bluuebunny/crossref_metadata_2025 huggingface dataset:
“What Bones And Bony Features Are You Teaching Your Students?” A Survey Of Skeletal System Coverage In Undergraduate Human Anatomy And Physiology Courses
Authors: Aryal Asmita , O'Loughlin Valerie D.
Published: May 2022

Undergraduate (baccalaureate) human anatomy (HA) and anatomy and physiology (A&P) courses are offered globally, but how similar is their content coverage for each organ system? There has been a dearth of published research on this subject. Previously, some of the authors (Reynolds et al., 2018; submitted) surveyed instructors of stand‐alone HA as well as A&P courses to document comparability and variability in skeletal muscular system coverage in these courses. We now focus our attention on skeletal system coverage in these courses. A study by Sparacino et al., (2019) evaluated the undergraduate nursing anatomy curriculum at four institutions in the United Kingdom, but the study did not list the specific bone and its features students needed to know.Thus, a representative sample is required to be able to assess the variations in skeletal system coverage in both stand‐alone HA and A&P courses across different institutions.

We have developed an online survey where undergraduate HA and A&P faculty can report on which skeletal elements (bones), and which features on these bones they require their students to know. The skeletal system coverage survey is made accessible to participants online via Qualtrics. Bones and their bony features listed in the survey are those listed and described in a typical undergraduate human anatomy textbook (Human Anatomy, 6th edition textbook by McKinley, O’Loughlin, and Pennefather‐O’Brien [2021]). This survey will also collect institutional demographic data such as the type of institution, course format, and geographic location of the institution, for comparison. We need to collect data from a large and representative sample of courses in order to accurately describe skeletal system coverage trends.

This poster will provide both an in‐depth justification for the need for such a survey and serve to recruit participants to complete the survey. Additional recruitment will occur via recruitment emails to AAA (American Association for Anatomy), HAPS (Human Anatomy and Physiology Society), and AACA (American Association of Clinical Anatomists) listservs, as well as at the HAPS 2022 annual meeting. Thus, we will meet to speak about the formation of this survey, collect survey data from willing AAA participants, and have a critical conversation about skeletal system coverage in undergraduate human anatomy and physiology courses.
Role of stress granule activation in cardiomyocyte function in sepsis
Authors: Wang Yaqiao , Wu Kehan , Liu Runmin , Wang Hao , Rui Tao
Published: May 2022

Sepsis is a life‐threatening organ dysfunction caused by a dysregulated host response to infection. It is the leading cause of mortality in Intensive Care Unit (ICU). Sepsis‐induced myocardial dysfunction (SIMD) is a major complication of the sepsis that contributes to high mortality of patients with sepsis. Stress granule (SG) is a cytoplasmic membraneless platform that is formed in response to cellular stresses. The role of SG in SIMD is unknown. The aim of present study is to understand whether activation of SG in cardiomyocytes (CM) plays a role in CM function in sepsis.

Methods

CMs were treated with lipopolysaccharide (LPS). CM SG activation (eIF2α phosphorylation) and TNFα production were assessed with Western blot and ELISA, respectively. CM function was evaluated with intracellular cAMP. ISRIB or over‐expression of G3BP1 was employed interfering SG.

Results

Challenging CMs with LPS results in eIF2α phosphorylation, an increase in TNFα production, and a decrease in intracellular cAMP in response to dobutamine. Pharmacologically (ISRIB) or genetically inhibition (G3BP1 knock down) of SG leads to an exaggerated increase in TNFα expression. Further, SG inhibition results in enhanced decrease in CM function (decreased intracellular cAMP) in CM with LPS. Over‐expression of G3BP1 in CMs prevents LPS‐induced CM TNFα production and improves CM contractility.

Conclusion

Our results indicate that the cardiomyocyte stress granule activation plays a protective role of myocardial function in sepsis.
Bile Duct Ligation Changes the Inhibitory Control of Vasopressin Neurons in Male Rats
Authors: Aikins Ato O. , Brock Courtney , Little Joel T. , Farmer George E. , Cunningham J. T.
Published: May 2022

Dilutional hyponatremia typically is caused by increased Arginine Vasopressin (AVP) secretion. This increases the risk of complications and mortality in patients with liver cirrhosis. Liver cirrhosis in male rats increases AVP release but the underlying mechanisms are still under investigation. In other models of chronic AVP release, changes in chloride homeostasis alters the inhibitory control of AVP neurons in the supraoptic nucleus (SON) causing GABA mediated excitation instead of inhibition. In the bile duct ligation (BDL) model of cirrhosis, changes in either intracellular chloride concentration or cell firing activity have not been directly tested. We hypothesize that liver cirrhosis increases intracellular chloride concentration leading to a decrease in GABA‐mediated inhibition of SON neurons from male rats.

To test this hypothesis, we injected an adeno‐associated virus with a vasopressin promoter and a ratiometric chloride indicator, AAV2‐0VP1‐ClopHensorN into the SON of adult male Sprague‐Dawley rats. The rats either received BDL surgery, which was used to model liver cirrhosis, or a sham ligation surgery. All BDL rats had significantly higher liver weight to body weight ratio (n = 10‐20, p < 0.05), lower plasma osmolality (n = 10‐20, p < 0.001) and lower hematocrit (n = 10‐20, p < 0.05) as compared to sham rats. All rats were anesthetized with isoflurane (2‐3%), their brains were collected, and slices containing the SON were prepared. Loose‐cell patch recordings were made at 31±1oC using standard artificial cerebrospinal fluid. Labeled AVP cells and unlabeled SON cells were tested with focal application of muscimol (100µM), a GABAA receptor agonist.

Cells from BDL rats showed impaired GABAA‐mediated inhibition in response to muscimol, as compared to those from sham rats. All putative AVP SON neurons from sham rats (4/4) showed decreased activity following muscimol while half of the putative AVP SON neurons (7/14) from BDL rats were excited (p < 0.001). Similar results were observed in experiments were the injections missed the SON. In these experiments, all SON neurons from sham rats were inhibited by muscimol (26/26) while thirty‐five percent of SON neurons from BDL rats (16/45) were excited by muscimol (p = 0.001). ClopHensorN based chloride imaging showed that muscimol application was associated with decreased intracellular chloride concentrations in putative AVP cells from BDL rats as compared to sham rats (n = 31‐32, p = 0.001). The results show that liver cirrhosis impairs the normal inhibitory response of SON neurons to GABAA receptor activation which could contribute to increased cell activity and dilutional hyponatremia in male rats.
The Flavonoid Troxerutin Prevents Tissue Damage and the Alteration of Cellularity Residing in Intestinal Mucositis by 5‐Fluorouracil
Authors: Pedroza Rafaella H. S. , Rocha Paulo V. P. , Carioca Levi C. , de Oliveira Vitoria C. , de Souza Pimentel Paulo V. , de Sousa Fideles Lázaro , Cerqueira Gilberto S. , de Miranda João A. L.
Published: May 2022

Intestinal mucositis, characterized by inflammatory and/or ulcerative processes, occurs due to cellular and tissue damage following treatment with 5‐fluorouracil (5‐FU). Troxerutin (TRX), a semi‐synthetic flavonoid extracted from Dimorphandra gardneriana, has been reported as a potent anti‐inflammatory agent. The present study aimed to evaluate the effect of TRX on 5‐fluorouracil‐induced intestinal mucositis in Swiss mice. The mice (25‐30g) were separated into 3 groups (n = 6): Saline group (NaCl 0.9%), 5‐FU group, TRX group (100mg/kg TRX, orally). All animals, except the Saline group, received 5‐FU at the concentration of 450 mg/kg in the first experimental protocol. Duodenal portions were removed for evaluation of mucositis by analysis of histopathological damage and cellularity of resident cells of the intestinal mucosa (mast cells and goblet cells). The results of the histopathological analysis demonstrated that 5‐FU promoted structural alterations of the intestinal mucosa (p <0.05), evidenced by the reduction of villi height, deepening of the crypts, and TRX treatment (100 mg/kg) prevented the 5‐FU‐induced histopathological changes (p <0.05). Regarding cellularity, TRX decreases 5‐FU‐induced mast cell infiltration (p <0.05), as well as decreases cell lineage degranulation and, regarding goblet cell count, mucus‐producing cells, TRX prevented the decrease of these cells (p <0.05), which is a problem promoted by the 5‐FU. Our findings suggest that TRX at a concentration of 100 mg/kg had chemopreventive effects on 5‐FU‐induced intestinal mucositis due to the prevention of histological damage and maintenance of resident cellularity in the intestinal mucosa.
The Effects of Sex on CB1 Expression in Male and Female Mice After Morphine Treatment
Authors: Lauer Alyssa , Bobeck Erin
Published: May 2022

The endocannabinoid system (ECS) has recently gained interest as a promising therapeutic target for chronic pain treatment. An emerging body of research has revealed that the ECS interacts with the endogenous opioid system in a variety of complex processes. However, pain research has historically excluded female animals and failed to consider sex differences. This project aims to investigate whether morphine administration has a sexually dimorphic effect on type 1 cannabinoid receptors (CB1) in the ventral tegmental area (VTA) and amygdala of mice. These brain regions have been selected due to their role in both addiction and the emotional aspect of pain, which appear to be sexually dimorphic phenomena. Using reverse transcriptase qPCR, this study aims to determine (i) whether chronic morphine treatment impacts CB1 expression in the VTA and amygdala and (ii) whether this treatment differentially impacts male and female animals.
Differential Expression of Microglial BDNF in Response to Acute Hyperglycemia and Hypoxia
Authors: Mussa Bashair , Srivastava Ankita , Bajbouj Khuloud , Shafarin Jasmin
Published: May 2022

Diabetes is one of the most common chronic disorders that affects the homeostasis of several systems including the central nervous system (CNS). Hyperglycemia, main feature of diabetes, is involved in pathogenesis of diabetic neuropathy. In addition, hyperglycemia increases mitochondrial oxygen consumption which leads to hypoxia and affects the neuronal survival, adversely. Brain‐derived neurotrophic (BDNF) is well known as neuroprotective factor which supports the survival of the neurons in the CNS. We hypothesize that microglial BDNF has a protective role against acute hyperglycemia and hypoxia. The objective of this study is to investigate the expression of BDNF in response to hyperglycemia and hypoxia. Murine microglial cell line BV‐2 cultured in Dulbecco's Modified Eagle Medium were used to investigate the effects of increased concentrations (30, 60, 90, 120 mM) at different durations (4, 8, 24 hours) on the viability of microglial cells. In addition, the effects of hypoxia on the survival rate of these cells was also assessed. Gene and protein expression of microglial BDNF was examined in response to increased concentrations of glucose and hypoxia as well as hypoxia inducible factor (HIF1‐alpha). Increased concentrations of glucose produced significant increase in gene expression of microglial BDNF and this response was concentration‐dependent (p < 0.001). On the other hand, gene expression of microglial BDNF was significantly decreased in response to hypoxia (4, 8, 24 hours) and this response was associated with reduced expression of HIF‐1 alpha (p < 0.05). In conclusion, the outcomes of this study demonstrated that expression of microglial BDNF was sensitive to increased concentration of glucose and decreased levels of oxygen. Further investigation will dissect the signaling pathways that are involved in this differential expression to gain a better understanding of the protective role of microglial BDNF against glucose and oxygen variability.
PLEKHA5 Regulates Mitotic Progression by Promoting APC/C Localization to Microtubules
Authors: Cao Xiaofu , Shami Shah Adnan , Sanford Ethan J. , Smolka Marcus B. , Baskin Jeremy M.
Published: May 2022

Progression through major cell cycle checkpoints involves spatiotemporal coordination of cytoskeletal dynamics with changes in protein activities, chromosomal dynamics, and signaling events. A major role of the ubiquitin–proteasome system is to orchestrate cell cycle progression through temporally controlled protein degradation programs. The anaphase‐promoting complex/cyclosome (APC/C) is the primary E3 ligase controlling all aspects of mitosis, including G2/M transition, early mitosis, the metaphase–anaphase transition, and mitotic exit, via polyubiquitination of important mitotic effector proteins. The temporal control of APC/C‐mediated ubiquitination in these processes is well established, including identities of ubiquitination substrates and accessory factors that modulate E3 ligase activity. Much less is known, however, about the spatial organization of APC/C function, including its potential interplay with the microtubule cytoskeleton, which serves as a signaling platform and whose dynamic behavior is central to advancement through mitosis. Here, we investigate and describe pleckstrin homology domain‐containing family A, member 5 (PLEKHA5) as a new regulator of APC/C function and the ubiquitination pathway in mitosis. We found that PLEKHA5 localized to the microtubule network and interacted with APC/C. PLEKHA5 knockdown antagonized mitotic entry and progression, causing a buildup of APC/C substrates implicated in both the G2/M and metaphase–anaphase transitions, in a manner dependent upon the PLEKHA5 interaction with APC/C. In vitro ubiquitination assays showed that APC/C isolated from mitotic cells lacking PLEKHA5 had lower catalytic activity and a decreased association with its key mitotic co‐activator CDC20. We investigated the dynamic localizations of the APC/C and the potential role of PLEKHA5 in its regulation by developing a TurboID proximity biotinylation tool to assess microtubule localization of endogenous proteins. Microtubule‐targeted TurboIDs revealed that PLEKHA5, APC/C subunits, and the co‐activator CDC20 all localized to microtubules both in interphase and in M phase. Importantly, upon PLEKHA5 knockdown, a pool of APC/C, but not CDC20, lost its microtubule localization, which could explain the decreased APC/C‐CDC20 association. We propose that PLEKHA5 functions as a novel APC/C adaptor to promote its subcellular localization to microtubules and enable APC/C to search for its co‐activator CDC20 more efficiently in one dimension along the microtubule network. In this model PLEKHA5 acts to coordinate the spatial regulation of APC/CCDC20 and facilitate its efficient polyubiquitination of key mitotic effector proteins to ensure proper progression of mitosis.
Remote ischemic preconditioning promotes cardioprotection through a central nervous system dependent mechanism
Authors: Amilcar Ashline Paul Manoucheca , Rosas‐Martínez Lorena , Torres‐Narváez Juan Carlos , Sánchez‐López Araceli , Cervantes‐Pérez Luz Graciela , Centurión David , Sánchez‐Mendoza Alicia
Published: May 2022

Remote ischemic preconditioning (rIPC) is the phenomenon of brief periods of ischemia and reperfusion applied to a distant organ promote cardioprotection against a subsequent ischemia/reperfusion injury (I/R). Although a neurohumoral hypothesis has been proposed, the cardioprotective mechanisms of rIPC have not been fully elucidated. We investigated whether rIPC could induce cardioprotection in a pithed Wistar rat model hypothesizing that the absence of central nervous system (CNS) structures would prevent its cardioprotective effect. rIPC protocol consisting of 3 cycles of 4 minutes of ischemia and 4 minutes of reperfusion was applied to the left posterior extremity of male Wistar rats. I/R was induced surgically by left coronary descending artery ligation for 30 minutes followed by 2h of reperfusion. Isolated rat hearts were perfused in a Langendorff system to measure coronary vascular resistance (CVR). Finally, the infarct size was evaluated by Evans blue and triphenyltetrazolium staining. rIPC reduced the infarct size of the ischemic hearts; however, this effect was abolished in pithed rats. rIPC also prevented the rise of CVR of ischemic hearts while there was no difference compared to pithed rats. We concluded that rIPC reduces the infarct size of hearts during I/R preserving the coronary artery tone through a CNS‐dependent mechanism.
Structural Characterization of LpqH (19 kDa surface antigen), an Immunomodulatory Lipoprotein of Mycobacterium tuberculosis
Authors: Chatterjee Shruti , Kundapura Shankar , Basak Aditya J. , Ramagopal Udupi A. , Samanta Dibyendu
Published: May 2022

Tuberculosis, caused by Mycobacterium tuberculosis, is predominantly a disease of the lungs acquired by inhaling mycobacteria from infected individuals via airborne droplets. As soon as Mycobacterium reaches the lungs, its first encounter happens to be with the resident alveolar macrophages which immediately can phagocytose, but cannot kill the pathogen. Thus, the pathogen‐laden macrophages now disseminate to other organs, aiding in the spread of infection in an undetected manner. Certain mycobacterial surface motifs known as pathogen‐associated molecular patterns (PAMPs), recognize specific pattern recognition receptors (PRRs) present on the host cell surface, thereby mediating these early interactions of the mycobacteria with macrophages. A major group of these bacterial PAMPs includes mycobacterial lipoproteins, of which, the 19 kDa surface antigen LpqH, have a critical role in shaping the host immunological response in tuberculosis. The protein pleiotropically modulates the host immune response by interacting with Toll‐Like Receptors (TLRs) such as TLR2 homodimers and TLR1/TLR2, TLR2/TLR4, and TLR2/TLR6 heterodimers. On one hand, it acts as an apoptosis‐inducing factor for macrophages whereas on the other, it shows that prolonged exposure of LpqH to TLRs eventually favours immune evasion, thereby allowing the dissemination of the pathogen. Despite the essential role of LpqH in tuberculosis,till date, the detailed structure‐function relation of this protein remains to be elucidated. We have determined the high‐resolution crystal structure of Mycobacterium tuberculosisLpqH at a resolution of 1.26Å and refined to Rwork and Rfree of 14.29% and 17.79%, respectively. It has been observed that the structural architecture of LpqH is indeed unique and has never been previously described. Furthermore, sequence identity between LpqH and other well‐known mycobacterial lipoproteins is very low, which further implies that the protein is indeed unique and its structural characterization is vital to the overall understanding of its pathogenesis in case of active tuberculosis.
OVLT Lesion attenuates hypertension and polydipsia in 2‐kidney, 1‐clip hypertensive Rats
Authors: Ruiz Lauar Mariana , Pestana‐Oliveira Nayara , Van Helden Dusty , Collister John P. , Osborn John W.
Published: May 2022

Hypertension (HTN) is a multifaceted condition that affects one‐third of the population of western countries. It is well established that renal ischemia results in secondary renovascular HTN. Experimental renovascular HTN is induced in rats by creating stenosis of the left renal artery, thereby decreasing perfusion pressure (2K1C‐HTN), leading to increased renin‐angiotensin system activity which may act centrally to increase arterial pressure, sympathetic nerve activity, and thirst. The organum vasculosum of the lamina terminalis (OVLT) is a circumventricular organ which is responsive to blood borne substances such as angiotensin II and has been implicated in the regulation of arterial pressure, sympathetic activity, and thirst. Previous studies have shown that OVLT lesion attenuates development of the DOCA‐model of hypertension, but does not modify the water intake (Collister et al., 2018). In the present study we tested the hypothesis that the OVLT is required for the development of 2K1C‐HTN as polydipsia is observed in this model. Male Sprague‐Dawley rats (n=3‐5/group) were randomly selected to receive either electrolytic lesion of the OVLT (OVLTx) or sham operation. After one week, they were instrumented with telemeters to measure mean arterial pressure (MAP). Two weeks after OVLTx, the left renal artery was stenosed by application of a silver clip. MAP was measured continuously for 6 weeks and, once a week, rats were housed in metabolic cages for 24 hours to measure water intake and urine volume. At the end of the protocol, MAP was significantly (p<0.001) lower in OVLTx‐2K1C (134±8 mmHg) compared to sham lesion‐2K1C (163±6 mmHg) rats. In addition, water intake was significantly (p<0.001) lower in OVLTx‐2K1C (42±2 ml/day) compared to sham lesion‐2K1C (73±2 ml/day) rats. Similarly, urinary volume excretion was reduced in OVLTx‐2K1C (25±2 ml/day) compared to sham lesion‐2K1C (52±5 ml/day) rats. We conclude that the integrity of OVLT is required for the development of 2K1C hypertension as well the polydipsia observed following renal artery stenosis.
Meniscus Bioprinting Technology Roadmap
Authors: Gaspar Paulo E. A. , Maia Isadora , Amaral Daniel C. , da Silva Barud Hernane , Rezende Rodrigo A. , Amaral Creusa Sayuri T. , Amaral André C.
Published: May 2022

Meniscus lesions represent a challenge for medicine in view of the high occurrence, the limited intrinsic regenerative potential of the meniscus and the economic and social impact associated with the restricted results of current treatments. One of the most promising alternatives is the development of meniscus engineered substitutes (MES) through 3D bioprinting. To define strategies for technologies development, processes, and products necessary for MES bioprinting, innovation management and planning tools, such as roadmaps (RMap), can be used. This research aimed to build an RMap to portray the scientific and technological contemporary status related to tissue engineering strategies for bioinks development (BI) for MES bioprinting. For this, the search for scientific publications were carried out in journals (Science Direct, PubMed, and Web Of Science) and patents (Google patents and EspaceNet) using as strings the combination Meniscus AND Scaffold AND Bioprinting AND Tissue Engineering. The search was restricted to original scientific articles and patents published between 2015 and 2021. The selected publications were carefully analyzed to identify the technologies developed (bioprinting process, biomaterials, and cells). The information was compiled and compared with the biomechanical performance criteria of a meniscus, to infer the degree of biomimicry and the potential of each innovation proposal. 9 articles and 4 patents were selected. Except for one publication using the inkjet printing method, all explored the extrusion method. Only 4 of the publications incorporated cellular contingent, predominantly the use of stem cells derived from fat and bone marrow. Regarding biopolymers, polycaprolactone (PCL) predominated, used alone or in association with collagen, gelatin methacrylate, alginate, poly(lactic‐co‐glycolic acid) and decellularized extracellular matrix. It is noteworthy that PCL generates a rigid matrix, technically incompatible with the biomechanical nature of the meniscus. It means that the proposals under development have at least one characteristic that could impede the future use of the under‐development technologies. This finding reinforces the importance of using technological planning techniques to support biotechnology research. The Rmap obtained describes the frontier technologies of BI knowledge for MES bioprinting, showing incompatibility between the MES biomechanical performance characteristics and the properties necessary to achieve faithful biomimicry concerning biological menisci. The expectation is that this Rmap will contribute to the development of new BI strategies, focusing on the selection and/or modification of biomaterials and cells, to achieve the biomimetic performance of MES.
Macromolecular Crowding Effects on Citrate Synthase Kinetics are Concentration‐Dependent
Authors: Ariola Ashton B.
Published: May 2022

Enzyme studies are usually conducted in dilute, in‐vitro conditions; however, enzymes operate within extremely crowded intracellular environments, where macromolecules such as proteins, carbohydrates, and lipids occupy up to 40 % of the available volume. This macromolecular crowding slows diffusion, alters protein structure, and has erratic consequences on enzyme kinetics. To study the potential impacts of crowding on enzyme behavior, Michaelis‐Menten kinetics were employed to monitor citrate synthase reaction rates under crowded conditions. The synthetic polymer dextran was used to mimic the crowded cell interior, and assays were conducted in the presence and absence of dextran at various sizes (10 kDa ‐ 550 kDa) and concentrations (25 g/L ‐ 300 g/L). Crowding decreased the maximum reaction rate, Vmax, for all dextran conditions; however, the severity of this decrease in reaction rate depended on the concentration of dextran. For dextrans at concentrations below their overlap concentrations (c*), crowding resulted in a 10% ‐ 20% decrease in Vmax. However, the reaction rate dropped 30% ‐ 50% at dextran concentrations above c*. The impact of mixtures of dextran sizes on citrate synthase kinetics is also being studied. Taken together, these results indicate that citrate synthase activity is sensitive to the overlap concentration of these dextran solutions.
The Glass Ceiling Thickens: How The COVID‐19 Pandemic Negatively Impacted Academic Medicine Faculty
Authors: McNulty Margaret A. , Lufler Rebecca S.
Published: May 2022

INTRODUCTION

Inequities faced by women in academic medicine before the COVID‐19 pandemic are well‐known, yet there are little formal data on exactly how the pandemic has affected faculty, particularly in academic medicine. The purpose of this cross‐sectional study was to elucidate the impact of the pandemic on faculty in academic medicine and identify whether it is disproportionately affecting populations, focusing specifically on responsibilities at home, work, and mental health according to gender identification, faculty rank, and faculty appointment.

METHODS

An online questionnaire was distributed in February 2021 to faculty at academic medicine centers. This questionnaire asked respondents to provide demographic data, answer questions about their responsibilities at home and work, and indicate whether and how the pandemic has affected both. Respondents were also asked questions on self‐care and mental health and to document how their institution(s) can assist faculty in mitigating struggles associated with the pandemic. Responses were analyzed via quantitative (Pearson’s chi‐square tests) and qualitative (thematic analysis) means.

RESULTS

Women, mid‐, and early‐career faculty were more likely to be responsible for the care of others (p=0.014), and the impact of the pandemic on caring for others was negative, especially for early career faculty (p=0.019). Research, teaching, and clinical practice were negatively impacted by the pandemic, with women feeling this more severely in clinical practice (p=0.003), as a result of an increased teaching load (p=0.042), and inadequate work environment (p=0.013). Women (p<0.01), early career‐faculty (p<0.01), and clinical faculty (p=0.029) were also more negatively impacted by the pandemic in the areas of self‐care and mental health. While there was no significant difference in who requested a leave of absence or tenure clock extension, early‐career and women faculty were more likely to fear retribution for such requests. Faculty provided actionable suggestions to combat these negative effects under five themes, including Flexible Expectations, Support, Mental Health, Compensation, and Communication.

CONCLUSION

Pre‐pandemic stress and burnout among faculty in academic medicine has been well documented, and this study demonstrates that academic medicine faculty, particularly women and early‐career faculty, continue to feel additional burdens secondary to the COVID‐19 pandemic. These data can be utilized by administrations and future studies to make informed decisions regarding policies and programs created to assist those populations who are most vulnerable to the effects of the pandemic.

SIGNIFICANCE

To the authors’ knowledge, this is the first study to provide both quantitative and qualitative data demonstrating a negative impact of the COVID pandemic on academic medicine faculty, particularly women and early career faculty. These data are necessary to create change to improve recruitment and retention of quality faculty.
Tac1‐expressing cells in the pre‐Bötzinger complex are potential targets to prevent opioid‐induced respiratory depression
Authors: Rousseau Jean‐Philippe , Scarpellini Carolina S. , Montandon Gaspard
Published: May 2022

Rational

Opioids are extensively used for their analgesic properties but present a variety of unwanted side effects, including tolerance, dependence and respiratory depression. The analgesic effect of opioids is due to activation of µ‐opioid receptors (MOR) in the central nervous system and no treatments are currently available to prevent respiratory depression without reducing their analgesic properties. The neural circuits and mechanisms regulating respiratory depression, sedation, and analgesia by opioids often overlap, therefore making challenging the identification of the mechanisms regulating respiratory depression. Neurons expressing tachykinin precursor 1 peptide (Tac1) located in the pre‐Bötzinger complex (preBötC) also co‐express neurokinin‐1 receptors (NK1R) and MORs. NK1R neurons are preferentially inhibited by opioids and play an essential role in mediating opioid‐induced respiratory depression.

Objective

Here, we tested the hypothesis that optogenetic activation of Tac1‐expressing preBötC cells in freely behaving mice will modulate breathing and prevent respiratory depression by opioids.

Methods

Using a Cre‐loxP recombination approach, we injected in Tac1 Cre recombinase mice the adeno‐associated virus containing the gene cassette of the excitatory channelrhodopsin‐2 ChETA flanked between loxP sites. A 200 µm optical fiber was then fixed above the preBötC for laser stimulation with blue light (wavelength: 480 nm). After a four weeks period to allow ChETA expression in targeted cells, the mouse was placed in a plethysmographic chamber and breathing was measured while laser stimulations were performed under baseline conditions (30 Hz). Following baseline measurements, the clinically relevant µ‐opioid drug fentanyl was administered by intraperitoneal injection (0.3 mg/Kg). About 5 minutes after fentanyl injection, targeted preBötC cells were activated using laser stimulations (30 Hz).

Results

Data show that stimulation of Tac1‐expressing cells increased respiratory frequency under baseline conditions. Fentanyl depressed breathing in 5 minutes and stimulation of Tac1 cells prevented opioid‐induced respiratory depression; an effect that was reversible.

Conclusion

Tac1‐expressing preBötC cells may constitute a target to prevent depression of breathing by opioids. These results will help identify the cells mediating opioid‐induced respiratory depression, an essential step toward the development of therapeutic targets to reduce the risk of opioids overdose and associated mortality.
Defining Dopamine‐Mediated Changes in NLRP1, NLRC5, NLRC4, and AIM2 Inflammasomes in Human Myeloid Cells
Authors: Canagarajah Joanna , Matt Stephanie , Gaskill Peter J.
Published: May 2022

Although the study of dopamine is most often associated with the regulation of reward and motor function, a growing body of research indicates that dopamine also acts as an immunomodulatory regulator. Our data show that dopamine can influence the secretion of inflammatory cytokines in myeloid cells such as macrophages and microglia. Specifically, we have shown that dopamine modulates inflammation in primary human monocyte‐derived macrophages (hMDM) through activation of the transcription factor NF‐κB and priming of the NLRP3 inflammasome, upregulating the expression of IL‐1 family cytokines (IL‐1β and IL‐18). However, the effect of dopamine on other proteins in the NLR family, including NLRP1, NLRC4, NLRC5, and AIM2, has not been defined. Each NLR protein is associated with a different inflammasome complex that has distinct activating stimuli, yet all inflammasomes mediate the secretion of IL‐1 family cytokines. We hypothesize that the effects of dopamine on different types of inflammasomes is distinct and may result in activation of discrete inflammatory pathways and/or differentially regulate cytokine secretion. To address this, we examined dopamine‐mediated changes in NLR proteins in both hMDM and a human microglial cell line (C20 cells). Our data indicate that in hMDM, dopamine upregulates expression of NLRC5, NLRC4, and AIM2 transcripts, while not changing NLRP1 mRNA expression. However, in microglia there was no expression of NLRC4 or AIM2, and dopamine downregulates NLRP1, while not changing NLRC5 mRNA expression. Ongoing work is using high‐content immunofluorescent imaging and Western blotting to analyze inflammasome protein expression in these myeloid cell models and determine how this correlates with NLR gene expression as well as dopamine receptor expression. Determining how dopamine affects NLR expression could improve our understanding of the role of dopamine and its receptors as drug targets in individuals with substance use disorders. Further, the activity of these inflammasomes is associated with a number of neuroimmunological diseases such as Parkinson’s, Alzheimer’s, multiple sclerosis and HIV that show disrupted dopamine signaling. Therefore, understanding the more specific immunologic actions of dopamine could initiate drug repurposing and the development of new therapeutic strategies based on manipulating dopamine’s effects on the immune system.
Exploring ATAD2 bromodomain function in the dynamic epigenetic landscape
Authors: Malone Kiera , Glass Karen
Published: May 2022

The ATPase family, AAA domain‐containing protein 2 (ATAD2) is a nuclear co‐regulator protein highly expressed in many unrelated cancers. ATAD2 overexpression is linked to poor outcomes in patients. ATAD2 contains a C‐terminal bromodomain that “reads” acetylated lysine post‐translational modifications (PTM) that occur on histone proteins present in the nucleosome core particle. The epigenetic landscape is dynamic and combinatorial, containing multiple types and numbers of modifications on the histone proteins at any given time. Yet, it is unknown how the presence of other modifications adjacent to acetylated lysine residues impact bromodomain protein recognition and function. We hypothesized that the presence of nearby post‐translational modifications including methylation and phosphorylation would modulate the ability of the ATAD2 bromodomain to recognize its acetylated lysine binding partners. Previously, we systemically screened for multiple PTM combinations recognized by the ATAD2 bromodomain using dCypher technology from EpiCypher. In our current study, we further characterized the interaction with these histone ligands using biophysical techniques including isothermal titration calorimetry (ITC), nuclear magnetic resonance (NMR), and X‐ray crystallography. Our results indicate that the histone binding activity of the ATAD2 bromodomain is impacted by the presence of nearby PTMs. Our results provide new insights on how the bromodomain functions to target the ATAD2 protein to chromatin, and how this interaction may be fine‐tuned via dynamic changes in the epigenetic landscape.
Ameliorative potential of D‐Ribose‐L‐Cysteine in male Wistar rats with metabolic syndrome
Authors: Ojetola Abodunrin A. , Fasanmade Adesoji A.
Published: May 2022

Metabolic Syndrome (MS) refers to cluster of cardiometabolic risk factors which include obesity, insulin resistance, dyslipidaemia, hyperglycaemia and hypertension. Although, oxidative stress is implicated in its aetiology, protocols in MS management have not included the use of antioxidants. D‐ribose‐L‐cysteine (DRLC) is a potent antioxidant which has been suggested to be useful in the management of dyslipidaemia. This study was designed to investigate the effects of DRLC in rats with MS.

Twenty male Wistar rats (190‐200g) were assigned into four groups (n=5). Groups I and II were fed with standard diet (SD) for eight weeks and were subsequently treated orally with: 10mL/kg distilled water and 250mg/kg DRLC daily for another eight weeks respectively. Groups III and IV were fed with a modified standard diet (MSD) consisting of 17% fructose and 20% lard with additional 10% fructose solution, given ad‐libitum for eight weeks to induce MS in rats. They were subsequently treated orally with: 10mL/kg distilled water (group III) and 250mg/kg DRLC (group IV) daily for another eight weeks respectively. Animals were weighed and Blood Pressure (BP) measured using rat tail cuff method. They were euthanized, and blood and heart were collected for biochemical analysis. Fasting Blood Glucose (FBG) was determined through glucose oxidase method. Serum C‐reactive Protein (CRP), Tumour Necrosis Factor alpha (TNFα) and insulin were determined through ELISA. Cardiac reduced Glutathione (GSH), Glutathione Peroxidase (GPx), Superoxide Dismutase (SOD), Xanthine Oxidase (XO), malondialdehyde; serum triglycerides, Total Antioxidants Capacity (TAC), High Density Lipoprotein (HDL) and Total Cholesterol (TC) were determined by spectrophotometry. Low Density Lipoprotein (LDL) was determined using Friedwald’s formula. Data were expressed as mean ± SEM and compared by ANOVA at p < 0.05.

Body weight, systolic BP, FBG and TNFα of group IV (255.8±10.8g, 167.8±8.6mmHg, 84.3±6.0mg/dL, 433.0±54.8pg/mL, respectively) were significantly reduced compared to group III (318.9±12.7g, 219.0±9.8mmHg, 140.9±9.9mg/dL, 577.8±31.8pg/mL, respectively). Also, insulin, TC, LDL and heart XO of group IV were significantly reduced compared to group III. The serum levels of CRP, triglycerides and malondialdehyde in group II were significantly reduced compared to group I. There were significant increases in serum TAC, cardiac GPx, SOD, GSH; and serum HDL of group IV (1.2±0.3mmol/mL, 2.4±0.5U/L, 1.1±0.1U/L, 11.3±9.7mM, 13.4±1.6mg/dL, respectively) compared to group III (0.6±0.2mmol/mL, 1.1±0.2U/L, 0.7±0.1U/L, 0.8±0.2mM, 7.7±1.4mg/dL, respectively).

The D‐ribose‐L‐cysteine ameliorated dyslipidaemia, hyperglycaemia and hypertension by decreasing oxidative stress and pro‐inflammatory cytokines in male rats with metabolic syndrome.
D230025D16Rik: A Novel Regulator of Muscle Cell Differentiation
Authors: Chaffer Tomer J. , Moamer Alaa , Leduc‐Gaudet Jean‐Philippe , Broering Felipe E. , Mayaki Dominique , Huck Laurent , Sandri Marco , Gouspillou Gilles , Hussain Sabah
Published: May 2022

Rationale

Loss of skeletal muscle mass and function is a common clinical feature and poor prognostic indicator in various diseases. A promising therapeutic approach of regenerative medicine for repair of skeletal muscle is through the delivery of cellular therapies targeting regulators of muscle regeneration (myogenesis). However, not all the proteins involved in this process have been identified. To address the gap in knowledge of proteins involved in myogenesis, we recently identified a novel gene, D230025D16Rik, which has never been characterized within the context of skeletal muscle growth and repair.

Methods

The role of D230025D16Rik in a C2C12 murine model of myoblast differentiation was assessed by using an siRNA loss‐of‐function approach. Differentiation of C2C12 myoblasts into myotubes in cells with intact D230025D16Rik and those with D230025D16Rik knockdown (KD) was determined by various experimental techniques, such as RT‐qPCR, microarray, immunofluorescence staining, and immunoblotting.

Results

D230025D16Rik has a mainly nuclear localization in myoblasts, whereas in myotubes, D230025D16Rik has both a nuclear and cytosolic localization. D230025D16Rik expression level is induced at the onset of differentiation (i.e., Day 1) and steadily increases as myoblasts fuse into myotubes. In response to D230025D16Rik KD, the expression of transcription factors MyoD1 and Myogenin and muscle‐specific transcripts Myosin Heavy Chain (MyHC) and Muscle Creatine Kinase (MCK) were upregulated such that a significant acceleration of myogenesis was observed. This observation was further confirmed by an analysis of transcriptomes of control and D230025D16Rik KD myoblasts that revealed an upregulation of myogenesis‐related transcripts in cells with D230025D16Rik KD. Phenotypically, we observe hypertrophic myotube formation at days 0 and 1 in response to D230025D16Rik KD, as indicated by immunofluorescence staining for MyHC. Finally, the presence of nuclear aggregates on D230025D16Rik KD myotubes indicates that D230025D16Rik is essential for proper myotube formation.

Conclusion

Altogether, these findings highlight, for the first time, the potential regulatory functions of D230025D16Rik during myogenesis.
Using Forward and Reverse Genetics to Understand Calcium Dysregulation in Enteric Viral Virulence
Authors: Gebert J. Thomas , Hyser Joseph
Published: May 2022

Acute gastroenteritis (AGE) remains the second leading cause of death among children under the age of 5 worldwide. AGE becomes fatal when pathogen‐associated upregulation of secretory activity across the intestinal epithelium causes severe volume depletion, hypovolemic shock, and multi‐system failure. While enteric viruses remain the most common cause of fatal AGE in kids, the drivers of their virulence (i.e. how they increase secretory activity) remain poorly understood. We recently found that cells infected with rotavirus, the most prevalent enteric virus in kids, release adenosine diphosphate (ADP) to induce coordinated signals known as “intercellular calcium waves,” which spread through uninfected neighboring cells. This dysregulates calcium signaling pathways, which enhances fluid secretion from uninfected cells and thereby contributes to volume depletion. Pharmacological blockade of calcium waves in infected mouse pups decreases disease severity, suggesting they are integral to rotavirus virulence. Understanding how rotavirus triggers intercellular calcium waves may allow us to design safer, more effective vaccines and therapeutics, but we still lack a mechanistic understanding of this process. Here, we report data showing that recombinant expression of a single rotavirus protein, non‐structural protein 4 (NSP4), is sufficient to activate intercellular calcium waves. Using the rotavirus reverse genetics system, we show that the diminished calcium wave phenotype associated with an attenuated strain of porcine rotavirus segregates with the NSP4 gene upon reassortment. Furthermore, we show that this attenuation is attributable to a single amino acid polymorphism at the c‐terminal end of NSP4. This implicates the c‐terminal end and a putative PDZ‐binding motif therein as key upstream mediators of intercellular calcium waves. These findings deepen our understanding of enteric virus pathogenesis, characterize a new role for rotavirus non‐structural protein 4, offer novel targets for anti‐secretory therapeutics, and expand foundational knowledge to support the development of improved live‐attenuated vaccines.
Preclinical Drug Development of a Novel Antiviral Target in Rotavirus
Authors: Duffy Emily‐Claire , Banks Lori D.
Published: May 2022

In 2021, rotavirus is the leading cause of severe gastroenteritis for children under the age of 5. While there is currently no cure for this virus, the use of medication is relied upon to alleviate pain from symptoms. A study conducted by the CDC in 2019 found that while only >1% of rotavirus infections under the age of 5 are fatal in high‐income countries, children from low‐income countries account for roughly 82% of all annual rotavirus‐related deaths. As a result, it is critical to developing new effective and affordable drugs to help improve accessibility in addition to decreasing the rates of infection. As the virally encoded ion channel (viroporin), non‐structural protein 4 (NSP4), is central to the replication and pathogenesis of rotavirus, it is a prime target for drug discovery studies. Similar viroporin proteins from HIV‐1 (Vpu) and Influenza A (M2) have been successfully targeted to treat their corresponding viral infections, but this method has not yet been tested for rotavirus infections. In this experiment, an E. coli‐based assay was developed to test known, antiviral ion channel inhibitors for their ability to bind and inhibit the channel activity of rotavirus NSP4. The subsequent data demonstrates that viroporin activity can be measured in a 96‐well format, rather than previous versions that employed 1‐2L cultures. As well, there is no perturbation of NSP4 protein expression or growth kinetics of the E.coli strains used in the assay. Ongoing experiments seek to test the anti‐viroporin activity of compounds representing a variety of core structures and mechanistic targets (Azelnidipine, Rimantadine hydrochloride, and 5‐(N,N‐Hexamethylene)amiloride). These compounds are being utilized to uncover the most effective molecular mechanism of NSP4 inhibition and to provide some biophysical information about the nature of inhibitor interactions with NSP4. These assays, along with drug binding kinetics being determined by thermofluor assay, will give us the first glimpse into NSP4 as a possible target for the treatment of rotavirus infections.
Renin inhibitor attenuates lipopolysaccharide‐induced anhedonia and upregulation of neuroinflammatory markers in mice
Authors: Alzarea Sami I. , Afzal Muhammad , Alharbi Metab , Alshammari Abdulrahman , Zafar Ameeduzzafar
Published: May 2022

Evidence suggests that the inhibition of renin‐angiotensin system can greatly decrease the neuroinflammation response. Neuroinflammation plays a pivotal role in the pathogenesis of depression characterized by the inability to experience pleasure in some patients. The aim of this study was to investigate the effects of aliskiren, a renin inhibitor, on lipopolysaccharide (LPS)‐induced anhedonia and upregulation of neuroinflammatory markers in mice. Adult healthy male albino mice were divided onto two groups (control vs LPS). Animals in control group received saline, whereas animals in LPS group received 0.5 mg/kg of systemic LPS injection. Pretreatment of aliskiren was done in all groups at different doses (1, 3, and 10 mg/kg). Sucrose preference test (SPT) and open field test (OFT) were used for the assessment of anhedonia and locomotor activity respectively in mice. Proinflammatory cytokines (TNF‐α, IL‐1β, and IL‐6) and glial cells markers (CD11b for microglia and GFAP for astrocytes) in the prefrontal cortex were determined via RT‐PCR. Results depicts that LPS was able to induce anhedonia and upregulate mRNAs of proinflammatory cytokines and glial cells markers after 24 h. Prophylactic treatment with aliskiren (10 mg/kg) significantly prevented LPS effects by increasing the consumption of sucrose in SPT. Locomotor activity in OFT did not show significant changes in aliskiren‐pretreated mice 24 h after LPS administration. Aliskiren (10 mg/kg) significantly reduce LPS‐induced upregulation of proinflammatory cytokines and glial cells markers in the prefrontal cortex. Taken together, the renin inhibitor aliskiren exerted beneficial effects in animal inflammation‐based model of depression characterized by appearance of anhedonia and upregulation of neuroinflammatory markers.