bluuebunny/crossref_metadata_2025 huggingface dataset:
Cervical spinal cord injury in a transgenic mouse expressing fluorophores in neurons, microglia, astrocytes, and oligodendrocytes
Authors: Sunshine Michael Daniel , Streeter Kristi A. , Gaire Janak , Otto Kevin J. , Fuller David Dwight
Published: June 2021

Spinal cord injury (SCI) most commonly occurs at cervical levels, and such injuries typically impair breathing. Reproducible cervical SCI models which facilitate high resolution evaluation of the tissue response to the injury will aide in the development and testing of therapeutics to improve respiratory function. These models can be evaluated by anatomical assessment of injury lesions. However, tissue processing using traditional immunohistochemistry is limited by the ability of antibodies to penetrate deep within the tissue sample. To address these issues, we studied the PRISM+ mouse, a genetically modified mouse with fluorophores tagging the four major cell types of the central nervous system endogenously express fluorophores, thereby eliminating the need for immunochemistry procedures. In the genetically modified mouse, neurons express YFP (512ex/529em), microglia express GFP (488ex/509em), astrocytes express DsRed (554ex/586em), and oligodendrocytes express CFP (435ex/485em). PRISM+ mice received a mid‐cervical (C3) dorsal‐lateral crush injury and tissues were harvested 3 days later. Initial histological evaluation of the injured spinal cords confirm an absence of neurons in the lesioned dorsal cord, as expected. However, YFP‐positive motor neurons were prominent in the ventral cord. The ipsilesional dorsal quadrant was clearly observed to have increased GFP‐positive microglial infiltration and lack of CFP‐positive oligodendrocytes. At this 3‐day post‐injury time point we detected no astroglial scar, and in fact there were fewer DsRed‐positive astrocytes in immediate vicinity of the lesion as compared to the rest of the spinal cord. Importantly, the fluorophores were robustly detected in all four cell types, and this enabled reliable and automated analysis of the histological sections. These data indicate that the Prism+ mouse provides a useful model for studying the histological response to SCI.

Support or Funding Information

This work was sponsored by T32‐HD043730 (MDS), 1F32NS095620‐01 (KS), SPARC OT2 OD023854 (DDF). 1 R01 NS080180‐01A1 (DDF), the University of Florida Preeminent Initiative, and by the Defense Advanced Research Projects Agency (DARPA) Microsystems Technology Office (MTO), under the auspices of Dr. Jack W. Judy (jack.judy@darpa.mil) and Dr. Doug Weber (Douglas.Weber@darpa.mil) as part of the Reliable Neural Technology Program, through the Space and Naval Warfare Systems Command (SPAWAR) Systems Center (SSC) Pacific grant No. N66001‐11‐1‐4013.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A New, Noninvasive Method of Measuring Pulmonary Gas Exchange: Results in Normal Subjects and Patients with Lung Disease
Authors: Wang Daniel L. , Prisk Kim , West John B.
Published: June 2021

Methods

The person breathes through a device that continually measures and displays the inspired and expired PO2 and PCO2. An oximeter is worn, and the arterial PO2 is calculated from the SpO2 and the oxygen dissociation curve, using the end–tidal PCO2 to account for the effect of PCO2 on the oxygen affinity of hemoglobin. The PO2 difference between the end‐tidal gas and the calculated arterial value is called the Oxygen Deficit. Since the calculation is only accurate when the arterial oxygen saturations are 94% or less, the normal subjects breathed 12.5% oxygen.

Results

Studies were made on 17 patients with a variety of pulmonary diseases who were attending an outpatient clinic. The mean oxygen deficit was 48.7 mm Hg, SE 3.1. This can be contrasted with a mean oxygen deficit of 4.0 mm Hg, SE 0.91 in a group of 30 normal subjects who were previously studied. The p value for the difference between the two groups was < 0.0001. Within the normal subjects, the mean oxygen deficit for the 19 younger participants (ages 22–31) was 2.02 mm Hg, SE 0.84 while that in the 11 older participants (ages 47–88) was 7.53 mm Hg, SE 1.55. The p value for the difference between the young and old groups was 0.0015. The mean oxygen deficit in younger normal subjects is remarkably small. This implies that there is very little difference between the calculated arterial PO2 and the alveolar PO2, consistent with little impairment of gas exchange. However, the mean oxygen deficit in older normal subjects is significantly higher consistent with the increased Alveolar‐arterial (A‐a) PO2 gradient with age, which has been well documented in the literature. Analysis of the factors determining the magnitude of the oxygen deficit showed that the end–tidal PO2 was the most important factor, followed by the end–tidal PCO2, and the calculated arterial PO2. Interestingly the SpO2 had a minor influence in spite of the fact that the oximeter reading is frequently used to guide therapy. The analysis emphasizes the value of measuring the composition of alveolar gas in determining ventilation‐perfusion ratio inequality. Remarkably, this factor is largely ignored in the classical index of impaired pulmonary gas exchange using the ideal alveolar PO2 to calculate the A‐a O2 gradient. We argue that ignoring the actual alveolar value gives an incomplete picture of gas exchange in the lungs because the traditional index is heavily biased by lung units with low ventilation‐perfusion ratios. Therefore, by using the new index of oxygen deficit, we derive a more comprehensive measurement of gas exchange in the lungs.

Support or Funding Information

Supported by MediPines Inc

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Development and implementation of a classroom‐based Authentic Research Experience, Physiological Models for Human Disease.
Authors: McGinnis Courtney L.
Published: June 2021

The National Academies and the American Association for the Advancement of Science (AAAS) have acknowledged that undergraduate education could be improved by more opportunities in authentic research (1, 2). In addition, there appears to be strong correlations between research‐based laboratories fostering an increased interest in STEM disciplines and improved grades, over traditional “cookbook” laboratories. This is especially true for underrepresented minorities and given our changing demographics at Quinnipiac, designing courses that better engage and support the student body is an essential component to our mission (3). Authentic research experiences are being implemented at many colleges and universities, as a means of giving students hands on experience conducting real science. Since there are limited research opportunities for our students to conduct faculty‐mentored research, developing a course where students can have an authentic research experience can address these shortages and obstacles (4). Additionally, in a senior exit survey completed by students within the College of Arts and Science, they expressed an interest in having more authentic lab experiences, more independent research opportunities and they wished they participated in these opportunities earlier in their career. To address some aspects of these shortages an authentic research course was developed to give students the opportunity to design and carry out experiments using one of four model organisms. Drosophila melanogaster (Fruit Fly), Caenorhabditis elgans (Roundworm), Dugesia tigrina (Planaria), or Danio rerio (Zebrafish), were all selected as model organisms because of their adaptability to undergraduate research and breadth of research topics. Student groups designed a novel experiment and generated a hypothesis, based on the current literature. The students were responsible for the animal husbandry of the models; C. elegans, D. tigrina and D. melanogaster throughout the course and performed experiments for over 50% of the semester. Once the experimental design and materials list were submitted students conducted experiments in weeks 4–6 and then analyzed their data. Following this data analysis, student groups either: repeated the same experiment, refined their experiment or generated a new question and then performed experimentation in weeks 8–11. Repeating experiments is a necessary component of scientific research, one that is typically lost in a traditional cookie‐cutter laboratory design. Students spent the remaining weeks of the semester working in their groups, analyzing data and preparing their oral presentation. The course has ran twice (Fall 2015 and Fall 2017) since it was developed and student survey response reports general satisfaction with the course.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Neuroscience drug and alcohol education has impact on students
Authors: Yim Rathana , Esperanza Samantha , Puder Barb
Published: June 2021

Prior research has shown the significant and positive impact that community outreach and involvement can have on a student's educational experience. The outreach component of this research project involved two elementary schools in Martinez (CA) and one middle school in Pleasant Hill (CA). Our neuroscience station – The Human Brain on Drugs – discussed the physiological effects of stimulants and depressants on the human brain. Each station began with a three‐minute pretest, ten‐minute poster presentation and activity, and three minute post‐test. Additionally, students were given a learning experience survey at their final station. After the outreach activity was finished, all qualitative and quantitative data were collected and analyzed using Microsoft Excel and scored for statistical analysis. The results from all three schools showed statistically significant increases in mean test scores from pretest to post test. Overall, the results provided some insight on the importance of educational outreach and its effectiveness to enhance a student's learning experience.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
EXPERT – EXercise for PEer Led Review and Teaching
Authors: Pizzimenti Marc A.
Published: June 2021

Organizing a formalized review session for students prior to an exam, particularly for a lab‐based course, often presents challenges to time, content, and opportunities for learning. Here we present a focused and efficient approach to organizing student participation for a laboratory examination review session using peer‐teaching to provide detailed overviews of concepts. In this exercise, each student group (e.g., 4 students) prepares for the activity by becoming an ‘expert’ for an assigned region of anatomy or a set of anatomical concepts. Students prepare for the activity by working through learning issues and/or clearly identifying anatomical structures on cadaveric donors in the lab. Through independent and group preparation, each group member contributes to and offers strategies on how the teaching of the region/concept will be presented. The general outline of the session divides the groups so that for the first round, half of the students are teaching, while the other half are learning by visiting other groups. Students that participate as ‘experts' remain at their table/station and repeat the topic every 8 minutes, as student learners rotate to each station. For the second round, the role of each student reverses (Fig. 1). Students spent an average of 2.3 h (± 1.5) preparing for the activity, while the total time for the activity was approximately 1.5 h. Student feedback (5‐point Likert scale) on the activity (n=146) was very positive. Students thought that the exercise was well‐organized (4.5 ±0.4) and worthwhile (4.6 ±0.4). They reported that they were able to become an ‘expert’ (4.4 ±0.4) in their assigned area and that the activity allowed them to determine their areas of strength (4.5 ±0.4) and weakness (4.5 ±0.4). Moreover, students reported that they were able to demonstrate their knowledge (4.5 ± 0.4) and felt that their classmates at other tables were prepared for the activity (4.4 ± 0.4). Although this activity was applied to the laboratory setting it could be easily modified for other learning/teaching environments and courses. This session incorporates active learning strategies, social support for learning, and provides an efficient method for group interaction.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Virtual Learning Modality for Neuroanatomical Education
Authors: Heise Natascha , Hall Heather A. , Garbe Brendan A. , Eitel Chad M. , Clapp Tod R.
Published: June 2021

One of the most difficult tasks for students in functional neuroanatomy is visualizing the routes of neuronal signaling through circuits and complete systems in 3 dimensions (3D). During class, instructors present 2 dimensional line diagrams and cross‐sections to help students visualize this anatomy on their own. Students are tasked to interpret these two‐dimensional images into 3D in order to understand a complete and functional system. This type of visualization and interpretation is necessary when applying pathologies and anticipating a resulting patient presentation. By using virtual reality (VR), students have the ability to build, visualize, and interact with an entire neuronal system. Here we describe a custom neuroanatomy VR program used by undergraduate students at Colorado State University (CSU). Virtual reality is a computer‐generated, 3D, artificial environment that allows users to interact in a simulated experience. We implemented the virtual reality software in the neuroanatomy laboratory where students had the option of interacting with the program. Students were asked to complete a survey that indicated their opinions about virtual reality in an educational setting. In addition, we collected data through faculty observations on the student's ability to solve novel neuroanatomical problems involving lesions and patient presentations. Results from opportunity sampling method, in conjunction with student surveys and faculty observations, suggested that students appreciated the introduction of VR into their curriculum and reported that VR helped them visualize and interpret the structure and function of the nervous system. Students also reported that the VR program reinforced anatomical structures and neuronal connections that are not visible on a gross specimen with the naked eye. In addition, students' ability to recognize the outcomes of neuroanatomical lesions increased. The implementation of a VR program created an authentic environment that enhanced student engagement and application of functional neuroanatomy. We believe that thoughtful deployment of virtual reality programming can improve student satisfaction and learning outcomes in neuroanatomy.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Development of an Inexpensive, Recordable, DIY Digital Ultrasound‐coupled Stethoscope for Medical Education
Authors: Resuehr David , Barger J. Bradley
Published: June 2021

For approximately 20 years, electronic and digital stethoscopes have been available. These devices were developed with at least three goals: improving auscultation of heart sounds, saving phonocardiograms in a digital format, and sharing heart sounds over the internet (telemedicine). In the last five years these devices have become inexpensive and widely available, such that research on their use in medical education has been published. The publication record demonstrates improved student learning when using digital stethoscopes for training of auscultation. Despite the utility of electronic stethoscopes, drawbacks exist, primarily cost of the devices; many of the commercially available devices are still nearly US$500. The project described here makes use of low cost stethoscopes and microphone systems to produce DIY digital stethoscopes appropriate for medical education with a total per‐unit cost of only ~US$20. Other advantages of the new system described here (termed “DigiStet”) include opensource hardware and software, meaning the device can be modified for other applications by the end user. In addition to describing several methods of constructing wireless digital stethoscopes, this project describes the use of the “DigiStet” to record heart sounds while simultaneously recording synchronized ultrasound images of the heart. We are unaware of any publications on this novel use of the digital stethoscope, i.e. recording heart sounds and ultrasound images simultaneously. These recordings are demonstrated to students in conjunction with the Wigger's Diagram to help first‐year medical students better visualize heart structure, function, action, and auscultation. We are planning on conducting a focus group with the first‐year students after their cardiology module to assess their opinions of the new technology, assess their knowledge acquisition, and look for improvements to future iterations of the DigiStet.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Construction of an Instrument for Evaluating the Accuracy and Utility of Virtual 3D Middle and Inner Ear Models in Students' Short‐ and Long‐term Retention
Authors: Meyer Edgar R. , Dehon Erin , Notebaert Andrew , Eby Thomas L. , Cui Dongmei
Published: June 2021

In medical education, 3D anatomical models are frequently used to help medical trainees master human anatomy. In the clinical setting, 3D models are used to guide residents and physicians in devising optimal surgical approaches to access specific anatomical regions and treat pathological structures. Since there are variations among 3D models, experts will be asked, using a list of generated survey questions, for their opinions on what anatomical structures and three‐dimensional relationships are most critical in 3D models of the middle and inner ear for the purpose of educating medical trainees. Experts will be faculty (physicians and scientists) who have proficiency pertaining to the middle and inner ear and who are involved in educating medical trainees on these anatomical regions. Their opinions will be solicited using email communications and/or interviews (in‐person or phone) with the generated survey questions. All of the expert responses will be compiled into a list of survey items to send to all of the experts. The experts will be asked to review each potential item in the list and rate the importance of each item in medical training using a Likert scale of 1 = not important to 4 = extremely important. Responses will be used to calculate a content validity index (CVI) and determine whether an item will be retained. The CVI for each item is the proportion of experts who rate the item as 3 or 4 (important) versus 1 or 2 (not important). As recommended in the literature, CVIs that are less than 0.83 will be dropped from the evaluation tool. This process of using two rounds of questionnaires targeting experts follows a Delphi method procedure which is also well established in the literature. The results of these surveys and statistical measures will be used to develop an instrument that can be used to evaluate the accuracy and utility of virtual 3D models of the middle and inner ear in medical training. Future studies will focus on the effectiveness of middle and inner ear models and on the evaluation of graduate and medical students' and residents' short‐ and long‐term retention of the 3D relationships pertaining to these regions.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Efficacy of Gross Anatomy Cadaver Online Video Learning Modules in First‐Year Physical Therapy Students
Authors: Poletti Mary , Rihm Jessica , Egan Casey , Day Leslie
Published: June 2021

The purpose of this study was to determine if online video modules (videos) identifying dissected cadaveric structures improve physical therapy students' learning and understanding of Gross Anatomy material assessed by performance on laboratory practicals. Current research in other disciplines suggests utilizing online resources will benefit students' learning of complex material. However, a gap in the research exists in relation to the benefits of video usage within the population of physical therapy students learning gross anatomy. A cohort of 110 first‐year Doctorate of Physical Therapy students were included in the study. Six interactive videos were created that taught the identification and relationships of multiple anatomic structures of varying levels of difficulty. Subjects viewed the videos prior to corresponding in‐person laboratory session. A likert‐scale and open‐ended questionnaire was administered to assess overall usefulness of videos and students' confidence with the material covered in the cadaveric videos. Video metadata was collected and analyzed to deduce student usage of the videos throughout the course. In addition, student practical exam scores were analyzed to compare accuracy of identifying structures discussed in the videos compared to structures not discussed in the videos of relative difficulty. The results provided evidence to support implementing interactive video modules to improve practical exam performance and confidence of material in a gross anatomy course for physical therapy students.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Cadaver vs. Microsoft HoloLens: A Comparison of Educational Outcomes of a Breast Anatomy Module
Authors: Yong Valeda , Sridharan Preethy , Ali Sajjaad Ahmad , Tingle Galen , Enterline Rebecca , Ulrey Lauren , Tan Lisa , Eastman Henry , Gotschall Robert , Henninger Erin , Griswold Mark , Wish‐Baratz Susanne
Published: June 2021

PURPOSE

Microsoft HoloLens (HL) mixed reality technology offers students a novel modality to visualize clinically important anatomical structures, such as the breast, which are uniquely challenging to discern with the naked eye in traditional cadaveric dissection. In this study, we developed a 3‐D anatomical model of the breast and integrated it into a dynamic, educational module on the HoloLens. Here, we report the educational outcomes and overall impressions of medical students learning breast anatomy through our module, as compared with traditional dissections.

METHODS

A mixed reality breast anatomy module was created using AutoDesk 3DSMax and integrated into the HoloLens device. 38 first‐year medical students were then recruited and divided into two groups: one participated in the HL module prior to dissecting the cadaveric breast, and the second dissected the cadaveric breast prior to the HL module. Before and after each teaching modality, participants answered seven comprehension‐based breast anatomy questions and a survey about their experience.

RESULTS

Our results show that scores on the comprehension‐based questions significantly improved more after the HL module than after the dissection (p=0.0209). Additionally, students appear to react more positively to the HL module than the dissection, regardless of which they were presented with first (p = 0.0008).

CONCLUSIONS

The present study demonstrates that students show better comprehension outcomes of breast structures using the HL. This is likely due to the ability to visualize structures that cannot be seen in the cadaver. Additionally, students appear to react more positively about learning breast anatomy through the HL module than through cadaveric dissection. Our results will be important in informing a future technology‐driven medical education, especially in using mixed reality to supplement what a traditional cadaveric dissection may lack.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effect of Interventions on Spatial Abilities in Anatomy Education: A Systematic Review and Meta‐Analysis
Authors: Langlois Jean , Bellemare Christian , Toulouse Josée , Wells George A.
Published: June 2021

Objective

Spatial abilities have been correlated to anatomy knowledge assessment using practical examination, 3D‐synthesis from 2D‐views, drawing of views, and cross‐sections in a previous systematic review. The objective of this study was to evaluate the effect of interventions on spatial abilities in the field of anatomy education.

Methods

Search criteria included ‘interventions', ‘spatial abilities' and ‘anatomy’. Keywords related to these criteria were defined. A literature search was conducted to August 3, 2017 in Scopus and EBSCOhost platform (Medline with Full Text, Cinahl Plus with Full Text, ERIC, Education Source, and PsycInfo). Citations were reviewed and those involving the field of anatomy education, an intervention, and a spatial abilities test were retained and the corresponding full‐text articles were reviewed for inclusion. Citations related to abstracts, literature reviews, books, book sections, and theses were excluded. Citations and full‐text articles were reviewed by two independent investigators. Methods in the field of anatomy education relating an intervention to spatial abilities test scores obtained before and after the intervention were identified as eligible. Eligible articles were reviewed in a systematic way and assessed for quality using Scottish Intercollegiate Guidelines Network‐50 assessment instruments. Effect sizes using standardized mean differences were calculated since different measurement scales for spatial abilities test scores were used. If I2 was greater than 50%, then heterogeneity was formally evaluated and random effects model was considered.

Results

Of the 2405 citations obtained, 52 articles were identified and reviewed, yielding 8 eligible articles. Instruction in anatomy and mental rotations training were found to improve spatial abilities. For the 7 studies retained for the meta‐analysis that included the effect of interventions on spatial abilities test scores, the pooled difference was 0.68 [95% CI (0.40; 0.95); n = 11] improvement with an I2 of 82%. For the 2 studies that included the practice effect on spatial abilities test scores in a control group, the pooled difference was 0.59 [95% CI (0.41; 0.76); n = 2] improvement with an I2 of 0%. In these 2 studies, the impact of the intervention on spatial abilities test scores was found valid despite the practice effect.

Conclusion

Evidence was found for improvement of spatial abilities in the field of anatomy education using instruction in anatomy and mental rotations training.

Support or Funding Information

Funding: none.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Infraorbital Foramen is Located between Nasospinale and Jugale
Authors: Russell Michelle L. , Koons Aaron W. , Bliss Kaitlyn N. , Mangus Kelsey R. , Zdilla Matthew J.
Published: June 2021

Identification of the infraorbital foramen (IOF) is important in infraorbital nerve block and the prevention of iatrogenic injury of the infraorbital nerve in maxillofacial surgeries. Though many methods have been proposed for identifying the location of the IOF, numerous inconsistencies exist among them. This study assessed the location of 1272 infraorbital foramina, from crania varied by age, sex, and geographic origin, relative to a midpoint of a line segment spanning from nasospinale to jugale (NS‐J). On average, the midpoint was located 1.92±1.85 mm (mean±SD) from the IOF. Most often, the midpoint was located either directly upon the IOF (Mode=0; 404:1272; 31.8%) or inferior to the IOF (403:1272; 31.7%). The NS‐J midpoint can be used to locate the infraorbital foramen in both females and males of varied geographic populations and ages regardless of craniofacial diversity. The results of this study may aid in the approach to infraorbital nerve block procedures and maxillofacial surgery.

Support or Funding Information

WV Research Challenge Fund [HEPC.dsr.17.06] and [HEPC.dsr.14.13]

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Extracellular Matrix Protection Factor‐2 Reduces Collagen Degradation in Primary Serum‐Free Cultures of Inflamed Human Gingival Fibroblasts
Authors: Chmielewski Sarah , Mattioli Patrisia , Isaac Tamika , Buckley Andrea Lynn , Seutter Sara , Gambardella Eric , Green Kevan , Shamseddin Seyed , Borghaei Ruth , Selim Abdulhafez , D'Angelo Marina
Published: June 2021

Some form of periodontal disease affects nearly everyone over the age of 30. The hallmarks of periodontal disease include inflammation of the gum tissue surrounding the teeth and concomitant collagen degradation in the extracellular matrix of gingival fibroblasts. Our laboratory has developed a novel enzyme inhibitor, Extracellular Matrix Protection Factor‐2 (ECPF‐2), that can reversibly interfere with the collagenase, matrix metalloprotease‐8 (MMP‐8), and its ability to digest its substrate, collagen type I. Serum‐free, primary cultures of human gingival fibroblasts (HGVF) isolated from tissue removed during oral surgery were established (passages 1 through 4). The tissue was collected from a patient population that included (a) non‐inflamed, non‐smokers, (b) inflamed non‐smokers, (c) inflamed previous smokers and (d) inflamed current smokers. The cell layer and associated extracellular matrix of these cultures was tested for collagen degradation utilizing an immunosorbence assay that measures degradation fragments generated when collagenase digests fibrillar collagen type I. Cells isolated from noninflamed tissue produced 2.56ng/culture degraded collagen, while inflamed tissue produced an average of 8.11 ng/culture. When smoking was considered as a factor, cultures of inflamed gingival fibroblasts prepared from a current smoker produced the most degraded collagen per culture (9.83 ng/culture current smoker, 7.90 ng/culture previous smoker and 6.64 ng/culture non‐smoker). Treatment for 24 hours with 5 ug ECPF‐2 reduced the amount of degraded collagen produced in gingival fibroblast cultures isolated from the inflamed current smoker by 15% (9.80 ng/culture untreated versus 8.45 ng/culture ECPF‐2 treated). These data support the therapeutic potential of ECPF‐2 to slow degradation of gingival tissue associated with periodontal disease

Support or Funding Information

Center for Chronic Disorders of Aging Pilot Grant and PCOM's Division of Research

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Comparison of 3D Ultrasound Imaging to Computed Tomography in Knee Osteophyte Depiction
Authors: Vendries Valeria , Ungi Tamas , Kunz Manuela , MacKenzie Leslie W. , Venne Gabriel
Published: June 2021

Background

Osteophytes (marginal bony outgrowths) are a common radiographic marker of osteoarthritis (OA) and joint degeneration. However, due to their variable morphologic composition, osteophytes are not accurately depicted using conventional imaging modalities. This represents problems for evaluating the anatomical changes of the osteoarthritic joint, and for the design of surgical interventions that rely on the accuracy of pre‐operative images. Studies have shown that ultrasound is a promising tool to detect articular changes such as the presence of osteophytes, and to monitor the progression of OA. Furthermore, 3D ultrasound (3DUS), a tool for volume rendering and surface representation, can potentially offer a means to quantify and depict osteophytes.

Objective

To compare osteophyte depiction in the knee joint using 3DUS and conventional Computed Tomography (CT) and to evaluate the ability of 3DUS at quantifying osteophyte surface depiction.

Methods

Eleven fresh‐frozen‐thawed human cadaveric knees were pre‐scanned for the presence of osteophytes according to a previously validated US semi‐quantitative grading system. Five knee sides with visible signs of OA were selected; 3DUS and CT images were obtained, segmented and digitally 3D reconstructed. The knees were dissected and Structured Light Scanner (SLS) images of the physical joint surface were obtained. Using a custom software, surface matching and Root Mean Square Error (RMSE) analyses were performed to assess the accuracy of each of the evaluated modalities in capturing the anatomy of the bone surface at the sites of osteophytes. 3DUS and CT models were compared to the SLS model, which was used as ground truth.

Results

The average RMSE for 3DUS to SLS and for CT to SLS model comparisons were 0.87 mm and 0.95 mm respectively. No statistical difference was found between 3DUS and CT (p=0.43). Comparative observation of imaging modalities set against each other suggests that 3DUS is superior in depicting osteophytes with cartilage and fibrocartilage tissue characteristics compared to CT.

Conclusions

3D Ultrasound can depict features of OA such as osteophytes together with their cartilaginous portion, which is not accurately represented using CT. It is feasible to compare 3DUS to conventional imaging for bone surface depiction within the knee joint. Lastly, 3DUS can provide useful information about not only the presence, but the extent of osteophytes.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Variation in Gantzer muscle origin, insertion, and innervation
Authors: Celuck Tyler J. , Andrews Reed C. , Pacurari Paula , Russell Michelle L. , Zdilla Matthew J. , Lambert H. Wayne
Published: June 2021

Gantzer muscles are inconsistent muscular variations in the anterior forearm. When present, Gantzer muscles usually arise from the flexor digitorum superficialis (FDS) and insert into the flexor pollicis longus (FPL) or, less often, the flexor digitorum profundus (FDP). The presence of Gantzer muscles can cause a compressive neuropathy affecting the anterior interosseous nerve (Kiloh‐Nevin syndrome), and these muscles must be considered when anterior forearm fasciotomies are performed for the management of acute compartment syndrome. While Gantzer muscles have been commonly reported, many studies neglect to report the innervation of the muscle and its relationship with nearby nerves. This study assessed 40 forearms from 20 cadaveric specimens to determine the prevalence of the origin and insertion of the Gantzer muscle. Furthermore, innervation was assessed along with the potential for nearby nerve compression. Gantzer muscles were present in 26 of 40 (65.0%) forearms, and all of the 22 intact muscles originated from the FDS, after noting four proximal attachments were damaged during student dissections. Concerning the 24 intact distal attachments of the Gantzer muscles, 20 (83.3%) muscles inserted into the FPL, two (8.3%) muscles inserted into the FDP solely, and one (4.2%) inserted at both the FDP and FPL. In addition, one forearm presented with duplicated and bifurcated Gantzer muscles with one tendon inserting into the FDP and the other one into the FPL. The innervation of the 21 Gantzer muscles was identified with the anterior interosseous nerve innervating 16 muscles (76.2%). In the five remaining Gantzer muscles, four (19.0%) were innervated solely by the median nerve, and one (4.8%) had a dual innervation from both of the two aforementioned nerves. In the one forearm with a bifurcated Gantzer muscle, the authors agreed upon an inherent anatomical predisposition for median nerve entrapment.

Support or Funding Information

West Virginia University Initiation to Research Opportunities (WVU – INTRO); WV Research Challenge Fund [HEPC.dsr.17.06] and [HEPC.dsr.14.13]

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Inhibition of Epstein‐Barr Virus by an Atypical Antipsychotic
Authors: Anderson Abbie , Weseli Joshua , Gorres Kelly
Published: June 2021

The Epstein‐Barr Virus (EBV) is a member of the herpes virus family and causes infectious mononucleosis. Epstein‐Barr Virus was the first virus discovered to cause cancer in humans. After infection with EBV, the virus maintains a lifelong dormant infection within the host. The virus's life cycle consists of two phases, the latent and the lytic phase. The latent phase allows the virus to lie dormant within the host without presenting any symptoms, while during the lytic phase the virus reproduces and spreads among cells. The virus switches between the latent and lytic phases in response to environmental stimuli, including some pharmaceuticals. We investigated the response of the virus to atypical antipsychotic drugs. Antipsychotic drugs are used to treat conditions such as schizophrenia and bipolar disorder. Atypical antipsychotics, also known as second generation antipsychotics, have a different chemical structure and are generally more effective than the typical (first generation) antipsychotics. The effects of varying concentrations of the drugs on the reactivation of EBV into the lytic cycle were tested. The degree of viral reactivation was measured by expression of the viral BZLF1 gene, a regulatory gene expressed during reactivation into the Epstein‐Barr Virus lytic cycle. Quantitative polymerase chain reaction (qPCR) monitored BZLF1 gene expression. Expression of the BZLF1 gene and viral reactivation was found to be inhibited. Understanding the conditions and cellular pathways that inhibit the lytic phase of the virus will help to better understand the virus's life cycle in order to develop treatments for cancers caused by Epstein‐Barr Virus.

Support or Funding Information

This work was funded by the UWL College of Science and Health, UWL Faculty Research Grants to KLG, and Undergraduate Research and Creativity grants to AGA.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Isolation, Purification, Modeling, and binding site analysis of the Salmonella Bacteriophage ɛ34 repressor for subsequent crystallization and structure‐function studies
Authors: Jackson Doba
Published: June 2021

The bacteriophage ɛ34 is one of many phages that infect the bacteria Salmonella which is the main cause of salmonellosis (food poisoning). The bacteriophage ɛ34 encodes a protein called the cI repressor that is like other phage repressors. It has a C‐terminal protease domain which is about 80% identical to the repressors of phage λ (which infects E. coli), but its DNA binding N‐terminal portion is only about 50% identical to its closest known relatives in phages λ, and Lex A. This means that the ɛ34 repressor may have the same pathway for inactivation as phage λ, but probably has a different operator binding specificity due to lower sequence homology of its DNA‐binding domain. The N‐terminal DNA‐binding domain is homologous to the Helix‐Turn‐Helix family of DNA‐binding domains.

The main goal of this project was to initiate a structure‐function study on the repressor protein of phage ɛ34. The ɛ34 Bacteriophage DNA was first isolated and purified. The gene encoding the bacteriophage ɛ34 repressor was amplified by PCR and cloned into a pET11a vector. The cI repressor, was overexpressed and purified by immobilized metal affinity chromatography and ion exchange chromatography. We also generated a homology model of this protein using the bacteriophage λ as a template in the SWISS‐MODEL homology modelling server. Our models predict a unique structure for the N‐terminal Helix‐Turn‐Helix domain. Finally, we have identified a potential binding site upstream of the cI gene on the Bacteriophage λ DNA. Crystallization trials are currently in progress. This project was done as a collaborative venture between two laboratories, Huntingdon College (Dr. Doba Jackson) and Alabama State University (laboratory of Dr. Robert Villafane). This project is in line with Dr. Robert Villafane's research into the Genomic analysis of bacteriophage ɛ34 of Salmonella enteric serovar Anatum.

Support or Funding Information

1Department of Chemistry & Biochemistry, Huntingdon College

2Department of Biology, Alabama State University

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Investigating Amino Acid Residues in The Active Site of Dibenzothiophene Monooxygenase (DszC)
Authors: Johnson Kellee , Mendez Gilberto , Gonzalez‐Osorio Liliana , Jirde Samatar , Palfey Bruce , Vey Jessica
Published: June 2021

Fossil fuel combustion releases sulfur dioxide into the atmosphere, which is a harmful air pollutant and precursor of sulfate aerosols that contribute to acid rain. It is necessary to reduce atmospheric sulfur levels to mitigate these effects. A current method, hydrodesulfurization, is costly and inefficient at removing sulfur from complex aromatic compounds. Biodesulfurization is a cost‐effective alternative that utilizes bacteria to remove sulfur. Rhodococcus erythropolis uses a four enzyme pathway to remove sulfur from dibenzothiophene. The first enzyme of the pathway, dibenzothiophene monooxygenase (DszC), requires molecular oxygen, reduced flavin, and the formation of an intermediate, C4a‐hydroperoxyflavin, to sequentially oxidize dibenzothiophene to dibenzothiophene sulfone. This study aims to elucidate the roles of DszC active site residues in catalysis and binding and to enable rational engineering of DszC for future application in fuel refinement. Active site residue mutants were created using polymerase chain reaction, followed by plasmid DNA isolation and sequence confirmation. Steady state activity assays of DszC, quantitated by reverse‐phase HPLC, compared the amount of product formed by the mutants relative to wild type‐DszC. The ability of the mutants to form the intermediate was studied using stopped‐flow spectrophotometry. In this study, seven active site residue mutants were created and the percent of product formation by each mutant relative to wild type‐DszC was: His92Asn (12%), His92Gln (20%), Tyr96Phe (12%), His388Ala (38%), His388Asn (39%), His391Val (0%) and Asp392Ala (3%). These results indicate that all of the residues analyzed are necessary for proper product formation. Additionally, the intermediate formation rate constant for Tyr96Phe (1.03 × 104 M−1 s−1) was similar to wild type‐DszC (1.69 × 104 M−1 s−1). These results indicate that Tyr96 is not critical for proper intermediate formation, but has another role in catalysis as evidenced by the low (12%) product formation. His92Asn and Asp392Ala were also analyzed using stopped‐flow spectrophotometry but showed no intermediate formation signifying that these residues are essential to proper intermediate formation. Overall, these results show the necessity of all the residues presented in product formation and the fundamental roles of residues His92 and Asp392 in intermediate formation.

Support or Funding Information

National Institute of Health (NIH SCORE 5SC2AI109500)

Research Corporation (CCSA 22672)

MBRS/RISE Program/National Institute of Health (Grant 2R25GM063787‐10)

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Teaching the Use of Automated Pipettes to Undergrads through Direct Visualization of Data: A Quantitative Chemical Analysis Lab Focusing on Technique, Skill, and Accuracy in Solution Chemistry
Authors: Sostarecz Audra Goach , Sturgeon Bradley E. , Distin Steven P.
Published: June 2021

The correct use of automated pipettes in undergraduate labs has been demonstrated with an activity primarily for second semester chemistry and biology majors. This lab involves the use of automated pipettes for transferring a specific amount of dye mixtures to a cotton bandanna. The data from this lab comprise a color wheel on a bandanna that displays the amount of dye transferred based on the volume set on the automated pipette by the student. This allows for the students to see their mistakes and does not allow them to restart. Student learning outcomes include the ability to choose the correct automated pipette for the volume needed; to set the correct volume on an automated pipette; to properly draw up the solution; and to use proper dispensing technique. Pre‐surveys assessing students' confidences in accurately using automated pipettes show that the average freshman student (60% of those surveyed) has not used automated pipettes in high school but has used them in their first semester chemistry course. The majority of those students also have confidence in their ability to choose the correct automated pipette; to draw up the liquid, and to dispense it properly. There was less confidence in setting the correct volume on the automated pipette. The results show that some students still have difficulties in setting volumes on automated pipettes correctly and in dispensing the liquid. However, the post survey suggests that setting the correct volume on the automated pipette was the most learned skill from performing the lab (26% to 88% increase in confidence by the students). After performing the lab, 96% of the class was confident in their ability to use automated pipettes. The learning outcomes were met along with the unintentional learning outcome of following directions. Automated pipettes are increasingly being utilized in our labs at our institution for both classroom lab activities and research lab activities. The ability to use automated pipettes accurately and precisely is a vital skill for chemistry, biochemistry, and biology majors with the increased use of this instrument in scientific jobs and in research. The use of these pipettes is a learned skill and it should not be assumed that students know how to correctly utilize them. Feedback from students supports this conclusion.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.