bluuebunny/crossref_metadata_2025_split huggingface dataset:
Use of Labeled Prosections for Self‐Guided Learning in Head & Neck Anatomy: Comparison to Traditional Instructional Approaches
Authors: Mears David 
Published: May 2022

AimThe purpose of this study was to evaluate the effectiveness of a self‐guided learning approach for head and neck anatomy using faculty‐prepared, labeled cadaveric prosections.IntroductionAt the onset of the COVID‐19 pandemic, when in‐person group learning activities were suspended for public health reasons, medical schools worldwide were forced to develop innovative ways to teach anatomy without traditional student‐led dissections. One approach that we implemented for head and neck anatomy at USU was to display faculty‐prepared, fully‐labeled prosections in the anatomy lab, and allow students to come in individually to learn from them. This self‐guided approach was positively received by students as a COVID‐19 contingency (spring 2020), so we decided to use a modified version of this strategy for some of the head and neck anatomy labs even after the local pandemic conditions had allowed in‐person dissection to resume (spring 2021). In this study, we quantitatively compared learning outcomes from self‐guided learning, with outcomes from two traditional instructional methods: student‐led dissection and faculty‐guided prosection demonstration.Approach/MethodsFor the 2021 Head and Neck Anatomy block, six cadaver laboratories were taught by traditional dissection (teams of four students with faculty preceptors present) and six were taught using labeled prosections. The prosections were put on display along with a detailed key to guide the students’ learning of the structures and key relationships. For the lab session, students in groups of four were assigned times to view the displays (5 minutes per specimen), with faculty on hand to answer questions. After the session, specimens were made available 24/7 for students’ independent review. Student performance on practical exam questions was compared for labs taught with labeled prosections versus the same labs taught by traditional dissection or faculty‐guided prosection demonstration from a prior year of the course (2019), using ANOVA and student’s t‐tests.ResultsLearner satisfaction with the self‐guided prosections was high, with 87% of students reporting that they found this instructional method to be helpful for learning anatomy. There were no significant differences in student exam performance for labs taught via labeled prosections compared to traditional dissection or faculty‐guided prosection demonstration.DiscussionWe found that self‐guided learning from labeled prosections was an effective, well‐received instructional method for head and neck anatomy. Implementing this approach for 6 of the 12 laboratory sessions in the Head and Neck Anatomy block resulted in a 25% decrease in curricular contact hours, and a 33% decrease in faculty time required to staff the laboratory sessions, with no detriment to learning outcomes. This approach could easily be extended to other aspects of the Gross Anatomy curriculum.
Blood Storage Solutions Effecting Advanced Glycated End Products (AGEs) N‐CEL and N‐CML on Red Blood Cell Membranes
Authors: Skrajewski Lauren , Geiger Morgan , Jacobs Monica , Spence Dana 
Published: May 2022

Blood transfusions are critical, often lifesaving, procedures that result in approximately 10.5 million transfusions per year in the United States. Donated red blood cells (RBCs) is stored for a maximum of 42 days in FDA approved storage solutions. These storage solutions contain high levels of glucose to promote the longevity of the cells donated. However, AS‐1 storage solution contains 111 mM glucose, which is about 20 times higher than normal blood concentrations (5‐6 mM). Hyperglycemia is often associated with diabetes (7‐9 mM glucose), and prolonged hyperglycemic conditions are known to cause various blood flow‐related complications (retinopathy, neuropathy, nephropathy, cardiovascular disease). There is evidence of adverse RBCs changes in hyperglycemic conditions during the time over which storage would occur. Specifically, increased glycation is linked to increased amounts of advanced glycated end products (AGEs).AGEs are formed from the glycation process, as higher glucose conditions produce a spontaneous non‐enzymatic reaction of free reducing sugars with free amino groups that form Amadori products, which undergo irreversible changes that lead to the formation of AGEs. These compounds promote oxidative stress and inflammation and have been linked to various chronic diseases, often linked with diabetes. RBCs release the necessary ATP for proper blood circulation, and there is previous evidence showing the high glucose storage decreases ATP release over 36 days. Also, there is a consistent ATP levels for the AS‐1N (5 mM glucose) storage solution over 36 days which are higher than the hyperglycemic storage solution.Decreased ATP release during storage may be linked to hyperglycemic storage conditions increasing AGE biomarkers on the RBC membrane. Two of these AGE adducts of carboxymethyl lysine (N‐CEL and N‐CML) are used to quantify glycation on the RBC membrane by studying adduct concentration changes during storage. Two solutions were prepared (AS‐1 and AS‐1N) and used for storage for 36 days, with aliquots of RBCs removed every 4‐7 days for analysis. The amount of irreversible glycation products, thiols, and total lysine groups were determined to evaluate the amount of modified species over time. LC‐MS was used to quantify these modified N‐CEL and N‐CML adducts on the RBCs using internal standards. Overall, our results probe the hyperglycemic conditions on RBCs and examine the effects of increased glycation on cellular release of components from these stored cells.
Immune Phenotypes of Oligodendroglial‐Lineage Cells in MDD and in Response to Chronic Stress‐Induced Microglial Inflammation
Authors: Madeira Miguel M. , Kokkosis Alexandros G. , Hage Zachary , Tsirka Stella E. 
Published: May 2022

BackgroundMajor Depressive Disorder (MDD) is a complex and heterogenous psychiatric disorder affecting more than 300 million people worldwide. Though the mechanisms underlying MDD are not fully understood, a subset of depressive patients can manifest chronic neuroinflammatory responses. In addition, brain‐imaging studies in MDD patients have provided evidence of white matter reductions and such changes are hypothesized to lead to disruption of Oligodendroglial‐Lineage cells (OLN) homeostatic mechanisms, resulting in destabilization of emotional/cognitive circuitry. The present goal of the study is to investigate whether these histopathological alterations are linked with chronic inflammation through single‐nucleus transcriptomics of MDD patients and in a depression mouse model.MethodsBioinformatics: Raw counts were downloaded from GEO (GSE144136), and expression objects created using Seurat. OLN were subsetted, clustered (using UMAP), and annotated based on their expression of early and late OLN markers. Pseudotime trajectory analysis was conducted using Moncle3. In vivo: The Repeated Social Defeat Stress (RSDS) paradigm (10 days) was used to induce depressive‐like behavior in 8–12‐week‐old male CX3CR1‐GFP+ and CSPG4‐EGFP+ mice. Behavioral tasks (BH) were performed to stratify the defeated mice to susceptible (S; depressive‐like) and resilient (R; non‐depressive) to stress groups. The study focuses on the MDD‐affected prefrontal cortex (mPFC) area (Fig).ResultsSeparate clustering of MDD and Control samples yielded four similar clusters labeled based on their gene expression as: OPCs, Committed OPCs, Immature Oligodendrocytes (OLs), and Mature OLs. One MDD‐specific cluster was also obtained and labeled as Immune OLs (ImOL), given its expression of immune genes, such as: ARGHAP24, ADAM28, LPAR6, C3, CD74, and P2RY12. Further analysis of the ImOL cluster revealedexpression of markers associated with later phases of OLN progeny. Finally, a predictive model of pseudotime trajectory revealed alterations in the progression of OLN progeny in MDD samples. We then sought to identify this novel subset of ImOL in the RSDS model of depression. MHCII, C3 and P2RY12 were examined in conjunction with OLN markers (CSPG4, O4, CNP), with significant changes observed in S groups. Interestingly, microglial phagocytosis of myelin elements (CX3CR1‐GFP, MHCII, CNP), was significantly increased in S groups post RSDS, rendering the ImOL populations as potential recruiters of innate immunity.ConclusionsTogether, our computational data in MDD patients and depression mouse model, show that OLN can swiftly respond to chronic stress, and adopt an immune‐like phenotype, potentially contributing to the white matter disturbances observed in MDD.
Antagonism of Na,K‐ATPase signaling by pNaKtide can attenuate systemic inflammatory response in murine sepsis model induced by cecal ligation and puncture
Authors: Pereira Duane G. , Pillai Sneha S. , Lakhani Hari Vishal , Wang Jiayan , Sodhi Komal 
Published: May 2022

IntroductionOxidative stress and associated inflammatory processes are crucial in the development and progression of sepsis, resulting in an immune response that leads to multiple organ failure or dysfunction. Hence, the strategies to limit this systemic inflammatory response might result in the development of an effective sepsis therapy. Previous studies from our lab have demonstrated the role of Na/K‐ATPase signaling in exacerbating systemic oxidative stress and inflammation and the potential role of Na/K‐ATPase signaling antagonist, pNaKtide, in ameliorating several pathophysiological abnormalities.HypothesisThe aim of this study is to demonstrate the role of Na,K‐ATPase signaling in exacerbating oxidant stress and inflammatory response noted in a murine model of sepsis. Furthermore, we aim to demonstrate the effect of systemic administration of Na,K‐ATPase signaling antagonist, pNaKtide, in experimental sepsis using cecal ligation and puncture (CLP) model, as a drug intervention against septic shock.MethodMurine sepsis was induced by CLP in male C57BL6 mice with or without pNaKtide (25 mg/kg body wt) which was administered, intraperitoneally, 24 hours before CLP procedure. Sham surgery was performed without the ligation and puncture, which served as controls for the study. All mice were assessed for Murine Sepsis Score (MSS) at baseline (before Sham or CLP), at 4hours, 8hours and 24 hours after Sham or CLP. Mice were euthanized after 24 hours of Sham or CLP surgery and tissues were collected for morphological and biochemical assessment. Blood was used to measure plasma levels of inflammatory cytokines.ResultsSystemic administration of pNaKtide demonstrated improved MSS at 24 hours following CLP surgery, as compared to CLP mice without pNaKtide. Histological assessment of lung tissues by H&E staining showed significantly less congestion, edema, infiltration of inflammatory cells and hemorrhage in CLP mice administered with pNaKtide, as compared to CLP alone. pNaKtide also improved systemic inflammatory cytokines assessed by plasma levels of TNFα and MCP1, as well as improved mRNA expression of inflammatory and macrophage infiltration markers, in lung tissues of CLP mice.ConclusionOur study demonstrates that antagonism of Na,K‐ATPase signaling by pNaKtide may attenuate CLP‐induced sepsis by inhibition of inflammatory milieu noted in this pathophysiological condition. Hence, Na,K‐ATPase signaling may serve as a viable clinical target for therapeutic intervention of sepsis and associated inflammatory mechanisms.
The impact of cyclooxygenase inhibition with ketorolac on regional cerebral perfusion in healthy adults
Authors: Muer Jessica D. , Didier Kaylin D. , Wannebo Brett M. , Carter Katrina J. , Hagen Scott A. , Eldridge Marlowe , Al‐Subu Awni M. , Livett Timothy A. , Wieben Oliver , Schrage William G. 
Published: May 2022

IntroductionNon‐selective cyclooxygenase (COX) inhibition tests the role of COX on cerebrovascular control. Most studies using COX inhibitors (typically Indomethacin) demonstrate robust decreases in cerebral blood flow (CBF) (20‐42%), but have focused primarily on middle cerebral artery (MCA) velocity, which does not quantify CBF or provide insight into regional differences in the influence of COX on basal CBF control. The purpose of this study was to determine global and regional microvascular effects of COX inhibition to gain a better understanding of mechanisms underlying resting CBF control.HypothesisWe hypothesize that microvascular CBF will be attenuated after ketorolac infusion globally and in discrete regions.MethodsYoung healthy adults (M=4, F=2) were included in the study (age: 24 ± 3, BMI: 22.4 ± 0.7). Participants completed two Arterial Spin Labeling (ASL) scans. ASL scans were completed before and after a 3‐minute intravenous infusion of ketorolac (0.495 mg/kg, mean dose 33.9 ±2.8 mg). ASL analysis was completed in SPM12. ASL analysis included smoothing grey matter, co‐registering and normalizing raw data, brain masking and region of interest processing. A two‐way repeated measures ANOVA with a Tukey’s post hoc were computed in SigmaPlot13.ResultsGlobal cerebral blood flow during room air (58±15mL/100g/min) did not change after ketorolac infusion (57±24mL/100g/min)(p=0.83). Regional cerebral blood flow for pre‐ketorolac infusion and post‐ketorolac infusion were not significantly different((PRE mL/100g/min, POST mL/100g/min(p‐value)): Frontal lobes(Left:66±15, 65±23(p=0.87),Right: 68 ±15, 66 ±25(p=0.62)),Temporal lobes(Left: 56±12, 54±22(p=0.82); Right: 56±14, 54±24(p=0.61)),Parietal lobes (Left:56±12, 57±25(p=0.95); Right: 59±16, 59±26(p=0.99)), Occipital lobes(Left: 51±15, 54±30(p=0.58); Right: 50±16, 54±32(p=.44))).DiscussionOur findings suggest ketorolac does not alter microvascular perfusion in healthy adults, suggesting not all COX inhibitors act on cerebral vasculature equally. These data add to previous work by comprehensively interrogating the entire brain rather than flow (or velocity) through single arteries.ConclusionThere were no effect of ketorolac on global or regional cerebral blood flow in healthy adult
Cholinergic‐mediated Circulatory Responses at the Onset of Isometric Exercise: Does Sex Matter?
Authors: Samora Milena , Teixeira André L. , Sabino‐Carvalho Jeann L. , Vianna Lauro C. 
Published: May 2022

IntroductionWe have shown that handgrip exercise evokes an initial decrease in systemic vascular resistance (SVR), which was abolished by cholinergic blockade (glycopyrrolate) and, thereby, augmented the blood pressure (BP) response. These findings suggest that a cholinergic mechanism is important for the BP and SVR responses at the onset of isometric handgrip exercise in humans. Noteworthy, this previous study included men only, and was not designed to investigate the potential existence of sex‐related differences in immediate BP and circulatory responses evoked at the exercise onset. Given this, the aim of this study was to test the hypothesis that women present an attenuated fall in SVR and increased pressor response compared to men at the onset of isometric handgrip exercise due to a blunted cholinergic vasodilation.MethodsTen healthy men (aged 21 ± 1 year) and 10 women (aged 24 ± 1 year) were recruited. Women were non‐oral contraceptives users and performed experiments in the early follicular phase of the menstrual cycle. Participants performed 20s bout of isometric handgrip exercise at 40% of maximal voluntary contraction. Beat‐to‐beat heart rate (HR) (ECG) and mean BP (finger photopletysmography) were continuously measured. Beat‐to‐beat stroke volume (SV) was obtained with the Modelflow method and matched with HR to estimate cardiac output (CO) and SVR. The exercise responses were expressed as a change from 10s immediately before the onset of voluntary contraction.ResultsThe BP response pattern appeared to be different between men and women at the onset of isometric exercise. While men presented a typical exercise‐evoked biphasic BP response (3s: Δ2 ± 2 mmHg; 10s: Δ1 ± 1 mmHg; 20s: Δ5 ± 3 mmHg), women showed a progressive increase in mean BP (3s: Δ1 ± 1 mmHg; 10s; Δ3 ± 1 mmHg; 20s: Δ6 ± 1 mmHg; P=0.681). Men demonstrated a decrease in SVR at 3s (Δ‐0.9 ± 0.3 mmHg.L‐1.min), 10s (Δ ‐1.6 ± 0.4 mmHg.L‐1.min) and 20s (Δ‐1.9 ± 0.7 mmHg.L‐1.min) following the exercise onset, and these responses tended to be attenuated in women (3s: Δ‐0.3 ± 0.3 mmHg.L‐1.min; 10s: Δ‐1.2 ± 0.4 mmHg.L‐1.min; 20s: Δ‐0.6 ± 0.5 mmHg.L‐1.min; P=0.131). There were no sex differences in HR, SV and CO responses to exercise.ConclusionThese findings suggest a lack of sex differences in immediate BP and circulatory responses at the onset of isometric handgrip exercise, indicative of similar cholinergic‐mediated vasodilation in skeletal muscle between men and women.
BRD4‐MK2 signaling: target for Crohn’s Disease‐associated fibrosis
Authors: Chulkina Marina , McAninch Steven , Li Yi , Koltun Walter , Yochum Gregory , Tian Bing , Zhou Jia , Beswick Ellen J. , Brasier Allan R. , Pinchuk Irina V. 
Published: May 2022

IntroductionFibrosis is the major complication of Crohn’s disease (CD). No targeted therapies are currently available to revert CD‐associated fibrosis. While inflammatory mechanisms in CD have been extensively investigated, understanding the pathogenesis of fibrosis is relatively limited. Mesenchymal cells are the major effectors in profibrotic processes in CD. A dramatic increase in α‐SMA expressing subsets of mesenchymal cells, also known as myofibroblasts (MF), is among the main hallmarks of profibrotic changes in CD. Mitogen‐activated protein kinase‐activated protein kinase 2 (MK2) is involved in the increase of α‐SMAexpression by MFs. Along with an increase in MK2 activity in CD we also observed an increase in the bromodomain‐containing Protein 4 (BRD4), an important epigenetic reader. BRD4 is critical for the expression of enhancer‐mediated profibrotic genes and, thus, is a potential novel regulator of MK2. However, the presence of mechanistic link between BRD4 and MK2 activity with respect to profibrotic activation of MFs in CD is unknown. Herein, we tested hypothesis that BRD4 mediated increase in MK2 activity is critical to the profibrotic reprogramming of MFs in CD and this pathway has potential for novel anti‐fibrotic therapy.MethodsHuman normal (N)‐ and CD‐derived MFs were analyzed using RNAseq, cytokine/chemokine multiplex array, FACS, and immunofluorescent microscopy. The T cell transfer murine colitis model relevant to CD was also used in this study.ResultsAn Increase in BRD4 and MK2 expression was observed in primary human CD‐ when compared to N‐MFs and were associated with the expression of the profibrotic genes including Col1α2, α‐SMA, TNC, Fn1, and TGF‐β1. Inhibition of MK2 activity in CD‐MFs with the MK2 specific inhibitor PF‐3644022 and murine MK2 KO MFs showed significantly reduced basal and TGF‐β1‐induced profibrotic gene expression. Use of the BRD4 specific inhibitor ZL0454 significantly reduced MK2 mRNA expression in CD‐MFs Additionally, ex vivo treatment of the CD tissues with ZL0454 also abrogated MK2 expression. Therapeutic use of the BRD4 inhibitor ZL0454 (10 mg/kg, daily for 10 days) in T cell transfer murine colitis significantly decreased MK2 expression and downstream profibrotic responses as measured by collagen deposition and expression of fibrosis‐related genes.ConclusionsOur data suggests that BRD4 positively regulates MK2 expression in CD. Further, BRD4‐MK2 signaling is critical to profibrotic activation of mesenchymal cells in CD and is a potential therapeutic target to reverse CD‐associated fibrosis.
Superior Pharmacokinetic Profile of an Innovative, Next Generation Curcumin Formula
Authors: Rai Deshanie , Morde Abhijeet , Padigaru Muralidhara , Acharya Manutosh 
Published: May 2022

IntroductionCurcumin is the major bioactive component of turmeric or Curcuma longa L., a widely used spice, food colorant, preservative as well as medicinal agent in Indian and Chinese remedies. Numerous studies have demonstrated its potential benefits for heart health, immunity, diabetes, arthritis as well as joint health and mobility. Currently, curcumin‐based products in capsules, ointments, tablets, and cosmetics are marketed worldwide. One important drawback of using curcumin is its poor bioavailability and hence the need to use it at high doses to achieve measurable plasma levels and efficacy – this limits both consumer compliance and product formulation. Curcumin’s poor solubility, low absorption from the gut, rapid metabolism and rapid systemic elimination contributes to its poor bioavailability. To help overcome this major limitation of curcumin, we developed a proprietary curcumin formulation (Curcuwin Ultra+) (CU+) to enhance its bioavailability. We hypothesized that the formulation would protect the curcuminoids from degradation at intestinal alkaline pH conditions, and through this mechanism, significantly increase the availability of curcuminoids in blood and at the tissue level.GoalOur goal was to measure the pharmacokinetic profile of CU+ vs. a standard turmeric extract (STE).MethodA randomized, double‐blind, crossover study design involving 24 healthy volunteers under fasting conditions was implemented. Subjects received a single dose of 250 mg CU+, 500 mg CU+ and 1900 mg of STE. Blood samples before dosing, and at various time points post dosing up to 24 hours were collected. Total curcuminoids were measured using a validated LC‐MS/MS method. Formulations were compared based on Cmax, AUC0‐6, AUC0‐24, Tmax, t1/2, and relative absorption.ResultsMaximum serum concentration (Cmax) and total systemic exposure (AUC0‐6 and AUC0‐24) for total curcuminoids were significantly higher (p<0.05) for both doses of CU+ vs. STE. For example, AUC0‐6 was 144 and 149 times greater for CU+ at 250 mg and 500 mg respectively compared to 1900 mg of STE. Additionally, CU+ showed 40% faster absorption as measured by Tmax vs. the STE (p<0.05). No adverse events were observed.ConclusionsThese pharmacokinetic data demonstrates the ability of this curcumin formulation to deliver superior bioavailability vs. STE, and, at doses that are considerably lower than STE and several other commercially available curcumin products. This has important relevance to consumer compliance in terms of lowering curcumin dose and frequency of dosing. Since maximizing bioavailability influences the therapeutic effects of a compound, the next phase of our research program is to determine if this enhanced bioavailability can be translated to an efficacious benefit of CU+.