DCs) are activated in hypertension producing cytokines that drive T cell activation. Moreover, DCs from hypertensive mice promote hypertension when adoptively transferred into normotensive mice. The specific human subsets of antigen presenting cells (APCs) that promote human hypertension have yet to be described at a single cell level. We isolated total peripheral blood mononuclear cells (PBMCs) from normotensive (n=5) and hypertensive (n=4) human subjects. We sorted for HLA‐DR+ and CD19‐ cells by magnetic bead separation. Single cell sequencing was performed on a 10x Chromium platform. After performing quality controls, we found a total of 10 clusters that were expressed in both normotensive and hypertensive human subjects. KEGG Pathway analysis on all HLA‐DR+ myeloid cells revealed that pathways associated with oxidative phosphorylation, toll‐like receptor signaling and interleukin (IL)‐17 signaling were increased in hypertensive subjects. Conversely, pathways associated with antigen processing and presentation and cell adhesion molecules were enriched in normotensive subjects. Hypertensive subjects had a significant increase in CD14+ CD36+ monocytes compared to controls (p=0.009). Conversely, CD14+ CD36‐ monocytes and plasmacytoid DCs were significantly reduced in hypertensive subjects (p=0.002 and p=0.0008, respectively). Of particular interest were a cluster of pro‐inflammatory monocytes with expression of CD14 CCL3, IL1B, CD83, TNF, and SGK1. Within this cluster, CXCL8, the gene encoding interleukin (IL)‐8, and CCL3 were significantly increased in hypertensive subjects compared to controls (adjusted p value = 3.0E‐106and 2.0E‐107, respectively). Further studies are needed to investigate CXCL8 (IL‐8) and its role in myeloid cell activation and human hypertension.
MANGO: pipeline for identifying genomic translocations with key biological significance in inflammatory breast cancer
Authors: Morales Carlos S. , Peterson‐Peguero Esther , Driscoll Emilia
Published: May 2022

Genomic instability in cancer is essential for aberrant gene expression and translocation events that potentiate oncogenesis. Thus gene fusions, nonsense mutations and gene deregulation are key contributors to oncogenic phenotypes. Certain genomic translocations have been linked to cancer subtypes, such BCR—ABL1 for chronic myeloid leukemia (CML) serving as potential onco‐biomarkers. Inflammatory breast cancer (IBC) is the rarest and most lethal subtype of breast cancer with a 5‐year survival rate below 50%. Ethnic disparities for breast cancer exist; negative estrogen (ER) and progesterone (PR) hormone receptor status is more frequent in Hispanic and African American women. In addition, inflammatory breast cancer lacks defined molecular markers limiting its early onset detection which leads to poorer overall prognosis. Thus, we hypothesized that profiling IBC translocations, including its subtypes, show valuable insights to better understand this complex disease. We developed MANGO, a pipeline that leverages STAR, STAR‐Fusion, FusionInspector, and Trinity to identify, visualize and assemble gene fusions from IBC sequencing data. Using the SUM149PT cell line as a model for IBC, we identified and experimentally validated a model fusion gene ZDHHC5—EPB41L5. Fusion unaware sequence assemblies cross‐validated the fusion and its multiple splice sites, one of which encodes a fusion peptide. Differential gene expression between SUM149PT and SUM190PT, an IBC cell line without evidence for the identified translocation, showed loss of expression of ZDHHC5 but no change in EPB41L5, suggesting the latter retained its expression in its new genomic localization. Furthermore, in‐silico functional predictions for experimental results reveals an in‐frame fusion peptide with transmembrane signals in EPB41L5. Genomic translocation mapping reveals triple‐negative IBC diverges from traditional breast cancer translocation patterns. In short, MANGO serves as a powerful tool to identify biologically significant translocations involved in IBC and other cancer classifications.
Longitudinal profiling of the Plasma Glycome from Normal and Alzheimer’s Disease individuals
Authors: Arnett Bryson P. , Hawkinson Tara , Young Lyndsay , Clarke Harrison , Buoncristiani Michael , Gentry Matthew , Sun Ramon
Published: May 2022

Alzheimer’s disease (AD) is an incurable dementia known for its neurodegenerative pathologies including amyloid beta plaques, hyperphosphorylated neurofibrillary tau tangles, and brain glucose hypometabolism. One key metabolic pathway that also utilizes glucose is glycan synthesis, which is also known to be disrupted in AD. Glycosylation is an essential protein modification which requires glucose flux through the hexosamine pathway. Prior research in our lab has demonstrated glycosylation abnormalities can be detected in the peripheral blood of early‐stage AD patients. Recently, our lab has developed effective workflows to analyze glycogen and glycan content in plasma utilizing multiple mass spectrometers, including a gas chromatography mass spectrometer (GCMS) and a matrix‐assisted laser desorption/ionization‐mass spectrometer imaging (MALDI‐MSI). In collaboration with Biobank at the University of Kentucky, led by Dr. Pete Nelson, blood plasma samples were collected at various stages of dementia status including AD (N=2), MCI (N=2), and cognitively normal controls (N=3) over a course of 11 years. These samples were subject to metabolite extraction in preparation for GCMS and MALD‐MSI analysis. Utilizing these workflows, we were able to show change in abundance of several metabolite and glycan profiles overtime in both AD and MCI patients compared to our control group. Future analyses will require larger patient cohorts and full metabolic profiling to determine glycosylation differences at various cognitive stages of disease.
Metabolomic characterization of Pseudomonas protegenssecretions for identification of biopesticides against Bacterial Panicle Blight of rice
Authors: Balboa Samantha J. , Vierengel Maia , Gates Kristen , Rojas Clemencia , Hicks Leslie
Published: May 2022

Bacterial Panicle Blight (BPB) is a rice disease caused by Burkholderia glumae that has become a global concern in reducing rice yields. Currently, there are no effective chemical treatments or resistant cultivars to control BPB. We have observed that bacterial secreted fractions from the antagonistic microorganism Pseudomonas protegens PBL3 inhibits B. glumae infections in rice plants. However, the responsible antimicrobial molecules are unknown. Genome comparison between P. protegens PBL3 and the reference strain P. protegensCHA0, a well‐characterized microbial biological control agent, revealed the presence of 14 known biosynthetic gene clusters (BGCs) in P. protegensPBL3. From this genomic data, it is likely that P. protegensPBL3 secretes metabolites with activity against BPB that are not produced by P. protegensCHA0.

Cultures of P. protegensPBL3 and the reference strain, P. protegensCHA0, were grown in M9 minimal media supplemented with 1% myo‐inositol. The metabolite secretome of P. protegens PBL3 and CHA0 were harvested in log phase and analyzed with a Waters Acquity H‐Class HPLC coupled to a Thermo Q‐Exactive HF‐X. In addition, active PBL3 fractions against B. glumaewere analyzed via LC‐MS to confirm the presence of bioactive compounds. The resulting data were searched against the Global Natural Products Social Molecular Networking (GNPS) spectral library. For potential antimicrobial peptide identification, data were searched against the Mascot database. To validate these results, known bioactive metabolites in P. protegensCHA0 were identified. By utilizing LC‐MS/MS based metabolomics, several compounds were uniquely identified in P. protegensPBL3 that could be used as biocontrol agents against BPB in rice.
Folding state as the Decision Maker in Detecting Misfolded Proteins by Hsp70
Authors: Karunanayake Chamithi S. , DelVecchio Rian , Page Richard C.
Published: May 2022

Maintaining cellular proteostasis is challenging due to the constantly changing and crowded cellular environment. Mutations, transcriptional and translational errors, and chemical and pathological stresses in a cell lead to protein misfolding and eventual formation of insoluble toxic aggregates. These aggregates typically result in loss of native protein function and are often associated with conformational diseases such as Alzheimer's, Parkinson's, Huntington's, and various types of cancer. The cellular protein quality control systems are essential for creating a stable cellular environment by resolving the misfolded proteins to uphold a stable proteome. The triage decision for a misfolded protein is regulated by Heat shock protein 70 (Hsp70), the central hub of protein quality control machinery, along with the E3 ubiquitin ligase C‐terminus of Hsc70‐interacting protein (CHIP). Hsp70 itself helps misfolded “client” proteins to refold and directs protein clients to downstream refolding pathways, whereas Hsp70 interaction with CHIP can lead to CHIP‐mediated ubiquitination, ultimately directing the proteins to proteasomal degradation. Understanding the molecular mechanisms that enable the Hsp70/CHIP complex to triage client proteins would allow for the identification of new avenues of treatment for the above‐mentioned diseases. With an objective to illustrate how the varying degrees of folding in a model client protein dictate the interaction between Hsp70 and the client, we have designed a spectrum of client proteins we named folding sensor TPRs (FSTPRs). We initially hypothesized that the more unfolded the client higher the affinity towards Hsp70. Circular dichroism (CD) spectroscopy was used to assess the folding states of each FSTPR variant, and biolayer interferometry (BLI) was utilized to quantify the affinities of each FSTPR towards the Hsp70 substrate‐binding domain (SBD). During our BLI experiments, we observed a range of threshold unfolding percentages that correspond to the highest affinity. Above and below the threshold range, the affinity decreased. This behavior was recapitulated with a disulfide‐locked FSTPR that is folded in the oxidized state and predominantly unfolded in the reduced state. Even though support from further data is necessary, these studies provide us preliminary insight into how Hsp70 interacts with a misfolded client and the role played by folding state in dictating Hsp70/client interactions.
Core Concepts as a Framework to Assess the Progression of Student Learning in a Ume Curriculum
Authors: Averill David , Waite Gabi N. , Sharma Prerna , Mamillapalli Sireesha , Carbe Christian
Published: May 2022

PURPOSE

Conceptual frameworks have demonstrated better learning outcomes in undergraduate medical education (UME). We propose developing a conceptual framework based upon core concepts (CCs) in biomedical sciences to link the progression of student learning to program objectives of a recently revised UME curriculum.

METHODS

Our project involves three steps. Step 1 identifies clinically relevant CCs pertinent to learning objectives of basic science disciplines. Step 2 tags multiple choice questions (MCQs) allowing us to follow students’ progression of conceptual learning across formative and summative assessments and to identify conceptual learning gaps for continuous program improvement. Step 3 will develop enterprise‐grade dashboards to monitor the learner’s conceptual understanding across UME.

RESULTS

For Step 1, we identified 7‐10 clinically relevant CCs in the disciplines of physiology, biochemistry, and genetics. We adapted CCs published by discipline‐specific organizations and by Aquifer, Inc. to our clinical presentation‐based curriculum. The selection process and outcomes will be presented at the meeting. For step 2, we established concept categories in Examsoft™ where our MCQs are housed and are already tagged by discipline and clinical presentation. We will present results of ANOVA‐based analysis of student performance data. For Step 3, we will incorporate CCs with One45 Analytics‐based dashboards which draw upon data from Examsoft™ and our Learning Management System. This is a collaboration with our Department of Assessment and Evaluation. A beta version of the dashboard will be presented at the meeting.

CONCLUSIONS

This study presents student performance data on concept tagged MCQs. These data will track student learning as well as the efficacy of conceptual clinical presentation frameworks implemented in our revised curriculum. Together with high‐quality systemic dashboards, we can monitor conceptual learning and basic science application in our UME curriculum.
SMAD3 is a Mediator of TGF‐β ‐induced Barrier Dysfunction and Hyperpermeability in Microvascular Endothelial Cells
Authors: Tharakan Binu , Hunter Felicia A. , Muthusamy Saravanakumar , Waithe O'lisa Y. , Rao Veena N. , Reddy E. Shyam P. , Childs Ed W.
Published: May 2022

The endothelial transcription factor ETS (Erythroblast‐transformation specific)‐related gene (ERG) regulates canonical TGFβ‐SMAD signaling, by repressing SMAD3 activity. SMAD proteins function as important mediators of intracellular signal transduction of TGF‐β. Following their phosphorylation by TGF‐β receptor‐I, Receptor‐regulated SMADs (including SMAD2 and SMAD3 proteins) form a heteromeric complex with co‐SMAD and then translocate into the nucleus where they bind and regulate the expression of multiple target genes. Recent studies from our lab have demonstrated that ERG is a critical regulator of permeability in human lung microvascular endothelial cells (HLMECs) where ERG gene activation provided protection to the barrier. We have hypothesized that the protective effects of ERG on barrier integrity may be mediated via inhibiting TGF‐β/SMAD3 signaling pathway. To study the role of SMAD3 in microvascular endothelial barrier functions and permeability, HLMECs were grown on Transwell inserts and other cell culture platforms. Monolayer permeability was evaluated based on FITC‐albumin extravasation and measurement of its fluorescence intensity. TGF‐β (1‐50 ng/ml) induced a dose‐dependent increase in monolayer hyperpermeability. TGF‐β‐induced hyperpermeability was attenuated by a SMAD3 pharmacological inhibitor (SIS3) or CRISPR/cas9‐mediated knockdown of SMAD3. Immunoblot analysis demonstrated highly efficient SMAD3 knockdown by SMAD3 CRISPR/Cas9. TGF‐β treatment resulted in significant increase in matrixmetalloproteinase‐9 (MMP‐9) activity and the effect was decreased by SMAD3 inhibition. MMP‐9 is known to proteolytically breakdown the adherens junction proteins in microvascular endothelial cells, resulting in hyperpermeability. These results demonstrate that SMAD3 is a mediator of TGF‐β signaling‐induced loss of endothelial barrier integrity and permeability in microvascular endothelial cells. It also suggests that one of the downstream effects of TGFβ‐SMAD signaling is the activation of MMP‐9 which mediates loss of adherens junctional integrity.
Cord Blood Metabolomics and Autism Spectrum Disorder
Authors: Schickling Brandon , Gumusoglu Serena , Santillan Donna A. , Santillan Mark K.
Published: May 2022

Introduction

Autism spectrum disorder (ASD) has increased dramatically in prevalence and incidence in recent years (1:59 in the US, CDC). With prenatal and/or genetic origins, early detection and intervention are critical to improve outcomes. Few studies have assessed perinatal biomarkers for later development of ASD. We hypothesize that cord blood plasma‐based metabolomics will differentiate infants who would later be diagnosed with ASD and typically developing (TD) controls.

Methods

Banked cord blood plasma samples and clinical data from the Iowa Maternal Fetal Tissue Bank were used. Samples were analyzed via gas chromatography and mass spectrometry (GC‐MS) and relative MS signal intensity of a maximum of was quantified ratiometrically. All metabolites were normalized to internal controls (succinate) to account for effects of extraction, derivatization, and/or loading. Ratiometric levels of each metabolite were compared by t‐test between ASD and control groups. Results P<0.01 are reported given the pilot nature of this small study. Metabolite set enrichment analysis (MSEA) was performed against human libraries. Significantly altered metabolites were compared to a tertiary sample of cord blood from individuals later diagnosed with attention deficit hyperactivity disorder (ADHD) to determine specificity for ASD.

Results

91 metabolites were detected and analyzed. Homocysteine (fold change [FC]: 1.411; p=0.017), Myristic acid (FC: 1.221; p=0.084), Pentadecanoic acid (FC: 1.256; p=0.093) were increased in ASD samples, while L‐Isoleucine (FC: 0.858; p=0.047), L‐Threonine (FC: 0.7131; p=0.068), O‐Phosphoethanolamine (FC: 0.704; p=0.070), and 2‐Hydroxybutyric acid (FC: 0.615; p=0.073) were decreased. Of these, only pentadecanoic was also significantly changed in ADHD samples (FC: 1.514; p=0.002). MSEA revealed enrichment of homocysteine degradation (p=0.017), catecholamine biosynthesis (p=0.048), threonine and 2‐Oxobutanoate degradation (p=0.068), Phosphatidylcholine Biosynthesis (p=0.070), and Phosphatidylethanolamine Biosynthesis (p=0.070) in ASD cord blood samples.

Conclusion

We found that cord blood plasma metabolomics reveals intriguing and specific differences between ASD and control samples. While some of differences are supported by evidence, these alterations have not previously been extended to the prenatal period. Future work will investigate concordance with maternal blood metabolomics to determine maternal‐fetal pathways.
Applying Virtual Screening for Discovery of Novel Inhibitors of Wolbachia Endosymbiont of Brugia malayi Enoyl‐Acyl Carrier Protein Reductase (FabI)
Authors: Chang Annmarie , Beckham Josh
Published: May 2022

Lymphatic filariasis, more commonly known as elephantiasis, is a tropical parasitic disease caused by nematodes. The parasite infects the lymph nodes, and the disease is endemic in 80 countries, affecting hundreds of millions of people worldwide. A potential target for lymphatic filariasis is the enoyl‐(acyl‐carrier‐protein) reductase (FabI) protein of the Wolbachia endosymbiont of Brugia malayi, a filarial nematode. FabI is a significant, and relatively underutilized, drug target due to its essential role in catalyzing the last step of fatty acid elongation in the bacterial fatty acid biosynthesis pathway. The antiparasitic drug diethylcarbamazine is currently used to treat lymphatic filariasis; however, there are no known antibiotics specifically targeting FabI in the Wolbachia endosymbiont of Brugia malayi, necessitating a search for novel inhibitors of this target using virtual screening. Given that there is no published crystal structure of Wolbachia FabI in the Protein Data Bank (PDB), a 3D homology model was created for molecular docking purposes from 3K2E, an enoyl‐(acyl‐carrier‐protein) reductase from Anaplasma phagocytophilum. A pairwise alignment of both sequences resulted in 65% identity match. Using the molecular docking program Genetic Optimization for Ligand Docking (GOLD), which utilizes a genetic algorithm for ranking, several libraries of compounds were virtually screened against the Wolbachia FabI protein and assigned a fitness score of predicted binding affinity based on binding interactions and conformations. Control ligands were GOLD docked first, resulting in a range of scores from 46.09 to 93.24. Tens of thousands of compounds were then screened from libraries including HitFinder9 and Zinc ChemBridge. The top 10% scoring compounds were screened a second time, and the resulting top 10% were saved; ultimately, the top 1% scoring compounds were analyzed. HitFinder9 and Zinc ChemBridge yielded similar results, with top 1% score ranges of 74.45 to 107.63 and 73.63 to 105.16, respectively. Neither library significantly exceeded the other in terms of high scoring compounds. Top chemical compounds from each library can be investigated further for inhibitory activity of Wolbachia FabI in enzyme assays to determine potential viability as novel inhibitors.
Short‐duration Aerobic Exercise Improves Chemotherapy Induced Physical Dysfunction in Mice
Authors: Halle Jessica L. , Counts Brittany R. , Carson James A.
Published: May 2022

Chemotherapy regimens for colorectal cancer cause body weight loss, fatigue, and weakness that may persist long after treatment completion. While increased physical activity can help to mitigate these effects, low adherence to standard exercise prescriptions limits the therapeutic potential. Therefore, it is necessary to determine if an exercise threshold is required to improve chemotherapy toxicity. Gaps remain in our understanding of the extent of chemotherapy toxicities after treatment completion, and if a low dose of exercise is sufficient to improve chemotherapy‐induced dysfunction.

Purpose

To examine the effects of exercise dose on body composition and physical function in mice during the recovery from Folfox chemotherapy.

Methods

Male C57BL/6J mice (12wks) were injected with 4 cycles (1 cycle= 1 injection every other wk) of Folfox (FOL; 5FU 30mg/kg, Oxaliplatin 6mg/kg, Leucovorin 90mg/kg) or PBS. Mice were sacrificed 0wks (n=8) or 4wks (n=16) after the 4th cycle. Subsets of mice performed 14 sessions (6d/wk, 5% grade) of short‐ (15min/d; n=10) or long‐duration (60min/d; n=10) treadmill exercise during recovery. Body composition by MRI, treadmill run time to fatigue (RTF), and grip strength tests were performed pre, post, and after 4wks recovery.

Results

FOL treatment attenuated body mass gain (PBS 10%, FOL 3%) independent of food intake (p=0.586) or cage activity (p=0.801). Fat mass accumulation was decreased (PBS: 56%, FOL: ‐3%) with no changes in lean mass during treatment. During recovery, body mass remained suppressed (‐8%; p=0.022), and RTF (‐43%; p<0.01) and grip strength were reduced (‐9%; p=0.045) in FOL mice compared to PBS. Treadmill exercise improved body weight loss (3%; p=0.041) independent of fat mass. Long‐ compared to short‐duration exercise increased lean mass (p<0.01) in both treatment groups. Both short‐ and long‐duration exercise improved RTF (93%; p<0.01) without improving grip strength.

Conclusion

Repeated cycles of Folfox chemotherapy can induce persistent deficits in physical function and fat mass. Two weeks of short‐duration aerobic exercise was sufficient to reverse chemotherapy induced fatigue without improving strength. Future studies should examine if the combined effects of aerobic and resistance exercise can improve strength.
Hepatic Neddylation Deficiency Triggers Fatal Liver Injury via Inducing NF‐κB‐inducing Kinase and Oxidative Stress
Authors: Chen Weiqin , Xu Cheng , Zhou Hongyi , Jin Yulan , Robicsek Anna , Dong Kunzhe , Zhou Jiliang , Barrett Amanda , Su Huabo
Published: May 2022

The conjugation of NEDD8 to target proteins, termed neddylation is aberrant in many pathological diseases. Its relevance to liver function and failure remains poorly understood. Here, we first identified dysregulated expression of neddylation enzymes (e.g. NAE1, a regulatory subunit of the only NEDD8 E1 enzyme) in human HBV‐induced acute liver failure (ALF). Embryonic–onset deletion of NAE1 in hepatocytes led to severe hepatomegaly, formation of numerous cysts, inflammation, and fibrosis, which ultimately resulted in ALF in mice. Adult–onset hepatic deletion of NAE1 caused ALF with 100% mortality within 30 days post deletion. Mechanistically, hepatic neddylation deficiency triggers glutathione deficiency, oxidative stress, mitochondrial dysfunction, and reprogramming of mature hepatocytes to fetal‐like cells, potentiating liver injury. Importantly, we identified NF‐κB‐inducing kinase (NIK), a stress kinase aberrantly activated in chronic and acute liver diseases, is a novel neddylation substrate. Neddylation of NIK promotes its ubiquitination and degradation; and inhibition of neddylation caused NIK overactivation, accentuating hepatocyte damage. Notably, oral N‐acetylcysteine (NAC) administration attenuated lethal liver injury caused by hepatic NAE1 deletion, with improved liver function and mortality. Therefore, hepatic neddylation is essential in maintaining postnatal and adult liver homeostasis, and the novel neddylation targets/pathways may endow us with new insights into therapeutically intervening ALF progression.
Chronic, episodic developmental nicotine exposure alters GABAergic synaptic transmission to hypoglossal motor neurons at a critical developmental age in rats
Authors: Wollman Lila B. , Flanigan Emily G. , Fregosi Ralph F.
Published: May 2022

The adverse effects of nicotine exposure on brain structural and functional development is attributed to its action on the nicotinic acetylcholine receptors (nAChRs). nAChRs are expressed post‐synaptically on hypoglossal motor neurons (XIIMNs), the neurons that innervate the tongue. We recently showed that chronic, episodic developmental nicotine exposure alters the intrinsic properties of these neurons, which may be one mechanism to explain impaired tongue muscle function that is commonly observed in nicotine‐ exposed human and animal neonates. Importantly, along with the intrinsic properties of XIIMNs, the balance of excitatory and inhibitory fast‐synaptic inputs to these neurons is important in dictating the strength of motor output to the tongue. nAChRs are also located on the soma, dendrites, and axon end terminals of excitatory and inhibitory interneurons that synapse onto XIIMNs, activation of which increases action potential firing and neurotransmitter release. Here, we tested the hypothesis that chronic, episodic developmental nicotine exposure alters GABAergic transmission, a major source of fast‐synaptic inhibition, to XIIMNs. XIIMNs were recorded in brainstem slices from control and nicotine‐ exposed rats in two age groups: postnatal days (P)1‐5 and P10‐12, which is considered a critical period of brain development. In voltage‐ clamp mode, the frequency of GABAergic spontaneous inhibitory post‐synaptic currents (sIPSCs) was recorded at baseline and in response to activation of nAChRs with acute, bath‐applied nicotine. The baseline frequency of GABAergic sIPSCs was not different between control and nicotine‐exposed neurons at either age. In the P1‐5 age group, acute nicotine increased the frequency of GABAergic sIPSCs in both control and nicotine‐exposed neurons. However, at ages P10‐12, acute nicotine increased the frequency of GABAergic sIPSCs in nicotine‐exposed neurons, but had no effect in control neurons. These findings indicate that chronic, episodic nicotine exposure alters GABAergic synaptic transmission to XIIMNs at a critical developmental age, and may contribute to excitatory/inhibitory imbalance that underlies impaired control of the muscles of the tongue.
Cadaveric Clinical Integration: Integration of Basic Science and Medical Science
Authors: Day Leslie , Greene Ericka , Patham Bhargavi
Published: May 2022

Introduction

Cadaveric dissection is an integral part of many medical schools and provides an opportunity for students to visualize anatomical structures through active learning and critical thinking. Often during dissection, several pathological features and medical interventions can be identified, but are not regularly explored due to time or knowledge constraints. To encourage horizontal integration of content, a self‐directed learning (SDL) activity was implemented across four concurrent courses, gross anatomy (MGA), Foundations of Medicine which includes histology and physiology (FOM), Practice of Medicine that includes physical examination (POM) and Engineering Innovation (ENG). The goal of this longitudinal experience was to foster integration of basic science with clinical science thereby increasing metacognitive and lifelong learning skills.

Methods

49 students enrolled in the Engineering‐Medicine dual degree program at Texas A&M College of Medicine were divided into 7 groups, based on the cadaveric dissection assignments in MGA. Over the 16‐week MGA course, students documented the medical interventions and pathology found during dissection. During most dissection labs, a surgeon related to the field was present to answer questions. In addition, at three time points in the semester an anatomical pathologist performed a modified autopsy on the cadavers based on students request. This self‐directed, problem‐based learning (PBL) activity culminated in a forensic clinical summary, reconstructed comprehensive patient encounter 1 year prior to death, and 15‐minute oral presentation of their findings of their assigned cadaver. To assess potential changes in learning behavior, students completed a 46‐question, 8 subcategory modified Motivated Strategies for Learning Questionnaire (MSLQ) related to the activity. In addition, student’s provided verbal and written feedback on their perceptions of the project.

Results

47 out of 49 students successfully completed all aspects of the project. The was a positive trend and strong correlation in 7 out of 8 MSLQ categories, including intrinsic motivation, metacognition, task value, critical thinking, self‐efficacy, peer learning and help seeking. A paired t‐test showed a decrease in student's extrinsic motivation (t(46) = 1.831, p = 0.037) as related to the project. Students reported an overall high satisfaction with the project. The student presentations demonstrated high level of integration of basic science with clinical medicine.

Conclusion/Implication

This innovative multi‐disciplinary activity increased the value of cadaveric dissection, integration of content across courses, and promoted self‐directed learning of integrated clinicopathology material.
Subarcuate Fossa Anatomy as an Indicator of Future Locomotor Behavior in Newborn Primates
Authors: Militello Molly L. , Bryson Emily R. , Smith Timothy D. , DeLeon Valerie B.
Published: May 2022

Previous studies have demonstrated a link between subarcuate fossa size and locomotor behavior in primates. This fossa contains the petrosal portion of the paraflocculus, a portion of the brain associated with mobility and motor control. Studies have determined that the size of the fossa is larger in primate species that have more complex forms of locomotion. This study tested whether size of the subarcuate fossa in newborns predicts adult locomotor behavior. The sample included adult and newborn individuals of species representing strepsirrhine, tarsier, and platyrrhine primates. MicroCT images were used to create virtual endocasts in 3DSlicer software. The subarcuate fossa was isolated, and the volume was measured for each specimen.

Results demonstrated that newborns from species with higher levels of locomotor agility exhibit a larger subarcuate fossa at the time of birth than newborns from species with lower levels of locomotor agility. This confirms that the subarcuate fossa is an indicator of the future locomotion of the primate. Comparison of subarcuate fossa volume between newborn and adult scans from the same species indicated that there is little growth in the fossa from birth to adulthood in most primates. These results are consistent with an expectation that the size of the subarcuate fossa is highly constrained by the semicircular canals, which are not expected to increase in size through growth. The possibility exists that the size of the semicircular canals is the object of selection, and the size of the subarcuate fossa is a secondary effect. These findings show that this aspect of a newborn primate’s anatomy expresses information about its future behavior prior to any experience with locomotor action.
Ligand‐Dependent Reversal Kinetics of Mu Opioid Receptor Agonists by the Novel Antagonist, Methocinnamox
Authors: Smith Hudson R. , Chavera Teresa S. , Berg Kelly A. , Clarke William P.
Published: May 2022

Naloxone (NLX) is a competitive antagonist at the mu opioid receptor and is currently the only pharmacological intervention available to treat an opioid overdose. Unfortunately, NLX is limited by its surmountability and its rapid metabolism/clearance can lead to a reemergence of narcotic effects (renarcotization). Methocinnamox (MCAM) is a long‐acting antagonist at mu receptors both in vivo and in vitro. In addition to its orthosteric antagonism, we have found that MCAM differentially modulates orthosteric properties (affinity and/or efficacy) of mu agonists via an allosteric mechanism (Zamora, Smith, et al. (2021). Pharmacology Research & Perspectives. https://doi.org/10.1002/prp2.887). To further explore the interactions between MCAM and the human mu opioid receptor, we compared the rate by which MCAM and NLX reverse cAMP inhibition by the mu receptor agonists DAMGO and fentanyl. Treatment with either DAMGO or fentanyl (~ 30 x EC50) inhibited forskolin (FSK)‐stimulated intracellular cAMP levels. When administered 20 min after agonist, NLX (400 nM; ~100 x Ki) reversed DAMGO‐ and fentanyl‐mediated inhibition of cAMP production at a similar rate (25.4% ± 2.5 min‐1 vs 26.8% ± 5.5 min‐1, respectively). Since reversal by NLX was driven mostly by dissociation of the agonists, the similar rate of reversal by NLX indicate that DAMGO and fentanyl dissociate from the receptor at similar rates. Reversal by MCAM (100 nM; ~ 100 x Ki) however was ligand‐dependent. The rate of reversal of DAMGO‐mediated inhibition by MCAM was 11.7% ± 1.3 min‐1, whereas reversal of fentanyl was slower (4.1% ± 0.9 min‐1; P<0.001). The ligand‐dependence of MCAM’s reversal kinetics on DAMGO vs fentanyl’s inhibition of cAMP production is consistent with the reported allosteric action of MCAM, and may be explained by modulation of ligand dissociation by MCAM. Together, these data suggest in addition to its orthosteric antagonism, MCAM may allosterically modulate the binding of mu agonists.
Essential oil metabolites can regulate adrenal androgen production by inhibition of CYP17A1 activities
Authors: Sharma Katyayani , Lanzilotto Angelo , Pandey Amit V.
Published: May 2022

Endocrine‐disrupting chemicals (EDCs) can affect human steroid metabolism. Previous clinical case reports have shown that some Essential Oils (EOs) like lavender oil and tea tree oil may act as potential EDCs and are linked to prepubertal gynecomastia in boys and premature thelarche in girls due to regular exposure to lavender based fragrances among Hispanic population. These studies suggested role of EOs in steroid metabolism in humans. We have screened a range of EO metabolites for effects on androgen production by CYP17A1.

For preliminary screening, human adrenal NCI H295R cells were treated with 10 µM of test compounds for 24 hours. The test compounds had been extracted and purified from natural resources and are found as major components in EOs. For CYP17A1 activity, the conversion of radiolabelled substrate, 17‐Hydroxy‐pregnenolone to Dehydroepiandrosterone was determined using tritiated water release assay.

Eucalyptol, Dihydro‐β‐Ionone, (‐)‐α‐pinene were extracted from eucalyptus, rose and pine resin. Out of about 50 test compounds, eucalyptol, Dihydro‐β‐Ionone & (‐)‐α‐pinene showed 20% to 40% inhibition of DHEA production. Rest of the compounds showed either no or low inhibition. Some compounds were also tested for effects on CYP19A1 (aromatase) activity where upto 30% inhibition was observed.

EOs are often used in various beauty and hygiene products as they have few known side‐effects. However, prolonged exposure to these products may result in steroid imbalance. Due to their anti‐androgenic activity, these compounds should be studied further as chemical leads for the treatment of hyperandrogenic disorders such as Prostate cancer and Polycystic ovary syndrome.
Mechanistic Insights into the Role of TRIM28 in the Regulation of Skeletal Muscle Size
Authors: Steinert Nathaniel D. , Lin Kuan‐Hung , Hermanson Jake B. , Hornberger Troy A.
Published: May 2022

Protein homeostasis plays a critical role in the regulation of skeletal muscle size, and the maintenance of skeletal muscle size contributes significantly to disease prevention and quality of life. Over the past few decades, it has become widely accepted that skeletal muscle size is controlled by the net balance between the rates of protein synthesis and protein degradation. Despite this assertion, the mechanisms that modulate this balance and lead to changes in muscle size remain incompletely defined. Nevertheless, advancements have been made. For instance, a recent study from our lab revealed that the myofiber‐specific loss of a transcriptional intermediary factor named TRIM28 led to a significant reduction in basal myofiber size and attenuated the increase in myofiber size that occurs in response to mechanical overload. Moreover, rigorous follow‐up studies indicate that the TRIM28 knockout‐induced deficits in myofiber size are not driven by a decrease in the rate of protein synthesis, but that this phenotype is instead associated with elevated levels of canonical markers of protein degradation. Together, these observations led to our central hypothesis that TRIM28 confers its effects on myofiber size via the regulation of protein degradation. Thus, in an effort to gain further insight into this possibility, we performed a deep RNA‐sequencing analysis, the results of which led to the identification of Mettl21c and Mettl21e as two genes whose expression were significantly downregulated by the loss of TRIM28. We were intrigued by this discovery because recent reports have implicated Mettl21c and Mettl21e in the regulation of protein degradation, and it has been shown that the loss of these proteins in muscle promotes aberrant protein degradation and reduces muscle size. Combined with our initial observations, it became apparent that Mettl21c and Mettl21e might be important parts of the pathway via which TRIM28 regulates protein degradation to control myofiber size. To test this theory, we generated expression plasmids encoding HA‐Mettl21c and HA‐Mettl21e, and then used electroporation to demonstrate that the expression of Mettl21c and Mettl21e in myofibers is sufficient to induce a robust hypertrophic response in control animals, and that the hypertrophic effect of Mettl21c, but not Mettl21e, is conserved in muscles lacking TRIM28. Collectively, these findings provide evidence that TRIM28 regulates myofiber size via the regulation of protein degradation, and that Mettl21c, but not Mettl21e, might play an important role in the pathway via which TRIM28 confers this effect.
The CHST11‐CP‐metal ionic axis contributes to NSCLC immune escape and metastasis
Authors: Li Li‐Jie , Chang Wei‐Min , Hsiao Michael
Published: May 2022

Escaping of immune surveillance is essential for tumor metastasis. Recently, intratumoral metal ionic balance plays an important role in controlling cancer immune checkpoint protein expression and tumor immune escape. Copper (Cu) promotes the expression of PD‐L1. In contrast to Cu, heme biosynthesis disturbs PD‐L1 expression. Heme protein is a kind of metalloproteins with ferrous iron (Fe2+)‐porphyrin core. Here, we found CHST11 served as a pro‐metastatic gene in non‐small cell lung cancer (NSCLC). We further identified the expression of ceruloplasmin (CP) was regulated by CHST11. CP is not only the major blood Cu carrier protein but also a Cu‐dependent ferroxidase in iron metabolism. It may hint a crosstalk mechanism between lung cancer metastasis and tumor immune surveillance through the CHST11‐CP axis. CP was also associated with poor prognosis in patients with NSCLC. Both ectopic CP expression and high intracellular ionic stimulations, like ferric iron and copper, promoted NSCLC progression, such as in vitro migration and invasion, as well as in vivo tumorigenesis abilities in Nod‐SCID mice. The ionic chelator treatments repressed in vitro metastatic potential and, interestingly, regulated the gene expressions of immune checkpoints in NSCLC cells. By using microarray analysis, we found CHST11 controlled CP expression through interferon‐gamma (IFNγ), Janus kinase (JAK), and signal transducer and activator of transcription (STAT) pathways. Treatment with ruxolitinib, a JAK inhibitor, suppressed the expressions of CP and immune checkpoint proteins. In conclusion, our findings prove that the CHST11‐CP axis plays an important role in both NSCLC immune escape and metastasis through IFNγ‐JAK‐STAT pathway. The combination of an iron chelator and ruxolitinib may provide a potential translation application that protects patients with NSCLC from cancer immune escape and metastasis.
Brain Shape Provides a Structural Mechanism for Postorbital Septation in Primates
Authors: DeLeon Valerie B. , Franklin Katheryn P. , Cunningham Andreana S. , Summers Siobhan A. , Bryson Emily R. , Vinyard Christopher J. , Smith Timothy D.
Published: May 2022

Postorbital septation is a character that unites haplorhine primates. As a result, evidence of postorbital septation is expected for fossil primates that may be ancestral to both modern tarsiers and modern anthropoids. Our previous work on newborn primates has indicated that postorbital septation in tarsiers is secondary to growth of their large eyes. If this is the case, postorbital septation is convergent in tarsiers and anthropoids, and therefore should not be expected in their fossil ancestors. Here, we used morphometric analysis of the cranium and endocast of newborn and adult primates to show that postorbital septation in the order Primates may be explained by shape of the brain.

We used a sample of over thirty species of modern primates representing all major clades, with individuals representing both newborn and adult stages. The newborn stage is most informative for understanding spatial relationships of soft tissues during formation of the skull. MicroCT image data were used to create virtual reconstructions of the cranium and endocast as an estimate of brain shape. Fixed landmarks were used to capture the shape of the cranium, and sliding semilandmarks represented the contour of frontal and temporal lobes on the endocast. As expected, morphometric analysis showed a significant effect of phylogeny on cranial and endocast shape in both newborns and adults. Notably, the phylogenetic effect was apparent in only the contours of the frontal and temporal lobes, independent of cranial shape. We present 3D models of the endocast and cranium of newborn primates to illustrate the spatial relationship of brain and facial structures as a mechanism for postorbital septation in anthropoid primates.
Effects of Salvia Hispanica on Metabolic Syndrome and Bone in Mouse Osteoblastic Cells – a miRNA analysis
Authors: Garcia‐Rodriguez Natasha , Nangavaram Mahathi , Banu Jameela
Published: May 2022

Metabolic Syndrome and bone diseases are high priority human diseases which, scientists have tried to better understand in order to aid those suffering from their detrimental effects. Therapies for Metabolic Syndrome and bone diseases available today come with several side effects and do not alleviate the conditions fully; thus, it is beneficial to find alternative medicines and treatments which have the potential to provide less or optimal side effects. Salvia hispanica is a medicinal and edible plant known for its high nutritional value and therapeutically potential which shows promise in possessing properties useful in developing alternative therapies or treatments for metabolic syndrome and bone diseases. miRNA expression patterns can be disease‐specific, reflect the developmental and state of tumors or cancers but equally important they are essential biological processes which, may lead to various pathological and occasionally malignant outcomes. Mouse MC3TC cells were cultured, differentiated, and treated with aqueous extract of the seeds of S. hispanica for 48 hours. The cells were then harvested and sent for miRNA analysis. The data revealed that the S. hispanica seed extract up‐regulated let‐7e‐5p, let‐7a‐5p, let‐7j, and miRNA‐151‐5p. Moreover, the data revealed that several miRNAs were downregulated including miRNA‐181a‐5p, miRNA‐503‐5p, miRNA106b‐5p, miRNA‐152‐5p, miRNA‐140‐3p, miRNA‐33‐5p, miRNA‐214‐3p, miRNA‐99b‐5p, miRNA‐30c‐2‐3p, miRNA199a‐5p. The knowledge extrapolated regarding the miRNAs and the metabolic pathways that are impacted by Salvia hispanica will help us identify the benefits of S. hispanica in treating Metabolic Syndrome and bone diseases.
Stratification by Race Reveals Disparate Vascular Toxicity in Response to Anti‐Cancer Therapies
Authors: Hader Shelby N. , Terwoord Janée D. , Norwood Toro Laura E. , Gutterman David D. , Beyer Andreas M.
Published: May 2022

Introduction

Cardiovascular toxicity is the primary reason for halting anti‐cancer therapy (CTx) or limiting the dose, and cardiovascular side effects of CTx represent a major clinical problem with implications for survival and quality of life. Ongoing work in our laboratory suggests that CTx induces microvascular endothelial dysfunction, which is a well‐established predictor of future CV events. CV disease in general and CTx‐induced adverse events are significantly more prevalent in Black patients (B) relative to non‐Hispanic White patients (NHW). Despite clinical evidence of these health disparities, little to no data exist to explain the underlying causes. Therefore, we tested the novel hypothesis that CTx‐induced microvascular dysfunction is augmented in B, which may explain the increase in cardiotoxic events in this population compared to NHW.

Methods

Fresh human tissues were obtained as de‐identified surgical discard specimens. Peripheral adipose arterioles (100–200 µm) from healthy B and NHW were incubated ex vivowith the CTx agents, doxorubicin (Dox), trastuzumab (TZM), or paclitaxel (PTX) for 16–20 h and used for isolated arteriole preparations. Endothelium‐dependent vasodilation in response to flow (Flow‐Mediated Dilation, FMD; 5–100 cmH2O pressure gradient) or acetylcholine (ACh; 10−9 to 10−5 M) and smooth muscle‐dependent dilation to papaverine (100 µM) were evaluated. Data are presented as % max. diameter ± SEM, N. Statistical significance was established with two‐way ANOVA RM.

Results

Under control conditions, there was no difference in microvascular FMD in healthy B (ACh: 89.8±4.0%, n=5; 85.8±1.8%, n=6) vs. NHW (ACh: 86.2±3.2%, n=6; 78.7±5.9%, n=8). Ex vivo exposure to Dox impaired endothelial function in arterioles from both B and NHW (ACh: 45.0±10.5%, n=4 and 34.2±9.8%, n=7; FMD: Figure 1A). TZM impaired endothelial function to a greater extent in arterioles from B (ACh: 36.1±7.1%, n=3; FMD: Figure 1B) compared to NHW (ACh: 44.4±8.6%, n=8; FMD: Figure 1B). Interestingly, PTX did not affect endothelial function in NHW (ACh: 91.5±1.6%, n=4; FMD: Figure 1C), but impaired endothelial function in arterioles from B (ACh: 54.7±12.9%, n=3; FMD: Figure 1C). Max dilator capacity to the endothelium‐independent dilator papaverine was not changed in either group (not shown).

Conclusion

Our findings reveal disparate effects of CTx agents on microvascular endothelial function from B and NHW patients. This raises the possibility that microvascular dysfunction may contribute to the known disparities in adverse CV clinical outcomes of cancer patients and potentially other CV pathologies (e.g., disparities in adverse events associated with PTX‐eluting stents.) Future work should differentiate between biological/genetic vs. social/environmental contribution to observed microvascular responses to CTx.