bluuebunny/crossref_metadata_2025 huggingface dataset:
Melanoma Impairs Mouse Heart Function Which Short‐Term Exercise Cannot Restore
Authors: Uurasmaa Tytti‐Maria , Streng Tomi , Alkio Milla , Karikoski Marika , Heinonen Ilkka , Anttila Katja
Published: May 2022

The aim of this study was to investigate whether melanoma impairs intrinsic heart function in mice and whether short‐term voluntary running wheel exercise could reduce the possible negative effects. Additionally, we investigated whether changes in cell size, capillary density, calcium channel levels, metabolic enzyme activities or oxidative stress could explain the possible changes in heart function. Advanced melanoma has been shown to cause cardiac muscle wasting and influence heart function in vivo, we hypothesized that melanoma would also impair intrinsic heart function, which has not been investigated previously. We also hypothesized that voluntary short‐term exercise could reduce the effects of melanoma on cardiac function since exercise training is known to improve cardiovascular function and reduce the negative effects of some cancers.

Male mice were divided into untrained tumor‐free group (control) and untrained melanoma group and trained melanoma group. The mice did voluntary running‐wheel exercise until predetermined tumor size after which their hearts were isolated. The cardiac function was measured in retrograde perfusion at a constant pressure in multiple oxygen levels using Langendorff apparatus. The molecular and tissue level markers were measured afterwards.

The melanoma animals were not cachectic as indicated lack of body weight loss. The rate of pressure production, pressure amplitude and rate of pressure decline were all significantly lower in the isolated hearts of the melanoma animals as compared to tumor‐free animals. However, the heart function did not differ between the untrained and trained melanoma groups. Furthermore, there were no differences between the groups in the calcium channel levels, reactive oxygen species, catalase activity, lipid peroxidation or citrate synthase and lactate dehydrogenase activity. However, mice from both melanoma groups had significantly lower superoxide dismutase activity as compared to tumor‐free animals, which might reduce the heart’s ability to respond to possible changes in oxidative stress. Exercise trained animals had higher capillary density, but their cell size and heart weights did not significantly differ from the untrained groups. Running wheel exercise did not affect the final tumor growth either even though there was tendency at the beginning of the experiment that running wheel exercise could slow down the tumor growth. One reason for this could be that when melanoma proceeded, the mouse running activity reduced significantly.

In conclusion, melanoma is an aggressive cancer that impairs intrinsic heart function even when no cachexia is present. The aggressiveness of melanoma also prevented the short‐term voluntary exercise from alleviating the changes caused by melanoma. Future studies should combine the training to medical cancer treatment in order to estimate whether exercise training could be beneficial as adjunct therapy in melanoma treatment.
Axillary Arch (Langer’s Arch) in an 80‐Year‐Old White Male Cadaver
Authors: Granite Guinevere , Sheridan Ariana , Wind Gary
Published: May 2022

Recognizing the presence of an Axillary arch (Langer’s arch or muscle) can be important for clinical associations, such as when patients present with compression neuropathies and syndromes related to the upper extremity. Its presence is also relevant during surgical interventions involving the axillary fossa, such as sentinel node biopsy axillary lymph node dissection and pectoralis muscle flaps. While neglecting to identify variations rarely leads to an increase in mortality, it is essential to identify for clinical relevance. It is similarly important to anatomical instructors in the education of future medical providers. During cadaveric dissection of fifty cadavers, we observed a unilateral Axillary arch (Langer’s arch or muscle) found on the left side of an 80‐year‐old White Male cadaver inserting onto the tendon of the pectoralis major muscle. This anatomical variation is commonly referenced in the literature for its clinical significance. Operating surgeons, vascular interventionalists, and oncologists should be aware of this anatomical variant as it occurs while treating diverse patient populations.
Polarized Macrophages As Potential Anti‐Endometrioid agent
Authors: Artemova Daria , Vishnyakova Polina , Fomina Mariia , Lokhonina Anastasia , Elchaninov Andrey , Fatkhudinov Timur
Published: May 2022

Endometriosis is an estrogen‐dependent chronic inflammatory disease characterized by the presence of endometrial stroma and glands outside the endometrium. The gold standard in the treatment of endometriosis is laparoscopy but after that, the relapse rate is about 50%. Thus, the search for new effective non‐invasive methods of endometriosis therapy is an actual problem. Endometriosis has many properties similar to tumors including the fact that macrophages with an anti‐inflammatory phenotype (M2) prevail in the foci of endometriosis. It is already known that activated macrophages with a pro‐inflammatory phenotype (M1) have an antitumor effect. In this regard, we hypothesized that increasing the proportion of M1 macrophages could help in the treatment of endometriosis. The objectives of our work were (1) to assess the expression profile of macrophages in the foci of endometriosis in mice model of endometriosis; (2) to test the efficacy of administration of reprogrammed M1 macrophages as a possible anti‐endometrioid agent in the mouse.

We studied the phenotypic profile of macrophages in the foci of endometriosis and the anti‐endometrioid activity of M1 macrophages on in vivoallogeneic model of mouse endometriosis induced in ovariectomized mice after intraperitoneal transplantation of the mouse uterus accompanied by 17β‐estradiol therapy. Analysis of the phenotypic profile of macrophages in the foci of endometriosis was carried out using immunohistochemical staining, western blot analysis, and qPCR. We obtained a reproducible model with lesions that had characteristic morphology. It was shown that macrophages with an anti‐inflammatory arginase1+ phenotype prevail in the foci of endometriosis compared with the intact endometrium. After verifying endometriosis development in mice, the experimental group was injected with M1‐polarized macrophages, while the control group was injected with unpolarized macrophages, and then the animals were monitored for two weeks. Macrophage cell line RAW264 was incubated with lipopolysaccharide (100 ng/ml) for 24 h to obtain M1‐polarized macrophages. M1‐polarization was verified by significant up‐regulation of MARCO, iNOS markers using western blot and qPCR analysis. After treatment of animals with M1 macrophages, we found that the number of foci and their sizes significantly decreased (p<0.05) in comparison with control animals.

In our work, we proved the predominance of anti‐inflammatory macrophages in the foci of endometriosis and obtained the first results of a positive effect of M1 macrophages on the regression of foci of endometriosis.
Phenotypic profiling of mGlu7 overexpressing mice and its implications for neurodevelopmental disorders such as Rett syndrome
Authors: Buch Aditi , Fisher Nicole , Frietas Geanne , Vermudez Sheryl , Weiss Kelly , Gogliotti Rocco , Niswender Colleen
Published: May 2022

Neurodevelopmental disorders (NDDs) are characterized by deficits in communication, cognition, attention, social behavior and/or motor control. Previous studies have pointed to the involvement of genes that regulate synaptic structure and function in the pathogenesis of these disorders. One such gene, GRM7, encodes the metabotropic glutamate receptor 7 (mGlu7), a G protein‐coupled receptor that regulates presynaptic neurotransmitter release. Mutations and polymorphisms in GRM7 have been associated with (NDDs) in clinical populations; however, limited preclinical studies have evaluated mGlu7 in the context of this specific disease class. Rett syndrome (RTT) is an NDD caused by mutations in the Methyl‐CpG Binding Protein 2 (MECP2) gene. The cognitive deficits seen in RTT model mice have been previously correlated with a deficit in long‐term potentiation (LTP) at Schaffer collateral‐CA1 (SC‐CA1) synapses in the hippocampus. mGlu7 is abundantly expressed presynaptically at SC‐CA1 synapses and its role in the regulation of GABA release is imperative for the induction of LTP. Previous studies from our lab have shown that mGlu7expression is reduced in autopsy samples from patients diagnosed with RTT and in mouse models of RTT. Our lab has also shown that abnormal phenotypes in RTT mice can be corrected with mGlu7 positive allosteric modulators (PAMs). Despite the beneficial effects of mGlu7 potentiation, it is important to understand how an increase in mGlu7expression affects phenotypes seen in NDDs like Rett syndrome.

Here, we hypothesize that an excess of either of the two mGlu7 isoforms, mGlu7a and mGlu7b, in mice is sufficient to alter phenotypes. mGlu7 was globally overexpressed in mice by crossing a Grm7(a/b) Flx/+male with a CMV‐Cre+/‐ female. Starting at 6 weeks of age, behavioral assays were conducted for motor, cognitive, and stereotypic phenotypes. After the behavioral phenotyping, tissue was collected and processed for expression studies. Our data indicate abnormal phenotypes in motor, cognitive, and stereotypic assays. In particular, mGlu7 over‐expressing mice, regardless of isoform, display hyperactivity in a locomotor assay, associative memory deficits in a contextual fear conditioning assay, and increased repetitive behaviors as assessed by hindlimb clasping.

In conclusion, our study indicates that overexpressing mGlu7 in mice leads to abnormal phenotypes. We are now assessing synaptic plasticity phenotypes, including abnormal long‐term potentiation in mGlu7 overexpressing mice, and determining the effect of mGlu7 allosteric modulators in affecting these cognitive aberrations. Overall, these studies aim to capitalize on previous studies demonstrating the role of mGlu7 in the manifestation of phenotypes associated with NDDs.
Revisiting Small Molecule Inhibitors of Amyloid‐β Aggregation
Authors: Moore Kendall B. E. , Pirela James P. T. , Fortin Jessica S.
Published: May 2022

Alzheimer’s disease (AD) is associated with amyloid formation and accumulation in the brain contributing to neurodegeneration. To date, there are no suitable mitigation strategies that stop or slow AD progression. Novel therapeutic approaches applicable at different stages of AD amyloid formation are direly needed. The long‐term goal of this research is to establish effective treatment strategy for AD. Our overall objective is to develop and validate a new proprietary class of anti‐amyloidogenic molecules to impede the formation of Aβ toxic oligomers. The specific aims of our research are 1) to provide a relevant library of small molecules, 2) to identify the key chemical moieties driving their effect in interfering with oligomer formation (structure‐activity relationship). The potency of at least three different series of small molecules to reduce the formation of amyloid fibrils and toxic oligomers was assessed in vitrousing biophysical methods such as Thioflavin T (ThT) fluorescence and transmission electron microscopy (TEM). Based on the screening of 30 newly synthesized molecules, we discovered KN‐21‐48 and MO‐19‐49 as amyloid inhibitors of Aβ1‐40. A preliminary structure‐activity relationship study indicates that compounds KN‐21‐48 and MO‐19‐49 can serve as a scaffold for further modifications in the design of more potent novel small molecules targeting amyloidogenesis in AD. This project may lead to a better understanding of AD by providing a model compound that may confirm or deny the role of amyloidosis in disease progression.
Sex differences in aortic hemodynamics and associations with white matter hyperintensities in middle‐aged and older adults
Authors: Pearson Andrew G. , Miller Kathleen B. , Corkery Adam T. , Eisenmann Nicole A. , Howery Anna J. , Chin Nathaniel A. , Johnson Sterling C. , Barnes Jill N.
Published: May 2022

Elevated blood pressure (BP) in midlife increases the risk of developing white matter hyperintensities (WMH) in the brain. WMH are associated with increased risk of cognitive decline and dementia. Structural changes of the large central arteries occur prior to changes in brachial BP. We have previously reported positive associations between aortic hemodynamics and WMH in normotensive postmenopausal women. The purpose of this study was to evaluate sex differences in aortic hemodynamics and associations with WMH burden in middle‐aged and older adults. Ninety‐nine participants (age = 63 ± 4y), including n = 35 men and n = 64 postmenopausal women participated in this study. Aortic hemodynamics were derived from radial artery pressure waveforms measured using applanation tonometry. Aortic systolic blood pressure (aSBP), aortic diastolic blood pressure (aDBP), augmented pressure (AP), aortic augmentation index (AIx), and aortic transit time were calculated. WMH lesion volume and intracranial volume (ICV) were measured using a FLAIR and T1 scan on a 3T MRI scanner, respectively. WMH fraction was calculated as (WMH lesion volume/ICV)*100 and cubic root transformed to reduce skewness. Age‐adjusted multiple linear regressions were used to determine the influence of aortic hemodynamics on WMH fraction. There were no sex differences in aSBP or aDBP. AP was greater in women compared with men (16 ± 5 mmHg vs. 11 ± 5 mmHg; P < 0.001). AIx was also greater in women compared with men (28 ± 6 % vs. 18 ± 9 %; P < 0.001). Aortic transit time was lower in women compared with men (140 ± 11 ms vs. 151 ± 12 ms; P < 0.001). There were no sex differences in WMH fraction. There was a positive association between aSBP and WMH fraction in both men (r = 0.37, P = 0.015) and women (r = 0.36, P = 0.017). In addition, aDBP was positively associated with WMH fraction in men (r = 0.47, P = 0.003) and women (r = 0.36, P = 0.017). AP was positively associated with WMH fraction in men (r = 0.34, P = 0.027), but not women (r = 0.24, P = 0.296). AIx was positively associated with WMH fraction in men (r = 0.46, P = 0.004), but not women (r = 0.22, P = 0.570). There were no significant associations between aortic transit time and WMH fraction. In conclusion, AP and AIx were greater in women compared with men. The positive associations between AP and WMH fraction and AIx and WMH fraction were only apparent in men. These findings suggest a sex‐specific association between augmented pressure and augmentation index and WMH burden in middle‐aged and older adults.
Development of an FM interferential electrical stimulator for swallowing rehabilitation
Authors: Morimoto In
Published: May 2022

Dysphagia (disorder of swallowing) can develop due to structural, functional, psychological, and/or medical causes. The prevalence of dysphagia in the elderly is 7%~22% per 100,000 people, and its incidence increases with age. In Japan, pneumonia is the fifth leading cause of death, with aspiration pneumonia accounting for 70% of these cases. Treatment options for dysphagia have been proposed, including transcranial direct current stimulation (tDCS), transcranial magnetic stimulation (rTMS), peripheral neuromuscular stimulation (NMES), pharyngeal electrical stimulation (PES), and drug therapy. Among other things, an electrical stimulator using peripheral neuromuscular stimulation has been developed in the USA and is the most commonly used therapy in medical practice. This device aims to improve the muscle strength required for swallowing by applying a low‐frequency electric current to the muscles. In addition, a rehabilitation device using transcutaneous electrical sensory stimulation (TESS) without muscle contraction has been developed in Japan, which can increase the frequency of swallowing without discomfort by activating the sensation in the throat with a medium frequency.

In this study, a rehabilitation device will be developed using burst modulated square waves, which are high‐frequency square waves modulated by low‐frequency, devised by Hernández Arrieta. The high‐frequency component of burst‐modulated square waves can lower the impedance of the skin and induce muscle contraction with less pain than direct current or low‐frequency stimulation.

In our study, we developed a multi‐point stimulation electrode for the forearm. We applied 125 patterns of electrical stimulation with 25 electrodes to five healthy subjects and were able to produce 69 different hand postures. However, because different hand postures were sometimes expressed in each experiment even when the stimulus position was the same, we sought to use machine learning models to robustly estimate the stimulus position and hand posture. The results show that the Gaussian naïve Bayes classifier has the best prediction accuracy for the thumb and index finger, where the number of bends is low, and the decision tree algorithms LightGBM and GradientBoosting classifiers for the middle, ring, and little fingers, where the number of bends is high.

In the future, to use this electrical stimulator for swallowing, it is necessary to consider the shape of the electrodes to improve discomfort during stimulation, the stimulation position to stimulate appropriate muscles, and sufficient stimulation parameters to activate the swallowing muscle groups.
Discovering the Developmental Basis of Trachea‐Esophageal Birth Defects: Evidence for Endosome‐opathies
Authors: Edwards Nicole , Zhong Guojie , Ahimaz Priyanka , Kenny Alan , Kingma Paul , Wells James , Shen Yufeng , Chung Wendy , Zorn Aaron
Published: May 2022

The trachea and esophagus (TE) arise from a common foregut tube during embryonic development. Disruptions in TE morphogenesis cause congenital trachea‐esophageal defects (TEDs) such as esophageal atresia, tracheoesophageal fistula and tracheoesophageal clefts. TEDs occur in approximately 1 in 3500 births, but their etiology is poorly understood. We have established the www.CLEARconsortium.org; a multidisciplinary team of clinicians, geneticists, bioinformaticians, stem cell and developmental biologists using patient genome sequencing, animal models and iPSC‐derived human organoids to discover the genetic and developmental basis of trachea‐esophageal birth defects. Using the complementary advantages of Xenopus and mouse models we have defined the conserved molecular and cellular mechanisms that regulate normal TE morphogenesis. We show that downstream of Hedgehog/Gli signaling endosome‐mediated epithelial remodeling regulates TE morphogenesis which when disrupted results in tracheoesophageal clefts similar to human Pallister Hall syndrome patients. Proband‐parent trio genome sequencing identified an enrichment of potential damaging de novo variants in genes encoding membrane/vesicular‐trafficking proteins, suggesting a common “endosome‐opathy” pathway. Ongoing CRISPR mutagenesis screens in Xenopus tropicalis assessing candidate causative variants from patients confirms that the endosome protein Itsn1 is essential for TE morphogenesis, suggesting that the ITSN1 variant is likely pathogenic in the patient. Finally, leveraging results from animal models we have generated multi‐lineage human esophageal organoids from iPSCs with patient mutations to identify how mutations impact human esophageal differentiation. Together these results significantly advance our understanding of TEDs with the goal of revealing phenotype‐genotype associations that will inform prognosis and clinical treatment.
Is it Possible to do Quality Science in Anatomy? Brazilian Scientists say Yes!
Authors: Fazan Valéria Paula S.
Published: May 2022

In October 2021, the AD Scientific Index published a list of the Latin America Top 10.000 Scientists.This list was carefully elaborated to be a reference for the scientific performance and the added value of the scientific productivity of individual scientists. The total and last 5 years’ values of the i10 index, h‐index, and citation scores in Google Scholar were used. Individual scientists were classified by 12 subjects (Agriculture & Forestry, Arts, Design and Architecture, Business & Management, Economics & Econometrics, Education, Engineering & Technology, History, Philosophy, Theology, Law/Law and Legal Studies, Medical and Health Sciences, Natural Sciences, Social Sciences, and Others). Within these main subjects, a further classification included the authors' citation of their research field, such as organic chemistry, software architecture, anatomy, exercise physiology, neurosciences, and many others. Of the 10.000 scientists listed, a total of 7656 are Brazilian, most of them (2134) from the University of São Paulo. We searched the subject “Anatomy” on the list and found 33 mentions. From these, 29 are Brazilian scientists, 9 of them working with “plant anatomy” or related botanic fields, leaving a total of 20 names listed as working either on human or animal anatomy. We cross‐checked these 20 names with the actual list of the Brazilian Society of Anatomy members and found that 18 listed scientists are members of the Brazilian Society of Anatomy (Sociedade Brasileira de Anatomia – SBA). Despite being little in number, Brazilian Anatomists have been expressively contributing to the Anatomical Sciences, having h‐index above 26 and over 64,600 citations. The complete list with names, Institutions and field of interest will be presented.
Acute Cannabigerol Administration Lowers Blood Pressure in Mice
Authors: Vernail Victoria L. , Bingaman Sarah S. , Raup‐Konsavage Wesley M. , Vrana Kent E. , Arnold Amy C.
Published: May 2022

Cannabigerol (CBG) is a cannabinoid compound that is synthesized from Cannabis sativa L. and acts as a substrate for both Δ9‐tetraydrocannabinol (Δ9‐THC) and cannabidiol (CBD) formation. Given that it does not exhibit psychoactive effects, emerging research has focused on CBG as a potential therapeutic for health conditions including algesia, epilepsy, anxiety, and cancer. While CBG can bind to cannabinoid receptors CB1 and CB2, it has also been described as an agonist at α2‐adrenoreceptors (A2‐AR), which when activated inhibit the release of norepinephrine from α‐adrenergic neurons. This raises the concern that CBG could act at A2‐AR to reduce norepinephrine release to cardiovascular end organs, such as the heart and kidneys, causing a reduction in blood pressure. Despite this possibility, there are no reports examining cardiovascular effects of CBG. In this study, we tested the hypothesis that acute CBG administration can lower blood pressure. To test this, six male C57BL/6J mice underwent surgery at 8‐10 weeks of age to implant a radiotelemetry probe, which allows for continuous measurement of blood pressure, heart rate and locomotor activity in conscious, freely moving mice. Following 10 days of recovery, baseline measurements were obtained and then mice were randomized to receive intraperitoneal injections of CBG (3.3, 5.6, and 10 mg/kg) and vehicle in a crossover design, with at least one‐week washout between treatments. Changes in blood pressure, heart rate, and locomotor activity were measured for two hours post‐injection. We found that acute CBG significantly lowered blood pressure compared with vehicle (‐12±5 mmHg vehicle vs. ‐28±2 mmHg at 10 mg/kg CBG; p=0.018), with no apparent dose responsiveness at the doses used in this study (‐22±2 mmHg at 3.3 mg/kg CBG; ‐28±4 at 5.6 mg/kg CBG). The greatest blood pressure reduction was seen at 90‐minutes post‐CBG administration, which is consistent with reports for peak plasma concentrations of this compound in rodents. The blood pressure lowering effects of CBG occurred in the absence of changes in heart rate or locomotor activity. These overall findings suggest acute CBG may lower blood pressure in phenotypically normal young adult male mice, which may provide caution for the potential of hypotension as an adverse effect of CBG administration. Additional studies are needed to determine if the blood pressure lowering effects of CBG are via an A2‐AR mechanism.
Surgical Faculty’s Perspectives on the Unknown Utility of Alcohol‐Preservation in Surgical Resident Education
Authors: Leake Michael L. , Marek Mason , Ejaz Aslam , Chen Xiaodong (Phoenix) , Balta Joy Y.
Published: May 2022

Under ideal circumstances, surgical skills training should emulate live surgical conditions as closely as possible. While new anatomically accurate models and virtual/augmented reality simulators make surgical anatomical knowledge acquisition more accessible, research shows that these resources are limited in their ability to fully reflect the live human anatomy, and thus the animal and human body donor models remain the gold standards for surgical simulation (Stefanidis et al., 2013, Venne et al., 2020). Previously, there have been studies comparing the porcine and human body donor models. However, very little is known about alcohol‐preserved human body donors in the context of surgical training and education (Balta et al., 2015). The objective of our study was to examine the alcohol‐preservation technique and to further elucidate its utility in general surgery resident education. Utilizing semi‐structured interviews, we analyzed qualitative data obtained from the accounts of surgical faculty who experienced both the porcine and alcohol‐preserved models at The Ohio State University. Results indicated that the porcine and alcohol‐preserved human body donor models were preferred for different aspects of surgical training. While the porcine model was preferred for its ability to teach tissue‐handling and to simulate live blood flow, the alcohol‐preserved model was unanimously preferred for its anatomical relevance. Surgical faculty specifically noted that while there is anatomical overlap between the two models with the anatomy of the rectum, “the small bowel and colon are dramatically different, which does not give the full breadth of colon resection needed for the human model.” The alcohol‐preserved model was also preferred for its ability to teach advanced surgical procedures in low‐stake environments, with a surgical faculty guiding them in a stepwise fashion, and the model was regarded as superior in all aspects as compared to formalin‐fixed human body donors. From the results, it was concluded that the alcohol‐preserved human body donor model showed unique utility in the context of surgical resident education, simulating live operating conditions more accurately as compared to the formalin‐fixed model, and being more anatomically accurate than the porcine model. The findings of this study reflect a significant step in improving surgical resident training and education. Surgical technical proficiency is intimately linked with a surgeon’s amount of experience, and by elevating surgical resident preparation, patient surgical outcomes can likely be improved. Future directions of our research involve the continued refinement of the alcohol‐ and soft‐preservation techniques in order to more accurately simulate live operating conditions, to further improve surgeon training and preparation, translating to better future patient surgical outcomes.
Anatomical Gift Programs Continue to Meet Donor Demands of Anatomy Educators During the COVID‐19 Pandemic: Here’s How
Authors: Cope Janet , Bennett Cynthia M. , Zimmerman Monica , Balilionis Gytis , Person Dianne
Published: May 2022

INTRODUCTION

Schools of higher education across the US closed all operations and in‐person classes during the COVID ‐19 pandemic, significantly affecting laboratory‐based anatomy dissection courses and the anatomical gift programs (AGP) that support them. The purpose of this study was to investigate how the COVID‐19 pandemic affected AGP operations in the US. The researchers hypothesized that programs would modify donation acceptance criteria and that anatomy education demands for donors would be reduced during the first year of the pandemic.

METHODS/ RESULTS

The researchers sent a Qualtrics survey to 135 AGP directors with a response rate of 28% (n=38). Respondents were from 26 states and included 28 AGPs operating for 25 years or longer and ten programs operational for 15 years or less. Six AGP’s (16%) stopped accepting donors for varying lengths of time secondary to the COVID‐19 pandemic during its first year, while 32 programs (84%) continued operations. Of the programs that remained operational, 81% (n=26) made modifications to donor acceptance policies, while 19% (n=6) maintained existing acceptance policies. Schools that made policy modifications for donor acceptance criteria reported changes, including heightened screening for symptoms of COVID‐19 at the time of donor registration and upon death and requiring a negative COVID test for the anatomical donor near the time of death. In addition, COVID‐19 infection was added to the existing list of disqualifying conditions for many programs. Some sites excluded donors from large group facilities (hospitals, long‐term care), and some declined any donors with respiratory infections at the time of death. Several programs (13%) stopped accepting those donors who were not registered with their AGP prior to the onset of the COVID pandemic.

During COVID‐19, the demand for donors in anatomy education decreased significantly (p<0.05). Reasons for the reduced donor demand as reported by respondents included school closings and adjusted (smaller, shorter in time) laboratory sessions. These actions were taken because of concern for student/staff safety, reduced availability of appropriate personal protective equipment, and safety‐related student/faculty spacing needs.

CONCLUSION

Most AGPs were operational during COVID‐19 and able to maintain donor needs for anatomy education. Further research is warranted into the long‐term effects on AGPs as the pandemic persists.
Effects of Dietary Pattern on Average Cortical Bone Density and Estimated Thickness of the Proximal Ulna
Authors: Armon Austin , Erceg Fayja , Lochner Bailey , Rodgers Hannah , Zelazny Kaya G.
Published: May 2022

There is an established relationship between diet and phenotype in various human body systems. Vegetarian diets may contain lower quantities of calcium and protein compared to other diets, causing speculation that this dietary pattern may contribute to lower bone mineral density (BMD) or osteoporosis. Nutritional intake is a major element in a person’s overall Bone Mineral Density (BMD), but little is known about how the upper extremity responds to changes in dietary pattern. There is also conflicting evidence for the effects of a vegetarian diet on bone, with some studies suggesting that lower quantities of calcium and protein compared to other diets contributes to lower bone mineral density (BMD), while others find that the lower acidic load found in the vegetarian diet combined with a higher intake of potassium have a calcium‐sparing effect with positive effects on bone health. This study tests how vegetarian and convenience diets affect Cortical Bone Thickness (CBT) and Average Cortical Density (ACD) of the olecranon process in humans, a site of fracture in the upper limb.

Computed tomography (CT) scans with varying dietary pattern were obtained from the New Mexico Decedent Image Database and segmented in 3D Slicer. Cortical thickness was measured by calculating the difference between periosteal and endosteal section contours. Density was approximated by comparing the average radiodensity after applying a threshold of 500 HU to isolate cortical bone. MANOVA and post‐hoc ANOVA testing were conducted in R Studio to test differences between dietary groups. There was a statistically significant effect of bone length (p = 0.001) and diet (p = 0.014) on both proximal ulnar bone properties measured (approximated cortical thickness and density). Post‐hoc testing found that only bone length had a significant effect on cortical thickness (p = 0.003) while diet, but not bone length or sex, had a significant effect on average cortical density (p = 0.008).

These results provide evidence of detrimental effects of a convenience diet on bone density, and possible evidence for beneficial effects of a vegetarian diet. Convenience foods are high in saturated fats, sodium, and refined sugars, which have been shown to have negative effects on bone mass and bone microarchitecture in rodents. Our results suggest that similar effects may be seen in the cortical bone of humans, providing additional evidence for the negative effects of convenience food on health. Previous studies have found that nutritional acidic load, which may be lowered by a vegetarian diet, is positively correlated with fracture risk and negatively correlated with BMD. Our results support these findings and add to the growing evidence for the benefits of a vegetarian diet.
Deregulation of PiwiL1 and associated RNAs in cervical cancer: is HPV oncogene responsible?
Authors: Kunnummal Midhunaraj , Angelin Mary , Chandrika Sivakumar Krishnankutty , Ali Jamshaid , Raveendran Pooja S. , Issac Joby , Das Ani V.
Published: May 2022

Although HPV infection is the major cause of tumorigenesis in cervical cancer; interestingly it has been noted that HPV infection does not always lead to cervical cancer. This suggests that there could be some unknown factor that directly or indirectly interacts with these HPV oncoproteins leading to tumorigenesis. Our study aims to understand whether any correlation exists between the Piwi homologs with HPV oncoproteins in cervical cancer. To begin with, the expression pattern of Piwi proteins were evaluated in cultured cervical cancer (C33A, SiHa, and CaSki) cell lines. We observed that all the Piwi variants were differentially expressed in all the selected cervical cancer cell lines with PiwiL1 showing the highest expression in CaSki cells, suggesting a positive correlation with HPV oncoproteins. On investigating this correlation further, we found that PiwiL1 was co‐expressed and physically interacted with HPV oncoproteins E6 and E7. Not only E6 and E7 physically interact with PiwiL1, but also accentuated its expression when over‐expressed in HaCaT cells. Since PiwiL1 protein is known for its overexpression in many cancers, we wanted to know how this molecule is regulated in cancer. For this, we have carried out an in silico analysis of the PiwiL1 promoter to find the possible binding TFs using multiple online tools such as Alibhabha, AllegenPromo, and TFsitescan. We further shortlisted the TFs based on their function in cancer, interestingly we found both p53 and E2F in the list. Further analysis of these selected factors suggested that PiwiL1 promoter activity was significantly upregulated in the presence of E2F, whereas it was downregulated with p53. The binding of these transcription factors and their differential regulation of PiwiL1 promoter could be one of the reasons for its over‐expressed status in cancer. In addition to this, it is also known that the Piwi‐piRNA complex act as guiding signals for factors that play a significant role in tumorigenesis. Therefore, we have constructed a pipeline to predict the piRNAs from a small RNA Sequencing Data (SRP119662) obtained from the NCBI database, we identified 2274 piRNAs from both control and tumor samples, out of this, only 9 piRNAs were found to be significantly differentially expressed compared to control. Even though in silico analysis predicted 6 piRNAs to be significantly upregulated and 3 piRNAs to be significantly downregulated, the qPCR analysis was not entirely in accordance to the prediction. Out of the 3 down‐regulated piRNAs, hsa_piR_007863 showed elevated levels in CaSki in comparison to HaCaT while hsa_piR_002320 which was predicted to be upregulated in tumor showed lower expression. Our preliminary data suggest that PiwiL1 protein and associated piRNAs may have an important role in HPV‐associated tumorigenesis, whose exact molecular mechanism needs to be further evaluated.
Direct and Sequential Bioactivation of Pemigatinib to a Reactive Iminium Ion Intermediate Culminates in Mechanism‐Based Inactivation of Cytochrome P450 3A
Authors: Tang Lloyd Wei Tat , Wei Wan , Verma Ravi Kumar , Koh Siew Kwan , Zhou Lei , Fan Hao , Chan Eric Chun Yong
Published: May 2022

Mechanism‐based inactivation (MBI) is a unique phenomenon in drug metabolism that arises when an enzyme metabolically activates a drug to a chemically reactive intermediate and primes it for irreversible binding, leading to an irrevocable loss of catalytic activity that is only restored upon de novo synthesis of the implicated enzyme. Our laboratory recently established the MBI of cytochrome P450 3A (CYP3A) by the fibroblast growth factor receptor (FGFR) inhibitors erdafitinib and infigratinib. Serendipitously, our preliminary data has also revealed that pemigatinib (PEM) – another clinically approved FGFR1‐3 inhibitor – similarly elicited time‐dependent inhibition of CYP3A. This was rather unexpected as it was previously purported that PEM did not pose any metabolism‐dependent liabilities due to the absence of glutathione‐related conjugates in metabolic profiling experiments conducted in human liver microsomes. Consequently, we posited that PEM is bioactivated by CYP3A to a hard electrophile, thereby allowing it to elude glutathione trapping and detection in the aforementioned reactive metabolite screen. Here, we confirmed that PEM inhibited both CYP3A isoforms in a time‐, concentration‐, and cofactor‐dependent manner consistent with MBI – with KI, kinact, and partition ratio of 8.69 and 11.95 μM, 0.108 and 0.042 min−1, and ~44 and ~47 for CYP3A4 and CYP3A5 respectively, when rivaroxaban was employed as the probe substrate. While the rate of inactivation was diminished by coincubation with an alternative substrate (testosterone) or direct inhibitor of CYP3A (ketoconazole), the inclusion of nucleophilic trapping agents afforded no such protection. Furthermore, the lack of catalytic activity recovery following dialysis and oxidation with potassium ferricyanide coupled with the absence of a spectrally resolvable peak in the Soret region collectively implied that the underlying mechanism of inactivation was not elicited via the formation of pseudoirreversible metabolite‐intermediate complexes. Utilizing cyanide trapping and high‐resolution mass spectrometry, we illuminated the direct and sequential oxidative bioactivation of PEM and its major O‐desmethylated metabolite at its distal morpholine moiety to a reactive iminium ion intermediate that could covalently modify the CYP3A apoprotein and/or prosthetic heme and culminate in its MBI. Finally, structural determinants gleaned from our in silicocovalent docking simulations suggested that the discrepancies in MBI potencies between both CYP3A isoforms might be alluded to the propensity for PEM to interact more favourably with an accessible active‐site serine residue in position 312 of CYP3A4 as compared to CYP3A5. Further physiologically based pharmacokinetic modelling studies are currently underway to better discern the clinical implications of our findings.
Importance of Membrane Boundary Residues on the Rotor Ring of E. coli ATP Synthase
Authors: Altman Anna , Keppler Nicholas , Steed Ryan
Published: May 2022

ATP synthase is the primary producer in of adenosine triphosphate (ATP), the energy carrier molecule used in a variety of cellular processes. ATP synthase functions by using the proton gradient present across the cellular membrane to power a series of conformational changes resulting in the synthesis of ATP by a rotary mechansim. E. coli ATP synthase has two major parts, F1 and Fo, connected by the γ/ε stalk. Fo is membrane embedded and consists of subunits a, b2, and ten copies of subunit c, typically referred to as the c‐ring (or rotor). Protons enter through a half channel in the a subunit (stator) and are passed to the c‐ring, which rotates in a way that has protons exit on the low energy side of the membrane through a second half‐channel. Previous studies have shown that Cys substitutions at the membrane boundary of c‐ring (Phe54, Ile55, Tyr73, Phe76) can severely impact the function of the synthase, suggesting that these positions are important to the structure and/or function of the motor. Therefore, we have generated various substitutions at these residues to determine what chemical properties are needed in this region to support function. While the cytoplasmic boundary residues (54‐55) are surprisingly tolerant of mutation, substitutions at the periplasmic boundary residues (73‐76) reveal that reductions in both side chain size and hydrogen bonding capability impair activity of ATP synthase. These data help to characterize the interaction between the membrane‐embedded rotor and stator subunits.
Role of Potassium Channel Function and Long‐term Exercise Training in Soleus Resistance Arterioles
Authors: Caldwell Jacob , Reid‐Foley Emily , Ballehr Kelsey , Restrepro Alexis , Wahl John , Maraj Joshua , Park Hyerim , Delp Michael , Muller‐Delp Judy
Published: May 2022

Introduction

Nuclear factor (erythroid‐derived 2)‐like 2 (Nrf2) expression has been linked to potassium (K+) channel function. We hypothesized that myogenic responses are enhanced by exercise training and also modulated by K+ channel function but would be impaired in arterioles from skeletal muscle of trained rats in which Nrf2 (Nrf2 KO) is deleted.

Methods

Exercise training was performed in male wildtype (WT) Sprague‐Dawley rats, and their littermates lacking the Nrf2 gene (Nrf2 KO). Rats were exercise trained using a mixed moderate and high‐intensity interval treadmill program for 10‐12 weeks. A WT cage‐confined control group was also studied to compare the impact of exercise training. After exercise training or sedentary cage‐confinement, soleus resistance arterioles were isolated and cannulated for the assessment of vascular reactivity. Vasodilatory responses to low concentrations of KCl [10 and 20mM} were assessed. Vasoconstrictor responses to higher concentrations of KCl [30–100mM] and myogenic responses to increasing intraluminal pressure (pressure 0cm H2O – 130cm H2O) were also assessed. The contribution of inward rectifying K+ channels (Kir) to vasodilation during low concentrations of KCl was assessed by treatment with barium chloride (BaCl2, 100mM).

Results

Soleus resistance arteriole myogenic responsiveness was reduced in trained Nrf2 KO as compared to WT (p<0.01). Trained WT rats produced a significantly higher myogenic responsiveness than sedentary WT controls (p<0.05). Vasodilation to low concentrations of KCl was reduced in soleus arterioles from trained Nrf2 KO rats as compared to arterioles from trained WT rats (p<0.01). Exercise training did not alter vasodilation to low KCl concentrations in WT rats (p>0.05). Constriction to higher concentrations of KCl was similar in arterioles from trained WT and Nrf2 KO rats. Inhibiting Kir channels with BaCl2 eliminated the genotype‐ and training‐related differences in myogenic responses and vasodilation to KCl.

Conclusion

Our data indicate that Nrf2 may be a critical regulator of vascular adaptation that occurs in skeletal muscle arterioles in response to long‐term exercise training. Elimination of differences in myogenic responsiveness by BaCl2 suggest that Nrf2‐mediated adaptation of the myogenic responsiveness may involve Kir channels.
Bioinformatics and 3D Structural Analysis of the Coronavirus Main Protease Active Site Diversity
Authors: Wu Wu Amy , Lubin Joseph H. , Olivas Cassandra , Zardecki Christine , Balogun MaryAgnes , Bacorn Mickayla L. , Khare Sagar D. , Burley Stephen K.
Published: May 2022

Coronaviruses (Coronaviridae) such as SARS‐CoV‐2 (severe acute respiratory syndrome coronavirus) and MERS‐CoV (Middle East respiratory syndrome coronavirus) have been the source of recent outbreaks and global health concerns. While vaccines have been essential for controlling the SARS‐CoV‐2 (COVID‐19) pandemic, it is uncertain whether they will be effective against future coronavirus strains. Therefore, identification or design of a broad‐spectrum drug that targets highly conserved regions of the main protease of multiple coronavirus strains is essential in the long term. As part of a virtual summer research experience with the RCSB PDB, bioinformatics tools were employed to predict and construct 3D models of the coronavirus main protease (MPro) using SARS‐CoV‐2 as the template, with a focus on mutational trends and active sites. This study focused on the active sites of MPro, a cysteine protease essential for viral assembly and replication. Sequence alignments and structure modeling of MPro structures has identified conserved regions across multiple coronavirus strains. Inhibition of MPro halts coronavirus replication, making it an ideal drug target, and studies of MPro may foster and accelerate the discovery of high affinity broad‐spectrum drugs.
Multi‐Omics Integration and the Development of Gestational High Altitude Induced Pulmonary Arterial Hypertension
Authors: Leslie Eric , Gheorghe Ciprian , Brito Alex , Anti Nana O. , Jones Breanna , Hughes Lorelei , Sanchez Kayla , Dasgupta Chiranjib , Lopez Vanessa , Grapov Dmitry , Bosviel Remy , Blazenovic Ivana , Murray Sam , Lee Michael , Torres Roberto , Juarez Rucha , Yellon Steve , Zhang Lubo , Fiehn Oliver , Newman John , La Frano Michael R. , Wilson Sean M.
Published: May 2022

High altitude (> 2500 meters) during pregnancy reduces oxygen delivery to the fetus (i.e., gestational hypoxia), causing abnormal fetal lung development. The pulmonary arteries become thicker and less functional, increasing the risk of pulmonary hypertension, which is a multiorgan disease that promotes right heart failure and under five‐years mortality rate. Our previous work showed endoplasmic reticulum stress and metabolomic alterations in fetal sheep pulmonary arteries indicative of an unregulated inflammatory response after gestational hypoxia. To understand the relationships between inflammation, vascular remodeling, and pulmonary arterial hypertension development, we examined the omic links between the pulmonary arteries and plasma exosomes in fetal sheep exposed to high altitude (3801 meters) for the majority of the pregnancy (110+ days out of 138‐141 days of pregnancy). Specifically, metabolomics and proteomic analyses from the pulmonary arteries and transcriptomic analysis of plasma exosomes were integrated from fetal sheep to understand the mechanisms driving this multiorgan disease. Sixteen miRNAs and 75 proteins were differentially expressed between normoxic and hypoxic samples. Gestational hypoxia resulted in significant changes in 90 metabolites in fetal pulmonary arteries. These results show exosomal microRNA are linked with pulmonary artery proteins and metabolites important to cell membrane composition which might explain their role in the facilitation of changes in vascular remodeling, inflammation, and cell signaling. Top canonical pathways noted in omics data integration were acute phase response signaling, atherosclerosis signaling, FXR/RXR activation, intrinsic prothrombin activation pathway, and LXR/RXR activation. This multi‐omics study illustrates the intricacy of complex adaptations that occur in the fetal sheep lung in response to gestational hypoxia.