bluuebunny/crossref_metadata_2025 huggingface dataset:
Brain 11β‐hydroxysteroid dehydrogenase activity: which enzyme?
Authors: Formenti Silmara , Gomez-Sanchez Celso E , Colombari Eduardo , Gomez-Sanchez Elise P
Published: June 2021

We studied 11β‐HSD activity in rat kidney (positive control) and brain homogenates and subcellular fractions. Samples were incubated with 0.5 mM NAD+ or NADP+ and [1,2 3H] corticosterone for 1h at 37° C. Steroids were extracted, separated by TLC, and the radioactivity represented by corticosterone and 11‐dehydrocorticosterone read by a bioscanner. Conversion of corticosterone to dehdrocorticosterone by homogenates was significantly increased by NADP+, but not NAD+: Cerebel: Control‐24.1±0.4%, NADP+‐60.9±0.8%, NAD+‐21.9±0.7%; Hypothal: Control‐8.2±0.1%, NADP+‐23.6±0.8%, NAD+‐ 7.3±0.0%; Hippoc: Control‐24.2±0.9%, NADP+‐62.1±0.6%, NAD+‐21.9±0.2%, p<0.001 in all). Conversion was significantly greater isolated in microsomes compared to nuclei and mitochondria, and for NADP+ vs NAD+: Cerebel microsomes: NAD+‐5.4±1.7%, NADP+‐34.3±4.6%, p<0.001; Hypothal microsomes: NAD+‐0%; NADP+‐9.2±1.8%, p<0.001; and Hippoc microsomes: NAD+ 0.6±0.6% vs NADP+ 23.1±1.4%, p<0.001). Thiram inhibited NAD+‐dependent HSD activity in 11‐β HSD2‐ transfected CHO cells; it did not in the brain. These data suggest a novel NADP+‐dependent 11β‐HSD that may allow aldosterone to activate some brain mineralocorticoid receptors.

Support: VA Medical Research Funds, NIH HL27255 & HL75321, CAPES & FAPESP
The role of exercise on oxidative stress and inflammation in the aging brain
Authors: Chirico Erica N , Patsouris David , Geloen Alain , Rieusset Jennifer , Abidi Ramy , Canet-Soulas Emmanuelle , Pialoux Vincent
Published: June 2021

Brain inflammation and oxidative stress are implicated in certain pathologies such as stroke and Alzheimer's disease, and become more prominent with age and/or metabolic diseases like atherosclerosis. Exercise training has been shown to reduce oxidative stress and inflammation, providing protection in many diseases. We hypothesize that exercise training could improve oxidative stress and inflammation in ApoE−/− mice, specifically in the brain. A 12‐week exercise or sedentary program was enforced in young (8 weeks) and old (66 weeks) ApoE−/− mice on high fat/cholesterol diet. Old mice had significantly higher levels of oxidative stress and a decrease of antioxidant superoxide dismutase in the brain than the young mice. The old exercise trained mice had less oxidative stress and inflammation than the old sedentary mice, as demonstrated by an decrease in MDA, TNFα, NF‐κB, and IL‐1β. Plasma oxidative stress was positively correlated with brain oxidative stress and inflammation. In conclusion, these results suggest that the overall increase in systemic oxidative stress and inflammation inherent in atherosclerosis could lead to further brain oxidative stress and inflammation, which is further exacerbated by aging and a sedentary lifestyle. In addition, the decrease in oxidative stress through exercise training could improve systemic and brain inflammation in this animal model atherosclerosis.
The tragedy of taste and smell dysfunction in the United States – it affects millions
Authors: Henkin Robert I
Published: June 2021

As many as 21 million people in the U.S. suffer with taste and smell dysfunction. These symptoms occur after a viral‐type illness (~300,000 yearly), after brain injuries (~600,000 yearly), associated with allergic rhinitis (~200,000 yearly), following x‐irradiation to the head and neck (~50,000 yearly), after chemotherapy (~30,000 yearly) and with neurodegenerative disorders (>500,000 patients). These symptoms alter quality of daily life including appetite alteration with subsequent malnutrition, accidental exposure to gas or spoiled food, social isolation and depression due to persistence of these symptoms. When patients access their physicians for medical assistance they are told there is little known about these problems, there is no known treatment and they should learn to live with these symptoms. This leaves patients with a double problem – presence of these symptoms and inability to obtain help or hope for their evaluation or treatment. This tragedy has been averted in many patients based upon biochemical and clinical studies in which these symptoms were evaluated, their etiologies defined (related to biochemical changes in several body functions) and, based upon these evaluations, effective treatment initiated to correct these dysfunctions. This information must be promulgated to both patients and physicians so this tragedy can end and patients can receive effective evaluation and treatment.
Behavioral, physiological, and morphological outcomes under dynamic oxygen environments in zebrafish
Authors: Marks Chris , Moore Francisco B.G. , Kaut Kevin P. , Bagatto Brian
Published: June 2021

We investigated the relative contributions of genotype, early oxygen, and later oxygen environments on zebrafish behavior, physiology, and morphology. Four male and four female zebrafish were crossed in all possible combinations resulting in 389 offspring. These offspring were divided among four treatments: normoxia for 90 days, hypoxia for 90 days, normoxia for 30 days followed by hypoxia for 60 days, and hypoxia for 30 days followed by normoxia for 60 days. The effects of early oxygen environment, later oxygen environment, and genotype were then assessed with respect to zebrafish behavior, morphology, and physiology. Fish were tested in an arena where they could shoal with conspecifics before, during, and after the introduction of a novel stimulus. Blood glucose and size were also measured. Early oxygen environment influenced fish size, time spent swimming, and reactivity to a novel stimulus. While there was minimal genetic variation for the physiological, morphological, and behavioral outcomes, environmentally induced plasticity was predominate. Our results demonstrate the existence of critical windows of development for determining behavioral, physiological, and morphological outcomes. We also demonstrate the flexibility of extended quantitative genetic designs in addressing proximate sources of variation in complex environments.
A new model for studying focal and generalized chronic seizures in anesthetized rats
Authors: Naggar Isaac , Stewart Mark
Published: June 2021

Seizure activity in anesthetized animals can be recorded in a number of different animal models. One such model uses urethane as an anesthetic and kainic acid as a convulsant. In addition, it is known that animals can be made epileptic using kainic acid by causing a period of status epilepticus and waiting for epileptogenesis to occur. We report an animal model to study seizure activity in kainic acid‐induced epileptic rats under urethane anesthesia. Sprague‐Dawley rats were injected with sufficient kainic acid to cause a period of status epilepticus. At ≥8 months post status epilepticus, at a time when behavioral seizures were evident, the rats were anesthetized with urethane. EEG recordings from CA1 and CA3 were performed together with recordings of EKG, BP, and vagus nerve. All rats exhibited abnormal spiking in the EEG and 42% (5/12) experienced spontaneous focal seizures while under anesthesia. Spontaneous generalized seizure activity occurred in a subset of these animals. In 56% (5/9 tested), focal and generalized seizures could be triggered with brief trains of electrical stimuli. Vagus nerve activity was found to be increased during focal and generalized seizures. These results demonstrate a new animal model to study seizures, especially focal seizures of limbic cortical origin, in anesthetized epileptic rats with a variety of applications, including the study of autonomic activity during seizures.
Histamine enhances excitability and may augment neuroprotection in the Syrian hamster hippocampus
Authors: Ibanez Giancarlo Gutierrez , Chau Roni , Hamilton Jock Stevens , Horowitz John M , Horwitz Barbara A
Published: June 2021

Previous studies showed that the neuromodulator histamine (HA) increases hibernation bout duration and, in non‐hibernating species, attenuates ischemic neuronal damage. We tested the hypotheses that: (1) HA directly enhances CA1 and CA3 Syrian hamster (Mesocricetus auratus) pyramidal cell excitability and (2) HA promotes recovery from an anoxic insult. HA (10μM) was applied directly to hippocampal slices and evoked responses (ERs) were recorded before, during, and after HA perfusion at 30ºC. ERs were also recorded before, during, and after a 15 min anoxic insult (95%N2/5%CO2). We found that HA increased ERs recorded from the CA1 region (23% increase, P< 0.05) and shortened the latency to the ER peaks (16–26% decrease, P<0.01) in 7 slices from 5 hamsters. In the CA3 region, HA had similar, though smaller, effects on latency and amplitude. Following anoxia, in the presence of HA, 5 slices recovered faster, 2 slower, and 1 was unaffected vs. slices without HA. Data are consistent with both hypotheses and suggest that HA has multiple, synergistic effects on pyramidal cells. Moreover, the observed HA‐induced enhanced excitability of pyramidal cells supports the view that these cells play a key role in the neural pathway regulating hibernation bout duration. The increased rate of recovery suggests that HA also has a neuroprotective role, ensuring reliable signal processing in a hibernating hamster.
Effect of human amnion epithelial cells on stroke outcome in mice
Authors: Sobey Christopher , Taylor Kimberly , Lim Rebecca , Wallace Euan , Drummond Grant , Broughton Brad
Published: June 2021

We tested whether stem cells derived from the placenta (human amnion epithelial cells; hAECs) may represent a novel effective therapy for stroke. Male C57Bl6 mice were subjected to 0.5 h middle cerebral artery occlusion‐reperfusion, and injected with 1×106 fluorescently‐labelled hAECs or saline (vehicle) i.v. at either 1 h (acute treatment) or at 72 h (delayed treatment) post‐stroke and brains were removed after 3 or 14 d, respectively. Neurological and motor function as well as infarct damage were significantly improved in mice treated with hAECs vs. vehicle 1 h post‐stroke. Furthermore, immunofluorescence of the key marker of apoptosis, cleaved caspase‐3, revealed that less apoptosis occurred in the infarct core of mice treated with hAECs. Moreover, mice that received delayed post‐stroke hAEC treatment had improved survival to 14 d compared with vehicle‐treated mice, although functional outcomes did not differ between surviving animals from each treatment group. Thus, early post‐stroke i.v. delivery of hAECs reduces brain injury and functional deficit, and delayed post‐stroke hAEC treatment improves 14 d survival. hAECs could be a viable therapy for promoting recovery following stroke.
A novel signaling pathway for regulation of ATP‐binding cassette transporter A1 expression
Authors: Yang Hong , Chen Xinping , Zhang Hongfeng , Lin Xinghua , Okoro Emmanuel U , Zhou LiChun , Guo Zhongmao
Published: June 2021

The objective of this study is to characterize the signaling pathway for regulation of ATP‐binding cassette transporter A1 (ABCA1) expression. Quantitative real‐time RT‐PCR assay and Western blot analysis revealed that treatment of macrophages with reelin, apolipoprotein E (ApoE) or ApoE‐containing lipoproteins increased ABCA1 mRNA and protein expression. In addition, these reagents significantly increased the level of phosphorylated disabled‐1 (Dab1), phosphatidylinositol 3‐kinase (PI3K), protein kinase C‐æ (PKC‐æ), and specificity protein 1 (Sp1), increased the amount of Sp1 bound to the ABCA1 promoter, and induced PKC‐æ binding with Sp1. Mutation of the Sp1 binding site in the ABCA1 promoter suppressed the ABCA1 promoter activity induced by reelin, ApoE or ApoE‐containing lipoproteins. The upregulation effect of these reagents on ABCA1 expression was suppressed by: 1) knockdown of Dab1, very low‐density lipoprotein receptor (VLDLR) and apolipoprotein E receptor 2 (ApoER2) with siRNAs, 2) inhibition of PI3K and PKC with kinase inhibitors, 3) overexpression of kinase‐dead PKC‐æ, and 4) inhibition of Sp1 DNA‐binding with mithramycin A. These results, together with the known function of these signaling proteins, suggest that ABCA1 expression can be upregulated by activation of the VLDLR/ApoER2‐Dab1‐PI3K‐PKCæ‐Sp1 signaling cascade.

Grant support: SC1HL101431 (HY) and R01HL089382 (ZMG).
Short‐term bed rest increases inflammation as evidenced by elevated TLR4, NFκB1 and IL6 expression in skeletal muscle of older adults
Authors: Drummond Micah J , Timmerman Kyle L , Markofski Melissa M , Walker Dillon K , Dickinson Jared M , Jamaluddin Mohammad , Rasmussen Blake B , Volpi Elena
Published: June 2021

Aging is characterized by increased systemic and tissue inflammation. However, it is currently unknown if a period of inactivity exacerbates muscle inflammation in older adults. We hypothesized that short‐term bed rest would increase muscle levels of TLR4 and NFκB expression and circulating cytokine levels in older adults. Six healthy, older adults participated in a strict bed rest model for 7 consecutive days. Serum samples and muscle biopsies were taken before and day 7 of bed rest. Serum samples were analyzed for cytokine expression. Muscle samples were analyzed using western blotting and qPCR for TLR4, HSP60, Iκβα protein expression and IL6, NFκB1, IL1β, TNFα, TNFRSA1, SOCS3, IL10, and IL15 mRNA expression. Participants lost ~4% leg lean mass due to bed rest. Additionally, the cytokine, interferon gamma was significant elevated in serum samples after bed rest (P<0.05). In the muscle, TLR4 protein expression and IL6, NFκB1, IL10 and IL15 mRNA expression were increased after bed rest (P<0.05). We conclude that short‐term bed rest increases IL6 expression perhaps through a TLR4/NFκB mechanism, and may contribute to muscle loss observed following a period of bed rest. An increase in the anti‐inflammatory cytokines, IL10 and IL15 may represent a compensatory response to reduce inflammation. Funding: K01AG038556, R01AG018311, R01AR049877, 1UL1RR029876, P30AG024832
PhenoMiner: an interactive tool for physiologists integrating phenotype data using multiple ontologies
Authors: Dwinell Melinda , Shimoyama Mary , Nigam Rajni , Liu Weisong , Tutaj Marek , De Pons Jeff , Wang Shur-Jen , Smith Jennifer , Lowry Timothy , Hayman G. Thomas , Laulederkind Stanley , Petri Victoria , Jayaraman Pushkala , Worthey Elizabeth , Munzenmaier Diane , Jacob Howard
Published: June 2021

Aim

The Rat Genome Database (RGD, http://rgd.mcw.edu) has been developing tools and resources to aid researchers in integration of genomic and phenomic data derived from rat research for the past 10 years. Following the development of the Phenotypes and Models Portal, a tool was created to integrate phenotype data sets from multiple sources. The new PhenoMiner tool delivers an interactive tool to query phenotype data by strain, phenotype, measurement method and experimental conditions.

Methods

RGD was established primarily as a genomic resource but has gathered and curated physiological data for many years. The Phenotypes and Models Portal provides both new types of phenotype data, as well as different ways to access existing data in RGD. PhenoMiner was developed to integrate data collected from multiple laboratories, using a wide variety of techniques, and across many inbred, transgenic and knockout strains of rats. Data are integrated through the use of four ontologies (rat strain, clinical measurement, measurement method, experimental conditions).

Results

The more than 12,000 records can be viewed graphically or in tabular format, allowing for easy comparison of phenotypes across strains.

Conclusion

PhenoMiner promises to be an effective and user‐friendly method to access, retrieve, and compare physiological data and identify ideal disease models for further studies.
Teaching Dental Histology: Using Traditional Vs. Virtual Microscopy
Authors: Klinkhachorn Penprapa S. , Palmer Bruce , Martin Karen , Ressetar Holly
Published: June 2021

The Dental Histology Course at WVU has recently been updated to incorporate virtual microscopy, in addition to the traditional microscopy labs, with the goal of engaging students in an interactive learning environment with computer‐assisted and self‐directed instruction. We have developed a virtual microscopy website featuring an extensive collection of digitized oral histology specimens. The virtual slides have been annotated and are cross‐referenced to the lab guide for use in conjunction with the hands‐on microscopy laboratories. Response to the new virtual microscopy technology among both the students and faculty has been overwhelmingly positive. On the Likert scale of 1 to 5 (with 5 = strongly agree), the students liked the online lab guide with labeled digitized images (4.5). The ability to scan across tissues and adjust zoom is similar to the traditional microscope experience and is preferable to static images. They also preferred being tested using digital images (4.6). Most students agreed that the online resource helped them significantly, especially when studying for the oral histology exams. This resource facilitates self‐directed learning and is accessible to at any time (4.3). Virtual microscopy offers a more progressive and interactive mode of learning that preserves the interpretation and problem‐solving skills taught by traditional microscopy. This new teaching tool has resulted in better performance among our dental students who are now scoring above the national average in the Anatomical Sciences portion of the National Board Dental Exam.
Rethinking the postdoctoral training experience: Fellowships In Research and Science Teaching (FIRST)
Authors: Mercante Jeffrey , Ricks Ernest , Eisen Arri , Haynes John K , Eaton Douglas
Published: June 2021

FIRST is a collaborative project in postdoctoral training at Emory University, Morehouse School of Medicine, Morehouse College, Spelman College and Clark Atlanta University. FIRST is funded as part of the Institutional Research and Career Development Awards in the National Institute of General Medical Sciences to promote research‐oriented teaching at minority‐serving institutions (MSIs) and enhance research skills in developing scientists. FIRST was developed to foster cross‐institutional relationships and increase the number of underrepresented minorities in life sciences careers. FIRST is distinct in that it combines outstanding postdoctoral training at a top‐tier research institution with a mentored undergraduate teaching experience at an MSI. Fellows benefit from this approach where they develop broad skills in two complementary and critical aspects of the academic environment. Compared to traditional postdocs, the over 100 current and former FIRST fellows publish at a similar rate as their peers, are more successful in obtaining permanent academic positions, especially at MSIs, and receive equivalent external funding. Finally, the program fosters a supportive social and intellectual network of scholars during and beyond the three years of the fellowship, while providing strong role models for young African‐Americans in the formative years of their science education.
Implementation of Competencies for Anatomy in the First Year of Medical Training
Authors: Herrera Patricia M , Cortes Teresa , Petra Ileana , Aburto Monica , Reynaga Jesus , Soto Antonio
Published: June 2021

To assess the outcome of students that underwent the exercises to implement certain competencies at the beginning level.

It is quite a challenge to develop teaching and evaluation strategies for the implementation of competencies in a traditional medical school that accepts 1200 students yearly, used to the traditional systems, and grouped in classes of 40 students each with teacher having limited experience in competence based learning.

We established three experimental and three control groups. We used three different exercises for a particular theme and tested before and after; compared the results of the experimental and control groups with T Student and Chi Square. The first strategy was to use a case history of cranial fracture, skulls to study in small groups, group discussion and evaluate written communication in a composition. The second strategy was to use body painting to identify the cervical triangles and their content. The third strategy was a case history of deafness and the study of an on line presentation of the ear, group discussion and the projection of ear implant techniques. The fourth strategy was a case history of rheumatic fever, the study of heart anatomy, group discussion.

Results showed a marked improvement in grades achieved on all quizzes and examinations of the experimental groups.
Two‐system Theory and Fasciology
Authors: Yang Chun , Dai Jing-xing , Yuan Lin
Published: June 2021

Objective

In light of developmental biology of the fascia‐like connective tissue, the fascia network is homologous with the extracellular matrix, the middle lamella, and the mesenchyme, which common function is to sustain stability of the internal environment. Therefore, we proposed a new theory of two systems, supporting‐storing system and functional system, to investigate the relationship between the fascial network and meridians.

Methods

Based on the digital datasets of Virtual Chinese human (VCH) bodies, 3‐dimensional (3‐D) structures of virtual meridians and fascia connective tissue gathering areas were constructed and compared with each other. Three‐D structures of fascia connective tissue gathering areas were also constructed based on CT and MRI images of living human subjects, and compared with meridians.

Results

Three‐D structures of fascia connective tissue gathering areas in the VCH bodies, CT and MRI images showed beads‐on‐string patterns. The reconstructed fascial lines co‐localized precisely with the traditional meridian lines. When 3‐D structures of all fascia connective tissues throughout the body were constructed, a body‐shaped connective tissue network appeared.

Conclusions

The fascial network throughout the human body is one of the anatomical bases for the acupoints and meridians of traditional Chinese medicine. The discipline that studies the supporting‐storing system and the functional system and their mutual relationship is called fasciology.

Grant Funding Source : National basic research programm (973 programm grant number:2007CB512705)
Early Anatomical Identification Markers for Duchenne Muscular Dystrophy in a Subadult Subject
Authors: Harris Jasmine H. , Godwin Ellen , Marquez Samuel
Published: June 2021

Duchenne Muscular Dystrophy (DMD) readily affects gait and posture in subadult populations afflicted with the disease. This study used instrumented motion analysis (IMA) to identify how gait and posture changes respond to DMD disease. 3‐D motion analysis was performed with the Vicon Motion Capture System on a 5‐year‐old boy suspected with DMD. Thirty‐nine reflective markers were placed on specific anatomical landmarks according to the Plug‐in‐Gait Model used with the Vicon system. The child walked across the 25 feet gait analysis laboratory for 10 trials. A single representative trial was selected for analysis. Kinematic parameters of gait were compared to those of a typically developing child. IMA identified specific changes in joint kinematics in the child with DMD as compared to the typically developing child. Changes include: increase in anterior pelvic tilt and hip flexion in swing, genu recurvatum in stance, plantar flexion on initial contact with ground, and lack of dorsiflexion in swing. These gait deviations are commonly found in boys with DMD. The results show the ability to identify these changes translating in the early diagnosis of DMD as they represent a specific pattern of walking that is related to the progression of weakness observed. Identification of anatomical gait deviations can assist in the development of treatment interventions to assist the child to be ambulant as long as possible.

Grant Funding Source: None
Study of structural failure of the blood‐gas barrier and the epithelial‐epithelial cell contacts in rested and exercised chicken, Gallus domesticus
Authors: Maina John N , Jimoh Adekunle S
Published: June 2021

In birds, high pulmonary capillary blood pressure (PCBP) greatly challenges the structural integrity of the blood‐gas barrier (BGB). Among air‐breathing vertebrates, birds interestingly have thinnest BGBs. To reconcile presence of thin BGB with high PCBP, failure of the BGB and the epithelial‐epithelial cell contacts (E‐ECCs) in rested and exercised chickens was investigated. The number of red blood cells (nRBCs) and protein concentration (PC) were determined in the lavaged fluid. The numbers of E‐ECC breaks (nE‐ECCBs) and those of the BGB (nBGBBs) were counted in the regions supplied by the four branches of the pulmonary artery (PA). The blood lactate concentration increased 4‐fold between rest and highest level of treadmill exercise (speed 2.95 m/s) while the nRBCs and the PC increased gradually, tending to level at higher workloads. Indicating their relative fragility, the nE‐ECCBs surpassed the nBGBBs. In resting birds, there were no breaks in the cranial region. In exercised birds, differences in number of breaks only occurred at 0.66 m/s where the lowest and highest counts respectively occurred in the cranial‐ and caudomedial regions whereas at all other levels of exercise, the numbers of breaks were comparable.

Funded by the National Research Foundation (NRF)
Analyis of the forelimb of Felis catus in comparison to Smilodon fatalis
Authors: Ostrom Kelly , Naples Virginia L
Published: June 2021

Much is known about the anatomy of the domestic cat, and this species is studied extensively as a model of structure and function in anatomy laboratories. Nevertheless, knowledge of the smaller muscles remains unpublished. Musculoskeletal studies of living species permit identification of the corresponding features of extinct animals. This comparative anatomical method was used to expand what is presently known about the forelimb muscles of the saber‐tooth cat Smilodon fatalis. Comparing the forelimb anatomy of the domestic cat to the saber‐tooth was necessary to understand the habits, movement patterns, and hunting strategies of the extinct species. S. fatalis, a dirk‐toothed cat that lived during the Pleistocene, was a deadly ambush predator. This sabercat had a heavy muscular build, making it more powerful when compared with many other cats. Complete, accurate muscle maps have not been published for domestic cats or other felids. The muscle maps of Felis catus that were drawn in this study have proven beneficial for comparison with the extinct saber‐tooth. Using information from the muscle maps created for the domestic cat as a means of comparing relative sizes of muscle scars to the same regions of the corresponding bones of S. fatalis allowed creation of muscle maps for this saber tooth. These maps show that the musculature of F. catus and S. fatalis differ in relative proportions and movement patterns.

Grant Funding Source : Undergraduate
The shoulder musculature of Rhynchocyon cirnei
Authors: Grossman Ari , Holroyd Patricia A , Rovero Francesco
Published: June 2021

Traditionally sengis (elephant shrews, order Macroscelidea) have been studied in light of other insect‐eating mammals without a firm understanding of their phylogeny, and all prior work on their myology has been made to animals of similar size rather than similar ancestry. Sengis are now recognized as part of the Afrotheria. Afrotheres have a long evolutionary history and currently few morphological characters are known to unite them, in part due to a limited knowledge of their soft tissue anatomy within a phylogenetically‐meaningful context. One poorly studied afrothere is the giant sengi, Rhynchocyon. We conducted a dissection of the shoulder and arm musculature of R. cirnei. A single specimen preserved in ethanol was dissected using standard dissection techniques under a Zeiss microscope. Muscles were cleaned to determine origin(s), insertion(s) and extent. Each muscle was only reflected after it was documented and photographed. We also identified innervations of each muscle whenever possible. We confirmed prior observations on sengi myology and provide new data on innervations. Comparisons highlight the distinctiveness of sengis relative to other afrotheres where known but also highlight the need for considerable additional anatomical work to firmly establish which features are apomorphies and which may represent shared derived characters among afrotheres.

This work is supported by a Midwestern University intramural grant
Sensory contributions to the spinal accessory nerve in humans
Authors: Boehm Karl Edward
Published: June 2021

The purpose of this study was to determine the presence of microscopic sensory ganglia within the spinal portion of the accessory nerve (SCN XI). Ten cervical spinal cord segments were removed from human cadavers in the anatomy lab at A.T. Still University ‐ Kirksville College of Osteopathic Medicine. Fifteen SCN XI were removed from the cervical spinal cords in segments from each spinal level. The segments from each nerve were dehydrated and embedded in paraffin starting with cervical spinal level 1 (C1) in parallel descending order. Embedded samples were sectioned at 12 μm and placed on slides. Sections were stained using hematoxylin and eosin. The slides were examined using light microscopy and results were recorded digitally. Neuron cell bodies were observed in all of the specimens (100%). The total number of neuron cell bodies ranged from 37 to 531 per nerve with an average of 207 cell bodies per nerve. Neurons cell bodies were observed at spinal levels C1–C5 and non‐grossly visible ganglia were observed at levels C1–C4. Based on the somatic origin of CN XI we conclude that cell bodies observed in CN XI belong to psuedounipolar neurons, which are the most common peripheral afferent nerve cells. The results from this study indicate that CN XI contains a sensory component. Research support provided by the Graduate Program at A.T. Still University.

Grant Funding Source: Graduate Program at A.T. Still University.
IS EMBRYONIC PRESSURE OVERLOAD RESPONSIBLE FOR CAUSING ENDOCARDIAL FIBROLASTOSIS?
Authors: Pesevski Zivorad , Sedmera David
Published: June 2021

Endocardial fibroelastosis (EFE) is a diffuse thickening of the ventricular endocardium, causing myocardial dysfunction and presenting as unexplained heart failure in infants and children. It is believed to be caused by persistent and increased wall tension in the ventricles, possibly secondary to myocardial damage, mitral regurgitation, or both. The disease can be primary or secondary to various congenital heart diseases. Its frequent association with congenital aortic stenosis lead us to hypothesize that pressure overload is important in its pathogenesis. We have tested this in a chick model of conotruncal banding.

The eggs were incubated in a force draft incubator at 37.5 °C and high humidity. Conotruncal banding of chick embryonic hearts was performed at the ED4, leading to increased pressure in the ventricle. At ED12 modifications of myocardial architecture and fibrosis were studied by histology and immunoconfocal microscopy.

Histology of pressure‐overloaded hearts showed chamber dilatation and compact layer thickening. However, H&E with Alcian Blue staining did not reveal any significant fibrosis with the exception of cardiac skeleton and valves. Immunohistochemistry with anti‐periostin antibody and more sensitive pan‐collagen antibody clearly showed fine layer of subendocardial fibrous tissue that was more prominent in the pressure‐overloaded hearts.

It appears that increased pressure loading stimulates fibrous production in the subendocardium of the hearts.

Grant Funding Source: Supported by Ministry of Education, Youth and Sports VZ 0021620806 and Grant Agency of the Czech Republic P302/11/1308
Angles of the Branches of the Visceral Aorta: Implications for Complex Aneurysm Repairs
Authors: Belchos Jessica Helen , Johnson Marjorie , Forbes Thomas
Published: June 2021

Thoracoabdominal aortic aneurysms can require custom designed branched stent‐grafts. Visceral and renal artery orientations can change as more of this segment becomes aneurysmal. This study's purpose was to determine these orientations in infrarenal aneurysms as a baseline to compare to more complex proximal aneurysms. Aquarius TeraRecon 3D imaging software was used for morphological assessment of CT scans for 20 Infrarenal aneurysms. (15M, 5F, mean age=73.7). Four take off angles were measured. The clock‐face position (CFP) was noted in transverse sections for each. The direction of take off of the renal arteries was assessed (anterior, orthogonal and posterior) and maximal aortic aneurysm diameter was taken. The average take off angle was 63.06° in the Left Renal (LRA), 55.57° in the Right Renal (RRA), 60.07° in the Superior Mesenteric (SMA) and 56.82° in the Celiac Trunk (CT). The mode for CFP was 3:00 (n=8) in LRA, 10:00 (n=10) in RRA, 1:00 (n=9) in SMA and 1:00 (n=8) in CT. The mode for direction of take off angle was orthogonal (n=10) for LRA and anterior (n=12) for RRA. The mean maximal aneurysmal diameter was 57.95mm. The data suggests little variability in patients with Infrarenal aneurysms. This information will be used as a baseline as more proximal aneurysms are assessed.

Grant Funding Source: Departmental
High speed imaging of an aortic valve in a full‐functional reanimated human heart
Authors: Rolfes Christopher David , Bateman Michael G , Iaizzo Paul A
Published: June 2021

It is imperative to understand the nuances of a cardiac valve function, not only to optimize the designs of cardiac devices, but also for better identifying the early onset of disease. Commonly used clinical tools such as Doppler echo work well for diagnosis of a progressed disease. Yet, more subtle identifications of pre‐clinical functional alterations will likely require careful visualizations of both the valve shape and motion throughout the cardiac cycle. Here we employed a high speed camera system to capture valve motions during a regular heartbeat at 500 frames per second. Specifically, we visualized the aortic valve within a reanimated human heart, which was deemed non‐viable for transplantation, using Visible Heart methodologies. Normal conduction and contractile functions were confirmed with ECG and pressure monitoring such that a frame by frame assessment of valve function could be correlated with hemodynamics. Note, that such images are available on the Atlas of Human Cardiac Anatomy (www.vhlab.umn.edu/atlas) where they are freely downloadable for students, educators, patients, and device designers. As we obtain more of such images, this should provide unique insights as to how these valves function throughout the cardiac cycle and may become altered with disease. This project was funded in part by NIH training grant T32AR007612 and by Medtronic, Inc.
Individual and Combined Effect of Silicon and Calcium Ions on Osteoblast Gene Expression and Matrix Formation
Authors: Varanasi Venu Gopal , Dominia Lisa M , Jue Stephanie M , Lee Angela , Leong Kelly K , Loomer Peter M , Opperman Lynne A
Published: June 2021

This study tests the hypothesis that silicon and calcium ions combinatorially target gene expression during osteoblast differentiation. MC3T3‐E1 subclone 4 osteoblast progenitors were exposed to elevated Si4+ (Na2SiO3) and Ca2+ (CaCl2) ion treatments both individually (50 ppm) and combinatorially (50 ppm Si + 50 ppm Ca) above control media conditions (α‐MEM, 10% FBS, 1% pen‐strep). Cell density significantly increased with time (5 days), but no significant difference was observed between treatments. All treatments were additionally supplemented with ascorbic acid (AA, 50 ppm) to induce differentiation. Relative expression of collagen type 1 (Col1α1, day 1), core‐binding factor a (cbfa1 or Runx2, day 6), and osteocalcin (OCN, day 6) was studied based on the timeline of osteoblast differentiation. Increased Si or Ca ion treatments enhanced Col1α1, Runx2, and OCN expression. Increased Si + Ca ion treatments enhanced OCN expression alone with no affect on Col1α1 and suppression of Runx2 (no amplification). Collagen fiber bundles were dense and elongated, for extracellular matrices (ECM) exposed to Si ions as compared to other treatments. These results indicated that individual ions enhanced multiple osteogenic gene expression while combined ion treatments enhanced individual gene expression.

Grant Funding Source : NIH/NIDCR #5P30DE020742‐02
Combinatorics of Cis‐Regulatory Element Evolution in Stress Response Modules of Ascomycete Yeasts
Authors: García-González Aurian , Roy Sushmita , Konieczka Jay , Thompson Dawn , Regev Aviv
Published: June 2021

This study provides insight into the conservation of cis‐regulatory elements in the osmotic stress response (OSR) of seven ascomycete yeasts. The OSR is orchestrated by the MAP‐kinase HOG1 in Saccaromyces cerevisiae. HOG1 has a human homolog, p38, which regulates the OSR and is known to play a role in angiogenesis. Gene expression profiles under osmotic stress were measured for each ascomycete, only the most induced and most repressed set of genes were evaluated for conservation of cis‐regulatory elements. A programming tool, ModuleDigger, was used to identify cis‐regulatory modules (CRMs), each composed of a set of co‐occurring binding motifs. Species were sorted into two clades: pre‐ and post‐ whole genome duplication (WGD) event. Motifs for the transcription factors RCS1 and SIP4 were present genome‐wide in pre‐WGD duplication and in the fungal pathogens C. albicans and C. glabrata. The post‐whole genome duplication species shared three motif pair combinations (MIG1, MSN2/MSN4; MIG1, ADR1 and ADR1, MSN2/MSN4). These motifs correspond to stress‐responsive elements and carbon‐responsive elements, which suggests interacting biological pathways.
Internalization Pathways of Non‐Viral Vectors for Gene Therapy
Authors: Anghel Ersilia , Shi Julie , Pun Suzie
Published: June 2021

Gene therapy has the potential to alter the expression profile of genes that are responsible for disease development by introducing replacement genes or reducing mRNA transcripts through pathways such as RNA interference. However, current non‐viral gene delivery methods suffer from low in vivo transfection efficiency. By understanding the internalization pathways of non‐viral vehicles, it may be possible to optimize and direct them through specific pathways to increase gene transfer efficiency. In this work, we investigated internalization of gene carriers by treating HeLa and fibroblast Cos‐7 cells with two drugs that inhibit the clathrin‐ and caveolin‐mediated endocytic pathways. Transfection using PEI polymers on HeLa cells showed a dependent response with clathrin‐ mediated pathway‐inhibiting drugs only. Transgene expression in HeLa cells decreased when both clathrin‐mediated and caveloe‐mediated pathways were inhibited. Analyses of transfection studies in Cos‐7 cells are in progress. The effect of delivering these vehicles in serum media was also assessed. This work suggests that the internalization pathway for efficient gene delivery is cell type‐dependent.
Development of an integrated bioenergetics index [IBI] function for biophotonic therapy
Authors: Einstein George Przybyl , Tulp Orien L , Karam David
Published: June 2021

Bioenergetics forms the basis for ultraviolet phototherapy [UVPT] and processes of cellular metabolism and energy production in health and disease. To quantify the contribution of UVPT on bioenergetics a mathematical formula has been derived and applied to quantify immune‐linked mechanisms that modulate metabolism in normal and disordred states. Energy transfer processes are quantifiable events and are essential for processes that contribute to life, the pathophysiology of disease, and senescence. The effectiveness of UVPT can be explained in part by examining the effect of quanta of UV light on the chemical structure of bacteria and cellular processes. The IBI may be expressed as follows:

IBI= C1∙C2∙F1( B/A; N/W;S/E;SM/M) ‐ F2[(VIR_L;Bact_L;TOX_L;FREE_RAD)/(NMicEl_∙NDE TOX NUT )], where determinants of genetic, physiologic, biochemical, nutritional, and other factors are assessed. Bacterial cells are smaller and more susceptible to UV energy than eukaryotic cells due to a 5‐fold greater content of photosensitive amino acids[PSAA], enabling bacteria to absorb greater quanta of UVPT energy than similarly exposed eurkaryotic host cells, effectively “marking” them for demise. In contrast, lower cell content of PSAA in eukaryotic host cells result in less damage during UVPT. These results are consistent with UVPT enhanced humeral and cellular responses to immunogenic stimuli. Institutional Support.
Investigation of the potential role of 6‐hydroxynicotinate monooxygenase in the modulation of virulence in Bordetella pertussis
Authors: Justice Samantha L. , Snider Mark J.
Published: June 2021

Bordetella pertussis, the causative factor of whooping cough, claims the lives of an estimated 300,000 each year. Previous studies exploring modulation of B. pertussis have shown that high levels of nicotinic acid (NA) modulate the expression of virulence– associated genes. Here we provide evidence supporting the hypothesis that 6‐hydroxynicotinate monooxygenase (NicC), an enzyme involved in NA catabolism in Bordetella, is implicated in a mechanism of modulation. Several NA derivatives, including 5‐ bromonicotinate, 6‐chloronicotinate and 2,5‐pyridine dicarboxylate (2,5‐PDCA), are known modulators of B. pertussis, are shown here, by fluorescence titration, to be bound by NicC. Kinetic inhibition studies further suggest that 6‐chloronicotinate and 2,5‐ PDCA non‐competitively inhibit NicC. Real time PCR will be used to quantify pertussin toxin expression in wt‐B. pertussis and a B. pertussis (ΔnicC) strain in which the nicC gene is knocked out, in the presence and absence of these known modulators, to determine whether a mechanism of modulation involves NicC. This work was supported by a HHMI Undergraduate Science Education Program Award at the College of Wooster.
Crosslinking very long abortive transcripts (VLATs) to E. coli RNA polymerase: an approach to elucidate forward hyper‐translocation
Authors: Wieland Rebekah , Hsu Lilian M
Published: June 2021

On a typically strong E. coli promoter, as nascent RNA grows to 8–10 nt—accompanied by scrunching 6–8 bp of the enlarged transcription bubble—the initial DNA‐RNAP contacts disrupt, upstream bubble rewinds, promoter escape occurs, and the elongation phase ensues. On some unique promoters, escape is accomplished only when the nascent RNA has grown to 16–19 nt in length. Such a promoter on the cusp of escape contains extremely high stress energy, which when utilized for upstream bubble rewinding and allowing promoter escape, can generate so much force as to propel the RNAP forward by several bases (Chander et al. 2007 Biochemistry 46, 12687). We call this process forward hyper‐translocation, during which the nascent RNA 3’‐OH is moved upstream into the RNA exit channel, out of register with the active site. The RNA molecule cannot be further elongated and is released as a VLAT. This mechanism can account for the GreB‐resistant nature of VLAT synthesis. To prove the occurrence of forward hyper‐translocation, I incorporated a 4‐thiouridine (S4U) residue at the 3′ end of a 19‐nt VLAT and conducted UV crosslinking experiments followed by SDS‐PAGE analysis. Our results showed UV‐, RNAP‐, and S4U‐dependent crosslinking of the VLAT to the β/β′ subunits. Using mass spectrometry, we are determining the location of the crosslink, and whether it coincides with the RNA exit channel. Supported by NSF‐RUI grant (MCB 0841452) to L. M. Hsu
Autoregulation of SUS1: a splicing dependent regulation of histone H2B ubiquitination
Authors: Hossain Munshi Azad , Venkataramanan Srivats , Johnson Tracy L
Published: June 2021

Pre‐messenger RNA splicing is an underappreciated mechanism for regulating gene expression in yeast (Saccharomyces cerevisiae), likely due to the paucity of intron containing genes in this single‐celled eukaryote. However, a growing number of studies suggest that introns play an important role in modulating gene expression under stress conditions. Recently, we have shown that environmental stress modulates splicing of the SUS1, a two‐intron gene in yeast. Sus1p, which is a part of deubiquitination module within the SAGA complex, plays an important role in maintaining proper cellular levels of ubiquitinated H2B. The first intron of SUS1 contains non‐canonical 5’SS and branch point sequences, which leads to retention of the first intron and subsequently limits the production of SUS1 mRNA. Here, we show that increased accumulation of the first intron containing transcript leads to a decrease in SUS1 mRNA production. Interestingly, the first intron containing transcript can be translated to produce truncated Sus1 protein which appears to play a role in SUS1 autoregulation. This study hints at an autoregulatory mechanism of control of SUS1 splicing and demonstrates an important role for regulated splicing in yeast to control histone modification.

National Science Foundation awards (NSF CAREER MCB‐0448010 and MCB‐1051921); National Institutes of Health (GM085474)
Evidence for a secondary pathway for fibril growth during amyloid fibril formation by tau
Authors: Ramachandran Gayathri , Udgaonkar Jayant B.
Published: June 2021

The aggregation of the natively disordered protein, tau to form lesions called neurofibrillary tangles is a characteristic feature of several neurodegenerative disorders including Alzheimer's disease. The focus of this work is to understand the mechanism of aggregation and the heterogeneity inherent in the amyloid fibril formation reaction of tau. This is achieved by aggregating the protein in vitro using the inducer heparin in a near‐physiological buffer and studying the kinetics of aggregation using biophysical and biochemical probes. For tau, the fibril formation mechanism in the presence of inducers has been described in previous work as ligand‐induced nucleation‐dependent polymerization. We find however that a complete explanation of the kinetic data obtained by monitoring the evolution of ThioflavinT fluorescence requires us to invoke the existence of a secondary pathway for fibril growth. In general, secondary pathways may be of three kinds: fibril fragmentation, lateral growth/branching and secondary/heterogeneous nucleation on a fibril surface. We provide both kinetic and accessory evidence for the conclusion that the dominant secondary pathway for fibril growth during the aggregation of the functional, repeat domain of tau is fragmentation.

This work was funded by the Tata Institute of Fundamental Research and by the Department of Biotechnology, Government of India.
Identification of a Ubiquitin‐Binding Domain (UBD) in GTP cyclohydrolase 1
Authors: Zhao Yu , Zhu Huai-Ping , Zou Ming-Hui
Published: June 2021

GTP cyclohydrolase 1 (GCH1), the rate‐limiting enzyme in de novo synthesis of tetrahydrobiopterin (BH4), is critical in maintaining the levels of intracellular BH4, an essential co‐factor for several key enzymes including endothelial nitric oxide synthase (eNOS). GCH1 is reduced in diabetes and hypertension, likely due to increased proteasomal degradation. However, the molecular mechanism by which the ubiquitin‐proteasome system targets GCH1 is unknown. GCH1 were co‐immunoprecipitated with Lys48‐Linked polyubiquitin chains in endothelial cells and 293 cells. Truncation of the amino acids 95–142 abolished the binding of GCH1 with polyubiquitin chains, suggesting that amino acids 95–142 is essential for GCH1 binding to polyubiquitin chains. Further, direct mutagenesis of isoleucine (Ile131) in the hydrophobic patch of UBD of GCH1 attenuated its ubiquitin binding. Finally, proteasome inhibition with MG132 or lactacystin caused the accumulation of GCH1‐ubiquitin complex in cultured endothelial cells and 293 cells. Taken together, our results suggest that the amino acids 95–142 with a hydrophobic patch Ile131 constitute a novel UBD in GCH1, through which GCH1 non‐covalently binds to polyubiquitin chains resulting in proteasome degradation of GCH1.